Possible Risk Factors for Candida Esophagitis in Immunocompetent Individuals.
Candida esophagitis (CE) is a condition typically diagnosed in patients who are immunocompromised. Risk factors leading to the development of CE in immunocompetent patients have not been entirely elucidated. This study set out to identify risk factors associated with the development of CE in immunocompetent patients. This study was a single-center retrospective chart review. Patients diagnosed with CE confirmed by endoscopic biopsy or brushings at our hospital between 2007 and 2017 were reviewed. The medical histories, endoscopy reports and pathology results were noted. Abdominal pain, heartburn, dysphagia and odynophagia were the common indications for endoscopy. A total of 241 patients were identified as having been diagnosed with CE by endoscopic brushing or biopsy. Of these patients, 161 were excluded due to the presence of immunocompromising and 80 patients were included who had no underlying immunocompromising conditions. Eighty patients with CE satisfied the inclusion criteria. The mean age of patients at the time of diagnosis was 39.8 years old (95% CI: 34.9 - 44.7). The incidences in men and women were similar in this study (49% women and 51% men). Of these patients, 56 (70%) (95% CI: 59-80%; P < 0.005) were taking proton pump inhibitors (PPIs). Fifteen patients (19%) had a previous upper endoscopy with evidence of reflux esophagitis, and they were all treated with PPIs and subsequently found to have CE on repeat upper endoscopy with a mean of 21.6 months of PPI treatment. There were 16 (20%) patients without any attributable risk factor and were completely healthy. CE is an opportunistic infection typically seen in immunocompromised. We report incidence of CE in immunocompetent patients. In our cohort of immunocompetent patients, PPI use was the most common risk factor associated with the development of CE. This could be related to hypochlorhydria resulting from PPI use. However, the cause remains unclear in some patients.


Racial Variation in the Utility of Urinary Biomarkers PCA3 and T2ERG in a Large Multicenter Study.
To our knowledge it is unknown whether urinary biomarkers for prostate cancer have added utility to clinical risk calculators in different racial groups. We examined the utility of urinary biomarkers added to clinical risk calculators for predicting prostate cancer in African American and nonAfrican American men. Demographics, PCPT (Prostate Cancer Prevention Trial) risk scores, data on the biomarkers data PCA3 (prostate cancer antigen 3) and T2ERG (transmembrane protease serine 2 and v-ets erythroblastosis virus E26 oncogene homolog gene fusion), and biopsy pathology features were prospectively collected on 718 men as part of EDRN (Early Detection Research Network). Utility was determined by generating ROC curves and comparing AUC values for the baseline multivariable PCPT model and for models containing biomarker scores. PCA3 and T2ERG added utility for the prediction of prostate cancer and clinically significant prostate cancer when combined with the PCPT Risk Calculator. This utility was seen in nonAfrican American men only for PCA3 (AUC 0.64 increased to 0.75 for prostate cancer and to 0.69-0.77 for clinically significant prostate cancer, both p <0.001) and for T2ERG (AUC 0.64-0.74 for prostate cancer, p <0.001, and 0.69-0.73 for clinically significant prostate cancer, p = 0.029). African American men did not have an added benefit with the addition of biomarkers, including PCA3 (AUC 0.75-0.77, p = 0.64, and 0.65-0.66, p = 0.74) and T2ERG (AUC 0.75-0.74, p = 0.74, and 0.65-0.64, p = 0.88), for prostate cancer and clinically significant prostate cancer, respectively. Limitations include the small number of African American men (72). The post hoc subgroup analysis nature of the study limited findings to being hypothesis generating. As novel biomarkers are discovered, clinical utility should be established across demographically diverse cohorts.


Role of Nrf2 in host defense against influenza virus in cigarette smoke-exposed mice.
Influenza virus is a common respiratory tract viral infection. Although influenza can be fatal in patients with chronic pulmonary diseases such as chronic obstructive pulmonary disease, its pathogenesis is not fully understood. The Nrf2-mediated antioxidant system is essential to protect the lungs from oxidative injury and inflammation. In the present study, we investigated the role of Nrf2 in protection against influenza virus-induced pulmonary inflammation after cigarette smoke exposure with both in vitro and in vivo approaches. For in vitro analyses, peritoneal macrophages isolated from wild-type and Nrf2-deficient mice were treated with poly(I:C) and/or cigarette smoke extract. For in vivo analysis, these mice were infected with influenza A virus with or without exposure to cigarette smoke. In Nrf2-deficient macrophages, NF-κB activation and the induction of its target inflammatory genes were enhanced after costimulation with cigarette smoke extract and poly(I:C) compared with wild-type macrophages. The induction of antioxidant genes was observed for the lungs of wild-type mice but not those of Nrf2-deficient mice after cigarette smoke exposure. Cigarette smoke-exposed Nrf2-deficient mice showed higher rates of mortality than did wild-type mice after influenza virus infection, with enhanced peribronchial inflammation, lung permeability damage, and mucus hypersecretion. Lung oxidant levels and NF-κB-mediated inflammatory gene expression in the lungs were also enhanced in Nrf2-deficient mice. Our data indicate that the antioxidant pathway controlled by Nrf2 is pivotal for protection against the development of influenza virus-induced pulmonary inflammation and injury under oxidative conditions.


Functional neural circuits for mental timekeeping.
Theories of mental timekeeping suggest frontostriatal networks may mediate performance of tasks requiring precise timing. We assessed whether frontostriatal networks are functionally integrated during the performance of timing tasks. Functional magnetic resonance imaging (fMRI) data from 31 healthy adults were collected during performance of several different types of discrete interval timing tasks. Independent component analysis (ICA) was used to examine functional connectivity within frontostriatal circuits. ICA identifies groups of spatially discrete brain regions sharing similar patterns of hemodynamic signal change over time. The results confirm the existence of a frontostriatal neural timing circuit that includes anterior cingulate gyrus, supplementary motor area, bilateral anterior insula, bilateral putamen/globus pallidus, bilateral thalamus, and right superior temporal gyrus and supramarginal gyrus. Several other distinct neural circuits were identified that may represent the neurobiological substrates of different information processing stages of mental timekeeping. Small areas of right cerebellum were engaged in several of these circuits, suggesting that cerebellar function may be important in, but not the primary substrate of, the mental timing tasks used in this experiment. These findings are discussed within the context of current biological and information processing models of neural timekeeping.


Some theoretical limitations of the optical power spectrum analyzer.
Following conversion of a time signal to intensity transmittance variations on film, a lens can be used in collimated coherent light to project a light intensity pattern that is an estimate of the power spectrum of the data contained on the film. To record a general biplanar signal on a film a transmittance bias level is required. The effect of this bias is defined as noise, and signal-to-noise ratios are computed for single frequency signals. Both the unapodized aperture and a cosine-shaped apodization function are considered. The effect of the size of the aperture used to record the signal on film is shown, as is the effect of the recording film speed variations. Film thickness variations reduce the performance of the analyzer; this effect is also computed.


The relationship between intellectual impairment and mood disorder in the first year after stroke.
In a community-based study of patients with a first-ever stroke, intellectual impairment (as defined by scores on a common screening test for dementia, the Mini-Mental State Examination) was found in 26% at 1 month post-stroke, and in 21% at 6 and 12 month follow-up. Low scores on the screening test were associated with greater age, physical disability before the stroke, larger stroke lesion volumes as measured on CT scan, and non-stroke changes such as atrophy and white matter low attenuation on the CT scan. There was a negative correlation between scores on the Mini-Mental State Examination and symptom levels on two measures of mood disorder. However, there was no evidence of a specific relationship between major depression and low scores on the Mini-Mental State. We examined various aspects of the relationship between mood symptoms and low scores on the Mini-Mental State, but found no evidence to support the suggestion that this relationship represented an example of depressive pseudodementia. We discuss the significance of our findings for clinical psychiatry and neuropsychology.


%ggBaseline: a SAS macro for analyzing and reporting baseline characteristics automatically in medical research.
Demographic tables are widely used to report baseline characteristics in medical research. However, the traditional copy-paste production method is time-consuming and frequently generates typing errors. Current available statistical tools are still far away from ideal, because they are difficult to understand and they lack flexibility. A user-friendly, dynamic, and flexible tool is needed for researchers to automate the creation of demographic tables. In this paper, we introduce a SAS macro, %ggBaseline, that automatically analyzing and reporting baseline characteristics with the final production of publication-quality demographic tables. The macro provides optional parameters that allow for the full customization of desired demographic tables. Since %ggBaseline allows for the quick creation of reproducible and fully customizable tables, it can be beneficial to academics, clinical trials and medical research studies by making the presentation and formatting of results faster and more efficient.


Sexual orientation after prenatal exposure to exogenous estrogen.
Thirty women aged 17 to 30 years with documented prenatal exposure to the nonsteroidal synthetic estrogen diethylstilbestrol (DES) were compared to thirty women of similar demographic characteristics from the same medical clinic who had a history of abnormal Pap smear findings. A subsample of the DES women were also compared to their DES-unexposed sisters. Sexual orientation in its multiple components was assessed by systematic semistructured interviews. In comparison to both control groups, the DES women showed increased bisexuality and homosexuality. However, about 75% of the DES women were exclusively or nearly exclusively heterosexual. Nonhormonal and hormonal interpretations of these findings are discussed.


Evolution and Adaptation of the Avian H7N9 Virus into the Human Host.
Influenza viruses arise from animal reservoirs, and have the potential to cause pandemics. In 2013, low pathogenic novel avian influenza A(H7N9) viruses emerged in China, resulting from the reassortment of avian-origin viruses. Following evolutionary changes, highly pathogenic strains of avian influenza A(H7N9) viruses emerged in late 2016. Changes in pathogenicity and virulence of H7N9 viruses have been linked to potential mutations in the viral glycoproteins hemagglutinin (HA) and neuraminidase (NA), as well as the viral polymerase basic protein 2 (PB2). Recognizing that effective viral transmission of the influenza A virus (IAV) between humans requires efficient attachment to the upper respiratory tract and replication through the viral polymerase complex, experimental evidence demonstrates the potential H7N9 has for increased binding affinity and replication, following specific amino acid substitutions in HA and PB2. Additionally, the deletion of extended amino acid sequences in the NA stalk length was shown to produce a significant increase in pathogenicity in mice. Research shows that significant changes in transmissibility, pathogenicity and virulence are possible after one or a few amino acid substitutions. This review aims to summarise key findings from that research. To date, all strains of H7N9 viruses remain restricted to avian reservoirs, with no evidence of sustained human-to-human transmission, although mutations in specific viral proteins reveal the efficacy with which these viruses could evolve into a highly virulent and infectious, human-to-human transmitted virus.


Crystallographic study of azurin from Pseudomonas putida.
Azurin from Pseudomonas putida is a blue copper protein which functions as an electron carrier. Two crystal forms of azurin were grown, one in the presence and the other in the absence of zinc acetate; each belongs to space group P21 and contains two molecules per asymmetric unit. The zinc-free crystals have cell dimensions a = 43.25, b = 50.65, c = 54.60 A, beta = 107.79 degrees, while the crystals grown from zinc-containing solution have cell dimensions a = 40.76, b = 51.22, c = 54.96 A, beta = 103.12 degrees. The latter crystals were found to have four zinc ions incorporated into the crystal lattice. Both crystal structures were solved by the molecular-replacement method using the program MERLOT. The search model was the structure of azurin from Alcaligenes denitrificans. The crystallographic R factor for native azurin is 0.169 (Rfree = 0. 257) from 8 to 1.92 A resolution, while that for zinc azurin is 0. 181 (Rfree = 0.248) from 10 to 1.6 A resolution; for each structure the root-mean-square deviation in bond lengths from ideal values is 0.007 A. In both crystal structures the Cu atom forms three strong bonds in the equatorial plane, two with Ndelta1 from His46 and His117, and one with the thiolate S atom of Cys112. Two longer axial approaches are made by the Sgamma from Met121 and the carbonyl O atom from Gly45. This results in a distorted trigonal bipyramidal co-ordination around the Cu atom. It further confirms the presence of a weak fifth bond to the copper in P. putida azurin, as with other azurin structures described at high resolution. The Ndelta1 atom of His35 is protonated, as it is in the low-pH form of azurin from Pseudomonas aeruginosa but unlike the low-pH form of the azurins from Alcaligenes denitrificans or Alcaligenes xylosoxidans. In each crystal form the two molecules of azurin in the asymmetric unit are related by a local twofold axis and form a dimer stabilized by the interaction of a pair of hydrophobic patches surrounding the partially exposed His117 side chain. In the other known azurin crystal structures, analogous dimer formation is observed, but with different relative orientations of the molecules. The four zinc ions introduced during crystallization of zinc azurin are bound to the protein and participate in five- and sixfold ligand coordination with no affect on the copper binding site. The zinc ligands are Ndelta from His, carboxylate O atoms from Asp and Glu, Ogamma from Ser and water molecules. One of the zinc ions, located on a non-crystallographic twofold axis, links the dimers of the asymmetric unit into continuous chains parallel to the crystallographic (-101) direction and is primarily responsible for the altered unit-cell parameters. Two of the other zinc ions bind to His83, one in each molecule.


A holistic approach to environmental health research.
One of the most formidable questions facing the environmental health research community today is how to translate basic fundamental research into a product (e.g., disease outcome) that meets the needs of its stakeholders--the medical community, regulatory agencies, and ultimately, the citizens of our nation. Historically, a successful research program could be defined as one that received continuous funding, produced high-quality publications, and was respected by scientists in related fields. However, it is now apparent that this is not sufficient for attaining improved public health--the ultimate goal of these research efforts. Research results must be transferred in a more active way to the communities and professionals who have need of this information. The link must be recognized, and the roles of the stakeholders in the entire research process must be acknowledged to ensure full impact of the research endeavors.


Use of opioid medications for chronic noncancer pain syndromes in primary care.
To define the spectrum of chronic noncancer pain treated with opioid medications in 2 primary care settings, and the prevalence of psychiatric comorbidity in this patient population. We also sought to determine the proportion of patients who manifested prescription opioid abuse behaviors and the factors associated with these behaviors. A retrospective cohort study. A VA primary care clinic and an urban hospital-based primary care center (PCC) located in the northeastern United States. A random sample of VA patients ( n=50) and all PCC patients ( n=48) with chronic noncancer pain who received 6 or more months of opioid prescriptions during a 1-year period (April 1, 1997 through March 31, 1998) and were not on methadone maintenance. Information regarding patients' type of chronic pain disorder, demographic, medical, and psychiatric status, and the presence of prescription opioid abuse behaviors was obtained by medical record review. Low back pain was the most common disorder accounting for 44% and 25% of all chronic pain diagnoses in the VA and PCC samples, respectively, followed by injury-related (10% and 13%), diabetic neuropathy (8% and 10%), degenerative joint disease (16% and 13%), spinal stenosis (10% and 4%), headache (4% and 13%) and other chronic pain disorders (8% and 22%). The median duration of pain was 10 years (range 3 to 50 years) in the VA and 13 years in the PCC sample (range 1 to 49 years). Among VA and PCC patients, the lifetime prevalence rates of psychiatric comorbidities were: depressive disorder (44% and 54%), anxiety disorder (20% and 21%), alcohol abuse/dependence (46% and 31%), and narcotic abuse/dependence (18% and 38%). Prescription opioid abusive behaviors were recorded for 24% of VA and 31% of PCC patients. A lifetime history of a substance use disorder (adjusted odds ratio [OR], 3.8; 95% confidence interval [CI], 1.4 to 10.8) and age (adjusted OR, 0.94; 95% CI, 0.89 to 0.99) were independent predictors of prescription opioid abuse behavior. A broad spectrum of chronic noncancer pain disorders are treated with opioid medications in primary care settings. The lifetime prevalence of psychiatric comorbidity was substantial in our study population. A significant minority of patients manifested prescription opioid abusive behaviors, and a lifetime history of a substance use disorder and decreasing age were associated with prescription opioid abuse behavior. Prospective studies are needed to determine the potential benefits as well as risks associated with opioid use for chronic noncancer pain in primary care.


Association between parents' and children's use of oral health services.
Several parental factors influence children's use of oral health services. Some localized studies have shown that children's dental use patterns correlate positively with those of their parents. The objective of this study was to investigate associations between parents' and children's oral health-seeking behaviors among a representative sample of US children. We used the 2007 National Health Interview Survey to analyze a sample of children aged 2 to 17 years, matched with 1 parent. Using logistic regression, we examined associations between parents' and children's use of dental services and deferred dental care because of cost. The sample included 6107 child-parent pairs. Overall, 77% of children and 64% of parents had a dental visit in the previous 12 months. Adjusting for sociodemographic and use variables, children were more likely to have a dental visit when their parents also had a dental visit (adjusted odds ratio: 3.36 [95% confidence interval: 2.71-4.18]), compared with children of parents who did not have a dental visit. In addition, compared with children of parents who did not defer seeking dental care, children of parents who deferred their dental care because of cost were more likely to have care deferred because of cost as well (adjusted odds ratio: 12.47 [95% confidence interval: 9.09-17.11]). Parental oral health-seeking behaviors for themselves may have an important effect on oral health-seeking behaviors on behalf of their children, regardless of the child's insurance status. Comprehensive strategies to eliminate barriers that target parents and not just children may help to address children's underuse of oral health services.


Anthropoid versus strepsirhine status of the African Eocene primates Algeripithecus and Azibius: craniodental evidence.
Recent fossil discoveries have demonstrated that Africa and Asia were epicentres for the origin and/or early diversification of the major living primate lineages, including both anthropoids (monkeys, apes and humans) and crown strepsirhine primates (lemurs, lorises and galagos). Competing hypotheses favouring either an African or Asian origin for anthropoids rank among the most hotly contested issues in paleoprimatology. The Afrocentric model for anthropoid origins rests heavily on the >45 Myr old fossil Algeripithecus minutus from Algeria, which is widely acknowledged to be one of the oldest known anthropoids. However, the phylogenetic position of Algeripithecus with respect to other primates has been tenuous because of the highly fragmentary fossils that have documented this primate until now. Recently recovered and more nearly complete fossils of Algeripithecus and contemporaneous relatives reveal that they are not anthropoids. New data support the idea that Algeripithecus and its sister genus Azibius are the earliest offshoots of an Afro-Arabian strepsirhine clade that embraces extant toothcombed primates and their fossil relatives. Azibius exhibits anatomical evidence for nocturnality. Algeripithecus has a long, thin and forwardly inclined lower canine alveolus, a feature that is entirely compatible with the long and procumbent lower canine included in the toothcomb of crown strepsirhines. These results strengthen an ancient African origin for crown strepsirhines and, in turn, strongly challenge the role of Africa as the ancestral homeland for anthropoids.


Nano plasmon polariton modes of a wedge cross section metal waveguide.
Optical plasmon-polariton modes confined in both transverse dimensions to significantly less than a wavelength are exhibited in open waveguides structured as sharp metal wedges. The analysis reveals two distinctive modes corresponding to a localized mode on the wedge point and surface mode propagation on the abruptly bent interface. These predictions are accompanied by unique field distributions and dispersion characteristics.


Taxonomic and Metabolic Incongruence in the Ancient Genus Streptomyces.
The advent of culture independent approaches has greatly facilitated insights into the vast diversity of bacteria and the ecological importance they hold in nature and human health. Recently, metagenomic surveys and other culture-independent methods have begun to describe the distribution and diversity of microbial metabolism across environmental conditions, often using 16S rRNA gene as a marker to group bacteria into taxonomic units. However, the extent to which similarity at the conserved ribosomal 16S gene correlates with different measures of phylogeny, metabolic diversity, and ecologically relevant gene content remains contentious. Here, we examine the relationship between 16S identity, core genome divergence, and metabolic gene content across the ancient and ecologically important genus Streptomyces. We assessed and quantified the high variability of average nucleotide identity (ANI) and ortholog presence/absence within Streptomyces, even in strains identical by 16S. Furthermore, we identified key differences in shared ecologically important characters, such as antibiotic resistance, carbohydrate metabolism, biosynthetic gene clusters (BGCs), and other metabolic hallmarks, within 16S identities commonly treated as the same operational taxonomic units (OTUs). Differences between common phylogenetic measures and metabolite-gene annotations confirmed this incongruence. Our results highlight the metabolic diversity and variability within OTUs and add to the growing body of work suggesting 16S-based studies of Streptomyces fail to resolve important ecological and metabolic characteristics.


Reassessment of the lethal London fog of 1952: novel indicators of acute and chronic consequences of acute exposure to air pollution.
This article develops and assesses novel indicators of respiratory and other morbidity and mortality following London's lethal smog in the winter of 1952. Public health insurance claims, hospital admission rates for cardiac and respiratory disease, pneumonia cases, mortality records, influenza reports, temperature, and air pollutant concentrations are analyzed for December-February 1952-1953 and compared with those for the previous year or years. Mortality rates for the smog episode from December 1952 to February 1953 were 50-300% higher than the previous year. Claims that the smog only elevated health risks during and immediately following the peak fog 5-9 December 1952 and that an influenza epidemic accounted fully for persisting mortality increases in the first 2 months of 1953 are rejected. We estimate about 12,000 excess deaths occurred from December 1952 through February 1953 because of acute and persisting effects of the 1952 London smog. Pollution levels during the London smog were 5-19 times above current regulatory standards and guidelines and approximate current levels in some rapidly developing regions. Ambient pollution in many regions poses serious risks to public health.


Cytoskeletal interactions of synapsin I in non-neuronal cells.
Synapsin I is a neuronal phosphoprotein involved in the localization and stabilization of synaptic vesicles. Recently, synapsin I has been detected in several non-neuronal cell lines, but its function in these cells is unclear. To determine the localization of synapsin I in non-neuronal cells, it was transiently expressed in HeLa and NIH/3T3 cells as an enhanced green fluorescent protein fusion protein. Synapsin I-enhanced green fluorescent protein colocalized with F-actin in both cell lines, particularly with microspikes and membrane ruffles. It did not colocalize with microtubules or vimentin and it did not cause major alterations in cytoskeletal organization. Synapsin Ia-enhanced green fluorescent protein colocalized with microtubule bundles in taxol-treated HeLa cells and with F-actin spots at the plasma membrane in cells treated with cytochalasin B. It did not noticeably affect F-actin reassembly following drug removal. Synapsin Ia-enhanced green fluorescent protein remained colocalized with F-actin in cells treated with nocodazole, and it did not affect reassembly of microtubules following drug removal. These results demonstrate that synapsin I interacts with F-actin in non-neuronal cells and suggest that synapsin I may have a role in regions where actin is highly dynamic.


Free-space optical crossbar network integrated in a single block of LiNbO3 crystal.
A free-space optical crossbar network integrated in a single block of LiNbO(3) crystal is proposed, which consists of stages of 2×2 switches making use of the electro-optic effect of crystal and in-between routing devices for permutation based on double refraction and internal double reflection on interfaces. Two basic configurations are suggested. A control algorithm for the crossbar network is discussed, which may control a nonblocking interconnection between any input and output. The integrated crossbar network is low energy loss, nonblocking, easy to assemble, and insensitive to environment. A 3×3 crossbar network is designed and the experiment is demonstrated.


[Dependency levels and health care services' utilization in psychiatric hospitals in Aragon (Spain)].
To determine the characteristics of patients living in psychiatric hospitals in Aragon, to assess their dependency levels, and to analyze health care services' utilization by these patients. We performed a cross-sectional study between July 1 and November 31. The questionnaire used was the Resident Assessment Instrument- Mental Health (RAI-MH). The sample consisted of 437 patients living in public psychiatric hospitals in Aragon. These hospitals provide care to chronically mentally ill patients and to patients undergoing rehabilitation. The Resource Utilization Group (RUG-I) system was used to classify patients by their dependency levels for activities of daily life (ADL). Of the 437 patients, 259 (59.3%) were men with a mean age of 62.2 years. A total of 82.1% of the patients were classified as RUG-I group 1. Patients in groups 1 and 2 required more formal health care services. At least one visit by a psychiatrist was required by 25.3% of patients in group 1 and by 15.2% of those in group 2 compared with no visits by the other groups. Nursing interventions were more frequently required by patients in the more dependent groups. All of the of patients in groups 2 to 8 needed daily physical assistance for ADL vs. 26.3% of those in group 1. In the multivariate analysis, predictive variables were the hospital and type of unit. There is wide variation in health care services' utilization by patients living in psychiatric hospitals, which is related to dependency levels. Many psychiatric patients do not need formal psychiatric care. Health care professionals should assess the real needs of patients to provide each of them with appropriate care.


Pseudo-outbreak of Mycobacterium fortuitum on a Human Immunodeficiency Virus Ward: transient respiratory tract colonization from a contaminated ice machine.
Infection Control surveillance revealed an abnormally high number of sputum samples growing Mycobacterium fortuitum from a single ward that houses the human immunodeficiency virus service. We investigated the outbreak using a retrospective case-control study and molecular epidemiology. A total of 47 patients were identified with >/=1 sputum sample that grew M. fortuitum. No significant demographic or clinical variables were found to be associated with colonization. Environmental investigation demonstrated that the M. fortuitum isolated from patients was identical to the ice machine isolates by pulsed-field gel electrophoresis. Fortunately, there has been no evidence of progressive pulmonary disease developing in our patients after >/=6 months of follow-up. All cases were thought to represent transient colonization and not infection. The ice machine was disconnected and cleaned successfully with vinegar and then bleach. Only when a 0.5-micrometer filter was installed were no further patients colonized or water cultures found to be positive.


Usable Science for Managing the Risks of Sea-Level Rise.
Sea-level rise sits at the frontier of usable climate climate change research, because it involves natural and human systems with long lags, irreversible losses, and deep uncertainty. For example, many of the measures to adapt to sea-level rise involve infrastructure and land-use decisions, which can have multigenerational lifetimes and will further influence responses in both natural and human systems. Thus, sea-level science has increasingly grappled with the implications of (1) deep uncertainty in future climate system projections, particularly of human emissions and ice sheet dynamics; (2) the overlay of slow trends and high-frequency variability (e.g., tides and storms) that give rise to many of the most relevant impacts; (3) the effects of changing sea level on the physical exposure and vulnerability of ecological and socioeconomic systems; and (4) the challenges of engaging stakeholder communities with the scientific process in a way that genuinely increases the utility of the science for adaptation decision making. Much fundamental climate system research remains to be done, but many of the most critical issues sit at the intersection of natural sciences, social sciences, engineering, decision science, and political economy. Addressing these issues demands a better understanding of the coupled interactions of mean and extreme sea levels, coastal geomorphology, economics, and migration; decision-first approaches that identify and focus research upon those scientific uncertainties most relevant to concrete adaptation choices; and a political economy that allows usable science to become used science.


Accidental Swallowing of a Molar Band.
Accidental ingestion or aspiration of foreign bodies is considered as a medical emergency in dentistry. Despite their rare occurrence, accidental ingestions are associated with various complications and morbidity, thereby necessitating prevention of their incidence along with early and effective management. Herein, we report a case of accidental swallowing of an orthodontic molar band in a patient with unilateral cleft lip and palate and its management.


Factors associated with fragility fractures in women over 50 years of age: a population-based household survey.
To analyze the prevalence of and factors associated with fragility fractures in Brazilian women aged 50 years and older. This cross-sectional population survey, conducted between May 10 and October 31, 2011, included 622 women aged >50 years living in a city in southeastern Brazil. A questionnaire was administered to each woman by a trained interviewer. The associations between the occurrence of a fragility fracture after age 50 years and sociodemographic data, health-related habits and problems, self-perception of health and evaluation of functional capacity were determined by the χ2 test and Poisson regression using the backward selection criteria. The mean age of the 622 women was 64.1 years. The prevalence of fragility fractures was 10.8%, with 1.8% reporting hip fracture. In the final statistical model, a longer time since menopause (PR 1.03; 95%CI 1.01-1.05; p<0.01) and osteoporosis (PR 1.97; 95%CI 1.27-3.08; p<0.01) were associated with a higher prevalence of fractures. These findings may provide a better understanding of the risk factors associated with fragility fractures in Brazilian women and emphasize the importance of performing bone densitometry.


The Norwegian tenecteplase stroke trial (NOR-TEST): randomised controlled trial of tenecteplase vs. alteplase in acute ischaemic stroke.
Alteplase is the only approved thrombolytic agent for acute ischaemic stroke. The overall benefit from alteplase is substantial, but some evidence indicates that alteplase also has negative effects on the ischaemic brain. Tenecteplase may be more effective and less harmfull than alteplase, but large randomised controlled phase 3 trials are lacking. The Norwegian Tenecteplase Stroke Trial (NOR-TEST) aims to compare efficacy and safety of tenecteplase vs. alteplase. NOR-TEST is a multi-centre PROBE (prospective randomised, open-label, blinded endpoint) trial designed to establish superiority of tenecteplase 0.4 mg/kg (single bolus) as compared with alteplase 0.9 mg/kg (10% bolus + 90% infusion/60 minutes) for consecutively admitted patients with acute ischaemic stroke eligible for thrombolytic therapy, i.e. patients a) admitted <4½ hours after symptoms onset; b) admitted <4½ hours after awakening with stroke symptoms c) receiving bridging therapy before embolectomy.Randomisation tenecteplase:alteplase is 1:1. The primary study endpoint is favourable functional outcome defined as modified Rankin Scale 0-1 at 90 days. Secondary study endpoints are: 1) haemorrhagic transformation (haemorrhagic infarct/haematoma); 2) symptomatic cerebral haemorrhage on CT 24-48 hours; 3) major neurological improvement at 24 hours; 4) recanalisation at 24-36 hours; 5) death. NOR-TEST may establish a novel approach to acute ischaemic stroke treatment. A positive result will lead to a more effective, safer and easier treatment for all acute ischaemic stroke pasients.NOR-TEST is reviewed and approved by the Regional Committee for Medical and Health Research Ethics (2011/2435), and The Norwegian Medicines Agency (12/01402). NOR-TEST is registered with EudraCT No 2011-005793-33 and in ClinicalTrials.gov (NCT01949948).


Modulation of cytokine-induced cardiac myocyte apoptosis by nitric oxide, Bak, and Bcl-x.
-Cytokine-induced NO production depresses myocardial contractility and has been shown to be cytotoxic to cardiac myocytes. However, the mechanisms of cytokine-induced cardiac myocyte cell death are unclear. To analyze these mechanisms in detail, we treated neonatal cardiac myocytes in serum-free culture with a combination of the macrophage-derived cytokines interleukin-1beta, tumor necrosis factor-alpha, and interferon-gamma. These cytokines caused a time-dependent induction of cardiac myocyte apoptosis, but not necrosis, beginning 72 hours after treatment, as determined by nuclear morphology, DNA internucleosomal cleavage, and cleavage of poly(ADP-ribose) polymerase, reflecting caspase activation. Apoptosis was preceded by a >50-fold induction of inducible NO synthase mRNA and the release of large amounts (5 to 8 nmol/ microgram protein) of NO metabolites (NOx) into the medium. Cell death was completely blocked by an NO synthase inhibitor and attenuated by antioxidants (N-acetylcysteine and DTT) and the caspase inhibitor ZVAD-fmk. Cytokines also mediated an NO-dependent, sustained increase in myocyte expression of the Bcl-2 homologs Bak and Bcl-x(L). The NO donor S-nitrosoglutathione also induced apoptosis and cell levels of Bak, but not of Bcl-x(L). All effects of cytokines, including poly(ADP-ribose) polymerase cleavage, could be attributed to interleukin-1beta; interferon-gamma and tumor necrosis factor-alpha had no independent effects on apoptosis or on NOx production. We conclude that cytokine toxicity to neonatal cardiac myocytes results from the induction of NO and subsequent activation of apoptosis, at least in part through the generation of oxygen free radicals. The rate and extent of this apoptosis is modulated by alterations in the cellular balance of Bak and Bcl-x(L), which respond differentially to cytokine-induced and exogenous NO and by the availability of oxidant species.


Ceramide is responsible for the failure of compensatory nerve sprouting in apolipoprotein E knock-out mice.
Apolipoprotein E (apoE) is a key transporter of the cholesterol and phospholipids required for membrane synthesis and nerve growth. We now report a virtual absence in apoE knock-out (KO) mice of normal nerve growth factor (NGF)-driven compensatory sprouting of undamaged cutaneous nociceptive nerves. In contrast, NGF-independent regeneration of crushed axons was unaffected. Essentially similar results came from aged wild-type mice. In apoE KO mice, the endogenous sprouting stimulus was suspect, because NGF administration induced normal sprouting; nevertheless, NGF increased normally in denervated skin, transported normally in the axons, and led to phosphorylation of trkA, erk1, and erk2. However, sprouting was restored in apoE KO mice (although not in aged mice) by fumonisin B1, an inhibitor of ceramide synthesis. A shotgun analysis revealed a wide array of changes in individual ceramide species in DRG neurons of apoE KO mice, and the changes for ceramide species OH_N15:0 made it a candidate inhibitor of sprouting (increased in apoE KO mice and normalized by fumonisin B1). Nevertheless, the unknown effects of individual ceramide species on sprouting, as well as the variability of their changed levels in apoE KO mice and how these were affected by fumonisin B1, support a different conclusion. We suggest that absence of apoE expression alters the balance among ceramide species to one that collectively inhibits compensatory sprouting, whereas fumonisin B1 establishes a new balance that allows sprouting. Nontoxic ceramide modulators might usefully promote sprouting and circuitry repair in neurodegenerative disorders in which ceramide species are perturbed, adding to the benefits of reducing ceramide-induced neuronal apoptosis.


Effects of a proven error on evaluations of witness testimony.
Witnesses frequently make an error when reporting events they have observed. Although some error in witness reports is to be expected and does not mean the testimony as a whole is flawed, an important question is how such an error affects judgments of credibility of the witness. In 2 experiments we investigated the impact of a single demonstrated (probative or nonprobative) detail inaccuracy on judgments of the likely reliability of witness memory. Potential mediators (witness dishonesty and forgetfulness) were examined to explain the relationship between inaccuracy and perceived reliability of the witness's memory report. The presence of a single inaccuracy affected observers' judgments of the reliability of the other elements of testimony and the testimony as a whole. There was also some evidence that the less probative the detail the more other elements of the reported account of the event were questioned. The mediation analyses showed that a single testimonial error contributed to the witness being perceived as dishonest or forgetful, attributions that in turn shaped perceptions of witness credibility. These findings suggest that legal professionals should be cautious when highlighting an isolated testimonial error given the potential for it to suggest more widespread testimonial unreliability. (PsycINFO Database Record


Telomere shortening associated with chromosome instability is arrested in immortal cells which express telomerase activity.
Loss of telomeric DNA during cell proliferation may play a role in ageing and cancer. Since telomeres permit complete replication of eukaryotic chromosomes and protect their ends from recombination, we have measured telomere length, telomerase activity and chromosome rearrangements in human cells before and after transformation with SV40 or Ad5. In all mortal populations, telomeres shortened by approximately 65 bp/generation during the lifespan of the cultures. When transformed cells reached crisis, the length of the telomeric TTAGGG repeats was only approximately 1.5 kbp and many dicentric chromosomes were observed. In immortal cells, telomere length and frequency of dicentric chromosomes stabilized after crisis. Telomerase activity was not detectable in control or extended lifespan populations but was present in immortal populations. These results suggest that chromosomes with short (TTAGGG)n tracts are recombinogenic, critically shortened telomeres may be incompatible with cell proliferation and stabilization of telomere length by telomerase may be required for immortalization.


Glucose, not glutamine, is the dominant energy source required for proliferation and survival of head and neck squamous carcinoma cells.
Tumor metabolism is an essential contributor to disease progression and response to treatment. An understanding of the metabolic phenotype of head and neck squamous cell carcinoma (HNSCC) will allow the development of appropriate antimetabolic strategies for this tumor type. A panel of 15 HNSCC cell lines was assayed for glucose and glutamine dependence and sensitivity to metabolic inhibitors. In addition, broad-spectrum metabolomic analysis using mass spectrometry/liquid chromatography was combined with individual measurements of reducing potential, adenosine triphosphate, and lactate production to characterize cellular metabolic phenotypes. HNSCC energy and reducing potential levels closely mirrored extracellular glucose concentrations. Glucose starvation induced cell death despite the activation of secondary energetic pathways. Conversely, glutamine was not required for HNSCC survival and did not serve as a significant source of energy. 2-deoxyglucose (2-DG) and its fluorinated derivative decreased glycolytic and Krebs cycle activity, cellular energy, and reducing potential and inhibited HNSCC cell proliferation. 2-DG effects were potentiated by the addition of metformin, but not by inhibitors of the pentose phosphate pathway or glutaminolysis. Despite dependence on glucose catabolism, the authors identified a subset of cell lines with relative resistance to starvation. Exploration of 1 such cell line (HN30) suggested that the presence of wild-type p53 can partially protect tumor cells from glucose starvation. HNSCC tumor cells are dependent on glucose, not glutamine, for energy production and survival, providing a rationale for treatment strategies that target glucose catabolism. However, antimetabolic strategies may need to be tailored to the tumor background, more specifically, p53 status.


Resistance to downy mildew in pearl millet is associated with increased phenylalanine ammonia lyase activity.
Phenylalanine ammonia lyase (PAL) activity was studied in pearl millet cultivars with different levels of resistance to the downy mildew disease caused by Sclerospora graminicola, an important oomycete pathogen. PAL activity was elevated in resistant host cultivar and decreased in susceptible cultivars following downy mildew pathogen infection. The enzyme activation varied between cultivars and was correlated with the degree of resistance to downy mildew disease. The induction of PAL as a response to pathogen inoculation was further corroborated by a time-course study in seedlings and cultured cells of pearl millet. The level of PAL activity was highest at 1.5 h in cultured cells and 4 h in seedlings of resistant host cultivar after inoculation with Sclerospora graminicola. Further studies on PAL activity in different tissues of seedlings showed highest enzyme activity in the young growing region of the root of the resistant host cultivars. The accumulation of wall-bound phenolics and lignin was higher in the resistant cultivar seedlings as evidenced by phloroglucinol-HCl staining and p-coumaric acid assay. The temporal changes in lignin concentration and the concentration of soluble phenolics were greater in root tissues of resistant cultivars than in those of susceptible cultivars. Treatment of resistant seedlings with a PAL inhibitor, α-aminooxy-β-phenylpropionic acid, resulted in the enhancement of the enzyme activity, whereas in the presence of 1 mm trans-cinnamic acid the pathogen-induced PAL was completely inhibited. Treatment of pearl millet seedlings with exogenously applied PAL inhibitors induced downy mildew disease susceptibility in the resistant pearl millet cultivar, consistent with direct involvement of PAL in downy mildew resistance. Results are discussed with respect to the presumed importance of host phenolic compounds and lignin accumulation and its relation to PAL activation as a response to the pathogen infection.


Novel antigens in non-small cell lung cancer: SP17, AKAP4, and PTTG1 are potential immunotherapeutic targets.
Lung cancer is the leading cause of cancer deaths in both genders worldwide, with an incidence only second to prostate cancer in men and breast cancer in women. The lethality of the disease highlights the urgent need for innovative therapeutic options. Immunotherapy can afford efficient and specific targeting of tumor cells, improving efficacy and reducing the side effects of current therapies. We have previously reported the aberrant expression of cancer/testis antigens (CTAs) in tumors of unrelated histological origin. In this study we investigated the expression and immunogenicity of the CTAs, Sperm Protein 17 (SP17), A-kinase anchor protein 4 (AKAP4) and Pituitary Tumor Transforming Gene 1 (PTTG1) in human non-small cell lung cancer (NSCLC) cell lines and primary tumors. We found that SP17, AKAP4 and PTTG1 are aberrantly expressed in cancer samples, compared to normal lung cell lines and tissues. We established the immunogenicity of these CTAs by measuring CTA-specific autoantibodies in patients' sera and generating CTA-specific autologous cytotoxic lymphocytes from patients' peripheral blood mononuclear cells. Our results provide proof of principle that the CTAs SP17/AKAP4/PTTG1 are expressed in both human NSCLC cell lines and primary tumors and can elicit an immunogenic response in lung cancer patients.


CSF prolactin determination in patients following operation for pituitary tumor.
Serial cerebrospinal fluid (CSF) and plasma prolactin concentrations were determined from patients during prolactin stimulatory testing with thyrotropin-releasing hormone or during pneumoencephalographic stress. Six patients had been operated on for suprasellar extension of pituitary tumor and one had been irradiated for suprasellar extension of a pituitary tumor. Prior to testing, four patients had had no clinical evidence of tumor recurrence and 3 patients had had tumor recurrence. One of the recurrent tumors had again extended into a suprasellar location. Basal CSF prolactin was undetectable in all patients who had had no recurrence. In 3 of the 4 patients without recurrence, however, prolactin became detectable in CSF during stimulatory testing. CSF prolactin values also increased during stimulatory testing in the patient with suprasellar recurrence of the tumor. A basal CSF-to-plasma prolactin ratio was 0.1 or less in all patients without recurrence. In the 2 patients with recurrence but without suprasellar extension, the CSF-to-plasma prolactin ratio was 0.18 or less. The patient with suprasellar recurrence had a strikingly elevated CSF-to-plasma prolactin ratio of 1.1. Thus, an increase of CSF prolactin during stimulatory testing does not necessarily indicate suprasellar recurrence of a pituitary tumor. However, an elevated CSF-to-plasma prolactin ratio appears to remain a valid indicator of suprasellar extension despite prior pituitary surgery.


Detoxification of olive mill wastewaters by Moroccan yeast isolates.
A total of 105 yeast strains were isolated from Moroccan olive oil production plants and evaluated for their ability to grow in olive oil mill wastewaters (OMW). The 9 isolates that grew best on OMW were selected for further study to evaluate their effect on removal of organic pollutants and OMW phytotoxicity (barley seed germination test). The results showed that at least four yeast isolates effectively lowered the toxicity of this effluent in addition to providing very useful materials in terms of both yeast biomass (6 g/l DW) and an irrigation fluid. This group of yeast isolates significantly reduced the concentration of total phenols (44% removal) and Chemical Oxygen Demand, COD (63% removal). The best germination rate of 80% for undiluted OMW was obtained for strain Candida holstii that also increased the pH from 4.76 to 6.75. Principal component analysis of the results obtained for the best yeast strains confirmed the importance of COD and total phenol reduction along with increase of organic nitrogen and final pH for the improvement of germination rates and phytotoxic reduction. This study has highlighted the potential of indigenous yeasts in detoxification of olive mill wastewaters.


Tropical oils: nutritional and scientific issues.
Individually and in combination with other oils, the tropical oils impart into manufactured foods functional properties that appeal to consumers. The use of and/or labeling in the ingredient lists give the impression that these oils are used extensively in commercially processed foods. The estimated daily intake of tropical oils by adult males is slightly more than one fourth of a tablespoon (3.8 g), 75% of which consists of saturated fatty acids. Dietary fats containing saturated fatty acids at the beta-position tend to raise plasma total and LDL-cholesterol, which, of course, contribute to atherosclerosis and coronary heart disease. Health professionals express concern that consumers who choose foods containing tropical oils unknowingly increase their intake of saturated fatty acids. The saturated fatty acid-rich tropical oils, coconut oil, hydrogenated coconut oil, and palm kernel oil, raise cholesterol levels; studies demonstrating this effect are often confounded by a developing essential fatty acid deficiency. Palm oil, an essential fatty acid-sufficient tropical oil, raises plasma cholesterol only when an excess of cholesterol is presented in the diet. The failure of palm oil to elevate blood cholesterol as predicted by the regression equations developed by Keys et al. and Hegsted et al. might be due to the dominant alpha-position location of its constituent saturated fatty acids. If so, the substitution of interesterified artificial fats for palm oil in food formulations, a recommendation of some health professionals, has the potential of raising cholesterol levels. A second rationale addresses prospective roles minor constituents of palm oil might play in health maintenance. This rationale is founded on the following observations. Dietary palm oil does not raise plasma cholesterol. Single fat studies suggests that oils richer in polyunsaturated fatty acid content tend to decrease thrombus formation. Anomalously, palm oil differs from other of the more saturated fats in tending to decrease thrombus formation. Finally, in studies comparing palm oil with other fats and oils, experimental carcinogenesis is enhanced both by vegetable oils richer in linoleic acid content and by more highly saturated animal fats. The carotenoid constituents of red palm oil are potent dietary anticarcinogens. A second group of antioxidants, the tocotrienols, are present in both palm olein and red palm oil. These vitamin E-active constituents are potent suppressors of cholesterol biosynthesis; emerging data point to their anticarcinogenic and antithrombotic activities. This review does not support claims that foods containing palm oil have no place in a prudent diet.


"…and How Are the Kids?" Psychoeducation for Adult Patients With Depressive and/or Anxiety Disorders: A Pilot Study.
Depressive and anxiety disorders are highly prevalent and form a substantial burden for individuals and their family members. A recent study showed that approximately two-thirds of the children of patients with severe depressive and/or anxiety disorders develop one of these disorders themselves before 35 years of age. In the Netherlands, various preventive interventions are available for children of parents with mental illnesses. However, the actual reach of interventions is small (< 1% of all children participate). A major barrier appeared to be parents' hesitancy to let children participate in preventive programs. In order to address this barrier, we designed a pilot study to implement a psychoeducation program on parenting and mental illnesses. The main aim of this study was to investigate how a preventive family-focused approach can be embedded in routine adult psychiatric care. The pilot started in April 2017 and has ended in September 2018. It was conducted in the University Center Psychiatry (UCP) in Groningen, The Netherlands. This article describes the implementation process so far. The main intervention was a monthly organized group-psychoeducation called "parenting and a mental illness," which could be attended by parents currently treated in the UCP. In 18 months, implementation activities were divided in four phases; (1) Creating awareness, (2) Adoption of the intervention, (3) Implementation and evaluation, and (4) Continuation phase. The program development process was evaluated using both qualitative and quantitative data. Based on the pilot study we were able to make recommendations for the further implementation of this approach so that more parents can be reached in future. Further study with follow-up activities is needed to measure the effects of the psychoeducation, for instance on parenting functioning and the quality of the parent-child interaction.


Evaluation of the highest concentrations used in the in vitro chromosome aberrations assay.
There is controversy over the highest concentration to which an article should be tested in in vitro mammalian cell assays of genetic toxicity. Until recently, most guidelines specified the use of concentrations of up to 10 mM or 5,000 μg/ml (whichever is lower) when not limited by the toxicity of the test article to the cells used for the test. Several recent publications have called for lowering those limits. We examined concentration/response curves for in vitro chromosome aberrations assays. Data was extracted from two published databases to evaluate the lowest dose at which a positive response was reported. Concentration/response curves were simulated using Monte Carlo procedures on log normal distributions of the data. These curves were then used to predict the loss in assay sensitivity that would be incurred by arbitrarily lowering the highest concentration to which the assay is conducted. The simulations suggest that lowering the current high concentration limit from 10 mM would dramatically impact the sensitivity of the assay. In contrast, lowering the high concentration limit using the μg/ml scale, the most commonly applied scale in regulatory submissions, would not have a similar impact on assay sensitivity until the limit concentration was lowered to more than half of the current 5,000 μg/ml limit. This analysis suggests that the current limits of 10 mM and 5,000 μg/ml are not equivalent to one another and challenges the assumption that lowering the 10 mM limit will not decrease assay sensitivity.


Correlation not causation: the relationship between personality traits and political ideologies.
The assumption in the personality and politics literature is that a person's personality motivates them to develop certain political attitudes later in life. This assumption is founded on the simple correlation between the two constructs and the observation that personality traits are genetically influenced and develop in infancy, whereas political preferences develop later in life. Work in psychology, behavioral genetics, and recently political science, however, has demonstrated that political preferences also develop in childhood and are equally influenced by genetic factors. These findings cast doubt on the assumed causal relationship between personality and politics. Here we test the causal relationship between personality traits and political attitudes using a direction of causation structural model on a genetically informative sample. The results suggest that personality traits do not cause people to develop political attitudes; rather, the correlation between the two is a function of an innate common underlying genetic factor.


In vivo electroporation enhances the immunogenicity of an HIV-1 DNA vaccine candidate in healthy volunteers.
DNA-based vaccines have been safe but weakly immunogenic in humans to date. We sought to determine the safety, tolerability, and immunogenicity of ADVAX, a multigenic HIV-1 DNA vaccine candidate, injected intramuscularly by in vivo electroporation (EP) in a Phase-1, double-blind, randomized placebo-controlled trial in healthy volunteers. Eight volunteers each received 0.2 mg, 1 mg, or 4 mg ADVAX or saline placebo via EP, or 4 mg ADVAX via standard intramuscular injection at weeks 0 and 8. A third vaccination was administered to eleven volunteers at week 36. EP was safe, well-tolerated and considered acceptable for a prophylactic vaccine. EP delivery of ADVAX increased the magnitude of HIV-1-specific cell mediated immunity by up to 70-fold over IM injection, as measured by gamma interferon ELISpot. The number of antigens to which the response was detected improved with EP and increasing dosage. Intracellular cytokine staining analysis of ELISpot responders revealed both CD4+ and CD8+ T cell responses, with co-secretion of multiple cytokines. This is the first demonstration in healthy volunteers that EP is safe, tolerable, and effective in improving the magnitude, breadth and durability of cellular immune responses to a DNA vaccine candidate. ClinicalTrials.gov NCT00545987.


Transformation of Chronic Myeloid Leukemia to Acute Biphenotypic Leukemia.
Chronic myeloid leukemia (CML) is a myeloproliferative disorder with clonal proliferation of all myeloid cell lines. The disease typically manifests in three phases: chronic course followed by an accelerated phase and finally a terminal blast crisis. A blast crisis is defined as the presence of > 20% blasts in the peripheral blood or bone marrow. The blasts could be characterized as either myeloid (60-80% of cases) resulting in acute myeloblastic leukemia or lymphoid (20-30% of cases) resulting in acute lymphoblastic leukemia. In rare instances, a blast crisis could present with biphenotypic expression for both myeloid and lymphoid blasts. In such cases, about 6-10% of the time, the course of the disease is more aggressive and renders a poorer prognosis especially if there is evidence of extramedullary involvement. We present a case of a 41-year-old woman with history of CML who presented with acute biphenotypic blast crisis with extramedullary involvement in the context of aggressive chemotherapy. Literature review reveals < 10 reported cases of CML with biphasic transformation and only three cases of CML with acute leukemia on presentation. Most cases described are in pediatric patients with varied presentations and none involving extramedullary sites. Mortality rates in such cases are near 100% despite aggressive chemotherapy.


Direct measurement of CD8+ T cell responses in macaques infected with simian immunodeficiency virus.
The simian immunodeficiency virus (SIV) macaque model system has been used extensively to study AIDS pathogenesis and to test candidate vaccines for their ability to protect against homologous or heterologous challenge with pathogenic SIV or SHIV. Recent studies suggest that stimulation of HIV-1-specific CTL responses is important for effective vaccination against HIV-1. While quantitative measurements of SIV-specific cytotoxic T lymphocyte (CTL) responses have been facilitated by the use of tetrameric peptide complexes, this technique is currently limited to the study of Mamu-A*01-positive rhesus macaques. Furthermore, very few SIV-specific CTL epitopes have been identified, and there is limited identification of other MHC alleles in macaques. In this study, cytokine flow cytometry (CFC) was used to quantify SIV-specific CD8+ antigen-reactive T cells in macaques infected with SIV. We found a strong correlation (r = 0.96, P < 0.001) between CD8+ antigen-reactive T cells stained with the Mamu-A*01 p11C, C-M tetramer and production of intracellular TNF-alpha in the CFC assay. Furthermore, the CFC assay was used to identify a novel SIV-specific CTL epitope in Envelope (SIV Env, a.a. 486-494, sequence AEVAELYRL). The use of the CFC assay facilitates the study of antigen-reactive T cell responses in SIV infection and vaccination.


United States Census 2000 population with bridged race categories.
The objectives of this report are to document the methods developed at the National Center for Health Statistics (NCHS) to bridge the Census 2000 multiple-race resident population to single-race categories and to describe the resulting bridged race resident population estimates. Data from the pooled 1997-2000 National Health Interview Surveys (NHIS) were used to develop models for bridging the Census 2000 multiple-race population to single-race categories. The bridging models included demographic and contextual covariates, some at the person-level and some at the county-level. Allocation probabilities were obtained from the regression models and applied to the Census Bureau's April 1, 2000, Modified Race Data Summary File population counts to assign multiple-race persons to single-race categories. Bridging has the most impact on the American Indian and Alaska Native (AIAN) and Asian or Pacific Islander (API) populations, a small impact on the Black population and a negligible impact on the White population. For the United States as a whole, the AIAN, API, Black, and White bridged population counts are 12.0, 5.0, 2.5, and 0.5 percent higher than the corresponding Census 2000 single-race counts. At the sub-national level, there is considerably more variation than observed at the national level. The bridged single-race population counts have been used to calculate birth and death rates produced by NCHS for 2000 and 2001 and to revise previously published rates for the 1990s, 2000, and 2001. The bridging methodology will be used to bridge postcensal population estimates for later years. The bridged population counts presented here and in subsequent years may be updated as additional data become available for use in the bridging process.


CRISPR Systems Suitable for Single AAV Vector Delivery.
CRISPR (clustered regularly interspaced short palindromic repeats)/Cas gene editing is a revolutionary technology that can enable the correction of genetic mutations in vivo, providing great promise as a therapeutic intervention for inherited diseases. Adeno-associated viral (AAV) vectors are a potential vehicle for delivering CRISPR/Cas. However, they are restricted by their limited packaging capacity. Identifying smaller Cas orthologs that can be packaged, along with the required guide RNA elements, into a single AAV would be an important optimization for CRISPR/- Cas gene editing. Expanding the options of Cas proteins that can be delivered by a single AAV not only increases translational application but also expands the genetic sites that can be targeted for editing. This review considers the benefits and current scope of small Cas protein orthologs that are suitable for gene editing approaches using single AAV vector delivery.


Glycosylation Pattern and in vitro Bioactivity of Reference Follitropin alfa and Biosimilars.
Recombinant follicle-stimulating hormone (FSH) (follitropin alfa) and biosimilar preparations are available for clinical use. They have specific FSH activity and a unique glycosylation profile dependent on source cells. The aim of the study is to compare the originator (reference) follitropin alfa (Gonal-f®)- with biosimilar preparations (Bemfola® and Ovaleap®)-induced cellular responses in vitro. Gonadotropin N-glycosylation profiles were analyzed by ELISA lectin assay, revealing preparation specific-patterns of glycan species (Kruskal-Wallis test; p < 0.05, n = 6) and by glycotope mapping. Increasing concentrations of Gonal-f® or biosimilar (1 × 10-3-1 × 103 ng/ml) were used for treating human primary granulosa lutein cells (hGLC) and FSH receptor (FSHR)-transfected HEK293 cells in vitro. Intracellular cAMP production, Ca2+ increase and β-arrestin 2 recruitment were evaluated by BRET, CREB, and ERK1/2 phosphorylation by Western blotting. 12-h gene expression, and 8- and 24-h progesterone and estradiol synthesis were measured by real-time PCR and immunoassay, respectively. We found preparation-specific glycosylation patterns by lectin assay (Kruskal-Wallis test; p < 0.001; n = 6), and similar cAMP production and β-arrestin 2 recruitment in FSHR-transfected HEK293 cells (cAMP EC50 range = 12 ± 0.9-24 ± 1.7 ng/ml; β-arrestin 2 EC50 range = 140 ± 14.1-313 ± 18.7 ng/ml; Kruskal-Wallis test; p ≥ 0.05; n = 4). Kinetics analysis revealed that intracellular Ca2+ increased upon cell treatment by 4 μg/ml Gonal-f®, while equal concentrations of biosimilars failed to induced a response (Kruskal-Wallis test; p < 0.05; n = 3). All preparations induced both 8 and 24 h-progesterone and estradiol synthesis in hGLC, while no different EC50s were demonstrated (Kruskal-Wallis test; p > 0.05; n = 5). Apart from preparation-specific intracellular Ca2+ increases achieved at supra-physiological hormone doses, all compounds induced similar intracellular responses and steroidogenesis, reflecting similar bioactivity, and overall structural homogeneity.


A snail tale and the chicken embryo.
Some 25 years ago, a clone was identified that contained the chicken Slug sequences (now called Snail2 ). How could we anticipate at that time how much the chick embryo would help us to understand the ins and outs of cell migration during development and in disease? Indeed, the chick embryo helped us identify Snail2 as the first transcription factor that could induce the epithelial-mesenchymal transition (EMT), key for the migration of embryonic and cancer cells.


A Comparison of Outcomes for Cord Blood Transplantation and Unrelated Bone Marrow Transplantation in Adult Aplastic Anemia.
Earlier reports suggested that umbilical cord blood transplantation (UCBT) for aplastic anemia (AA) was feasible in alternative transplantation. To identify differences in outcomes of UCBT and HLA-matched or mismatched unrelated bone marrow transplantation (UBMT) in adults with AA, we analyzed registry data of the Japan Society for Hematopoietic Cell Transplantation and compared results of UCBT (n = 69) to 8/8-matched (n = 101), 7/8-matched (n = 65), or 6/8-matched (n = 37) UBMT. The transplantation period was from 2002 to 2012, and patients 16 years or older with AA were eligible. Median ages were 49, 35, 28, and 30 years for UCBT, 8/8-matched, 7/8-matched, and 6/8-matched UBMT, respectively. In multivariate analysis, risk of mortality was lower for 8/8-matched UBMT compared with that of UCBT (hazard ratio [HR], .55; 95% confidence interval [CI], .32 to .94; P = .029), adjusted for age and graft-versus-host disease (GVHD) prophylaxis, which were other associated factors. Mortality risks of 7/8-matched UBMT (HR, .55; 95% CI, .29 to 1.02) or 6/8-matched UBMT (HR, .67; 95% CI, .32 to 1.39) were not significantly different from those of UCBT. Risks of grade 3 or 4 acute and chronic GVHD were not different among the 4 groups. The most prevalent cause of death was graft failure in UCBT and 6/8-matched UBMT and infection in 8/8-matched and 7/8-matched UBMT. Under 40 years old,survival of UCBT was similar to that of UBMT (76%, 79%, 83%, and 83% for UCBT and 8/8-matched, 7/8-matched, and 6/8-matched UBMT, respectively, at 3 years), adjusted for transplantation period, which was another associated factor; however, for ages over 40 years, that of UCBT tended to be lower (47%, 64%, 64%, and 75% for UCBT, 8/8-matched, 7/8-matched, and 6/8-matched UBMT, respectively, at 3 years). To conclude, these data suggest that UCBT could be an alternative treatment option for younger adults when matched sibling or adequate UBMT donors are not available.


In situ preparation, electrical and surface analytical characterization of pentacene thin film transistors.
The fabrication of organic thin film transistors with highly reproducible characteristics presents a very challenging task. We have prepared and analyzed model pentacene thin film transistors under ultra-high vacuum conditions, employing surface analytical tools and methods. Intentionally contaminating the gold contacts and SiO2 channel area with carbon through repeated adsorption, dissociation, and desorption of pentacene proved to be very advantageous in the creation of devices with stable and reproducible parameters. We mainly focused on the device properties, such as mobility and threshold voltage, as a function of film morphology and preparation temperature. At 300 K, pentacene displays Stranski-Krastanov growth, whereas at 200 K fine-grained, layer-like film growth takes place, which predominantly influences the threshold voltage. Temperature dependent mobility measurements demonstrate good agreement with the established multiple trapping and release model, which in turn indicates a predominant concentration of shallow traps in the crystal grains and at the oxide-semiconductor interface. Mobility and threshold voltage measurements as a function of coverage reveal that up to four full monolayers contribute to the overall charge transport. A significant influence on the effective mobility also stems from the access resistance at the gold contact-semiconductor interface, which is again strongly influenced by the temperature dependent, characteristic film growth mode.


[Early determinants of random blood glucose among adults of the 1982 birth cohort, Pelotas, Southern Brazil].
To evaluate the effects of socioeconomic variables, birth weight, duration of breastfeeding and income changes on random blood glucose levels among young adults. This was a study on the birth cohort from 1982, when the 5,914 hospital births that occurred in the city of Pelotas (Southern Brazil) were identified and the mothers were interviewed. The children whose families lived in the urban area of the city were followed up several times. In 2004-5, 4,927 individuals of the cohort were interviewed and blood was collected from the fingertips of 3,730 of them for random blood glucose measurements. Associations between random blood glucose levels and skin color, family income at birth, maternal schooling, income change between 1982 and 2004-5, birth weight and duration of breastfeeding were evaluated. The mean blood glucose level was 97.3 +/- 15.1mg/dL, and it was greater among the men. None of the variables studied was associated with the men's blood glucose level. Among the women, maternal schooling, family income at 23 years of age and birth weight were inversely associated with blood glucose levels. However, birth weight lost its statistical significance in the multivariable analysis. Birth weight and duration of breastfeeding did not present any long-term effect on random blood glucose levels. Only maternal schooling level and present income presented associations with random blood glucose levels among the women.


Prognostic impact of B-cell lymphoma 6 in primary CNS lymphoma.
We investigated the prognostic significance of B-cell differentiation status and common B-cell differentiation markers in a post hoc analysis of 119 patients with primary CNS lymphoma (PCNSL) homogeneously receiving high-dose methotrexate (HDMTX)-based chemotherapy within the prospective G-PCNSL-SG1 trial. We evaluated protein expression of B-cell lymphoma 2 (BCL2), BCL6, CD10, and multiple myeloma oncogene 1/interferon regulatory factor 4 (MUM1/IRF4) by immunohistochemistry and analyzed the association with survival. The median follow-up of all patients was 67.5 months. Median progression-free survival (PFS) was 10.61 months (95% CI: 4.23-17.00). Median overall survival (OS) was 28.85 months (95% CI: 17.96-39.73). Eighty-nine tumors expressed BCL2 (92.7%), 24 (20.5%) expressed CD10, 60 (54.1%) expressed BCL6, and 87 (79.0%) expressed MUM1/IRF4. On the basis of the Hans algorithm, 80 tumors (73.4%) were classified to the non-germinal center B group, suggesting a post-germinal center origin of PCNSL. Expression of BCL6 (cutoff point 30%), but none of the other markers, was associated with shorter PFS (P = .047) and OS (P = .035). On multivariate analysis, BCL6 expression was associated with shorter PFS (hazard ratio: 1.95, 95% CI: 1.22-3.12, P = .005) but not OS (hazard ratio: 1.85, 95% CI: 0.71-4.80, P = .21). Classification according to Hans algorithm and expression status of the single B-cell markers BCL2, CD10, and MUM1/IRF4 did not correlate with prognosis. The findings are limited by the fact that only 23% of all G-PCNSL-SG1 patients could be included in the analysis. If validated in an independent cohort, BCL6 may assume clinical relevance as an unfavorable prognostic biomarker in PCNSL.


Implicit Frictional Boundary Handling for SPH.
In this article, we present a novel method for the robust handling of static and dynamic rigid boundaries in Smoothed Particle Hydrodynamics (SPH) simulations. We build upon the ideas of the density maps approach which has been introduced recently by Koschier and Bender. They precompute the density contributions of solid boundaries and store them on a spatial grid which can be efficiently queried during runtime. This alleviates the problems of commonly used boundary particles, like bumpy surfaces and inaccurate pressure forces near boundaries. Our method is based on a similar concept but we precompute the volume contribution of the boundary geometry. This maintains all benefits of density maps but offers a variety of advantages which are demonstrated in several experiments. First, in contrast to the density maps method we can compute derivatives in the standard SPH manner by differentiating the kernel function. This results in smooth pressure forces, even for lower map resolutions, such that precomputation times and memory requirements are reduced by more than two orders of magnitude compared to density maps. Furthermore, this directly fits into the SPH concept so that volume maps can be seamlessly combined with existing SPH methods. Finally, the kernel function is not baked into the map such that the same volume map can be used with different kernels. This is especially useful when we want to incorporate common surface tension or viscosity methods that use different kernels than the fluid simulation.


Childhood sexual abuse associated with dating partner violence and suicidal ideation in a representative household sample in Hong Kong.
This study investigated the prevalence and impact of childhood sexual abuse (CSA) on future intimate partner violence (IPV) in dating relationship in Hong Kong, China. A total of 1,154 Chinese adult respondents engaged in dating relationships were interviewed face-to-face about their CSA histories, childhood witnessing of parental violence, adult sexual victimization (ASV) by others and IPV victimization with their current dating partner. Self-reports also measured levels of suicidal ideation, self-esteem, and demographic details. Overall, 1.7% reported some form of CSA with a higher percentage being women. No gender differences were found in the prevalence of either ASV or IPV. Results showed that CSA had an independent effect on physical IPV and suicidal ideation. The odds of IPV were increased by behavioral and psychological factors of victims such as alcohol and drug abuse, sex with partner, and low self-esteem. The odds of suicidal ideation were also increased by drug abuse, childhood witnessing of parental psychological aggression, and low self-esteem. Clinical implications of results included screening for CSA victims and suicidal victims when treating IPV patients, tailoring treatment according to individual IPV victim's problems, correcting behaviors that are associated with risks of IPV, such as engagement in casual sex and substance abuse, and focusing not only on tangible services but also on the social and psychological aspects that are placing the victims at risk for IPV.


Eosinophil-derived IL-13 promotes emphysema.
The inflammatory responses in chronic airway diseases leading to emphysema are not fully defined. We hypothesised that lung eosinophilia contributes to airspace enlargement in a mouse model and to emphysema in patients with chronic obstructive pulmonary disease (COPD).A transgenic mouse model of chronic type 2 pulmonary inflammation (I5/hE2) was used to examine eosinophil-dependent mechanisms leading to airspace enlargement. Human sputum samples were collected for translational studies examining eosinophilia and matrix metalloprotease (MMP)-12 levels in patients with chronic airways disease.Airspace enlargement was identified in I5/hE2 mice and was dependent on eosinophils. Examination of I5/hE2 bronchoalveolar lavage identified elevated MMP-12, a mediator of emphysema. We showed, in vitro, that eosinophil-derived interleukin (IL)-13 promoted alveolar macrophage MMP-12 production. Airspace enlargement in I5/hE2 mice was dependent on MMP-12 and eosinophil-derived IL-4/13. Consistent with this, MMP-12 was elevated in patients with sputum eosinophilia and computed tomography evidence of emphysema, and also negatively correlated with forced expiratory volume in 1 s.A mouse model of chronic type 2 pulmonary inflammation exhibited airspace enlargement dependent on MMP-12 and eosinophil-derived IL-4/13. In chronic airways disease patients, lung eosinophilia was associated with elevated MMP-12 levels, which was a predictor of emphysema. These findings suggest an underappreciated mechanism by which eosinophils contribute to the pathologies associated with asthma and COPD.


Kinetic Multi-omic Analysis of Responses to SARS-CoV-2 Infection in a Model of Severe COVID-19.
The host response to severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is poorly understood due to a lack of an animal model that recapitulates severe human disease. Here, we report a Syrian hamster model that develops progressive lethal pulmonary disease that closely mimics severe coronavirus disease 2019 (COVID-19). We evaluated host responses using a multi-omic, multiorgan approach to define proteome, phosphoproteome, and transcriptome changes. These data revealed both type I and type II interferon-stimulated gene and protein expression along with a progressive increase in chemokines, monocytes, and neutrophil-associated molecules throughout the course of infection that peaked in the later time points correlating with a rapidly developing diffuse alveolar destruction and pneumonia that persisted in the absence of active viral infection. Extrapulmonary proteome and phosphoproteome remodeling was detected in the heart and kidneys following viral infection. Together, our results provide a kinetic overview of multiorgan host responses to severe SARS-CoV-2 infection in vivo. IMPORTANCE The current pandemic caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection has created an urgent need to understand the pathogenesis of this infection. These efforts have been impaired by the lack of animal models that recapitulate severe coronavirus disease 2019 (COVID-19). Here, we report a hamster model that develops severe COVID-19-like disease following infection with human isolates of SARS-CoV-2. To better understand pathogenesis, we evaluated changes in gene transcription and protein expression over the course of infection to provide an integrated multiorgan kinetic analysis of the host response to infection. These data reveal a dynamic innate immune response to infection and corresponding immune pathologies consistent with severe human disease. Altogether, this model will be useful for understanding the pathogenesis of severe COVID-19 and for testing interventions.


Staging of the intracerebral inclusion body pathology associated with idiopathic Parkinson's disease (preclinical and clinical stages).
The synucleinopathy known as idiopathic Parkinson's disease (IPD) is a multi-system disorder in the course of which only a few predisposed nerve cell types in specific regions of the human brain become progressively involved. The underlying neuropathological process (formation of proteinaceous intraneuronal inclusion bodies) intracerebrally begins in clearly defined induction sites and advances in a topographically predictable sequence. Components of the autonomic, limbic, and motor systems sustain especially heavy damage. During the presymptomatic stages 1 and 2, the IPD-related inclusion body pathology remains confined to the medulla oblongata and olfactory bulb. In stages 3 and 4, the substantia nigra and other nuclear grays of the midbrain and basal forebrain are the focus of initially subtle and, then, severe changes. The illness reaches its symptomatic phase. In end-stages 5 and 6, the pathological process encroaches upon the telencephalic cortex. IPD manifests itself in all of its dimensions, which under the influence of the supervening cortical pathology are subject to increasing complexity.


A randomized, double-blind trial of lisinopril and losartan for the treatment of cardiomyopathy in duchenne muscular dystrophy.
This study sought to compare the effectiveness and safety of an angiotensin converting enzyme inhibitor (ACE-I) (lisinopril) vs. an angiotensin receptor blocker (ARB) (losartan) for the treatment of cardiomyopathy (CM) in boys with Duchenne muscular dystrophy (DMD). Development of CM is universal in boys with DMD. ACE-I and ARB have both been suggested as effective treatment options. ARBs have been associated with skeletal muscle regeneration in a mouse model of DMD. The question of which, if either, is more effective for CM treatment in DMD remains. The purpose of this multicenter double-blind prospective study was to compare efficacy and safety of lisinopril versus losartan in the treatment of newly diagnosed CM in boys with DMD. Echocardiographic technician inter- and intraobserver variability were tested on 2 separate days on 2 different boys with DMD CM. Results were compared with paired t-testing. Twenty-two boys with newly diagnosed DMD CM (echocardiographic ejection fraction (EF) 10% EF drop. Three boys in the aCE-I group had 3 visits, due to study funding termination. Two were withdrawn because of low EF. All their data are included in the analysis for as long as they remained in the study. Mean EF's were similar at baseline (47.5%- ACE-I, 48.4%- ARB). After 1 year each group significantly improved to 54.6% and 55.2% respectively (p=.02). There was no difference between the 2 treatment groups at 1 year. Inter-observer and intra-observer reliability studies showed no differences between echocardiographers on serial examinations. EF improved equally in the two groups. There is no therapeutic difference in EF improvement between lisinopril and losartan over the one-year duration for treatment of boys with DMD-related CM. ClinicalTrials.gov NCT01982695.


Long non-coding RNA and epigenetic gene regulation of KSHV.
Kaposi's sarcoma-associated herpesvirus (KSHV/human herpesvirus 8) is a γ-herpesvirus linked to Kaposi's sarcoma (KS) and two lymphoproliferative disorders, primary effusion lymphoma (PEL or body-cavity B-lymphoma [BCBL]) and a subset of Multicentric Castleman's Disease. During lytic growth, pervasive viral transcription generating a variety of transcripts with uncertain protein-coding potential has been described on a genome-wide scale in β- and γ-herpesviruses. One class of such RNAs is called long non-coding RNAs (lncRNAs). KSHV encodes a viral lncRNA known as polyadenylated nuclear RNA (PAN RNA), a copious early gene product. PAN RNA has been implicated in KSHV gene expression, replication, and immune modulation. PAN RNA expression is required for optimal expression of the entire KSHV lytic gene expression program. Latent KSHV episomes are coated with viral latency-associated nuclear antigen (LANA). LANA rapidly dissociates from episomes during reactivation. Here we review recent studies suggesting that PAN RNA may function as a viral lncRNA, including a role in the facilitation of LANA-episomal dissociation during lytic replication.


Early adaptations to training: upregulation of alpha-myosin heavy chain gene expression.
Chronic exercise induces adaptations that increase the functional capacity of the cardiovascular system. Aside from ventricular growth, these adaptations include a shift in the MHC isoenzyme pattern to enhance ventricular contractility. It is unclear whether adaptations by the contractile elements are an early event and specific to exercise, or whether they progress as a function of cardiac growth. Examining early adaptations to training is also important because it is during this period when the greatest imbalance between increased demand and functional capacity exists, and it is likely that the mechanisms responsible for propagating changes in the myocardial phenotype are most active. To determine whether changes in left ventricular (LV) contractile elements are an early adaptation to chronic exercise. Rats were randomly assigned to sedentary control or exercise training groups for 1 or 10 wk of training. After training, the LV was analyzed for protein by Western blot or mRNA by Northern and real-time QRT-PCR analysis. Plantaris cytochrome oxidase activity was significantly (P < 0.05) increased by 1 wk (+28%) or 10 wk (+32%) of training. Training significantly increased LV myofibrillar alpha-MHC protein and alpha-MHC-mRNA after both training periods. No changes in myofibrillar beta-MHC protein or beta-MHC-mRNA were observed. After 1 wk of training, LV skeletal alpha-actin-mRNA was significantly increased, whereas no changes were found for ANF, glyceraldehyde dehydrogenase, or cytochrome oxidase IV. Gel mobility shift analysis determined that YY1 DNA binding was significantly decreased in LV extracts from trained animals, although no change in YY1-mRNA expression was observed. Increased myofibrillar alpha-MHC protein and alpha-MHC-mRNA expression are early events in the adaptation to chronic exercise and occur before significant cardiac growth. These adaptations enhance myocardial contractility and permit increases in maximal cardiac output during heavy exercise.


Bronchiolitis obliterans organizing pneumonia in three children with acute leukaemias treated with cytosine arabinoside and anthracyclines.
Bronchiolitis obliterans organizing pneumonia (BOOP) is a clinicopathological entity with well-defined diagnostic criteria, which can be idiopathic or produced by a variety of biological processes. We describe the occurrence of BOOP in three children, one with acute lymphoblastic leukaemia and two with acute promyelocytic leukaemia. In the three patients, BOOP developed 10-20 days after a course of therapy with cytosine arabinoside and anthracyclines. The possible relationships between the small conducting airway lesions, lung toxic reaction to the drugs and/or nonidentified infectious agents are discussed.


Catalytic and potentiometric characterization of E201D and E201Q mutants of Trypanosoma congolense trypanothione reductase.
Trypanothione reductase is a member of the structurally and functionally well-characterized family of flavoprotein reductases, which catalyze the reduced pyridine nucleotide dependent reduction of their disulfide, peroxide, or metal ion substrates. Trypanothione reductase is found in a wide variety of Trypanosoma species, where the enzyme serves physiologically to protect the organism from oxidative stress and assists in maintaining low intracellular levels of hydrogen peroxide. The redox potential of the flavin and the hydride ion transfer reaction of the pro-S hydrogen of NADPH to N5 of FAD have been proposed to be influenced by the presence of a conserved Lys-Glu (K60-E201) ion pair at the bottom of the nicotinamide binding pocket. We have evaluated this hypothesis by making modest substitutions for both the Lys and Glu residues using site-directed mutagenesis. Replacement of the K60 residue with an arginine led to a poorly expressed, and completely inactive, enzyme. Replacement of the Glu201 residue with either a glutamine (E201Q) or an aspartate (E201D) residue led to expressed enzymes which could be readily purified in > 20 mg amounts using protocols developed for the WT enzyme, and which had significant residual trypanothione-reducing activity. These enzymes have now been characterized to determine their redox potentials, catalytic activities, and nucleotide specificities. Relative to the WT enzyme, both E201D and E201Q exhibit ca. 5% of WT trypanothione-reducing activity using NADPH as reductant, but significantly enhanced quinone reductase activity. The oxidase activity of both mutants is enhanced by over 50-fold compared to that of the WT. The redox potential of the WT enzyme has been determined to be -273 mV, while both the E201D and E201Q exhibit more positive redox potentials (-259 and -251 mV, respectively).(ABSTRACT TRUNCATED AT 250 WORDS)


Novel interactions of domain III from the envelope glycoprotein of dengue 2 virus with human plasma proteins.
Blood cells and plasma are important media for the four serotypes of dengue virus (DENV1-4) spreading into an infected person. Thus, interactions with human plasma proteins are expected to be decisive in the course of the viral infection. Affinity purification followed by MS analysis (AP/MS) was used to isolate and identify plasma-derived proteins capable to interact with a recombinant protein comprising the domain III of the envelope protein of DENV2 (DIIIE2). The elution of the AP potently inhibits DENV2 infection. Twenty-nine proteins were identified using a label-free approach as specifically captured by DIIIE2. Of these, a direct interaction with C reactive protein, thrombin and Inter-alpha-inhibitor complexes was confirmed by ELISA. Results provide further evidence of a significant representation of proteins from complement and coagulation cascades on DENV2 interactome in human plasma and stand out the domain III of the viral envelope protein as participant on these interactions. A functional clustering analysis highlights the presence of three structural motifs among putative DIIIE2-binding proteins: hydroxylation and EGF-like calcium-binding- and Gla domains. Early cycles of dengue virus replication take place in human blood cells. Thus, the characterization of the interactome of dengue virus proteins in human plasma can lead to the identification of pivotal interactions for the infection that can eventually constitute the target for the development of methods to control dengue virus-caused disease. In this work we identified 29 proteins from human plasma that potentially interact with the envelope protein of dengue 2 virus either directly or through co-complex formation. C reactive protein, thrombin and Inter-alpha-inhibitor complexes were validated as interactors of the domain III of the envelope protein of dengue 2. Results highlight the presence of three structural motifs among putative DIIIE2-binding proteins: hydroxylation and EGF-like calcium-binding- and Gla domains. This finding together with the participation of domain III of the envelope protein on the interactions with human plasma proteins should contribute to a better understanding of dengue virus interactome in human plasma. Such knowledge can contribute to the development of more effective treatments to infected persons.


Histone acetylation and the cell-cycle in cancer.
A number of distinct surveillance systems are found in mammalian cells that have the capacity to interrupt normal cell-cycle progression. These are referred to as cell cycle check points. Surveillance systems activated by DNA damage act at three stages, one at the G1/S phase boundary, one that monitors progression through S phase and one at the G2/M boundary. The initiation of DNA synthesis and irrevocable progression through G1 phase represents an additional checkpoint when the cell commits to DNA synthesis. Transition through the cell cycle is regulated by a family of protein kinase holoenzymes, the cyclin-dependent kinases (Cdks), and their heterodimeric cyclin partner. Orderly progression through the cell-cycle checkpoints involves coordinated activation of the Cdks that, in the presence of an associated Cdk-activating kinase (CAK), phosphorylate target substrates including members of the "pocket protein" family. One of these, the product of the retinoblastoma susceptibility gene (the pRB protein), is phosphorylated sequentially by both cyclin D/Cdk4 complexes and cyclin E/Cdk2 kinases. Recent studies have identified important cross talk between the cell-cycle regulatory apparatus and proteins regulating histone acetylation. pRB binds both E2F proteins and histone deacetylase (HDAC) complexes. HDAC plays an important role in pRB tumor suppression function and transcriptional repression. Histones are required for accurate assembly of chromatin and the induction of histone gene expression is tightly coordinated. Recent studies have identified an important alternate substrate of cyclin E/Cdk2, NPAT (nuclear protein mapped to the ATM locus) which plays a critical role in promoting cell-cycle progression in the absence of pRB, and contributes to cell-cycle regulated histone gene expression. The acetylation of histones by a number of histone acetyl transferases (HATs) also plays an important role in coordinating gene expression and cell-cycle progression. Components of the cell-cycle regulatory apparatus are both regulated by HATs and bind directly to HATs. Finally transcription factors have been identified as substrate for HATs. Mutations of these transcription factors at their sites of acetylation has been associated with constitutive activity and enhanced cellular proliferation, suggesting an important role for acetylation in transcriptional repression as well as activation. Together these studies provide a working model in which the cell-cycle regulatory kinases phosphorylate and inactivate HDACs, coordinate histone gene expression and bind to histone acetylases themselves. The recent evidence for cross-talk between the cyclin-dependent kinases and histone gene expression on the one hand and cyclin-dependent regulation of histone acetylases on the other, suggests chemotherapeutics targeting histone acetylation may have complex and possibly complementary effects with agents targeting Cdks.


Cloning and expression of a Streptococcus sanguis surface antigen that interacts with a human salivary agglutinin.
Human saliva contains a high-molecular-weight glycoprotein (agglutinin) which binds to specific streptococci in a calcium-dependent reaction leading to the formation of bacterial aggregates. We report the cloning of a gene encoding a surface antigen from Streptococcus sanguis M5 and show that the expressed protein inhibits agglutinin-mediated aggregation and specifically binds the salivary agglutinin in a calcium-dependent fashion. Clones isolated from the immunological screening of S. sanguis M5 genomic libraries with polyclonal antibodies against whole cells were assayed for the ability to compete with S. sanguis for agglutinin. One clone, pSSP-5, expressed antigens of 165 and 130 kilodaltons (kDa) possessing this activity. A 3-kilobase-pair (kbp) insert fragment from this clone was used to screen a genomic library in lambda EMBL3 which resulted in the isolation of clone SSP-5A. This clone contained an insert of 17 kb and expressed proteins of 170 to 205 kDa that reacted with the anti-S. sanguis antibodies. Subcloning of a 5.3-kbp EcoRI-BamHI fragment from SSP-5A produced pEB-5, which expressed streptococcal components that were indistinguishable from SSP-5A. The streptococcal antigen was purified by gel permeation and ion exchange chromatography and shown to potently compete with S. sanguis M5 cells for agglutinin. The antigen also bound purified salivary agglutinin in the presence of 1 mM CaCl2. This binding was inhibited by EDTA. Both the SSP-5 antigen and a 205-kDa protein in surface protein extracts from S. sanguis M5 cross-reacted with antibodies directed against antigen B from S. mutans and SpaA from S. sobrinus 6715. These results indicate that a 205-kDa surface protein that is antigenically related to SpaA and antigen B is involved in the binding of salivary agglutinin to S. sanguis M5.