bluuebunny/crossref_metadata_2025 huggingface dataset:
ELISA for thyroglobulin in serum: recovery studies to evaluate autoantibody interference and reliability of thyroglobulin values
Authors: Erali M , Bigelow R B , Meikle A W
Published: January 2020

An ELISA for measuring thyroglobulin (Tg) in serum was developed with polyclonal antibodies to Tg on the solid phase and two monoclonal antibodies to Tg as the second antibodies. The assay has a detection limit of 1 microgram/L, a within-run imprecision (CV) of <7%, and a between-run CV of < 10%. Parallelism of the assay was shown in dilution studies, in which the percent observed/expected values for n = 5 autoantibody-containing samples gave a mean of 99% (SD 13.1 %); for n = 5 samples with undetectable autoantibody concentrations, the mean was 103% (SD 11.8%). The correlation of the ELISA with an RIA for Tg in 46 normal samples was ELISA = 1.11(RIA) + 0.52, S y/x = 2.23, SD intercept = 0.54, SD slope = 0.03, range = 0 to 53 micrograms/L, r = 0.980. Comparison of the ELISA with a reference laboratory RIA for 29 clinical samples gave a correlation of: ELISA = 1.53(RIA) - 0.48, S y/x = 9.00, SD intercept = 2.19, SD slope = 0.10, range = 0 to 98 micrograms/L, r = 0.950. To provide additional information concerning the reliability of the Tg measurement in samples containing autoantibodies to Tg, we developed a procedure for determining the percent recovery. A percent recovery greater than or equal to 80% indicates minimal interference by autoantibodies in this assay. The assay is straightforward to perform, results can be posted within 8 h, and the routinely good recovery of Tg in the presence of Tg autoantibodies indicates minimal autoantibody interference in this assay.
How to evaluate and implement new technologies in an era of managed care and cost containment
Authors: Koch D D
Published: January 2020

New technologies often enable clinical laboratorians to overcome the challenges we face. These technologies sometimes become available with amazing punctuality, addressing recent problems and keeping the physician-customer satisfied for another day. Traditionally, developments such as a new cancer marker have been implemented into routine use with little circumspection because the benefits diagnostically and financially were perceived to far outweigh the risk that the new test or technology was not as effective as described. The notion of subsequently removing older tests inferior to the new one has hardly been considered. The challenges confronting clinical laboratorians today, however, are affecting that typical pattern dramatically. Four of the most pervasive challenges are comprehensively examined in the 1995 Clinical Chemistry Forum. One of these key areas--the proper implementation of modern technology in the form of new tests or new approaches to producing clinical laboratory results--is the main topic of this review.
Keeping up with new technology: new approaches to diagnosis of Chlamydia infection
Authors: LeBar W D
Published: January 2020

Chlamydia trachomatis infections are among the most common sexually transmitted infections in the US today. One of the keys to the prevention of C. trachomatis infection rests on the ability to make this diagnosis on the basis of accurate laboratory testing. For many years the standard for diagnosis of C. trachomatis infections has been isolation in tissue culture. Numerous nonculture methods, including enzyme immunoassay, have been used as an alternative to cell culture. The performance characteristics of these tests have all been compared with a standard, cell culture, which at best will detect 90% of positive specimens. Nucleic acid amplification techniques, including PCR and ligase chain reaction, have been recently introduced. The advantage of these tests is their ability to detect 10-20% more positive specimens when compared with culture or confirmed nonculture methods performed with a single specimen. The sensitivity of amplified tests also allows us to test specimens from multiple sites (endocervix, urethra, urine), which expands our standard from an infected sample to detection of an infected patient. Tests based on amplified nucleic acid technology have greatly improved our ability to diagnose urogenital C. trachomatis infection. The use of an expanded standard will help us accurately define the true performance and clinical utility of nonculture Chlamydia diagnostic tests.
Use of medians and "average of normals" of patients' data for assessment of long-term analytical stability
Authors: Lott J A , Smith D A , Mitchell L C , Moeschberger M L
Published: January 2020

Using results from patients, we studied several methods for long-term (years) monitoring of the analytical stability of various chemistry tests. For each test, we obtained on a bimonthly basis histograms of > or = 200 to approximately 2000 data points and determined the median, mean, and mean of those data points in the reference range ("average of normal" or AON). Examining approximately 4 years' data, we found that the medians and AON are extremely stable for most tests, and that all observed shifts in the medians or in the AON could be explained by expected or unexpected changes in the laboratory. The means and peak values of the Erlang (gamma) distribution were less useful, owing to shifts caused by extreme outliers, especially for enzymes in serum. We assumed that the underlying patient population and testing patterns were stable. We recommend the use of medians or the AON of patients' data under defined conditions for monitoring the long-term analytical stability of certain tests involving serum or whole blood.
Clinical comparison of three labeled-antibody immunoassays of free triiodothyronine
Authors: Piketty M L , d'Herbomez M , Le Guillouzic D , Lebtahi R , Cosson E , Dumont A , Dilouya A , Helal B O
Published: January 2020

Three labeled-antibody immunoassays of free triiodothyronine (FT3) were studied in hyperthyroid patients, patients with nonthyroidal illness, and patients being treated with amiodarone; we also studied sera presenting known interferences (n for all groups = 465). The results were compared with those of a one-step labeled-analog assay. The precision of the two automated assays were similar to that of the manual assays. The three labeled-antibody FT3 assays demonstrated a satisfactory diagnostic performance for confirming hyperthyroidism and robustness to interference; nevertheless, two assays displayed unusual behavior in some patients with nonthyroidal illness, with chronic renal failure, or after amiodarone therapy.
Radioimmunoassay of leptin in human plasma
Authors: Ma Z , Gingerich R L , Santiago J V , Klein S , Smith C H , Landt M
Published: January 2020

Recent studies suggest that leptin, the ob gene product absent in ob/ob mice, is a negative regulator of adiposity. We developed an RIA to measure human leptin in plasma or serum. The minimum detectable concentration by the assay is 0.5 microg/L leptin and the limit of linearity is 100 microg/L. Recovery of leptin added to serum was 99-104% over by the linear range of the assay. The RIA agreed reasonably well with rough quantification by Western blot (RIA = 0.90 blot + 3.7 microg/L, Sy/x = 10.9 microg/L). CVs within- and between-run ranged from 3.4% to 8.3% and from 3.6% to 6.2%, respectively. Variation in plasma leptin concentrations in specimens collected on consecutive mornings was large (CVs of 10.9% and 22.5%). After an overnight fast, leptin concentrations were similar to those 1-2 h after 1-2 meals. Plasma leptin concentrations in specimens from 83 lean and obese adults correlated directly with body mass index (BMI; kg/m2): r = 0.72, P <0.001. Correlations were significantly improved by separating results by gender (men r = 0.84, women r = 0.87; p <0.001). The increase in leptin concentrations with increasing BMI was greater in women than in men (slope 2.53 vs 0.97 microg/L per unit BMI, respectively). Leptin concentrations determined in lean subjects (BMI between 18 and 25) were higher in women (7.36 +/- 3.73 microg/L) than in men (3.84 +/- 1.79 microg/L) (P <0.001). Plasma leptin varied little with age and no significant difference was observed between whites and blacks. We conclude that: (a) plasma leptin concentrations are accurately and precisely measured by this new RIA; (b) leptin concentrations vary little due to short-term fasting, age, or race; but (c) plasma leptin concentrations are gender specific.
Immunofluorometric assay for sensitive and specific measurement of human prostatic glandular kallikrein (hK2) in serum
Authors: Piironen T , Lövgren J , Karp M , Eerola R , Lundwall A , Dowell B , Lövgren T , Lilja H , Pettersson K
Published: January 2020

Prostate-specific antigen (PSA) and human prostatic glandular kallikrein (hK2) have 79% identity with the primary structure. When we used recombinant hK2 protein, only 7 of 23 monoclonal anti-PSA IgGs (monoclonal antibodies, MAbs) cross-reacted with hK2, which enabled us to design a novel immunofluorometric MAb-MAb assay for the specific detection of hK2. In the first incubation, an excess of MAb 2H11, which does not cross-react with hK2, is added to prevent both free and complexed PSA from reacting in subsequent immunoreactions. In the second incubation, biotinylated MAb H50, which cross-reacts with hK2 by an epitope overlapping with MAb 2H11, served to bind only hK2 to the microtitration wells coated with streptavidin. In the third step, Eu-labeled MAb H117, which cross-reacts with hK2, detected the immobilized hK2. The hK2 assay was calibrated with recombinant hK2. The detection limit of the assay was 0.1 microgram/L, and the cross-reactivity with recombinant PSA was < or = 0.7%. The concentration of hK2 was measured in serum samples from 334 males with total PSA concentrations ranging from 1 to 3400 microgram/L. Most of the samples (57%) had hK2 concentrations below the detection limit. The proportions of hK2 relative to total PSA were 0-2% in 79%, 2-5% in 14%, 5-10% in 4%, and >10% in 3% of the samples. Gel filtration of 10 serum samples with increased hK2 concentrations showed a single peak of hK2 immunoreactivity with an apparent molecular size of approximately 30 kDa, corresponding to that of recombinant hK2 and free PSA.
Prostaglandin D2 synthase: a component of human amniotic fluid and its association with fetal abnormalities
Authors: Melegos D N , Yu H , Diamandis E P
Published: January 2020

Prostaglandin D2 (PGD2) synthase is responsible for PGD2 production in the brain. Western blot analysis of human amniotic fluid and probing with a polyclonal antibody against prostate-specific antigen (PSA) revealed a strong immunoreactive band with a molecular mass of 25 kDa. The immunoreactive species, which does not react with monoclonal anti-PSA antibodies, was purified to homogeneity from 1 L of amniotic fluid through successive cycles of gel filtration and ion-exchange chromatography. Amino acid sequence analysis (15 cycles) revealed that the protein was highly homologous or identical to PGD2 synthase. On semiquantitative analysis, PGD2 synthase concentration appears to increase dramatically during gestational weeks 12-25 and then declines slowly until term. PGD2 synthase concentration in amniotic fluid was altered in many abnormal pregnancies, most notably its decrease in trisomic fetuses and fetuses with renal abnormalities.
Preparation of samples with equally spaced concentrations through mixing
Authors: Vaks J E
Published: January 2020

Linearity, interference evaluations of the performance of clinical chemistry systems, mathematical model selection for nonlinear calibration, and other assessments often involve several human sample pools with equally spaced analyte concentrations. Sequential mixing of equal volumes, first of the low and high pools to produce the middle pool, then of the low and middle pools to produce the mid-low pool, and of the high and middle pools to produce the mid-high pool, is recommended in the NCCLS EP7-P guideline for interference studies. Proportional mixing of the low and high pools to produce all of the required pool concentrations is recommended in the NCCLS EP6-P guideline for linearity studies. Mathematical analysis and computer simulation show that the sequential mixing is much more accurate and precise than the proportional mixing. Therefore, we recommend sequential mixing for clinical chemistry application.
Evidence for presence of a reduced form of digoxin-like immunoreactive factor (dihydro-DLIF) in mammalian tissues
Authors: Qazzaz H M , Jortani S A , Poole J M , Valdes R
Published: January 2020

Digoxin-like immunoreactive factor (DLIF) from adrenal glands is an endogenous ligand structurally related to the plant-derived cardiac glycoside digoxin. Cardiac glycosides regulate the activity of the sodium pump and thus play key roles in disease processes involving regulation of ion transport. We now report the discovery of an endogenous dihydro-DLIF analogous to dihydrodigoxin. We used HPLC, ultraviolet spectrophotometry, and cross-reactivity with two antibodies, one specific for digoxin and one for dihydrodigoxin, to support the hypothesis that dihydro-DLIF contains a chemically reduced lactone ring. The spectral absorbance maximum for dihydro-DLIF is at 196 nm, identical to dihydrodigoxin. DLIF and dihydro-DLIF are 975- and 2588-fold less immunoreactive than digoxin and dihydrodigoxin for their respective antibodies. The molar ratio of dihydro-DLIF to DLIF is approximately 5.3 in bovine adrenocortical tissue and approximately 0.38 in human serum. Dihydrodigoxin (reduced lactone ring) added to microsomes isolated from bovine adrenal cortex produced a 4.5-fold increase in digoxin-like immunoreactivity (oxidized lactone ring) after 3 h of incubation. The biotransformation is likely mediated by a cytochrome P-450 NADPH-dependent process. Our findings demonstrate the presence of a dihydro-DLIF in mammals and suggest a metabolic route for synthesis of endogenous DLIF in mammalian tissue.
Direct determination of plasma endothelin-I by chemiluminescence enzyme immunoassay
Authors: Iwata R , Hayashi T , Nakao Y , Yamaki M , Yoshimasa T , Ito H , Saito Y , Mukoyam M , Nakao K
Published: January 2020

A highly sensitive sandwich-type chemiluminescence enzyme immunoassay for plasma endothelin-I (ET-1), involving no extraction steps, has been developed. Two populations of polyclonal antibodies were used in the present study: One is specific to the C-terminus of the endothelin family of peptides; the other, a Fab' fragment against the N-terminal core region of ET-1, is coupled with horseradish peroxidase (HRP). Labeled HRP activity was measured by using an enhanced chemiluminescence reaction of luminol/hydrogen peroxide. The assay was sensitive enough to detect 0.5 ng/L (0.05 pg/well) of plasma ET-1 and had no significant cross-reactivities with other related peptides, including endothelin-3 and the endothelin precursor peptide, big ET-1. Reliability of the assay was confirmed by comparison with an extraction-based procedure and a commercial kit.
Monoclonal antibody assay for free urinary pyridinium cross-links
Authors: Gomez B , Ardakani S , Evans B J , Merrell L D , Jenkins D K , Kung V T
Published: January 2020

The pyridinium cross-links of collagen, pyridinoline (Pyd) and deoxypyridinoline (Dpd), provide structural integrity and rigidity to collagen fibrils in bone. During bone degradation (resorption), the cross-links are released into the circulation and eventually excreted in urine. Pyridinium cross-link measurements in urine have been shown to be sensitive and specific indicators of resorption by both established HPLC and newer enzyme immunoassay (EIA) techniques. We have developed a monoclonal antibody that preferentially binds to the non-peptide-bound free forms of Pyd & Dpd. We have incorporated the antibody conjugated to alkaline phosphatase in a competitive EIA by using Pyd-coated microtiter strip wells. After a 3-h incubation of sample and antibody-enzyme conjugate, color is developed for 1 h with p-nitrophenyl phosphate as the substrate. The intraassay (n = 52) CVs were 3.0-7.6%, and interassay (n = 8) CVs were 6.1-7.4%. Comparisons of the assay (y) with HPLC (x) and a polyclonal antibody-based EIA (x') gave regression equations of y = 0.46x + 4, r = 0.96, and y = 0.56x' + 8, r = 0.96. The EIA detected increased Pyd & Dpd concentrations in urine from postmenopausal women and patients with osteoporosis, hyperthyroidism, hyperparathyroidism, and Paget disease of bone. EIA concentrations also reflected the reduction in Pyd&Dpd excretion resulting from estrogen replacement in surgically menopausal women. Measurement of pyridinium cross-links with this simple EIA appears to provide an accurate index of the rate of resorption and may be useful for metabolic bone disease assessment and monitoring the effects of antiresorptive therapy.
Hair iron content: possible marker to complement monitoring therapy of iron deficiency in patients with chronic inflammatory bowel diseases?
Authors: Bissé E , Renner F , Sussmann S , Schölmerich J , Wieland H
Published: January 2020

Measurements of the concentration of iron in hair from 10 patients with chronic inflammatory bowel diseases and from 10 healthy controls showed that the iron concentrations were significantly (P < 0.05) lower in patients before iron intake than in controls. Three weeks after beginning iron treatment, the hair iron concentrations were found to be significantly correlated (r = 0.68; P < 0.05) to reticulocyte counts. Changes in the hair iron concentrations were accompanied by similar changes in the concentrations of the markers most commonly used to diagnose and monitor iron deficiency. The results suggest that quantification of hair iron may be useful to complement evaluations of the body iron status.
Titrating cardiovascular drugs
Authors: Marcus F I
Published: January 2020

Titrating cardiovascular drugs is important to ensure efficacy and to minimize the risk of toxicity. A serum assay is extremely useful to guide digoxin therapy. Assessment of the effect of warfarin on blood clotting should be used to adjust dose. Serum cholesterol and lipid measurements guide therapy with antilipemic agents. The antihypertensive drugs, beta blockers, calcium channel blockers, and vasodilators can be assessed by their clinical effects. There is no strict relation between serum concentration of antiarrhythmic drugs and their effects, nor is it clear that the long-term efficacy of these drugs can be assessed by surrogate end points.
Role of the clinical toxicologist in court
Authors: Chamberlain R T
Published: January 2020

The clinical toxicologist may play a role in court when issues arise concerning therapeutic drug monitoring, drug abuse, environmental chemicals, or toxic torts, where the traditional forensic toxicologist may not have expertise. Beyond being credible in court, the toxicologist's testimony must be based on good scientific evidence. The ruler for measuring good scientific evidence had previously been the Frye Test, or the general acceptance test. In 1993, however, the US Supreme Court established four balancing tests that should be used for the admissibility of scientific evidence. Although the ruling is binding only in federal courts, state courts are expected to follow. When testifying, the clinical toxicologist should be aware of other court rules and expectations. As with all testimony, objections from opposing counsel can be raised to disallow the presentation of evidence by a toxicologist. The toxicologist is usually used to establish causation of injury, whether from negligence, prenatal injury, or environmental chemicals. Several examples are presented.
Health consequences of short- and long-term postmenopausal hormone therapy
Authors: Weiss N S
Published: January 2020

Some women take an estrogen preparation for as long as several years to ease symptoms of the menopause. Such women appear to have little or no alteration in their risk of endometrial cancer, especially if they are also taking a progestogen, and no alteration in their risk of breast cancer. Similarly, the incidence of fractures is unaffected by relatively short-term hormone use. The risk of ischemic heart disease also is reduced among women who currently take estrogens (with or without a progestogen), but the influence of duration of use on this association is uncertain. Postmenopausal women who take estrogens for an extended period of time (e.g., a decade or more) incur a sharply increased risk of cancer of the endometrium. This is largely abated by use of a progestogen for at least 10 days per month. Such long-term estrogen use, whether accompanied by a progestogen or not, may increase the risk of breast cancer slightly, but this is an area of great controversy, at present unresolved. The incidence of both myocardial infarction and fracture is substantially reduced in long-term users of menopausal hormones.
Multiplex, fluorescent, solid-phase minisequencing for efficient screening of DNA sequence variation
Authors: Pastinen T , Partanen J , Syvänen A C
Published: January 2020

We developed a multiplex, solid-phase minisequencing method to detect multiple single-nucleotide polymorphisms in an undivided sample. The amplified DNA templates are first captured on a manifold. Then, with multiple minisequencing primers of various sizes, single-nucleotide extension reactions are carried out simultaneously with fluorescently labeled dideoxynucleotides. The size of the extended product, determined by using a DNA sequencing instrument, defines the site of the polymorphisms, and the incorporated nucleotide gives the identity of the nucleotide at each site. HLA-DQA1 typing was used as a model system to evaluate the method. The DR2 subgroup of the HLA-DRB1 gene was typed along with the DQA1 gene to demonstrate the feasibility of the method in analyzing multiple genes at multiple sites simultaneously. The method is generally applicable for screening any single-nucleotide polymorphisms or point mutations, and its manifold format allows practical handling of large numbers of samples.
Determination of serum and urinary aluminum by HPLC with fluorometric detection of Al-lumogallion complex
Authors: Lee B L , Chua L H , Ong H Y , Yang H G , Wu J , Ong C N
Published: January 2020

We describe a simple and sensitive HPLC method for quantifying aluminum (Al) in biological fluids by measuring the fluorescence of the Al-lumogallion complex (excitation wavelength 500 nm, emission wavelength 575 nm). Serum samples are deproteinized with 0.83 mol/L perchloric acid and centrifuged; the supernates are mixed with lumogallion reagent. Urine samples are pretreated with sodium hydroxide (2 mol/L) and methanol, kept for 1 h at -20 degrees C, and then centrifuged; the precipitate is resuspended in perchloric acid and mixed with lumogallion reagent, as for serum. The maximal fluorescence complex is formed after 1 h at pH 5 +/- 0.5. The HPLC mobile phase consists of (per liter) 100 mL, of 0.2 mol/L potassium hydrogen phthalate, 220 mL of acetonitrile, and distilled deionized water. The flow rate is 1 mL/min, and the injection volume is 5 microliters. The major aluminum species is eluted at 3.5 min, the lowest detection limit being 0.45 pg. We validated the method with samples collected from normal subjects and from workers occupationally exposed to aluminum. Comparing the results with those by traditional atomic absorption spectrometry of urinary aluminum suggests that the proposed method is reliable.
Interference of soluble TNF-alpha receptors in immunological detection of tumor necrosis factor-alpha
Authors: Moreau E , Philippé J , Couvent S , Leroux-Roels G
Published: January 2020

We have studied the interference of soluble tumor necrosis factor-alpha receptor p55 (sTNF-R55) on the quantification of tumor necrosis factor-alpha (TNF-alpha) in three immunoassays: the commercially available enzyme amplified-sensitivity immunoassay (EASIA) of Medgenix and the ELISA of Boehringer Mannheim, and an assay based on electrochemiluminescence (ECL) technology. TNF-alpha measurements by EASIA and ELISA were not affected by the presence of sTNF-R55, but results by the ECL method were clearly influenced by the receptor. We then performed a second set of experiments with the ECL system to examine what factors might influence the importance of interference from sTNF-R55 and soluble TNF-alpha receptor p75 (sTNF-R75), which influences TNF-alpha measurements the same as sTNF-R55. We found that extending the incubation time and increasing the concentration of capture antibody decreased the interfering capacity of sTNF-R55 and sTNF-R75. A clear insight into these phenomena is of utmost importance for correct interpretation of cytokine quantification.
Hormonal and genetic analysis of a patient with congenital adrenal hyperplasia
Authors: Summers R H , Herold D A , Seely B L
Published: January 2020

We describe a patient with signs and symptoms of virilization caused by 21-hydroxylase deficiency. The patient, a Hispanic woman, first sought medical attention at age 24, when she presented to a medical clinic with an uncomplicated urinary tract infection. At that time several signs of virilization were noted and she was referred to the endocrinology clinic. Evaluation revealed temporal balding, hyperpigmentation, acne, absent breast development, a muscular habitus, and clitoromegaly. Radiological studies revealed bilaterally enlarged adrenal glands and ovaries. Laboratory evaluation revealed markedly increased concentrations of 17-hydroxyprogesterone, androstenedione, and testosterone. The patient was diagnosed with congenital adrenal hyperplasia (CAH) and received hormone therapy. In her sister, encouraged to undergo testing for this autosomal recessive disorder, HLA testing demonstrated that certain haplotypes in this family were associated with CAH. The case highlights key steps in the laboratory diagnosis and genetics of CAH.
Two different approaches for developing immunometric assays of haptens
Authors: Grassi J , Créminon C , Frobert Y , Etienne E , Ezan E , Volland H , Pradelles P
Published: January 2020

To improve immunoassays of small haptens, we developed two different approaches for their measurement in a non-competitive format. We first devised two-site immunometric assays for small peptides (8-11 amino acids) by selecting two sets of antibodies specifically directed against C- and N-terminal moieties of the peptides. In each case, assay sensitivity improved substantially over that of the corresponding competitive assays. More interestingly, all of these new immunometric assays were much more specific than the competitive assays. In a second approach, we developed a new procedure, solid-phase-immobilized epitope immunoassay (SPIE-IA), in which a single monoclonal antibody uses the same epitope for capture and tracer binding and the hapten is covalently cross-linked to solid-phase proteins. To date, SPIE-IA have been successfully applied to the determination of haptens bearing primary amino groups, including substance P, thyroxine, leukotriene C4, endothelin, and angiotensin II. In each case, assay sensitivity was significantly improved.
Thrombus precursor protein (TpP): marker of thrombosis early in the pathogenesis of myocardial infarction
Authors: Carville D G , Dimitrijevic N , Walsh M , Digirolamo T , Brill E M , Drew N , Gargan P E
Published: January 2020

We have developed an enzyme-linked immunosorbent immunoassay for quantifying the immediate precursor proteins to intravascular thrombi. This thrombus precursor protein (TpP) assay identifies active thrombosis in several clinical conditions, including early myocardial infarction (MI). In a study of patients recruited for the GUSTO intervention study, MI patients had concentrations of TpP 4-20-fold that of controls; patients diagnosed without MI had concentrations similar to the control subjects. In a separate study of subjects presenting at the emergency room with chest pain, MI patients who presented early after the onset of chest pain had TpP concentrations significantly (P <0.01) higher than controls. Patients presenting late or diagnosed with other chest pain had concentrations within the reference range. The potential utility of the TpP assay as an aid for the diagnosis of thrombotic MI and other thrombotic conditions is described.
Chemiluminescence: sensitive detection technology for reporter gene assays
Authors: Bronstein I , Martin C S , Fortin J J , Olesen C E , Voyta J C
Published: January 2020

A series of enzyme-activated chemiluminescence-based assays of reporter gene expression, useful in many biomedical applications, has been developed. The chemiluminescence detection systems for beta-galactosidase, beta-glucuronidase (GUS), and secreted placental alkaline phosphatase (SEAP) reporter enzymes are all based on use of 1,2-dioxetane substrates. This detection technology also permits the combined luminescence detection of two different reporter enzymes in the same tube, e.g., a dual assay for beta-galactosidase and luciferase. The sensitivity of these chemiluminescence assays is several orders of magnitude greater than that of conventional colorimetric or fluorometric detection methods; e.g., the detection limit for beta-galactosidase by the chemiluminescence assay is 8 fg and by a fluorometric assay is 2 pg. Furthermore, chemiluminescence enables detection of beta-galactosidase, GUS, and SEAP enzyme concentrations over a dynamic range of more than five to six orders in magnitude. These assays offer highly sensitive, quantitative, rapid, nonisotopic detection of reporter enzymes that are widely used in both mammalian cells and plant cells.
ISO 9001 benefits and pitfalls: the path to successful certification
Authors: Krause M S
Published: January 2020

The lifeblood of a business is developing and commercializing new products with minimum cost and time and maximum quality. Implementation of a quality management system is often used to achieve these goals, and the ISO 9001 standard for a business quality system is rapidly becoming the model of choice. The existence of an ISO-compliant system is a key to meeting the forthcoming regulatory requirements in the European Union and the US Food and Drug Administrations's proposed good management practices. DuPont has demonstrated leadership in the achievement of ISO registration. I describe the path to these successful registrations along with key lessons from the experience. Elements of success are management commitment, adequate resources, education, communication, total organizational involvement, and auditing of system performance. For the system to flourish and provide benefits to the users, sufficient time must be allocated for the organization to change.
The burgeoning role of global health diplomacy to alleviate suffering of cancer patients in low- and middle-income countries
Authors: Adomako Melissa , Kamiar Alaei , Alshaikh Abdulla , Baines Lyndsay S , Benson Desiree , Bettcher Douglas W , Cheema Baljit , Corijn Lex , Fountain Evan , Gdaniec Bartosz G , Garonzik Elan , Harney Mary , Jindal Rahul M , Jones Kate , Kerr David , Mehjabeen Deena , Vahid Nick P , Okonetuk Emem , Pompeu Nathalia , Skosana Bongekile , Tan Soo , Thokwane Karabo , Welzel Tyson
Published: January 2020

The science of global health diplomacy (GHD) consists of cross-disciplinary, multistakeholder credentials comprised of national security, public health, international affairs, management, law, economics and trade policy. GHD is well placed to bring about better and improved multilateral stakeholder leverage and outcomes in the prevention and control of cancer. It is important to create an evidence base that provides clear and specific guidance for health practitioners in low- and middle-income countries (LMICs) through involvement of all stakeholders. GHD can assist LMICs to negotiate across multilateral stakeholders to integrate prevention, treatment and palliative care of cancer into their commercial and trade policies.
Validated Stability Indicating Chromatographic Methods for Quantification of Imidocarb Dipropionate; Application for the Determination of Its Residues in Bovine Meat and Milk Samples
Authors: Sedik Ghada AbdElHamid , Naguib Doha Mohamed , Morsy Fahima , Zaazaa Hala Elsayed
Published: January 2020

Background

Imidocarb dipropionate (IMD) is an immunomodulator agent commonly used for treatment of anaplasmosis in cattle.

Objective

Thus, two sensitive, specific, and precise stability-indicating chromatographic methods have been developed, optimized, and validated for its determination in presence of its acid, alkaline, and oxidative stressed degradation products.

Method

The first method is based on separation of IMD and its forced induced degradation products on reversed phase cyano column using isocratic elution system consisted of sodium acetate buffer–methanol–acetonitrile (55: 30:15, v/v/v), pH 4.6 at a flow rate of 1.2 mL/min, and UV detection at 254 nm. The second method utilized TLC combined with densitometric determination of the separated bands at 254 nm. The separation was achieved using silica gel 60 F254 TLC plates with a mixture of ethyl acetate–methanol–ammonia–water (8.5:1:0.5:0.2, v/v/v/v) as a developing system.

Results

HPLC analysis was applied in range of 0.25–40 µg/mL with LOD of 0.073 µg/mL. While densitometric measurements showed linearity in the range of 0.1–1.8 µg/band with LOD of 0.02 µg/band.

Conclusions

The suggested methods were validated in compliance with the ICH guidelines and were successfully applied for determination of IMD in its commercial veterinary formulations with good recoveries. Furthermore, the proposed HPLC method was extended to the determination of IMD residues in bovine meat and milk samples

Highlights

Bovine meat, HPLC, Imidocarb dipropionate, Milk, TLC.
Intradermal rabies pre-exposure vaccination schedules in older travellers: comparison of immunogenicity post-primary course and post-booster
Authors: Furuya-Kanamori Luis , Ramsey Lani , Manson Melanie , Gilbert Brian , Lau Colleen L
Published: January 2020

Background

Intradermal (ID) rabies vaccination for pre-exposure prophylaxis (PrEP) has become increasingly popular; however, there is limited evidence about the effectiveness of different ID PrEP schedules in travellers aged > 50 years or their response to ID boosters. This study aimed to compare across different ID vaccine schedules and age groups the proportion of travellers who were seropositive after (i) primary course of ID PrEP and (ii) a booster.

Methods

Travellers who received ID PrEP at a travel medicine clinic in South Australia from 2000 to 2016 were included. Three schedules were examined: 1IDx3 (1 × 0.1 ml on days 0, 7, 21–28), 2IDx2 (2 × 0.1 ml on days 0, 7) and 4IDx1 (4x0.1 ml on day 0). The 4IDx1 is a non-standard schedule that has been previously explored in research settings, but not endorsed by WHO for PrEP. Antibody titres of ≥0.5 IU/ml were considered seropositive. The proportion seropositive after a primary course or post-booster was estimated for each schedule and age category. Predictors of seronegative status after a primary course were examined using multivariable logistic regression models.

Results

Overall, 835 travellers (median age 37.5 years; 37.1% > 50 years) were included in the analyses of seropositivity after a primary course. Another group of 771 travellers (median age 45.9 years; 43.5% > 50 years) was included in the analyses of seropositivity post-booster. The proportion seropositive after primary course was 92.5% (95%CI: 90.5–94.1%) and highest with the 1IDx3 schedule (93.4%; 95%CI: 91.4–95.0%). After adjusting for age and timing of the serology, the odds of seronegative status were four times higher (OR 4.17; 95%CI: 1.43–12.18) with the 4IDx1 schedule compared to 1IDx3. Overall, 98.7% (95%CI: 97.6–99.3%) were seropositive post-booster. Of 46 travellers who received a booster ≥3 years after PrEP, all were seropositive post-booster.

Conclusions

In older travellers, the 1IDx3 schedule was the most effective, and a high proportion were seropositive post-booster even many years after a primary course.
Transcription Factor ZNF326 Upregulates the Expression of ERCC1 and HDAC7 and its Clinicopathologic Significance in Glioma
Authors: Wang Minghao , Han Qiang , Su Zhe , Yu Xinmiao
Published: January 2020

Previous reports that we have coauthored have shown that transcription factor ZNF326 can upregulate the expression of ERCC1 and HDAC7, and downregulate the expression of LTBP4 and ZNF383 in lung-cancer cells. However, whether tissue-specificity of the ZNF326 function exists in glioma tissue remains unclear. In this study, overexpression or knockdown of ZNF326 in glioma cells caused upregulation or downregulation, respectively, of the protein and micro RNA (mRNA) levels of ERCC1 and HDAC7. The levels of LTBP4 and ZNF383 were not significantly changed. Immunohistochemical results showed that ZNF326 was not only highly expressed in glioma but was also positively correlated with the expression of ERCC1 and HDAC7. Moreover, the expression of ERCC1 and HDAC7 was enhanced with the increase in tumor grade. However, there was no correlation between ZNF326 and the expression of LTBP4 and ZNF383. Therefore, the detection of ZNF326, ERCC1, and HDAC7 expressions was useful for identifying different grades of glioma.
Lung Cancer Screening at a Military Treatment Facility: A Retrospective Review
Authors: White Lindsey J , Kaur Antarpreet , Lapel Raechel T , E Boswell Gilbert , Luceri Robert E , Parrish John Scott , Seda Gilbert
Published: January 2020

Introduction

Lung cancer is the leading cause of cancer death among men and women, accounting for more fatalities than colon, breast, and prostate cancers combined. Smoking causes about 85% of all lung cancers in the United States and is the single greatest risk factor. In 2013, the US Preventive Services Task Force (USPSTF) published initial guidelines for low-dose computed tomography lung cancer screening (LCS) among patients 55–80 years old, with a 30-pack-year history, who are current smokers or who quit within the previous 15 years. Smoking prevalence is higher among military personnel compared to the civilian population, demonstrating a need for vigilant screening.

Materials and Methods

A retrospective review of Naval Medical Center San Diego’s (NMCSD) LCS data was conducted to examine screening numbers, lung cancer rates, and initial analysis of screening results. Patients were referred for screening if they met the USPSTF criteria. Between September 2013 and September 2018, 962 patients underwent LCS. A total of 1758 examinations were performed, including follow-up and annual surveillance examinations. The American College of Radiology’s Lung CT Screening Reporting and Data System (Lung-RADS) was used to classify lung nodules’ risk for malignancy.

Results

On this initial analysis, 42 enrolled patients received the diagnosis of lung cancer detected by screening. The initial calculated lung cancer rate is 4.4% (42/962) over the 5-year reporting period. The lung cancer rate among those patients with a Lung-RADS score of 3 or 4 was 31% (42/135). Thirty-seven patients were classified as having non-small cell lung cancer (NSCLC), while five were classified as having small cell lung cancer. Of the 37 NSCLC patients, 76% (28/37) were diagnosed at stage I and II, 11% (4/37) were diagnosed at stage III, and 13% (5/37) were diagnosed at stage IV. The total number of years a person smoked was a significant risk factor (P = 0.004), but not pack-years a person smoked (P = 0.052).

Conclusions

These preliminary results demonstrate the success of a Military Treatment Facility (MTF)-based LCS Program in the detection of early stage lung cancer. Earlier stage detection may result in better health outcomes for affected patients. In the population studied, duration of smoking proved to be more significant than pack-years in predicting lung cancer risk. These results validate the newly dedicated resources and continued efforts to strengthen the LCS program at NMCSD and across MTFs.
Distance mapping applied to four well-known planetary nebulae and a nova shell
Authors: Gómez-Gordillo Sebastian , Akras Stavros , Gonçalves Denise R , Steffen Wolfgang
Published: January 2020

Accurate distance estimates of astrophysical objects such as planetary nebulae (PNe), and nova and supernova remnants, among others, allow us to constrain their physical characteristics, such as size, mass, luminosity, and age. An innovative technique based on the expansion parallax method, the so-called distance mapping technique (DMT), provides distance maps of expanding nebulae as well as an estimation of their distances. The DMT combines the tangential velocity vectors obtained from 3D morpho-kinematic models and the observed proper motion vectors to estimate the distance. We applied the DMT to four PNe (NGC 6702, NGC 6543, NGC 6302, and BD+30 3639) and one nova remnant (GK Persei) and derived new distances in good agreement with previous studies. New simple morpho-kinematic shape models were generated for NGC 6543, NGC 6302, and NGC 6702, whereas for BD+30 3639 and GK Persei published models were used. We demonstrate that the DMT is a useful tool to obtain distance values of PNe, in addition to revealing kinematically peculiar regions within the nebulae. Distances are also derived from the trigonometric Gaia parallaxes. The effect of the non-negligible parallax offset in the second Gaia data release is also discussed.
Errors in Diagnostic Test Use and Interpretation Contribute to the High Number of Lyme Disease Referrals in a Low-Incidence State
Authors: Boyce Ross M , Speight Carly , Lin Jessica T , Farel Claire E
Published: January 2020

Lyme disease accounted for more than two-thirds (56 of 81, 69.1%) of all tick-borne disease referrals to a large, academic infectious diseases clinic in a low-incidence state. Deviations from diagnostic testing guidelines and errors in test interpretation were common (23 of 35, 65.7%), suggesting that frontline providers need additional clinical support.
Earnings Belief Risk and the Cross-Section of Stock Returns*
Authors: Brandon Rajna Gibson , Wang Songtao
Published: January 2020

We show in a theoretical asset pricing model incorporating heterogeneous beliefs that the expected excess return on a risky asset depends on its exposure to the risk arising from innovations in the average belief of investors about the expected return of a representative asset. Using the actual EPS data and the analyst EPS forecast data provided by I/B/E/S, we construct a market-wide average belief measure, which we call “the earnings belief measure.” We find that the average return on stocks with high sensitivity to earnings belief shocks is 7.14% per year higher than that on stocks with low sensitivity. This positive relationship holds after accounting for traditional risk factors, is prominent among large-cap stocks, and is invariant across sentiment levels.
Systematics of the Boana semilineata species group (Anura: Hylidae), with a description of two new species from Amazonian Ecuador
Authors: Caminer Marcel A , Ron Santiago R
Published: January 2020

The combination of genetic and phenotypic characters for species delimitation has allowed the discovery of many undescribed species of Neotropical amphibians. In this study, we used DNA sequences (genes 12S, 16S, ND1 and COI) and morphologic, bioacoustic and environmental characters of the Boana semilineata group to evaluate their phylogenetic relationships and assess their species limits. In addition, we included DNA sequences of several species of Boana to explore cryptic diversity in other groups. We found three Confirmed Candidate Species (CCS) within the B. semilineata group. Holotype examination of Hyla appendiculata shows that it is a valid species that corresponds to one of the CCS, which is here transferred to Boana. We describe the two remaining CCS. Our phylogeny highlights a number of secondary but meaningful observations that deserve further investigation: (1) populations of B. pellucens from northern Ecuador are more closely related to B. rufitela from Panama than to other Ecuadorian populations of B. pellucens; (2) we report, for the first time, the phylogenetic relationships of B. rubracyla showing that it is closely related to B. rufitela and B. pellucens; and (3) B. cinerascens and B. punctata form two species complexes consisting of several unnamed highly divergent lineages. Each of these lineages likely represents an undescribed species.
Staying in the club: Exploring criteria governing metacommunity membership for obligate symbionts under host–symbiont feedback
Authors: Venkateswaran Vignesh , Borges Renee M.
Published: January 2020

Metacommunity membership is influenced by habitat availability and trophic requirements. However, for multitrophic symbiont communities that are closely associated with host plants, symbionts and hosts may additionally influence each other affecting membership criteria in novel ways. For example, failure of beneficial services from a symbiont could elicit a response from the host that impacts the entire community. Understanding such host–symbiont feedback effects on symbiont community membership can be crucial for understanding symbiont community structure and function.

We investigate membership for a multitrophic insect symbiont community where symbionts colonize host inflorescences during specific developmental stages termed colonization windows. Inflorescences are host-derived organs and serve as habitat microcosms. Symbionts exhibit a diversity of interactions ranging from mutualism to parasitism. Hosts exhibit immediate feedback by aborting inflorescences not pollinated by mutualistic symbionts and habitats are consequently lost for all other symbiont species. Using relevant empirically measured microcosm parameters, we simulate symbiont dispersal from and colonization of other host inflorescences. We vary host densities and symbiont colonization window lengths, and track the persistence of each symbiont species in the metacommunity based on the temporal availability of the resource and the trophic position of the symbiont.

Since the persistence of the microcosm habitat is dictated by pollination performed by the mutualist, the mutualist fares better than all other symbionts. For prey, the length of colonization windows was positively related with colonization success and symbiont persistence. For predators, the cumulative length of the colonization windows of their prey dictated their success; diet breadth or prey colonization success did not influence the persistence of predators. Predators also had a greater host-plant density requirement than prey for persistence in the community. These results offer valuable insights into host density requirements for maintaining symbionts, and have implications for multitrophic symbiont community stability.

Synthesis. Factors influencing symbiont community membership can be unique when host–symbiont feedback impacts host microcosm development. Special constraints can govern symbiont community membership, function and structure and symbiont persistence in such metacommunities.
Will Gender Self‐Declaration Undermine Women's Rights and Lead to an Increase in Harms?
Authors: Sharpe Alex
Published: January 2020

This article considers and rejects claims that reform of the Gender Recognition Act 2004 (GRA) to allow gender self‐declaration will undermine non‐trans women's rights and lead to an increase in harms to non‐trans women. The article argues that these claims are founded on a mistaken understanding of the proper legal relationship between the GRA and the Equality Act 2010 (EA), and that the harm claim, in any event, lacks a proper evidential basis. The article considers three legal arguments made by gender critical feminists: that sex‐based exceptions under the EA cannot be invoked against trans women with a Gender Recognition Certificate (GRC), that the appropriate legal comparator for a trans woman non GRC‐holder in a discrimination case is a non‐trans man, and that section 22 of the GRA, which protects the privacy of GRC‐holders, undermines the ability of women's organisations to regulate access to women‐only spaces.