bluuebunny/crossref_metadata_2025 huggingface dataset:
Combined Use of Gas Chromatography, Mass Spectrometry, and Computer in Diagnosis and Studies of Metabolic Disorders
Authors: Jellum E , Stokke O , Eldjarn L
Published: January 2020

We describe a system for multicomponent analyses of biological materials. The methods include gas liquid chromatography (GLC) for separation purpose, mass spectrometry for identification and structure studies, and use of a computer for data handling. Urine and serum samples as well as biopsies and other biological materials can be analyzed. The peaks from the eight different GLC-systems used are identified by computer matching of the corresponding mass spectra against a library file of 17,000 reference spectra. The system is well suited for the diagnosis and study of inherited as well as other metabolic disturbances. About 40 of the known inborn errors can be detected. The application of the methods to the analysis of more than 700 patients has led to the discovery of three new inborn errors: methylmalonic aciduria, β-methylcrotonyl-CoA carboxylase deficiency, and pyroglutamic aciduria. The methods also represent a valuable tool in the study of drug metabolism.
Application of a New Peroxide Indicator Reaction to the Specific, Automated Determination of Glucose with Glucose Oxidase
Authors: Gochman Nathan , Schmitz Joan M
Published: January 2020

We describe an automated, colorimetric determination of glucose in biological fluids that combines the specificity of glucose oxidase and of a new peroxide indicator reaction. In the presence of peroxidase, 3-methyl-2-benzothiazolinone hydrazone oxidatively couples with N,N-dimethylaniline to form a stable, intensely colored, water-soluble indamine dye, the concentration of which is proportional to that of the third reactant, hydrogen peroxide. This reaction, used earlier to determine uric acid [Clin. Chem. 17, 1154 (1971)], is substantially less affected by negative interference of reducing substances than are previously described peroxide indicators. Results from use of AutoAnalyzers I and II and this method were compared with those from a manual spectrophotometric hexokinase/glucose-6-phosphate dehydrogenase procedure, and showed good correlation for specimens from patients. The automated methods are suitable for measuring glucose in serum, plasma from fluoride-or iodoacetate-preserved blood, urine (without ion-exchange pretreatment), or cerebrospinal fluid. They eliminate the problem of falsely high results caused by medication or reducing metabolites associated with uremia, in methods in which alkaline ferricyanide or copper—neocuproine is used.
Measurements of Nickel in Serum and Urine as Indices of Environmental Exposure to Nickel
Authors: McNeely Michael D , Nechay Maria W , Sunderman F William
Published: January 2020

Nickel was measured, by atomic absorption spectrometry, in serum and urine specimens from: (a) healthy hospital employees (age 19-62) who had resided >1 year in Sudbury, Ontario, and (b) healthy hospital employees (age 18-62) who had resided >1 year in Hartford, Connecticut. Subjects in groups a and b were matched according to age and sex. None of the subjects had occupational exposure to nickel. Nickel was analyzed in duplicate on a "blind" basis with specimens from groups a and b interspersed within each run. In population a, serum nickel concentrations averaged 4.6 ± 1.4 µg/liter (n = 25), urine nickel excretion 7.9 ± 3.7 µg/day (n = 19). In population b, serum nickel concentrations averaged 2.6 ± 1.0 µg/liter (n = 26), urine nickel excretion 2.5 ± 1.4 µg/ day (n = 20). These population means were all significantly different (P <0.001, t test). Serum and urinary nickel concentrations were not significantly influenced by age or sex, nor were they correlated with length of residence in either location. Nickel concentrations in municipal tap water averaged 200 ± 43 µg/liter in Sudbury (seven sites), and 1.1 ± 0.3 µg/liter in Hartford (five sites). Measurements of nickel in serum and urine are valid biological indices of environmental exposure to nickel.
Simplified Colorimetric Determination of Thiocyanate in Biological Fluids, and Its Application to Investigation of the Toxic Amblyopias
Authors: Pettigrew A R , Fell G S
Published: January 2020

A colorimetric procedure for determination of small amounts of cyanide and thiocyanate, involving the synthesis of a pyridine dyestuff by the reaction of pyridine and an aromatic amine, has been simplified for the estimation of thiocyanate alone in biological fluids. Replacement of benzidine with p-phenylenediamine in the colorimetric reaction has both improved the precision of the analytical procedure and avoided a carcinogenic hazard. This method has been used to follow the decrease in plasma thiocyanate associated with abstinence from cigarette smoking, and its subsequent increase upon resumption. It has also been used to measure the plasma and urinary thiocyanate concentrations of patients suffering from the particular toxic amblyopias—tobacco amblyopia and Leber’s hereditary optic atrophy— believed to be associated with cyanide toxicity, and to follow the increased thiocyanate concentrations that accompany significant improvements in the patients’ vision brought about by various treatments.
Anomalous Response of Macroamylase to Assay Temperature
Authors: Henderson Arthur R , King John , Imrie Clement W
Published: January 2020

An anomalous response to increased reaction temperature is described for two samples of macroamylase, as contrasted with normal serum and pancreatic amylases. A possible method of differentiating macroamylase by assays of activities at 45°C and 25°C is suggested. The ratios of activities found at these two temperatures for a normoamylasemic population are given
The Use of Laboratory Computers in Monitoring Kinetic Enzyme Assays
Authors: Hicks G Phillip , Ziesemer R A , Tietz Norbert W
Published: January 2020

An online computer program to monitor kinetic enzyme assays is described. The program analyzes the kinetic data in a manner similar to the way a technologist handles data in a manual procedure, taking into account the lag phase, substrate depletion phase, and linear portion of the rate curve. Thus, complete automation of even complex kinetic assays has been made practical. The program has been implemented for routine use in a clinical laboratory computer system ("LABCOM"), and the results correlate well with those obtained by established methods of manual data-handling procedures
Simplified, Rapid Measurement of [131I] Triiodothyronine Uptake, with Use of Competitive Charcoal Adsorption
Authors: Holub William R , Tortorella Catherine F , Ruhl Madelaine R
Published: January 2020

Hemoglobin-coated charcoal has been used as a competitive binder in developing a [131I] triiodothyronine uptake assay. Modifying the procedures of Herbert et al. [J. Lab. Clin. Med.66, 814 (1965)] and Braverman et al. [J. Amer. Med. Ass.199, 469 (1967)] and examining various optimum test parameters, we developed a rapid, simpler technique. Correlation with results of the "Triosorb-131" resin sponge assay (Abbott Laboratories) was 0.909. Inter-run precision for a lyophilized serum control was satisfactory (CV, 2.5%). Advantages of the present method include: rapid equilibration time (10 min), easily prepared and dispensed adsorbent, nonquantitative transfer of final reaction mixture, and a linear response to variation in concentrations of binding protein that is of use in the diagnosis of thyroid dysfunction
Ammonium Thymolphthalein Monophosphate as a New Substrate for Alkaline and Acid Phosphatase Determinations in Serum
Authors: Morin Leo G
Published: January 2020

A new substrate, ammonium thymolphthalein monophosphate, is proposed for use in alkaline and acid phosphatase determinations. The ammonium salt shares the advantages of the sodium salt, but is a more stable substrate and results in more stable working reagents. The substrate is buffered at pH 10.1 in diethanolamine for determining alkaline phosphatase activity in 5 min, and at pH 5.9 in citrate buffer for determining acid phosphatase activity in 30 min. The substrate is relatively specific for bone and liver alkaline phosphatase and prostatic acid phosphatase. Coefficients of variation were 2.5% and 6.6% for alkaline and acid phosphatase, respectively.
Bilirubin Binding Affinity of Serum: Comparison of Qualitative and Quantitative Sephadex Gel Filtration Methods
Authors: Kaufmann N A , Kapitulnik J , Blondheim S H
Published: January 2020

A simple, 30-min method of estimating the binding of bilirubin to serum protein was compared with a quantitative method. Good correlation was found between the two methods, both based on Sephadex gel filtration. The results indicate that the presence of loosely bound bilirubin can be determined with this simple, qualitative, semimicro-scale test. This determination is important for the clinical evaluation of jaundiced infants, since it may help to determine whether and when prophylactic therapy to prevent bilirubin encephalopathy is appropriate.
Coupling of Glucose-6-Phosphate Dehydrogenase and Hemoglobin S Screening to Automated Determinations of Blood Group and Type
Authors: Maples John S , Dickson Larry G , Uddin David E
Published: January 2020

Automated analyses for glucose-6-phosphate dehydrogenase (EC 1.1.1.49, GPD) and for hemoglobin S have recently been described. We modified these methods to couple them directly to the sample line of the 15-channel Technicon blood-typing analyzer. Rate of analysis is doubled (to 120 per hour) by using the reverse-washout cycle to decrease sample-to-sample interaction. Savings in reagents and technician time are significant.
Radioimmunoassay
Authors: Skelley D S , Brown L P , Besch P K
Published: January 2020

Principles of radioimmunoassay are discussed. Methods are outlined for preparing radiolabeled antigenic compounds and characterizing antibodies. The various techniques for separating bound and free antigen are also reviewed. Commercial suppliers of various components needed for radioimmunoassay are listed. Also presented is a selected list of normal values for hormonal and nonhormonal substances from humans.
Specific Analysis of Immunoglobulins. Techniques and Clinical Value
Authors: Heremans Joseph F , Masson Pierre L
Published: January 2020

We recommend that routine screening of sera for immunoglobulin disorders consist of a combination of electrophoresis in agarose and immunochemical quantitation of serum IgG, IgA, and IgM. Abnormalities detected by this procedure may be classified as (a) immune deficiencies, (b) polyclonal hyperimmunoglobulinemias, or (c) monoclonal immunoglobulin disorders. The analyst can contribute to the diagnosis of immune deficiency by detecting evidence for plasma protein losses in the urine, unsuspected monoclonal immunoglobulin disorders, and abnormalities in the immunoglobulin content and spectrum of secretions. Polyclonal hyperimmunoglobulinemia —seen in infections, autoimmune disease, liver disease, sarcoidosis, or other conditions—is frequently associated with an imbalance between the different immunoglobulin classes, and with the appearance of discrete inhomogeneities in the immunoglobulin zones of the electrophoretic spectrum. Factors involved in this process are listed, and examples are indicated. Monoclonal immunoglobulin disorders may not be detected if the tumor protein is masked by normal serum-protein fractions or misdiagnosed by confusion with nonimmunoglobulin components. Diagnosis of immunocyte malignancy is aided by detection of Bence Jones protein in the urine; measurements of serum IgG, IgA, and IgM concentrations; and monitoring the concentration of the abnormal component in the serum as the disease evolves.
Theoretical Considerations and Models of Red and White Cell Pentose Cycles
Authors: Hodges James M , Grümer H-D
Published: January 2020

The contribution of pentose cycle activity to glucose metabolism is usually estimated by cumbersome, time-consuming techniques such as isolating and degrading products of specifically labeled radioactive precursors, to localize the radioactivity in individual carbon atoms. Equations for the calculation of pentose cycle activity are based on models that assume either no recycling or complete recycling of pentose cycle intermediates, as well as incomplete metabolism of glucose by the cycle. A simpler and more reliable way to determine pentose cycle activity is needed. The present investigation is the development of an improved model for the evaluation of the relative extent of glucose metabolism via the pentose cycle.
Stable Reagents for Determination of Serum Triglycerides by a Colorimetric Hantzsch Condensation Method
Authors: Foster Lowell B , Dunn Ralph T
Published: January 2020

We describe a modified method for determining triglycerides in serum, in which alumina is used to adsorb interfering substances, and the glycerol content of triglycerides is determined colorimetrically. In contrast to all previous Hantzsch condensation procedures in which periodate and acetylacetone are used, the working periodate and the working acetylacetone reagents are stable for at least six months because they are differently formulated. The principle that governs this increased stability applies to all Hantzsch condensation methods for triglycerides.
Turbidimetric Procedure for Determination of Lipase Activity
Authors: Burlina Angelo , Galzigna Lauro
Published: January 2020

A method is described for turbidimetric assay of lipase activity. The substrate is a two-phase emulsion composed of a fairly homogenous dispersion of micelles of either triolein or olive oil. The de-emulsification of the substrate after lipase addition is a true measure of the enzymatic activity. Turbidity change is related to amount of the enzyme by use of a commercial standard, purified lipase from hog pancreas. The quantitative relationship between turbidity change and the number of micelles provides the theoretical basis for this type of assay.
Influence of Demographic Factors on Serum Concentrations of Seven Chemical Constituents in Healthy Human Subjects
Authors: Goldberg David M , Handyside Alan J , Winfield David A
Published: January 2020

Serum from 519 subjects attending a screening clinic and from the laboratory staff was used to study the influence of age, sex, body weight, social class, blood pressure, and smoking habit upon the concentration in serum of urea, uric acid, cholesterol, inorganic phosphate, total protein, calcium, and magnesium. Values for urea, uric acid, calcium, and magnesium were significantly higher in males. Age was a positive determinant of urea and cholesterol in both sexes, and of magnesium in females only. The correlation of uric acid with age was positive in females and negative in males, but in both sexes the correlation between uric acid and weight was strongly positive. Regression equations were developed to express the demographic and nondemographic contributions to the individual's blood chemistry values. From these, a set of demographically-corrected reference values were derived. When applied to a selected group of female hospital outpatients, these seemed to discriminate more accurately between subjects with and without disease than did conventional parametric limits.
An All Solid-State Electrochemical Electrolyte Analyzer
Authors: Smith Melvin D , Rogers Robert W , Genshaw Marvin A , Greyson Jerome
Published: January 2020

Ion-selective electrodes have been applied to analysis of blood electrolytes with some success. However, currently available ion-selective electrode systems tend to be elaborate and expensive. We describe a clinical electrolyte analyzer in which inexpensive ion-selective electrodes are used. The electrodes are in the configuration of thin wires and are all solid in construction. They may be dipped directly into undiluted 250-µl samples of serum or plasma, are nondestructive of the sample, and may be used with an expanded-scale pH meter, although a more sensitive electrometer is preferred. The complete electrolyte analyzer consists of an electrode holder, into which the electrodes are plugged, and a solid-state digital electrometer that displays units of electrolyte concentration. A discussion of the thermodynamics essential to the construction of reversible electrodes is also presented, as well as the results of a clinical study in which it is shown that data from a flame photometer and the electrochemical analyzer compare favorably.
The pathogenic role of coronary microvascular dysfunction in the setting of other cardiac or systemic conditions
Authors: Konst Regina E , Guzik Tomasz J , Kaski Juan-Carlos , Maas Angela H E M , Elias-Smale Suzette E
Published: January 2020

Coronary microvascular dysfunction (CMD) plays a pathogenic role in cardiac and systemic conditions other than microvascular angina. In this review, we provide an overview of the pathogenic role of CMD in the setting of diabetes mellitus, obesity, hypertensive pregnancy disorders, chronic inflammatory and autoimmune rheumatic disorders, chronic kidney disease, hypertrophic cardiomyopathy, and aortic valve stenosis. In these various conditions, CMD results from different structural, functional, and/or dynamic alterations in the coronary microcirculation associated with the primary disease process. CMD is often detectable very early in the course of the primary disease, before clinical symptoms or signs of myocardial ischaemia are present, and it portrays an increased risk for cardiovascular events.
Inhibition of N-type calcium channels in cardiac sympathetic neurons attenuates ventricular arrhythmogenesis in heart failure
Authors: Zhang Dongze , Tu Huiyin , Wang Chaojun , Cao Liang , Hu Wenfeng , Hackfort Bryan T , Muelleman Robert L , Wadman Michael C , Li Yu-Long
Published: January 2020

Aims

Cardiac sympathetic overactivation is an important trigger of ventricular arrhythmias in patients with chronic heart failure (CHF). Our previous study demonstrated that N-type calcium (Cav2.2) currents in cardiac sympathetic post-ganglionic (CSP) neurons were increased in CHF. This study investigated the contribution of Cav2.2 channels in cardiac sympathetic overactivation and ventricular arrhythmogenesis in CHF.

Methods and results

Rat CHF was induced by surgical ligation of the left coronary artery. Lentiviral Cav2.2-α shRNA or scrambled shRNA was transfected in vivo into stellate ganglia (SG) in CHF rats. Final experiments were performed at 14 weeks after coronary artery ligation. Real-time polymerase chain reaction and western blot data showed that in vivo transfection of Cav2.2-α shRNA reduced the expression of Cav2.2-α mRNA and protein in the SG in CHF rats. Cav2.2-α shRNA also reduced Cav2.2 currents and cell excitability of CSP neurons and attenuated cardiac sympathetic nerve activities (CSNA) in CHF rats. The power spectral analysis of heart rate variability (HRV) further revealed that transfection of Cav2.2-α shRNA in the SG normalized CHF-caused cardiac sympathetic overactivation in conscious rats. Twenty-four-hour continuous telemetry electrocardiogram recording revealed that this Cav2.2-α shRNA not only decreased incidence and duration of ventricular tachycardia/ventricular fibrillation but also improved CHF-induced heterogeneity of ventricular electrical activity in conscious CHF rats. Cav2.2-α shRNA also decreased susceptibility to ventricular arrhythmias in anaesthetized CHF rats. However, Cav2.2-α shRNA failed to improve CHF-induced cardiac contractile dysfunction. Scrambled shRNA did not affect Cav2.2 currents and cell excitability of CSP neurons, CSNA, HRV, and ventricular arrhythmogenesis in CHF rats.

Conclusions

Overactivation of Cav2.2 channels in CSP neurons contributes to cardiac sympathetic hyperactivation and ventricular arrhythmogenesis in CHF. This suggests that discovering purely selective and potent small-molecule Cav2.2 channel blockers could be a potential therapeutic strategy to decrease fatal ventricular arrhythmias in CHF.
Noise annoyance predicts symptoms of depression, anxiety and sleep disturbance 5 years later. Findings from the Gutenberg Health Study
Authors: Beutel Manfred E , Brähler Elmar , Ernst Mareike , Klein Eva , Reiner Iris , Wiltink Jörg , Michal Matthias , Wild Philipp S , Schulz Andreas , Münzel Thomas , Hahad Omar , König Jochem , Lackner Karl J , Pfeiffer Norbert , Tibubos Ana N
Published: January 2020

Background

Cross-sectional studies have shown that noise annoyance is strongly associated with mental distress, however, its long-term effects on mental health is unknown. We therefore investigated whether noise annoyance predicts depression, anxiety and sleep disturbance in a large, representative sample 5 years later.

Methods

We investigated longitudinal data of N = 11 905 participants of the Gutenberg Health Study, a population-based, prospective, single-centre cohort study in mid-Germany (age at baseline 35–74 years). Noise annoyance was assessed at baseline and 5-year follow-up (sources: road traffic, aircraft, railways, industrial, neighbourhood indoor and outdoor noise; and day vs. nighttime). Depression, anxiety and sleep disturbance were assessed using the Patient Health Questionnaire-9 and Generalized Anxiety Disorder-2. Participants suffering from depression, anxiety or sleep disturbance at baseline were excluded from the respective multivariate analyses of new onset at follow-up.

Results

General noise annoyance remained stable. Daytime noise annoyance predicted new onset of depressive, anxiety symptoms (also nighttime annoyance) and sleep disturbance (beyond respective baseline scores). Additional predictors were female sex, lower age and low socioeconomic status (SES). Regarding specific sources, daytime baseline aircraft annoyance predicted depression and anxiety. Sleep disturbance was most consistently predicted by neighbourhood annoyance (baseline and follow-up) and follow-up annoyance by aircraft (night) and road traffic (day and night).

Conclusions

We identified current and past noise annoyances as risk factors for mental distress and sleep disturbance. Furthermore, women, younger adults and those with lower SES are particularly susceptible to noise annoyance. Our results indicate the need to provide regulatory measures in affected areas to prevent mental health problems.
Functions of reactive oxygen species in apoptosis and ganoderic acid biosynthesis in Ganoderma lucidum
Authors: Zhu Jing , Wu Fengli , Yue Sining , Chen Chen , Song Shuqi , Wang Hui , Zhao Mingwen
Published: January 2020

Ganoderma lucidum is a medicinal fungus that is widely used in traditional medicine. Fungal PacC is recognized as an important transcription factor that functions during adaptation to environmental pH, fungal development and secondary metabolism. Previous studies have revealed that GlPacC plays important roles in mycelial growth, fruiting body development and ganoderic acid (GA) biosynthesis. In this study, using a terminal deoxynucleotidyl transferase-mediated dUTP nick end labelling (TUNEL) assay, we found that the apoptosis level was increased when PacC was silenced. The transcript and activity levels of caspase-like proteins were significantly increased in the PacC-silenced (PacCi) strains compared with the control strains. Silencing PacC also resulted in an increased reactive oxygen species (ROS) levels (∼2-fold) and decreased activity levels of enzymes involved in the antioxidant system. Further, we found that the intracellular ROS levels contributed to apoptosis and GA biosynthesis. Adding N-acetyl-cysteine and vitamin C decreased intracellular ROS and resulted in the inhibition of apoptosis in the PacCi strains. Additionally, the GA biosynthesis was different between the control strains and the PacCi strains after intracellular ROS was eliminated. Taken together, the findings showed that silencing PacC can result in an intracellular ROS burst, which increases cell apoptosis and GA biosynthesis levels. Our study provides novel insight into the functions of PacC in filamentous fungi.
Agricultural production systems can serve as reservoir for human pathogens
Authors: Schierstaedt Jasper , Grosch Rita , Schikora Adam
Published: January 2020

Food-borne diseases are a threat to human health and can cause severe economic losses. Nowadays, in a growing and increasingly interconnected world, food-borne diseases need to be dealt with in a global manner. In order to tackle this issue, it is essential to consider all possible entry routes of human pathogens into the production chain. Besides the post-harvest handling of the fresh produce itself, also the prevention of contamination in livestock and agricultural soils are of particular importance. While the monitoring of human pathogens and intervening measures are relatively easy to apply in livestock and post-harvest, the investigation of the prevention strategies in crop fields is a challenging task. Furthermore, crop fields are interconnected with livestock via fertilizers and feed; therefore, a poor hygiene management can cause cross-contamination. In this review, we highlight the possible contamination of crop plants by bacterial human pathogens via the rhizosphere, their interaction with the plant and possible intervention strategies. Furthermore, we discuss critical issues and questions that are still open.
Nutrient Removals by Pinestraw Harvesting in Slash Pine Plantations in Florida
Authors: Osiecka Anna , Minogue Patrick J , Miwa Masato , Lauer Dwight K
Published: January 2020

Pinestraw harvesting is an important industry in the southeastern United States. There is a need to understand how fertilization can be used efficiently to sustain or increase long-term pinestraw yields and avoid adverse environmental consequences. The effects of fertilization on needlefall nutrient concentrations, pinestraw yields, and nutrient removals on soils with contrasting soil nutrient sorption potential (Entisol vs. Ultisol) were compared using two midrotation slash pine plantations in North Florida. Diammonium phosphate was applied at 0, 144, 430, or 718 kg ha–1 in the spring of 2009 and 2010. Pinestraw was harvested annually in 2009–12. Needlefall mass, pinestraw yields, total Kjeldahl nitrogen concentrations in needlefall and pinestraw, and total Kjeldahl nitrogen, P, K, Ca, and Mg removals increased with fertilization. Diammonium phosphate at 718 kg ha–1 year–1 increased pinestraw yield over the control by 37 and 35 percent 2 years after the second fertilization, and by 11 percent (from 21.5 to 23.8 Mg ha–1) and 12 percent (from 25.0 to 28.1 Mg ha–1) over the 4-year control totals, at Entisol and Ultisol sites, respectively. Differences between sites were larger than fertilization response for most variables. Yields, nutrient concentrations, and removals were higher at the more fertile Ultisol than Entisol and, at both sites, higher than most reported in the literature.
Peripheral Blood Eosinophilia and Long-term Severity in Pediatric-Onset Inflammatory Bowel Disease
Authors: Prathapan Krishnapriya Marangattu , Ramos Rivers Claudia , Anderson Alyce , Koutroumpakis Filippos , Koutroubakis Ioannis E , Babichenko Dmitriy , Tan Xiaoqing , Tang Gong , Schwartz Marc , Proksell Siobhan , Johnston Elyse , Hashash Jana G , Dunn Michael , Wilson Annette , Barrie Arthur , Harrison Janet , Hartman Douglas , Kim Sandra C , Binion David G
Published: January 2020

Background

Peripheral blood eosinophilia (PBE) is a biomarker of an aggressive multiyear natural history in adults with inflammatory bowel diseases (IBDs). Additionally, PBE at diagnosis is associated with higher disease activity in pediatric-onset IBD. We sought to determine if PBE can function as a biomarker of long-term disease severity in pediatric-onset IBD patients who are followed into adulthood.

Methods

We analyzed a consented, prospective, natural history IBD registry at an adult tertiary center from 2009 to 2018. Prevalence of PBE was evaluated in both pediatric- and adult-onset IBD patients. Demographics, clinical characteristics, and health care utilization data were compared in patients with and without PBE.

Results

Among 2800 adult IBD patients, 23.4% had pediatric-onset disease. PBE was found in 34% of the pediatric-onset patients compared with 26.8% of the adult-onset IBD patients (P < 0.001). In the pediatric-onset IBD cohort, PBE was associated with higher rates of allergies (P < 0.0001), but not of asthma, allergic rhinitis, or primary sclerosing cholangitis. In the adult IBD patients with pediatric-onset disease, PBE was associated with higher rates of C-reactive protein elevation (P < 0.0001), erythrocyte sedimentation rate elevation (P < 0.0001), higher health care utilization, and higher average health care charges per year (P < 0.00001).

Conclusions

Peripheral blood eosinophilia was more prevalent in adult IBD patients with pediatric-onset compared with adult-onset disease. Among all IBD patients with long-term follow-up, PBE defined a subgroup with more severe illness. These data suggest that PBE may be a biomarker for a high-risk subgroup with high cost trajectory and long-term severity in pediatric-onset IBD that persists into adulthood.
The role of fractional flow reserve in coronary artery bypass graft surgery: a meta-analysis
Authors: Jayakumar Shruti , Bilkhu Rajdeep , Ayis Salma , Nowell Justin , Bogle Richard , Jahangiri Marjan
Published: January 2020

OBJECTIVES

Fractional flow reserve (FFR) measures the drop in perfusion pressure across a stenosis, therefore representing its physiological effect on myocardial blood flow. Its use is widespread in percutaneous coronary interventions, though its role in coronary artery bypass graft (CABG) surgery remains uncertain. This systematic review and meta-analysis aims to evaluate current evidence on outcomes following FFR-guided CABG compared to angiography-guided CABG.

METHODS

A literature search was conducted following PRISMA (Preferred Reporting Items for Systematic Reviews and Meta-Analyses) guidelines to identify all relevant articles. Patient demographics and characteristics were extracted. The following outcomes were analysed: repeat revascularization, myocardial infarction (MI) and all-cause mortality. Pooled relative risks were analysed and their 95% confidence intervals (CIs) were estimated using random-effects models; P-value <0.05 was considered statistically significant. Heterogeneity was assessed with Cochran’s Q score and quantified by I2 index.

RESULTS

Nine studies with 1146 patients (FFR: 574, angiography: 572) were included. There was no difference in MI or repeat revascularization between the 2 groups (relative risk 0.76, 95% CI 0.41–1.43; P = 0.40, and relative risk 1.28, 95% CI 0.75–2.19; P = 0.36, respectively). There was a significant reduction in all-cause mortality in the FFR-guided CABG group compared to angiography-guided CABG, which was not specifically cardiac related (relative risk 0.58, 95% CI 0.38–0.90; P = 0.02).

CONCLUSIONS

There was no reduction in repeat revascularization or postoperative MI with FFR. In this fairly small cohort, FFR-guided CABG provided a reduction in mortality, but this was not reported to be due to cardiac causes. There may be a role for FFR in CABG, but large-scale randomized trials are required to establish its value.
Injection Drug Use-Associated Candidemia: Incidence, Clinical Features, and Outcomes, East Tennessee, 2014–2018
Authors: Rossow John A , Gharpure Radhika , Brennan Julia , Relan Pryanka , Williams Sabrina R , Vallabhaneni Snigdha , Jackson Brendan R , Graber Caroline R , Hillis Sherry R , Schaffner William , Dunn John R , Jones Timothy F
Published: January 2020

Background

Injection drug use (IDU) is an established but uncommon risk factor for candidemia. Surveillance for candidemia is conducted in East Tennessee, an area heavily impacted by the opioid crisis and IDU. We evaluated IDU-associated candidemia to characterize the epidemiology and estimate the burden.

Methods

We assessed the proportion of candidemia cases related to IDU during January 1, 2014–September 30, 2018, estimated candidemia incidence in the overall population and among persons who inject drugs (PWID), and reviewed medical records to compare clinical features and outcomes among IDU-associated and non-IDU candidemia cases.

Results

The proportion of IDU-associated candidemia cases in East Tennessee increased from 6.1% in 2014 to 14.5% in 2018. Overall candidemia incidence in East Tennessee was 13.5/100 000, and incidence among PWID was 402–1895/100 000. Injection drug use-associated cases were younger (median age, 34.5 vs 60 years) and more frequently had endocarditis (39% vs 3%). All-cause 30-day mortality was 8% among IDU-associated cases versus 25% among non-IDU cases.

Conclusions

A growing proportion of candidemia in East Tennessee is associated with IDU, posing an additional burden from the opioid crisis. The lower mortality among IDU-associated cases likely reflects in part the younger demographic; however, Candida endocarditis seen among approximately 40% underscores the seriousness of the infection and need for prevention.
A novel tigecycline resistance gene, tet(X6), on an SXT/R391 integrative and conjugative element in a Proteus genomospecies 6 isolate of retail meat origin
Authors: He Dandan , Wang Liangliang , Zhao Shiyu , Liu Lanping , Liu Jianhua , Hu Gongzheng , Pan Yushan
Published: January 2020

Objectives

To characterize a novel tigecycline resistance gene, tet(X6), and a novel SXT-related integrative and conjugative element (ICE), ICEPgs6Chn1, found in a tigecycline-resistant Proteus genomospecies 6 strain, T60.

Methods

Strain T60 was identified by the VITEK 2 system, biochemical reactions and an SNP-based approach. The genetic profile of strain T60 was determined by WGS analysis. ICEPgs6Chn1 was analysed by PCR, conjugation experiments and bioinformatics tools. tet(X6) was characterized by cloning and protein structure prediction.

Results

Strain T60 was resistant to ampicillin, tetracycline, tigecycline, florfenicol, colistin and kanamycin, but susceptible to cefotaxime; it also exhibited high MICs of eravacycline (32 mg/L) and omadacycline (>64 mg/L). Only one chromosome was identified and tet(X6) was located in chromosomal ICEPgs6Chn1, a member of the SXT/R391 ICE family, of 114 368 bp and encoding the antimicrobial resistance genes floR, strB, strA, aph(3′)-Ia, aac(3)-IV, aph(4)-Ia, tet(X6) and sul2. The circular intermediate of ICEPgs6Chn1 was detected by PCR and sequencing, but conjugation experiments showed that it was not self-transmissible. Cloning of the novel gene tet(X6) and protein structure prediction revealed that Tet(X6) confers tigecycline resistance.

Conclusions

To our knowledge, this is the first report of a novel SXT/R391 ICE in a Proteus genomospecies 6 strain. Importantly, a novel high-level tigecycline resistance gene, tet(X6), emerged for the first time in the SXT/R391 element of Proteus genomospecies 6, revealing that ICEs may serve as an important platform for the accumulation of antibiotic resistance genes.
Impact of calmodulin inhibition by fluphenazine on susceptibility, biofilm formation and pathogenicity of caspofungin-resistant Candida glabrata
Authors: Ceballos Garzon Andrés , Amado Daniela , Robert Estelle , Parra Giraldo Claudia M , Le Pape Patrice
Published: January 2020

Background

In recent decades, Candida glabrata has emerged as a frequent cause of life-threatening fungal infection. In C. glabrata, echinocandin resistance is associated with mutations in FKS1/FKS2 (β-1,3-glucan synthase). The calmodulin/calcineurin pathway is implicated in response to antifungal stress and calcineurin gene disruption specifically reverses Fks2-mediated resistance of clinical isolates.

Objectives

We evaluated the impact of calmodulin inhibition by fluphenazine in two caspofungin-resistant C. glabrata isolates.

Methods

C. glabrata isolates were identified by ITS1/ITS4 (where ITS stands for internal transcribed spacer) sequencing and the echinocandin target FKS1/FKS2 genes were sequenced. Susceptibility testing of caspofungin in the presence of fluphenazine was performed by a modified CLSI microbroth dilution method. The effect of the fluphenazine/caspofungin combination on heat stress (37°C or 40°C), oxidative stress (0.2 and 0.4 mM menadione) and biofilm formation (polyurethane catheter) was analysed. A Galleria mellonella model using blastospores (1 × 109 cfu/mL) was developed to evaluate the impact of this combination on larval survival.

Results

F659del was found in the FKS2 gene of both resistant strains. In these clinical isolates, fluphenazine increased susceptibility to caspofungin and reduced their thermotolerance. Furthermore, the fluphenazine/caspofungin combination significantly impaired biofilm formation in an in vitro polyurethane catheter model. All these features participated in the increasing survival of infected G. mellonella after combination treatment in comparison with caspofungin alone.

Conclusions

In a repurposing strategy, our findings confirm that calmodulin could provide a relevant target in life-threatening fungal infectious diseases.
Cryptic Wolbachia (Rickettsiales: Rickettsiaceae) Detection and Prevalence in Culicoides (Diptera: Ceratopogonidae) Midge Populations in the United States
Authors: Covey Hunter , Hall Rafe H , Krafsur Alyssa , Matthews Megan L , Shults Phillip T , Brelsfoard Corey L
Published: January 2020

Culicoides midges vector numerous veterinary and human pathogens. Many of these diseases lack effective therapeutic treatments or vaccines to limit transmission. The only effective approach to limit disease transmission is vector control. However, current vector control for Culicoides midges is complicated by the biology of many Culicoides species and is not always effective at reducing midge populations and impacting disease transmission. The endosymbiont Wolbachia pipientis Hertig may offer an alternative control approach to limit disease transmission and affect Culicoides populations. Here the detection of Wolbachia infections in nine species of Culicoides midges is reported. Infections were detected at low densities using qPCR. Wolbachia infections were confirmed with the sequencing of a partial region of the 16S gene. Fluorescence in situ hybridization of Culicoides sonorensis Wirth and Jones adults and dissected ovaries confirm the presence of Wolbachia infections in an important vector of Bluetongue and Epizootic hemorrhagic disease viruses. The presence of Wolbachia in Culicoides populations in the United States suggests the need for further investigation of Wolbachia as a strategy to limit transmission of diseases vectored by Culicoides midges.
Society of Pediatric Psychology Workforce Survey: Updated Factors Related to Compensation
Authors: Kichler Jessica C , Valenzuela Jessica , Barker Dave , Noser Amy E , Brosig Cheryl L , Hilliard Marisa E , Christidis Peggy , Stamm Karen E , Wysocki Tim , Jelalian Elissa
Published: January 2020

Objective

The 2017 Society of Pediatric Psychology (SPP) Workforce Survey provides self-reported compensation by pediatric psychologists, identifies predictors of compensation, and establishes a better understanding of compensation within the context of gender and race/ethnicity minority status.

Methods

SPP members who attended the SPP Annual Conference (SPPAC; April 2017) were invited to complete the survey at the conference through electronic tablets provided on-site by the Workforce Survey Committee. The survey was subsequently distributed online to SPP members who did not complete the survey at SPPAC. The statistical analyses used for this salary data employed flexible semi-parametric models, cross-validation, and prediction models for both the overall sample and academic rank subgroups.

Results

Of 27 potential demographic and employment-related predictors from the 2017 SPP Workforce Survey, significant predictors of salary emerged within this sample: academic rank, time since obtaining doctoral degree, managing internal and external funds (of at least $50,000), years in primary employment position, obtaining Fellowship status in the American Psychological Association (APA), and managing other employees (at least 10 people). Given low response rates for males and individuals who identify as belonging to racial and ethnic minority subgroups, only limited, exploratory results are reported for these subgroups.

Conclusions

These findings suggest that not only is longevity in one’s career important but managing funds/personnel and obtaining professional designations are also predictors of higher salaries for pediatric psychologists, in general. Specific implications of salary according to the psychologist’s academic rank, gender, and racial/ethnicity group status are also explored.
Microdosimetric quantities of an accelerator-based neutron source used for boron neutron capture therapy measured using a gas-filled proportional counter
Authors: Hu Naonori , Tanaka Hiroki , Takata Takushi , Okazaki Keita , Uchida Ryohei , Sakurai Yoshinori
Published: January 2020

Boron neutron capture therapy (BNCT) is an emerging radiation treatment modality, exhibiting the potential to selectively destroy cancer cells. Currently, BNCT is conducted using a nuclear reactor. However, the future trend is to move toward an accelerator-based system for use in hospital environments. A typical BNCT radiation field has several different types of radiation. The beam quality should be quantified to accurately determine the dose to be delivered to the target. This study utilized a tissue equivalent proportional counter (TEPC) to measure microdosimetric and macrodosimetric quantities of an accelerator-based neutron source. The micro- and macro-dosimetric quantities measured with the TEPC were compared with those obtained via the the particle and heavy ion transport code system (PHITS) Monte Carlo simulation. The absorbed dose from events >20 keV/μm measured free in air for a 1-h irradiation was calculated as 1.31 ± 0.02 Gy. The simulated result was 1.41 ± 0.07 Gy. The measured and calculated values exhibit good agreement. The relative biological effectiveness (RBE) that was evaluated from the measured microdosimetric spectrum was calculated as 3.7 ± 0.02, similar to the simulated value of 3.8 ± 0.1. These results showed the PHITS Monte Carlo simulation can simulate both micro- and macro-dosimetric quantities accurately. The RBE was calculated using a single-response function, and the results were compared with those of several other institutes that used a similar method. However, care must be taken when using such a single-response function for clinical application, as it is only valid for low doses. For clinical dose ranges (i.e., high doses), multievent distribution inside the target needs to be considered.
Dynamic structural insights into the molecular mechanism of DNA unwinding by the bacteriophage T7 helicase
Authors: Ma Jian-Bing , Chen Ze , Xu Chun-Hua , Huang Xing-Yuan , Jia Qi , Zou Zhen-Yu , Mi Chen-Yang , Ma Dong-Fei , Lu Ying , Zhang Hui-Dong , Li Ming
Published: January 2020

The hexametric T7 helicase (gp4) adopts a spiral lock-washer form and encircles a coil-like DNA (tracking) strand with two nucleotides bound to each subunit. However, the chemo-mechanical coupling mechanism in unwinding has yet to be elucidated. Here, we utilized nanotensioner-enhanced Förster resonance energy transfer with one nucleotide precision to investigate gp4-induced unwinding of DNA that contains an abasic lesion. We observed that the DNA unwinding activity of gp4 is hindered but not completely blocked by abasic lesions. Gp4 moves back and forth repeatedly when it encounters an abasic lesion, whereas it steps back only occasionally when it unwinds normal DNA. We further observed that gp4 translocates on the tracking strand in step sizes of one to four nucleotides. We propose that a hypothetical intermediate conformation of the gp4–DNA complex during DNA unwinding can help explain how gp4 molecules pass lesions, providing insights into the unwinding dynamics of gp4.
CDK-Mediator and FBXL19 prime developmental genes for activation by promoting atypical regulatory interactions
Authors: Feldmann Angelika , Dimitrova Emilia , Kenney Alexander , Lastuvkova Anna , Klose Robert J
Published: January 2020

Appropriate developmental gene regulation relies on the capacity of gene promoters to integrate inputs from distal regulatory elements, yet how this is achieved remains poorly understood. In embryonic stem cells (ESCs), a subset of silent developmental gene promoters are primed for activation by FBXL19, a CpG island binding protein, through its capacity to recruit CDK-Mediator. How mechanistically these proteins function together to prime genes for activation during differentiation is unknown. Here we discover that in mouse ESCs FBXL19 and CDK-Mediator support long-range interactions between silent gene promoters that rely on FBXL19 for their induction during differentiation and gene regulatory elements. During gene induction, these distal regulatory elements behave in an atypical manner, in that the majority do not acquire histone H3 lysine 27 acetylation and no longer interact with their target gene promoter following gene activation. Despite these atypical features, we demonstrate by targeted deletions that these distal elements are required for appropriate gene induction during differentiation. Together these discoveries demonstrate that CpG-island associated gene promoters can prime genes for activation by communicating with atypical distal gene regulatory elements to achieve appropriate gene expression.
Germline cancer predisposition variants and pediatric glioma: a population-based study in California
Authors: Muskens Ivo S , de Smith Adam J , Zhang Chenan , Hansen Helen M , Morimoto Libby , Metayer Catherine , Ma Xiaomei , Walsh Kyle M , Wiemels Joseph L
Published: January 2020

Background

Pediatric astrocytoma constitutes a majority of malignant pediatric brain tumors. Previous studies that investigated pediatric cancer predisposition have primarily been conducted in tertiary referral centers and focused on cancer predisposition genes. In this study, we investigated the contribution of rare germline variants to risk of malignant pediatric astrocytoma on a population level.

Methods

DNA samples were extracted from neonatal dried bloodspots from 280 pediatric astrocytoma patients (predominantly high grade) born and diagnosed in California and were subjected to whole-exome sequencing. Sequencing data were analyzed using agnostic exome-wide gene-burden testing and variant identification for putatively pathogenic variants in 175 a priori candidate cancer-predisposition genes.

Results

We identified 33 putatively pathogenic germline variants among 31 patients (11.1%) which were located in 24 genes largely involved in DNA repair and cell cycle control. Patients with pediatric glioblastoma were most likely to harbor putatively pathogenic germline variants (14.3%, N = 9/63). Five variants were located in tumor protein 53 (TP53), of which 4 were identified among patients with glioblastoma (6.3%, N = 4/63). The next most frequently mutated gene was neurofibromatosis 1 (NF1), in which putatively pathogenic variants were identified in 4 patients with astrocytoma not otherwise specified. Gene-burden testing also revealed that putatively pathogenic variants in TP53 were significantly associated with pediatric glioblastoma on an exome-wide level (odds ratio, 32.8, P = 8.04 × 10−7).

Conclusion

A considerable fraction of pediatric glioma patients, especially those of higher grade, harbor a putatively pathogenic variant in a cancer predisposition gene. Some of these variants may be clinically actionable or may warrant genetic counseling.