bluuebunny/crossref_metadata_2025_split huggingface dataset:
Gene replacement ameliorates deficits in mouse and human models of cyclin-dependent kinase-like 5 disorder
Authors: Gao Yunan , Irvine Elaine E , Eleftheriadou Ioanna , Naranjo Carlos Jiménez , Hearn-Yeates Francesca , Bosch Leontien , Glegola Justyna A , Murdoch Leah , Czerniak Aleksandra , Meloni Ilaria , Renieri Alessandra , Kinali Maria , Mazarakis Nicholas D 
Published: January 2020

AbstractCyclin-dependent kinase-like 5 disorder is a severe neurodevelopmental disorder caused by mutations in the X-linked cyclin-dependent kinase-like 5 (CDKL5) gene. It predominantly affects females who typically present with severe early epileptic encephalopathy, global developmental delay, motor dysfunction, autistic features and sleep disturbances. To develop a gene replacement therapy, we initially characterized the human CDKL5 transcript isoforms expressed in the brain, neuroblastoma cell lines, primary astrocytes and embryonic stem cell-derived cortical interneurons. We found that the isoform 1 and to a lesser extent the isoform 2 were expressed in human brain, and both neuronal and glial cell types. These isoforms were subsequently cloned into recombinant adeno-associated viral (AAV) vector genome and high-titre viral vectors were produced. Intrajugular delivery of green fluorescence protein via AAV vector serotype PHP.B in adult wild-type male mice transduced neurons and astrocytes throughout the brain more efficiently than serotype 9. Cdkl5 knockout male mice treated with isoform 1 via intrajugular injection at age 28–30 days exhibited significant behavioural improvements compared to green fluorescence protein-treated controls (1012 vg per animal, n = 10 per group) with PHP.B vectors. Brain expression of the isoform 1 transgene was more abundant in hindbrain than forebrain and midbrain. Transgene brain expression was sporadic at the cellular level and most prominent in hippocampal neurons and cerebellar Purkinje cells. Correction of postsynaptic density protein 95 cerebellar misexpression, a major fine cerebellar structural abnormality in Cdkl5 knockout mice, was found in regions of high transgene expression within the cerebellum. AAV vector serotype DJ efficiently transduced CDKL5-mutant human induced pluripotent stem cell-derived neural progenitors, which were subsequently differentiated into mature neurons. When treating CDKL5-mutant neurons, isoform 1 expression led to an increased density of synaptic puncta, while isoform 2 ameliorated the calcium signalling defect compared to green fluorescence protein control, implying distinct functions of these isoforms in neurons. This study provides the first evidence that gene therapy mediated by AAV vectors can be used for treating CDKL5 disorder.
Acquisition of Antibiotic-Resistant Gram-negative Bacteria in the Benefits of Universal Glove and Gown (BUGG) Cluster Randomized Trial
Authors: Harris Anthony D , Morgan Daniel J , Pineles Lisa , Magder Larry , O’Hara Lyndsay M , Johnson J Kristie 
Published: January 2020

Abstract

Background
The Benefits of Universal Glove and Gown (BUGG) cluster randomized trial found varying effects on methicillin-resistant Staphylococcus aureus and vancomycin-resistant Enterococcus and no increase in adverse events. The aim of this study was to assess whether the intervention decreases the acquisition of antibiotic-resistant gram-negative bacteria.

Methods
This was a secondary analysis of a randomized trial in 20 hospital intensive care units. The intervention consisted of healthcare workers wearing gloves and gowns when entering any patient room compared to standard care. The primary composite outcome was acquisition of any antibiotic-resistant gram-negative bacteria based on surveillance cultures.

Results
A total of 40 492 admission and discharge perianal swabs from 20 246 individual patient admissions were included in the primary outcome. For the primary outcome of acquisition of any antibiotic-resistant gram-negative bacteria, the intervention had a rate ratio (RR) of 0.90 (95% confidence interval [CI], .71–1.12; P = .34). Effects on the secondary outcomes of individual bacteria acquisition were as follows: carbapenem-resistant Enterobacteriaceae (RR, 0.86 [95% CI, .60–1.24; P = .43), carbapenem-resistant Acinetobacter (RR, 0.81 [95% CI, .52–1.27; P = .36), carbapenem-resistant Pseudomonas (RR, 0.88 [95% CI, .55–1.42]; P = .62), and extended-spectrum β-lactamase–producing bacteria (RR, 0.94 [95% CI, .71–1.24]; P = .67).

Conclusions
Universal glove and gown use in the intensive care unit was associated with a non–statistically significant decrease in acquisition of antibiotic-resistant gram-negative bacteria. Individual hospitals should consider the intervention based on the importance of these organisms at their hospital, effect sizes, CIs, and cost of instituting the intervention.

Clinical Trials Registration
NCT01318213.
How to Choose Target Facilities in a Region to Implement Carbapenem-resistant Enterobacteriaceae Control Measures
Authors: Lee Bruce Y , Bartsch Sarah M , Hayden Mary K , Welling Joel , Mueller Leslie E , Brown Shawn T , Doshi Kruti , Leonard Jim , Kemble Sarah K , Weinstein Robert A , Trick William E , Lin Michael Y 
Published: January 2020

Abstract

Background
When trying to control regional spread of antibiotic-resistant pathogens such as carbapenem-resistant Enterobacteriaceae (CRE), decision makers must choose the highest-yield facilities to target for interventions. The question is, with limited resources, how best to choose these facilities.

Methods
Using our Regional Healthcare Ecosystem Analyst–generated agent-based model of all Chicago metropolitan area inpatient facilities, we simulated the spread of CRE and different ways of choosing facilities to apply a prevention bundle (screening, chlorhexidine gluconate bathing, hand hygiene, geographic separation, and patient registry) to a resource-limited 1686 inpatient beds.

Results
Randomly selecting facilities did not impact prevalence, but averted 620 new carriers and 175 infections, saving $6.3 million in total costs compared to no intervention. Selecting facilities by type (eg, long-term acute care hospitals) yielded a 16.1% relative prevalence decrease, preventing 1960 cases and 558 infections, saving $62.4 million more than random selection. Choosing the largest facilities was better than random selection, but not better than by type. Selecting by considering connections to other facilities (ie, highest volume of discharge patients) yielded a 9.5% relative prevalence decrease, preventing 1580 cases and 470 infections, and saving $51.6 million more than random selection. Selecting facilities using a combination of these metrics yielded the greatest reduction (19.0% relative prevalence decrease, preventing 1840 cases and 554 infections, saving $59.6 million compared with random selection).

Conclusions
While choosing target facilities based on single metrics (eg, most inpatient beds, most connections to other facilities) achieved better control than randomly choosing facilities, more effective targeting occurred when considering how these and other factors (eg, patient length of stay, care for higher-risk patients) interacted as a system.
Evaluation of a Cellulose-Acetate Electrophoresis System for Serum Protein Fractionation
Authors: Kaplan Alex , Savory John 
Published: January 2020

Abstract
A rapid system for the quantitative fractionation of serum proteins by electrophoresis on cellulose-acetate membranes was evaluated and found to be quite precise. The reproducibility (coefficient of variation) of the routine fractionation of a control serum carried out 40 times during a 15-week period was 2.4% for albumin and 14.2, 6.0, 6.1, and 5.2%, respectively, for the α1-, α2-, β-, and γ-globulin fractions.
Normal values are given for serum protein fractions (specimens from nonprofessional blood donors) obtained by cellulose acetate electrophoresis.
Evaluation of Gas-Liquid Chromatography in Assays for Blood Volatiles
Authors: Mather Alan , Assimos Angel 
Published: January 2020

Abstract
A simple screening by gas-liquid chromatography (GLC) can provide definitive answers in the detection and identification of a number of volatile substances, including acetone and the common alcohols. After identification, quantitative assay by an internal-reference technic yields highly specific values for ethyl alcohol concentration with a precision at least equal to (and for low levels, better than) that of conventional assays. The unique advantage of GLC is in its simultaneous quantitative assay of mixtures, some of which cannot be satisfactorily assayed or even recognized in any other way. The combination of speed and negligible sample volumes render the technic valuable for sequential studies on capillary blood samples and, potentially, for mass screening of large populations.
Determination of Urea in Blood and Urine with Diacetyl Monoxime-Glucuronolactone Reagent
Authors: Momose Tsutomu , Ohkura Yosuke , Tomita Jun 
Published: January 2020

Abstract
A simple method for determination of urea in blood and urine with diacetyl monoximeglucuronolactone reagent is presented. The stability of the color produced permits analysis of a large number of samples at one time. Deproteinization of blood is accomplished with sodium tungstate and alum.
Enzymatic Determination of Glucose in Urine by Automation Following Rapid Removal of Inhibitors by Ion-Exchange Resin
Authors: Logan J E , Haight D E 
Published: January 2020

Abstract
A commercial mixed-bed resin has been found effective for the rapid removal from urine of inhibitors of the glucose oxidase peroxidase system, thus allowing successful application of this specific method for glucose determination in urine specimens. The automated method of Hill and Kessler (10) for glucose determination was modified to make use of smaller quantities of the enzyme preparation and render the procedure less costly.
The percentage inhibition produced by graded amounts of uric acid and ascorbic acid was determined. Both compounds were found to be completely removed by the resin treatment. Normal levels of uric acid in urine inhibit the enzyme reaction markedly, whereas only large therapeutic doses of ascorbic acid would produce sufficient levels of this compound in urine for an inhibitory action.
On the Nature of the Inhibitor of Urinary Alkaline Phosphatase
Authors: Hilliard Stephen D , O'Donnell James F , Schenker Steven 
Published: January 2020

Abstract
The inhibition of urinary alkaline phosphatase has been shown to be caused principally by the inorganic phosphates which are naturally excreted in urine. An inorganic phosphate concentration of 15 mg./100 ml. resulted in approximately 50% inhibition, and that of 120 mg./100 ml. in an inhibition of about 90%. Evidence is presented which shows that the inhibition is of the competitive type.
Use of Bilirubin Standards
Authors: Meites Samuel , Traubert John W 
Published: January 2020

Abstract
A stable stock solution of bilirubin in chloroform may be used to standardize quantitative determination of bilirubin in serum as "azobilirubin." The molar absorptivity of azobilirubin (560 mµ, 1 cm.) is 70,300 ±280 (mean ± 1 S.D.) when determined from chloroform-methanol standards treated with a modified van den Bergh reagent in the Malloy and Evelyn procedure for total bilirubin. The absorptivity of azobilirubin prepared from bilirubin in serum standards is essentially the same. The molar absorptivity of bilirubin in serum (460 mµ, 1 cm.) is 49,100 ±690 (mean± 1 S.D.). Manufacturers of standard bilirubin should use the molar constants as criteria of quality. Two of three preparations differed by 6.8 and 8.5% from the molar absorptivity found in standards made by the authors. Reconstituted commercial standards are stable for 3 days when stored at 5°.
Determination of Glucose in Biologic Fluids with an Automated Enzymatic Procedure
Authors: Robin Michael , Saifer Abraham 
Published: January 2020

Abstract
An automated procedure for the enzymatic determination of "true" glucose with the glucose oxidase-peroxidase system is described. A threefold increase in sensitivity was obtained through measurement of the stable, pink chromogen obtained at high acid concentration. Good reproducibility and stoichiometry was obtained over the range of 50-400 mg./l00 ml. at a sampling rate of 40/hr. The normal range (70-105 mg./100 ml.) and the error of the method (±2.0%) are about the same as that obtained with the manual method of Saifer and Gerstenfeld (7).
Estrogens in Human Pregnancy Plasma
Authors: Kroman Henry S , Bender Sheldon R , Brest Albert N , Moskovitz Mark L , King Mark O 
Published: January 2020

Abstract
The concentration of the 3 classical estrogens, 17β-estradiol, estrone, and estriol, was determined in pooled pregnancy plasma by a combination of chemical extraction and gas chromatography. When pregnancy plasma was fractionated by gas chromatography, peaks representing the 3 estrogens were characterized by melting point determination, infrared analysis, two-dimensional thin-layer chromatography, and formation of the acetate derivatives with subsequent gas chromatographic separation.
The mean concentrations of estrogens per 100 ml. of pregnancy plasma were: estrone, 7.65 ± 1.60 µg.; 17β-estradiol, 3.39 ± 1.95 µg.; and estriol, 8.48 ± 2.34 µg.
A Titrimetric Method for Determining Organic Acids and Bases in Urine
Authors: Goldberg Gussie , Paul Wellington , Gonick Harvey 
Published: January 2020

Abstract
Organic acids and bases can be determined in a single procedure by first precipitating protein and removing carbonate with concentrated HCI, then adding Ca(OH)2 to precipitate phosphate and create an alkaline medium, removing ammonia by heat and suction, then finally titrating from pH 10.5 to 2.7 with dilute HCI.
Direct Measurement of Serum Iron and Binding Capacity
Authors: Goodwin Jesse F , Murphy Bramlett , Guillemette Marcel 
Published: January 2020

Abstract
Serum iron may be directly measured without protein precipitation, by incubation in an acetate buffer in the presence of a reducing agent and sulfonated bathophenanthro line. Mercaptoacetic and ascorbic acids have proved to be suitable reductants. Values obtained for serum iron by this procedure show excellent correlation with a trichloro acetic acid precipitation technic. Bilirubin interference may be compensated for. Hemoglobin interference is significant at values above 30 mg./100 ml. Lipid interference is not significant at concentrations below 1,960 mg./100 ml. The iron chelators, ethylenediaminetetraacetic acid (EDTA), diethylenetriaminepentaacetic acid (DTPA), and desferrioxamine do not interfere.
Both unsaturated iron-binding capacity (UIBC) and total iron-binding capacity (TIBC) may be estimated; values for both show good correlation. Serum iron, TIBC, UIBC, and % saturation values obtained from a group of children aged 4-12 years are presented. The advantages of the direct technic for serum iron, TIBC, and UIBC are simplicity and sensitivity.
The Colorimetric Determination of Iron in Biological Material with Reference to its Measurement During Chelation Therapy
Authors: Goodwin Jesse F , Murphy Bramlett 
Published: January 2020

Abstract
Iron may be determined in biologic material by a wet-ash method employing chloric and nitric acids and a non-ash method using trichloroacetic acid treatment. Bathophenanthroline or bipyridyl is utilized for color development, performed in a buffered medium with mercaptoacetic acid as reductant. The wet-ash technic is best for estimating total iron in urine, and the non-ash technic for nonhemoglobin-bound urinary iron and iron excretion during chelation therapy.
Recovery and precision studies indicate a high degree of reliability. Bilirubin, hemoglobin, and excess phosphates have no significant effect on iron estimation and interference of copper and cobalt may be eliminated.
Urinary iron excretion levels for 14 normal healthy children were 32-285 µg./24 hr. (mean, 113.45 ± 76 µg.); for 11 with sickle cell anemia, 115-1270 µg./24 hr. (mean, 685.4 ± 374.3); and for 2 with thalassemia major, 120-690 µg./24 hr. (mean, 363.3± 160.0).
Direct Photometric Analysis of Serum Calcium with Glyoxal Bis (2-Hydroxyanil)
Authors: Mager Milton , Farese Genevieve 
Published: January 2020

Abstract
A spectrophotometric method for the direct determination of microgram quantities of calcium in serum is presented. In this procedure, 0.02 ml. of serum is interacted with buffered glyoxal bis (2-hydroxyanil) solution (GBHA) at pH 12.6. The resultant red calcium-GBHA complex is diluted in methanol-acetone-water solution, and the absorbance determined at 520 mµ. The effects of pH, temperature, GBHA concentration, and alcohol-water ratio were evaluated, as was interference by serum constituents. The method is simple and precise, and the analytical results compare well with the chloranilate procedure for serum calcium.
Characteristics of a 17-Ketosteroid Reaction
Authors: Kraushaar L A , Epstein E , Zak B 
Published: January 2020

Abstract
An improved method for the determination of 17-ketosteroids is described, which involves an increase in sensitivity by the simple expedient of upsetting the equilibrium of the reaction in removing the products of reaction by precipitating the chromophore formed. This precipitation system appears to achieve about 1.6 times the color sensitivity for the same reaction carried out as a solution process. An alternative extraction step for color purification is described. Spectrums, variabilities in reagent parameters, and statistical evaluations are discussed.
An Enzymatic-Spectrophotometric Determination of Pyruvic and Lactic Acid in Blood
Authors: Rosenberg J C , Rush B F 
Published: January 2020

Abstract
Investigation of an enzymatic-spectrophotometric determination for lactic and pyruvic acid in blood has confirmed its suitability and reliability. The precision of both methods is high. No variability is encountered in the pyruvic acid analysis. Less than 5% variation is found with the lactic acid analysis. The recovery in both procedures approaches 100%.
The Quantitation of Major 17-Ketosteroid Fractions by Gas-Liquid Chromatography
Authors: Berrett Charles R , McNeil Crichton 
Published: January 2020

Abstract
A method for measuring the major urinary 17-ketosteroid fractions has been developed. This method includes extraction of the conjugated steroids, hydrolysis, extraction of the free steroids, purification of the extraction, and quantitation of the steroids contained in the purified extract by gas-liquid chromatography. Data are presented on the excretion of androsterone, etiocholanolone, and dehydroepiandrosterone by normal male and female subjects. The procedure is discussed with regard to accuracy, reproducibility, precision, time per analysis, and correlation with the level of total Zimmerman chromogens in the urine.
Phenolphthalein Monophosphate as a Substrate for Serum Alkaline Phosphatase
Authors: Babson Arthur L , Greeley Sharon J , Coleman Charles M , Phillips George E 
Published: January 2020

Abstract
A rapid and precise procedure for serum alkaline phosphatase employs phenolphthalein monophosphate as the substrate. This compound is readily hydrolyzed and directly provides a chromogen for colorimetric determination. The color produced is linear with respect to both enzyme concentration and incubation time and is stable. Reagent and serum blanks are negligible. Various factors which affect alkaline phosphatase activity have been evaluated. The importance of the proper selection of buffer species, concentration, and pH is emphasized.
The Automated Fluorometric Determination of Serum Magnesium
Authors: Klein Bernard , Oklander Morris 
Published: January 2020

Abstract
Two automated flow systems are presented for the fluorometric determination of serum magnesium. Each procedure is based on measurement of the fluorescence of magnesium 8-hydroxyquinoline-5-sulfonate. One method treats a serum dialysate; the other reacts serum directly with the aqueous reagent. Calcium interference is eliminated by sequestration.
Sequential Solvent Extraction of Urinary Corticosteroids and its Clinical Application
Authors: Bacchus Habeeb 
Published: January 2020

Abstract
A procedure which permits the measurement of corticosteroid metabolites in three solvent extracts of hydrolyzed urine is described. Carbon tetrachloride extracts the 11-desoxy, 17-hydroxycorticosteroids, and the 17-OH, 21-deoxycorticosteroids. These are quantitated by the Porter-Silber reaction and the method for 17-ketogenic steroids. The difference between 17-KGS and PSC is a measure of the total 17-OH, 21-deoxycorticosteroids. Subsequent treatment of the residual urine with methylene dichloride extracts compounds of the cortisol series, i.e., cortisol and cortisone in small amounts, THF and THE in larger amounts. These compounds are quantitated by either the Porter-Silber reaction or the 17-KGS procedure. Further treatment of the urine residue with ethyl acetate (employing the "salting out" procedure) results in the extraction of polar compounds including cortol, cortolone, 6 -hydroxycortisol, and its reduction product. This procedure was employed to study several clinical disorders.
Congenital virilizing adrenocortical hyperplasia is characterized by a marked increase in the 17-KGS in the carbon tetrachloride extract; in adult-onset virilizing adrenocortical hyperplasia this increase is also significant, but not as marked. In adrenal hyperactivity (e.g., Cushing's syndrome, ACTH infusion) there are increases in all solvent fractions studied. Inhibition of 11- hydroxylase activity induced by SU 4885 (Metopirone) is accompanied by an increase in the PSC in the carbon tetrachloride fraction. In hepatocellular disease and in pregnancy, the most noteworthy changes are in the increase in polar compounds extracted in the ethyl acetate fraction. The method described is attractive in its simplicity and is readily applicable in the clinical laboratory.
Colorimetric Method for Rapid Determination of Serum Arginase
Authors: Mellerup Brigitta 
Published: January 2020

Abstract
A method for the determination of serum arginase is given which combines the enzymatic formation of urea with the sensitive method of Coulombe (1) for measuring this substance. This procedure allows more accurate determinations in the normal range than do previous methods described and is convenient for clinical routine. Significant difference is found between the mean values of normal men and women, 3.9 units/L. for the former and 2.9 units/L. for the latter.
Fibrinogen Assay
Authors: Swaim William R , Feders Marija B 
Published: January 2020

Abstract
Precise, reproducible quantitation of fibrinogen from plasma using thrombin to convert it to fibrin requires careful attention to the technics of clot formation and syneresis. The precision of the method outlined indicates a coefficient of variation of 1.2% with replicate samples. The coefficient of variation in sequential studies of a single purified bovine fibrinogen pool was 2.8% and for a lyophilized commercial plasma it was 4.7%.
Microdetermination of Phosphatases Employing Phenolphthalein Diphosphate as Substrate
Authors: Fischl J , Segal S , Rabiah S 
Published: January 2020

Abstract
A micromethod is described for the determination of acid and alkaline phosphatases. Phenolphthalein diphosphate is used for substrate and may be prepared by a modified method of Huggins and Taltalay (1). The technics employed are simple, and the results bear a close relationship to units of Bodansky (2, 3). The substrates may be used for the preparation of test paper for a screening test.