bluuebunny/crossref_metadata_2025_split huggingface dataset:
Quantitative Real-Time Reverse Transcription-PCR Assay for Urokinase Plasminogen Activator, Plasminogen Activator Inhibitor Type 1, and Tissue Metalloproteinase Inhibitor Type 1 Gene Expressions in Primary Breast Cancer
Authors: Castelló Remedios , Estellés Amparo , Vázquez Carlos , Falcó Cristina , España Francisco , Almenar Sergio M , Fuster Carlos , Aznar Justo 
Published: January 2020

AbstractBackground: The plasminogen activation system and matrix metalloproteinases (MMPs) play a key role in the degradation of basement membrane and extracellular matrix in tissue remodeling, cancer cell invasion, and metastasis.Methods: Quantitative real-time reverse-transcription-PCR (RT-PCR) assays were developed to quantify urokinase-type plasminogen activator (uPA), plasminogen activator inhibitor type 1 (PAI-1), and tissue metalloproteinase inhibitor type 1 (TIMP-1) mRNA in 54 breast cancer tissues. Gene fragments were amplified in a LightCycler real-time PCR system using gene-specific primers and SYBR Green I. The results were normalized to β-actin mRNA. We also quantified antigen and functional concentrations of these components.Results: The intra- and interassay variabilities for mRNA quantification showed mean SDs for the crossing point of 0.12 and 0.15 cycles, respectively. PAI-1, uPA, and TIMP-1 mRNA and antigen concentrations and PAI-1 and uPA functional concentrations increased with tumor severity; the increase was statistically significant for PAI-1, uPA, and TIMP-1 mRNA and antigen concentrations and for uPA functional concentrations. Node-positive patients showed significantly higher PAI-1, uPA, and TIMP-1 mRNA and antigen concentrations than those who were node negative.Conclusions: Quantitative real-time RT-PCR is a highly sensitive, reproducible, and fast method for measuring gene expression of PAI-1, uPA, and TIMP-1 in breast cancer. These components may be involved in breast cancer development, and increased mRNA expression may be associated with a worse prognosis.
Development and Evaluation of a Practical ELISA for Human Urinary Lipocalin-Type Prostaglandin D Synthase
Authors: Oda Hiroshi , Shiina Yasuhiko , Seiki Kousuke , Sato Nobuyuki , Eguchi Naomi , Urade Yoshihiro 
Published: January 2020

AbstractBackground: Urinary excretion of lipocalin-type prostaglandin D synthase (L-PGDS) is significantly increased in patients with chronic renal failure, but its diagnostic potential in less advanced stages of renal diseases remains to be elucidated.Methods: Six mouse monoclonal antibodies (MAbs) were raised against recombinant human L-PGDS. We constructed a sandwich ELISA with two MAbs that recognized different epitopes with high affinities and assessed its assay performance and clinical utility with urine samples from healthy controls, diabetic patients, and patients with various renal diseases.Results: Western blot analyses with NH2-terminus-truncated L-PGDS mapped the epitopes to Ala23–Val28 (MAb-7F5 and -10A3), Ser52–Ala73 (MAb-9A6), Tyr107–Val120 (MAb-1B7 and -6F5), and Gly140–Pro155 (MAb-6B9). A sandwich ELISA was constructed with MAb-1B7 and -7F5, the Kd values of which were 3.6 and 3.9 nmol/L, respectively, for native L-PGDS. Recoveries were 91–111%, and intra- and interassay CVs were &lt;6% and &lt;9%, respectively. The ELISA showed parallelism of standard and urine samples and no significant interference by a variety of urinary constituents. Urinary L-PGDS excretion was significantly increased in patients with diabetic nephropathy, IgA nephropathy, and chronic glomerulonephritis even when serum creatinine was not increased. In patients with renal diseases, urinary L-PGDS was correlated with urinary albumin (r = 0.64; P &lt;0.0001), N-acetyl-β-d-glucosaminidase (r = 0.43; P &lt;0.001), and serum creatinine (r = 0.66; P &lt;0.0001). At a cutoff value of 284 mg/mol creatinine, the assay had sensitivities of 74% for diabetic nephropathy and 83% for chronic glomerulonephritis and a specificity of 93%.Conclusions: This ELISA system is suitable for measurement of urinary L-PGDS in a routine clinical assay and may be useful to detect less advanced stages of renal diseases.
Simultaneous Analysis of Nicotine, Nicotine Metabolites, and Tobacco Alkaloids in Serum or Urine by Tandem Mass Spectrometry, with Clinically Relevant Metabolic Profiles
Authors: Moyer Thomas P , Charlson Joel R , Enger Robert J , Dale Lowell C , Ebbert Jon O , Schroeder Darrell R , Hurt Richard D 
Published: January 2020

Abstract
Background: Assessment of nicotine metabolism and disposition has become an integral part of nicotine dependency treatment programs. Serum nicotine concentrations or urine cotinine concentrations can be used to guide nicotine patch dose to achieve biological concentrations adequate to provide the patient with immediate relief from nicotine withdrawal symptoms, an important factor in nicotine withdrawal success. Absence of nicotine metabolites and anabasine can be used to document abstinence from tobacco products, an indicator of treatment success.
Methods: The procedure was designed to quantify nicotine, cotinine, trans-3′-hydroxycotinine, anabasine, and nornicotine in human serum or urine. The technique required simple extraction of the sample with quantification by HPLC–tandem mass spectrometry.
Results: The procedure for simultaneous analysis of nicotine, its metabolites, and tobacco alkaloids simultaneously quantified five different analytes. Test limit of quantification, linearity, imprecision, and accuracy were adequate for clinical evaluation of patients undergoing treatment for tobacco dependency. The test readily distinguished individuals who had no exposure to tobacco products from individuals who were either passively exposed or were abstinent past-tobacco users from those who were actively using a tobacco or nicotine product.
Conclusions: Nicotine, cotinine, trans-3′-hydroxycotinine, nornicotine, and anabasine can be simultaneously and accurately quantified in either serum or urine by HPLC–tandem mass spectrometry with imprecision &lt;10% at physiologic concentrations and limits of quantification ranging from 0.5 to 5 μg/L. Knowledge of serum or urine concentrations of these analytes can be used to guide nicotine replacement therapy or to assess tobacco abstinence in nicotine dependency treatment. These measurements are now an integral part of the clinical treatment and management of patients who wish to overcome tobacco dependence.
Interlaboratory Variation of Plasma Total Homocysteine Measurements: Results of Three Successive Homocysteine Proficiency Testing Surveys
Authors: Hanson Naomi Q , Eckfeldt John H , Schwichtenberg Kerry , Aras Ömer , Tsai Michael Y 
Published: January 2020

Abstract
Background: Numerous studies have demonstrated that increased plasma total homocysteine (tHcy), whether measured after fasting or after a methionine load, is associated with increased risk for cardiovascular and thromboembolic diseases. However, little information is available regarding interlaboratory variation of tHcy measurements, especially at the increased tHcy concentrations observed after loading.
Methods: We conducted three Homocysteine Proficiency Testing Surveys at 6-month intervals. Sets of five plasma pools with endogenous tHcy concentrations ranging from 5 to 48 μmol/L were sent to participants. We received 11, 23, and 17 responses in the first, second, and third surveys, respectively. The following methods were used by participating laboratories: fluorescence polarization immunoassay (FPIA); HPLC with fluorescent detection (HPLC-FD), further subdivided by type of reduction/derivatization; HPLC with electrochemical detection (HPLC-ED); amino acid analyzer with ninhydrin detection; and liquid chromatography–electrospray tandem mass spectrometry (LC-MS/MS).
Results: In surveys 1 and 2, no notable differences among the mean tHcy values obtained by the different methods performed were observed. In survey 3, tHcy values obtained by the FPIA method were significantly lower (P &lt;0.05) at increased tHcy concentrations (34 μmol/L) compared with values obtained by HPLC-FD regardless of reduction/derivatization agents used. Our laboratory confirmed the observation that tHcy values obtained by the FPIA method differed from those obtained by HPLC-FD at increased tHcy concentrations by reanalyzing each pool 10 times by FPIA and HPLC-FD using tributylphosphine–ammonium 7-fluoro-2,1,3-benzoxadiazole-4-sulfonate (P &lt;0.001 for tHcy &gt;19 μmol/L). The mean among-method variations in surveys 1, 2, and 3 were 19%, 12%, and 9.6%, respectively. When results of the three surveys were combined, the mean among-method variation on 170 samples was 13%. Within-method variation was lowest for the FPIA method (4.4%), and ranged from 11–20% for HPLC methods.
Conclusions: Various degrees of imprecision and lack of correlation among tHcy methods indicate that there is a need to improve analytical precision, decrease analytical difference, and standardize tHcy measurements among laboratories.
Comparison of Anti-Transglutaminase ELISAs and an Anti-Endomysial Antibody Assay in the Diagnosis of Celiac Disease: A Prospective Study
Authors: Carroccio Antonio , Vitale Giustina , Di Prima Lidia , Chifari Nadia , Napoli Salvatore , La Russa Cristina , Gulotta Gaspare , Averna Maurizio R , Montalto Giuseppe , Mansueto Serafino , Notarbartolo Alberto 
Published: January 2020

Abstract
Background: Most studies of anti-transglutaminase (anti-tTG) assays have considered preselected groups of patients. This study compared the sensitivity, specificity, and predictive value of an immunofluorescence method for anti-endomysial antibodies (EmAs) and two anti-tTG ELISAs, one using guinea pig tTG (gp-tTG) and the other human tTG (h-tTG) as antigen, in consecutive patients investigated for suspected celiac disease (CD).
Methods: We studied 207 consecutive patients (99 men, 108 women; age range, 17–84 years) who underwent intestinal biopsy for suspected CD. Patients presented with one or more of the following: weight loss, anemia, chronic diarrhea, abdominal pain, dyspepsia, alternating bowel habits, constipation, pain in the joints, and dermatitis. At entry to the study, an intestinal biopsy was performed and a serum sample was taken for IgA EmAs, anti-gp-tTG, and anti-h-tTG.
Results: Intestinal histology showed that 24 patients had partial or total villous atrophy; in these patients the diagnosis of CD was confirmed by follow-up. The remaining 183 patients had villous/crypt ratios that were within our laboratory’s reference values and were considered controls. Serum EmAs, anti-gp-tTG, and anti-h-tTG were positive in all 24 CD patients; in the control group, none were positive for serum EmAs, but 15 of 183 (8.2%) were positive for anti-gp-tTG, and 6 of 183 (3.3%) were positive for anti-h-tTG. Sensitivity was 100% for all assays, whereas specificity was 100% for the EmA, 92% for the anti-gp-tTG, and 97% for the anti-h-tTG assay. The negative predictive value was 100% for all assays; the positive predictive value was 100% for the EmA, 80% [95% confidence interval (CI), 65–95%] for the anti-h-tTG (P = 0.03 vs EmA) and 60% (95% CI, 44–76%) for the anti-gp-tTG assay (P = 0.0002 vs EmA). Areas (95% CIs) under the ROC curves were 0.987 (0.97–1.0) for anti-h-tTG and 0.965 (0.94–0.99) for anti-gp-tTG. Most of the patients testing false positive for anti-tTG had Crohn disease or chronic liver disease.
Conclusions: Although both anti-tTG ELISAs showed optimum sensitivity, their lack of specificity yielded positive predictive values significantly lower than those for the EmA assay.
Quantitative Analysis of p40/p46 and p69/p71 Forms of 2′,5′-Oligoadenylate Synthetase mRNA by Competitive PCR and Its Clinical Application
Authors: Takahashi Akira , Iwasaki Yoshiaki , Miyaike Jiro , Taniguchi Hideaki , Shimomura Hiroyuki , Hanafusa Tadashi , Yumoto Yasuhiro , Moriya Akio , Koide Norio , Tsuji Takao 
Published: January 2020

AbstractBackground: 2′,5′-Oligoadenylate synthetases (2-5AS) are type I interferon (IFN)-induced proteins with antiviral capacity. Three major forms of 2-5AS with distinct enzymatic activities have been described in IFN-treated human cells. We measured distinct forms of 2-5AS mRNA to analyze the relationship with its enzymatic activity and response to IFN therapy in chronic hepatitis C.Methods: We established a method to quantify p40/p46 and p69/p71 forms of 2-5AS mRNA by use of reverse transcription followed by competitive PCR. The 2-5AS mRNA concentrations were measured in peripheral blood mononuclear cells from 40 patients with chronic hepatitis C and 28 control individuals.Results: Reconstitution experiments and comparison with Northern blot analyses revealed that our method accurately and linearly quantified 2-5AS mRNA. 2-5AS mRNA concentrations and 2-5AS enzymatic activity were correlated (P &lt;0.03). Our data demonstrated a correlation in 2-5AS mRNA between p40/p46 and p69/p71 (P &lt;0.02), indicating a similar regulation of the expression of these genes. Our data also demonstrated that pretreatment concentrations of 2-5AS mRNA correlated with responses to IFN therapy in chronic hepatitis C.Conclusions: Our method for measuring 2-5AS mRNA concentrations could provide an important marker for selecting patients for IFN therapy and may be useful for the development of more effective therapeutic strategies for chronic hepatitis C.
Modification of Fully Automated Total Iron-binding Capacity (TIBC) Assay in Serum and Comparison with Dimension TIBC Method
Authors: Yamanishi Hachiro , Iyama Shigeru , Yamaguchi Yoshihisa , Kanakura Yuzuru , Iwatani Yoshinori 
Published: January 2020

AbstractBackground: We previously reported the development of a fully automated assay for total iron-binding capacity (TIBC) in serum, using a multipurpose automated analyzer. However, this method requires four different reagents and is thus useful only with a limited number of available analyzers. We simplified our original assay and compared the analytical performance of the modified method with that of a commercial, fully automated TIBC assay (Dimension® TIBC assay).Methods: We simplified our original method to require only three reagents. Calibration was also altered and was performed with human transferrin standard solutions. An advantage of this method is that it does not require separation of excess unbound iron after the first step of transferrin saturation. Unbound iron is eliminated by formation of a complex with the chromogenic reagent ferrozine in the second step. Iron dissociated from transferrin by acidic pH reacts with ferrozine to form a colored complex in the final step, and the increase in absorbance at 570/660 nm is directly proportional to the TIBC measured. TIBC values were determined for 49 healthy individuals and 148 patients with this modified TIBC assay and with a commercial, fully automated TIBC method (Dimension clinical chemistry system), and calculation of TIBC based on the sum of the serum iron and unsaturated iron-binding capacity was performed for 97 patients.Results: The within-run CVs for the modified TIBC assay and the Dimension TIBC assay were &lt;4.8% and &lt;2.4%, and the between-run CVs were 1.2% and 1.7%, respectively. The dilution curves were linear for TIBC values up to at least 180 μmol/L with both methods. TIBC values obtained by our method were linearly correlated with serum transferrin concentrations (r = 0.984; Sy|x = 3.18 μmol/L; P &lt;0.001). The correlation between the values obtained with the present method (y) and those obtained with the Dimension TIBC method (x) was y = 1.04x + 1.19 μmol/L (r = 0.985; Sy|x = 2.47 μmol/L), and with the calculation method (x) was y = 1.18x + 2.62 μmol/L (r = 0.976; Sy|x = 3.27 μmol/L).Conclusions: Our modified, fully automated TIBC assay performed similarly to the Dimension TIBC assay and is adaptable for use with many multipurpose automated analyzers.
Skill-Biased Management: Evidence from Manufacturing Firms
Authors: Feng Andy , Valero Anna 
Published: January 2020

Abstract
This article investigates the link between management practices and workforce skills in manufacturing firms, exploiting geographical variation in the supply of human capital. Skills measures are constructed using newly compiled data on universities and regional labour markets across 19 countries. Consistent with management practices being complementary with skills, we show that firms further away from universities employ fewer skilled workers and are worse managed, even after controlling for a rich set of observables and fixed effects. Analysis using regional skill premia suggests that variation in the price of skill drives these relationships.
Learner Initiatives in the EFL Classroom: a Public/Private Phenomenon
Authors: Donald Shane 
Published: January 2020

Abstract
This paper examines how a teacher responds to ‘learner initiatives’ during classroom instruction. Learner initiatives refer to students making ‘uninvited’ contributions in class when not selected as the next speaker. This paper focuses on learners initiating an interactional sequence through asking the teacher a question. Using conversation analysis, this research describes two practices adopted by a teacher when responding to such learner initiatives. These practices shape how learners participate within learner-initiated interactional sequences and hence the opportunities that occur for learning the target language. The teacher utilizes recipient design to either better understand learner queries or explicitly answer learner questions as part of dealing with learner initiatives. This paper contributes to understanding of how learner initiatives are managed by teachers and has a role to play in teacher education by raising practitioner awareness of how this aspect of classroom interaction can be managed to further learner participation.
Association between depression and hypertension using classic and revised blood pressure thresholds
Authors: DeMoss Dustin S , Teigen Kari J , Claassen Cynthia A , Fisk Mandy J , Blair Somer E , Bakre Sulaimon A , Hurd Cheryl L , Rush Augustus J 
Published: January 2020

Abstract

Background
In a primary care population, the relationship between treatment of depression and hypertension (HTN) under the recently revised American College of Cardiology and American Heart Association HTN thresholds for diagnosing HTN is unknown.

Objective
To compare the association between changes in severity of co-occurring depression and HTN over time using the newly revised versus previous HTN guidelines.

Methods
In this retrospective cohort study, outpatients ≥18 years (n = 3018) with clinically significant depressive symptoms and elevated blood pressure at baseline were divided into a ‘revised’ guideline group (baseline blood pressure ≥130/80 mmHg), a ‘classic’ guideline group (≥140/90 mmHg) and a ‘revised-minus-classic’ group (≥130/80 and &lt;140/90 mmHg). Depressive symptom change was assessed using the Patient Health Questionnaire-9 (PHQ-9). Correlations between changes in PHQ-9 scores and HTN levels by group over a 6- to 18-month observation period were assessed using robust regression analysis.

Results
There were demographic and clinical differences between groups. A total of 41% of study subjects (1252/3018) had a visit during the follow-up period where additional PHQ-9 and HTN results were available. Depressive symptom change was unrelated to change in blood pressure in the revised and revised-minus-classic groups. The classic HTN group demonstrated a clinically insignificant change in systolic blood pressure for each unit change in PHQ-9 score (β = 0.23, P-value =0.02).

Conclusions
Although a statistically significant association between reduced HTN levels and improvement in depressive symptoms was demonstrated under classic HTN guidelines, there was no clinically meaningful association between treatment of depression and improved HTN levels under either guideline.
Relations of Executive Function and Physical Performance in Middle Adulthood: A Prospective Investigation in African American and White Adults
Authors: Leibel Daniel K , Williams Megan R , Katzel Leslie I , Evans Michele K , Zonderman Alan B , Waldstein Shari R 
Published: January 2020

AbstractObjectivesPrevious studies in older adults found robust associations between executive functions (EF) and physical performance, as well as sociodemographic variation in physical performance decline. To examine these associations earlier in the adult lifespan, we investigated relations of EF, race, and sex with age-related physical performance decline during middle adulthood.MethodParticipants were 2,084 urban-dwelling adults (57.2% female; 57.8% African American; 37.3% living in poverty; mean baseline age = 48.1) from the Healthy Aging in Neighborhoods of Diversity across the Life Span study. Mixed-effects regression was used to examine interactive relations among EF, race, sex, and age (indexing time) with change in dominant and nondominant handgrip strength and lower extremity strength over approximately 5 years. All analyses adjusted for poverty status, and subsequently adjusted for education, body mass index, hypertension, and diabetes.ResultsThere were no significant prospective associations between EF and decline in physical performance measures. Significant cross-sectional associations revealed that lower EF was associated with worse performance on all physical performance measures averaged across both time points (p &lt; .05). A significant two-way interaction of Sex × Age (p = .019) revealed that men experienced greater age-related decline in lower extremity strength than women.DiscussionFindings did not reveal prospective associations between EF and physical performance decline in middle adulthood. However, they identified robust cross-sectional associations between EF and physical performance, and unexpectedly greater decline in lower extremity strength in men than women. Ultimately, these findings may inform prevention and intervention strategies targeting groups at risk for poorer physical function status and decline.
How Can the Arts Influence the Attitudes of Dementia Caregivers? A Mixed-Methods Longitudinal Investigation
Authors: Windle Gill , Caulfield Maria , Woods Bob , Joling Karlijn 
Published: January 2020

Abstract

Background and Objectives
Arts activities can improve social connectedness and foster reciprocity between people living with dementia and their caregivers. The extent to which the arts can help shape attitudes towards dementia remains unclear. This paper explores the impact of a 12-week visual arts program “Dementia and Imagination” on the attitudes of family and professional caregivers through a mixed-methods longitudinal investigation, underpinned by a conceptual framework of the arts in dementia care.

Research Design and Methods
One hundred and forty-six family and professional caregivers were recruited across three settings in England and Wales (residential care homes, a county hospital, and community venues). Quantitative and qualitative data on caregivers’ attitudes and perceptions of the impact of the arts program were collected through interviews at three time points.

Results
Thematic analysis identified four themes shared across the caregivers: (1) Recognizing capabilities, (2) Social connectedness, (3) Improvements to well-being, and (4) Equality and personhood. Two further themes were distinct to family caregivers: (5) Duration of the effects of the art program, and (6) Enriched perspective. The final theme Inspiring professional development was distinct to the professional caregivers. The quantitative analysis found no effect for caregivers’ attitude change over time. Family caregivers scored significantly lower than professional caregivers at each of the three time points.

Discussion and Implications
Art programs have the potential to make visible the capabilities of the people living with dementia, enabling caregivers to see the person behind the condition. This study highlights practice implications for future implementation, such as the role of the arts in dementia care education.