We measured apolipoproteins (apo) A-I and B by rate immunonephelometry (rate INA) during Phase 1 of the National Health and Nutrition Examination Survey (NHANES) III. We also made the measurements by radial immunodiffusion (RID) in a 20% subset of the samples. Aliquots of this subset were also analyzed in the Northwest Lipid Research Laboratories by fixed-time INA calibrated to the World Health Organization (WHO)-International Federation of Clinical Chemistry (IFCC) First International Reference Materials for Apolipoproteins A-I and B. The CVs for the rate INA and RID measurements were: apoA-I, 4.5-7.7% and 2.5-7.6%, respectively; apoB, 2.3-5.3% and 2.3-6.4%, respectively. In NHANES III, rate INA values (x) can be transformed to WHO-IFCC Reference Material-based values (y) as follows: for apoA-I, y = 0.87x + 251.8 mg/L (r = 0.93, SEslope = 0.13, SEintercept = 17, n = 708); for apoB (mg/L), y = 1.068x + 112.8 mg/L (r = 0.98, SEslope = 0.08, SEintercept = 7, n = 646).

Recombinant adeno-associated viral (AAV) vectors have unique properties, which make them suitable vectors for gene transfer. Here we assess the liver transduction efficiency and biodistribution of AAV-pseudotyped capsids (serotypes) 1, 5, 6, and 8, combined with single-stranded and double-stranded genomic AAV2 structures carrying the luciferase reporter gene after systemic administration. The analysis was performed in vivo and ex vivo, in male and female mice. Gender-related differences in AAV-mediated transduction and biodistribution were shown for the four serotypes. Our data confirm the superiority of AAV8 over the rest of the serotypes, as well as a significant advantage of double-stranded genomes in terms of liver transduction efficiency, particularly in females. Regarding biodistribution, AAV5 displayed a narrower tropism than the other serotypes tested, transducing, almost exclusively, the liver. Interestingly, AAV1 and AAV8, in particular those having singlestranded genomes, showed high transduction efficiency of female gonads. However, no inadvertent germ line transmission of AAV genomes was observed after breeding single-stranded AAV8-injected female mice with untreated males. In conclusion, double-stranded AAV8 vectors led to the highest levels of liver transduction in mice, as demonstrated by luciferase expression. Nevertheless, the transduction of other organs with AAV8 vectors could favor the use of less efficient serotypes, such as AAV5, which display a narrow tropism.

Orotic acid (ORA) is an important biochemical marker for uridine monophosphate synthase deficiency, an autoso-mal recessive disease characterized by macrocytic hypo-chromic megaloblastic anemia, growth retardation, orotic aciduria, and crystalluria (1). In addition, patients with urea cycle diseases excrete increased amounts of urinary ORA (2). Thus, ORA aciduria is observed in patients with ornithine carbamoylasetransferase deficiency (OCTD), an X-linked disorder, and could reveal heterozygosity after a protein load, and in citrullinemia, argininosuccinic acid-uria, and argininemia (2, 3). Currently there are two widely accepted approaches to the determination of urinary ORA, stable-isotope-dilution gas chromatography–mass spectrometry (GC/MS), and ion-exchange HPLC methods with ultraviolet detection,

Me medicine vs. we medicine: reclaiming biotechnology for the common good, by Donna Dickenson, New York, Columbia University Press, 2013, 296 pp., £19.95 (hardback), ISBN 978-0-231-15974-6

“Is personalised medicine replacing public healthcare in ways that are ultimately detrimental to all of us?” This is the central question that Donna Dickenson sets out to answer in her most recent book Me Medicine vs. We Medicine. Presenting itself as a “reality check” for the much-publicized claims about the desirability –and often the inevitability – of revolution(s) and sundry “paradigm shifts” to be brought about through personalized medicine, the book centres on four case studies: direct-to-consumer genetic testing, private cord blood banking, pharmacogenomics and human enhancement technologies. These represent the “me medicine” of the title; the individualized self-knowledge and self-management offered by direct-to-consumer genetic (and genomic) testing, the patient-tailored drug regimens promised by pharmacogenetics (PGx), the evocation of parental duty to privately bank umbilical cord blood as a future self-repair kit of stem cells for individual children and the possibilities for self-improvement suggested by biomedical
enhancement.

This paper was my first as a graduate student of Professor Peter Perlmann at Stockholm University. At the time, RIAs were in full bloom, but they were too sophisticated for many areas of research and diagnosis because they required expensive equipment and used antigens and antibodies labeled with radioactive isotopes with short half-lives. We wanted something simpler with the same sensitivity. Techniques for labeling antibodies with enzymes had been described for immunohistochemistry (1), and we thought they could be useful for serologic assays as well. First, we gave the prospective enzyme immunoassay a name: enzymelinked immunosorbent assay, or ELISA (an important decision). Second, we chose to use an enzyme for which there was, at the time, a soluble and sensitive substrateproduct system, namely alkaline phosphatase. Third, we used the procedure of a standard RIA, i.e., a com

BACKGROUND High-sensitivity cardiac troponin T (hs-cTnT) is a biomarker used in diagnosing myocardial injury. The clinical utility and the variation of this biomarker over time remain unclear in hemodialysis (HD) and peritoneal dialysis (PD) patients. We sought to determine whether hs-cTnT concentrations were predictive of myocardial infarction (MI) and death and to examine hs-cTnT variability over a 1-year period. METHODS A total of 393 nonacute HD and PD patients (70% HD and 30% PD) were followed in a prospective observational study for new MI and death. RESULTS Median hs-cTnT was 57 ng/L (interquartile range, 36–101 ng/L) with no observed difference between HD and PD patients (P = 0.11). Incremental increases in mortality (P = 0.024) and MI (P = 0.001) were observed with increasing hs-cTnT quartiles. MI incidence increased significantly across quartiles in both HD and PD patients (P = 0.012 and P = 0.025, respectively), whereas mortality increased only in HD patients (P = 0.015)....

Total homocysteine is defined as the sum of all homocysteine species in plasma/serum, including free and proteinbound forms. In the present review, we compare and evaluate several techniques forthe determination of total homocystelne. Because these assays include the conversion of all forms into a single species by reduction, the redistribution between free and protein-bound homocysteine through disulfide interchange does not affect the results, and total homocysteine can be measured in stored samples. Total homocysteine in whole blood increases at room temperature because of a continuous production and release of homocysteine from blood cells, but artificial increase is low if the blood sample is centrifuged within 1 h of collection or placed on ice. Different methods correlate well, and values between 5 and 15 mol/L in fasting subjects are considered normal. Total homocysteine in serum/plasma is increased markedly in patients with cobalamin or folate deficiency, and decreases only when they are treated with the deficient vitamin. Total homocysteine is therefore of value for the diagnosis and follow-up of these deficiency states and may compensate for weaknesses of the traditional laboratory tests. In addition, total homocysteine is an independent risk factor for premature cardiovascular diseases. These disorders justify introduction of the total homocysteine assay in the routine clinical chemistry laboratory.

linear with respect to a lysate volume of 0 -300 L (r 2 ϭ 0.997). The influence of the substrate concentration on TPMT activity is shown in , B and C. K m s were 227 and 4.9 mol/L for 6-MP and SAM, respectively, and the maximum velocities (V max ) were 28.1 and 24.9 nmol/h per mL of packed cells. These values are similar to those reported previously using a radiochemical assay . The TPMT activity determined in a population of Caucasian subjects was 10.4 -41.7 nmol/h per mL of packed cells with a mean value of 28.9 nmol/h per mL of packed cells. These preliminary data are in close agreement with the results reported previously in an adult Caucasian European population (9, 13 ). One subject (2.4%) had TPMT in the intermediate range, 40 (97.6%) subjects had high TPMT activity, and no patient had low or undetect-

Background: The objectives of the study were to characterize the expression of the αand β-subunits of granulocyte-macrophage colony stimulating factor (GM-CSF) receptor in bovine cumulus cells and oocytes and to determine the effect of exogenous GM-CSF on cumulus cells expansion, oocyte maturation, IGF-2 transcript expression and subsequent competence for embryonic development.

We describe the preparation of a lyophilized material containing purified human pancreatic a-amylase and the certification of its catalytic concentration. The enzyme was purified from human pancreas by ammonium sulphate precipitation and chromatography successively on DEAE-Sephacel, CM-Sepharose and Sephadex G-75. The purified enzyme Nonstandard abbreviations: IFCC, International Federation had a specific activity of 52.9 kU/g protein and was >99% pure on polyacrylamide gel electrophoresis. Only trace amounts of lipase and lactate dehydrogenase were detected in the purified fraction. The purified pancreatic a-amylase had a molar mass of 57 500 g/mol and an isoelectric point at 7.1. The material was prepared by diluting the purified or-amylase in a matrix containing PIPES buffer 25 mmol/1, pH 7.0, sodium chloride 50 mmol/l, calcium chloride 1.5 mmol/1, EDTA 0.5 mmol/1 and human serum albumin 30 g/l, dispensing in ampoules and freeze-drying. The ampoules were homogeneous and the yearly loss of activity on the basis of accelerated degradation studies was less than 0.01% at -20°C. The certified value for or-amylase catalytic concentration in the reconstituted reference material is 555 U/1 _+ 11 U/I when measured by the specified method at 37°C. The material can be used to verify the comparability of results from different laboratories, for intra-laboratory quality control or for calibration of a-amylase catalytic concentration measurements.

The first generation of troponin T ELISA (TnT 1) can yield false-positive results in patients with severe skeletal muscle injury. Therefore, a cardiac-specific second-generation troponin T ELISA (TnT 2) was developed, in which the cross-reactive antibody 1B10 has been replaced by a high-affinity cardiac-specific antibody M11.7. No cross-reactivity of TnT 2 was observed with purified skeletal muscle troponin T (1000 micrograms/L) or in test samples from 43 marathon runners and 24 patients with rhabdomyolysis and highly increased creatine kinase. TnT 2 was increased > 0.2 microgram/L in 5 of 40 patients with renal failure and in 4 of 20 muscular dystrophy patients. The detection limit is 0.012 microgram/L. Day-to-day imprecision (CV) within the range 0.19-14.89 micrograms/L was < 5.8%. In 4955 patients without myocardial damage, 99.6% had TnT < 0.10 microgram/L. Assay comparison (TnT 1 vs TnT 2) over the whole concentration range (i.e., in 323 samples from AMI-suspected patie...

We presentevidence for the utilityof an improvedassay for the activity of lactate dehydrogenase (EC 1.1.1.27) isoen-zymes1 and 2 in serum, involvinginhibitionof the H-subunit of LD by pyruvate at pH 7.1. Resultscorrelate well with the LD-1/total LD ratio as evaluated by immunologicalassay and, as an indexto infarct, the method is superiorto either the change in CK-MB activityor to the LD-1 activity or to a combinationof these tests, as is the percentage of LD-1 to total LD activity. Moreover, the percentage inhibition ofLD activity by pyruvate may have an advantageover other methods of isoenzyme fractionation because of its smaller populationCV for patients with acute myocardialinfarction than is true of other methods. We also demonstrate how, usinga lineardiscriminantanalysis,we comparedthis method with alternative methods.We determinedthat evaluation of CK-MB isoenzymecontributesno informationin additionto that obtainedfrom the LD-1 isoenzyme.

In atomic force microscopy, the tip experiences electrostatic, van der Waals, and hydration forces when imaging in electrolyte solution above a charged surface. To study the electrostatic interaction force vs distance, curves were recorded at different salt concentrations and pH values. This was done with tips bearing surface charges of different sign and magnitude (silicon nitride, A1203, glass, and diamond) on negatively charged surfaces (mica and glass). In addition to the van der Waals attraction, neutral and negatively charged tips experienced a repulsive force. This repulsive force depended on the salt concentration. It decayed exponentially with distance having a decay length similar to the Debye length. Typical forces were about 0.1 nN strong. With positively charged tips, purely attractive forces were observed. Comparing these results with calculations showed the electrostatic origin of this force.

Many available methodologies for in vitro regeneration of commercial tomato varieties promote not only the production of normal shoots but also individual leaves, shoots without apical meristems and vitrified structures. All these abnormal formations influence and diminish the regeneration efficiency. At the basis of this phenomenon lies callus development. We optimized an alternative procedure by which the regeneration occurs without abnormal shoot formation. The portion including the proximal part of hypocotyls and the radicle was cultured on medium consisting of Murashige and Skoog salts, 4 mg/L thiamine, 100 mg/L mio-inositol and 3% sucrose. After twothree weeks, 60% explants showed adventitious shoot formation. No changes in the morphological characteristics of regenerated plants and fruits were observed as compared with parents. Karyotypic analysis of regenerated plants showed no variations in chromosome number. The optimized procedure offers the advantage of tomato plant regeneration avoiding callus formation, which enables normal plant recovery with an efficiency ranging from 1.45 ± 0.05 to 2.57 ± 0.06 shoots per explant in Campbell-28, Amalia, Lignon, and Floradel cultivars.

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The personalized medicine is an emergent and rapidly developing method of clinical practice that uses new technologies to provide decisions in regard to the prediction, prevention, diagnosis and treatment of disease. A continuous evolution of technology and the developments in molecular diagnostics and genomic analysis increased the possibility of an even more understanding and interpretation of the human genome and exome, allowing a "personalized" approach to clinical care, so that the concepts of "Systems Medicine" and "System Biology" are actually increasing. The purpose of this study is to evaluate the personalized medicine about its indications and benefits, actual clinical applications and future perspectives as well as its issues and health care implications. A careful review of the scientific literature on this field that highlighted the applicability and usefulness of this new medical approach as well as the fact that personalized medicine strategy is even more increasing in numerous fields of applications.

Metanephrines are O-methylated metabolites of catecholamines. We report the use of liquid chromatography with electrochemical detection to determine plasma concentrations of normetanephrine (NMN) and metanephrine (MN). Plasma NMN and MN in 32 normal volunteers and inpatients were compared with concentrations in 23 patients with pheochromocytoma. Metanephrines were adsorbed from plasma onto a cation-exchange column and eluted with ammoniacal methanol. The dried residue was dissolved in mobile phase and injected onto a reversedphase column. Recoveries of NMN and MN from 1 mL of plasma averaged 50-70%, and resultsvaried linearlywith quantity injected over a range of 0.13-55 pmol. The detection limit was 25 fmol for NMN and 50 fmol for MN. Intra-assay CVs were <5%. In normal volunteers and inpatients, plasma concentrations of NMN ranged between 0.12 and 0.73 nmol/L (mean 0.38 nmol/L), and MN between 0.06 and 0.63 nmol/L (mean 0.19 nmol/L). Plasma NMN concentrations were increased in all 23 patients with pheochromocytoma (range 1-172 nmol/L), whereas MN concentrations (range 0.10-382 nmol/L) were increased in only 9 patients. The assay method is reliable and sensitive and offers an approach to examine the extraneuronal metabolism of catecholamines. The method may also be useful in the diagnosis of pheochromocytoma. AddItional Keyphraaes: chromatography, reversed-phase catecholamine pheochromocytoma Normetanephrine (NMN) and metanephrine (MN), O-methylated metabolites of norepinephrine and epinephrine, are produced by the actions of catechol-Omethyltransferase (EC 2.1.1.6), an enzyme largely confined to extraneuronal tissues (1 )3 Measurements of NMN and MN concentrations in biological fluids may therefore provide a means to examine extraneuronal metabolism of catecholamines. Liquid chromatography and gas chromatography! mass spectrometry methods have been described for Clinical Neuroscience Branch, National Institute of Neurological Disorders and Stroke, and2 Hypertension-Endocrine Branch, National Heart, Lung, and Blood Institute, National Institutes of Health, Bethesda,MD. 'Address for correspondence: Building 10, Room 5N214, National Institutes of Health, Bethesda,MD 20892. 3Nonstandard abbreviations: MN, metanephrune;NMN, normetanephrine; MHPEA, 3-methoxy-4-hydroxyphenylethylamine; HMBA, 4-hydroxy-3-methoxybenzylamine;EHPEA, 3-ethoxy-4hydroxyphenylethanolamine;and FNMN, 6-fluoronormetanephrue.

The emergence of novel technologies allows researchers to facilitate the comprehensive analyses of genomes, transcriptomes, and proteomes in health and disease. The information that is expected from such technologies may soon exert a dramatic change in the pace of cancer research and impact dramatically on the care of cancer patients. These approaches have already demonstrated the power of molecular medicine in discriminating among disease subtypes that are not recognizable by traditional pathologic criteria and in identifying specific genetic events involved in cancer progression. This review

An electrophysiological phenomenon running up and down the spine, elicited by light pressure contact at very precise points and thereafter taking the external appearance of an undulatory motion of the spine, is analyzed from its standing wave, coherence, and synchronization-at-a-distance properties. This standing spinal wave can be elicited in both normal and quadriplegic subjects, which demonstrates that the neuronal circuitry is embedded in the spine. The latter, along with the inherent rhythmicity of the motion, its wave properties, and the absence of external sensory input once the phenomenon is elicited reveal a Central Pattern Generator (CPG). The major investigative tool is surface electromyographic (sEMG) wavelet signal analysis at various points along the paraspinal muscles. Statistical correlation among the various points is used to establish the standing wave phenomenon on a specific subband of the Daubechies wavelet decomposition of the sEMG signals. More precisely, ∼10 Hz coherent bursts reveal synchronization between sensory-motor loops at a distance larger, and a frequency slower, than those already reported. As a potential therapeutic application, it is shown that partial recovery from spinal cord injury can be assessed by the correlation between the sEMG signals on both sides of the injury. ;15(5):461-64. PMID: 19450165 Reorganizational Healing, (ROH), is an emerging wellness, growth and behavioral change paradigm. Through its three central elements (the Four Seasons of Wellbeing, the Triad of Change, and the Five Energetic Intelligences) Reorganizational Healing takes an approach to help create a map for individuals to self-assess and draw on strengths to create sustainable change. Reorganizational Healing gives individuals concrete tools to explore and use the meanings of their symptoms, problems, and life-stressors as catalysts to taking new and sustained action to create a more fulfilling and resilient life. 1-15

Skin cancer has been the most common and represents 50% of all new cancers detected each year. If detected at an early stage, simple and economic treatment can cure it mostly. Accurate skin lesion segmentation is critical in automated early skin cancer detection and diagnosis systems. In this paper, we propose an Evolution Strategies (ES) based segmentation algorithm to identify the lesion area within an ellipse. The method is applied to 51 XLM and 60 TLM images which have manual segmentation from dermatologists as TRUTH. Unlike most segmentation methods, the proposed ES-based segmentation method can detect the lesion automatically without setting parameters and initial values by trial and error. The method is also compared to algorithms developed in . The ES-based method gives comparable accuracy for easily segmented images and much better results for images with either higher noise level, less prominent edge information, or very small size lesions.

BACKGROUND: ⌬ 9-Tetrahydrocannabinol (THC), 11hydroxy-THC (11-OH-THC), and 11-nor-9-carboxy-THC (THCCOOH) have been reported in blood from frequent cannabis smokers for an extended time during abstinence. We compared THC, 11-OH-THC, THCCOOH, cannabidiol, cannabinol, THC-glucuronide, and THCCOO-glucuronide blood and plasma disposition in frequent and occasional cannabis smokers. METHODS: Frequent and occasional smokers resided on a closed research unit and smoked one 6.8% THC cannabis cigarette ad libitum. Blood and plasma cannabinoids were quantified on admission (approximately 19 h before), 1 h before, and up to 15 times (0.5-30 h) after smoking. RESULTS: Cannabinoid blood and plasma concentrations were significantly higher in frequent smokers compared with occasional smokers at most time points for THC and 11-OH-THC and at all time points for THCCOOH and THCCOO-glucuronide. Cannabidiol, cannabinol, and THC-glucuronide were not significantly different at any time point. Overall blood and plasma cannabinoid concentrations were significantly higher in frequent smokers for THC, 11-OH-THC, THCCOOH, and THCCOO-glucuronide, with and without accounting for baseline concentrations. For blood THC Ͼ5 g/L, median (range) time of last detection was 3.5 h (1.1-Ͼ30 h) in frequent smokers and 1.0 h (0-2.1 h) in 8 occasional smokers; 2 individuals had no samples with THC Ͼ5 g/L. CONCLUSIONS: Cannabis smoking history plays a major role in cannabinoid detection. These differences may impact clinical and impaired driving drug detection. The presence of cannabidiol, cannabinol, or THCglucuronide indicates recent use, but their absence does not exclude it.

In this study, we have investigated the evidence of fetal heart rate asymmetry and how the fetal heart rate asymmetry changes before and after 35 weeks of gestation. Noninvasive fetal electrocardiogram (fECG) signals from 45 pregnant women at the gestational age from16 to 41 weeks with normal single pregnancies were analysed. A nonlinear parameter called heart rate asymmetry (HRA) index that measures time asymmetry of RR interval time-series signal was used to understand the changes of HRA in early and late fetus groups. Results indicate that fetal HRA measured by Porta's Index (PI) consistently increases after 35 weeks gestation compared to foetus before 32 weeks of gestation. It might be due to significant changes of sympatho-vagal balance towards delivery with more sympathetic surge. On the other hand, Guzik's Index (GI) showed a mixed effect i.e., increases at lower lags and decreases at higher lags. Finally, fHRA could potentially help identify normal and the pathological autonomic nervous system development.

We aimed to evaluate the in vitro osteogenic and osteoinductive potentials of BioS-2P and its ability to promote in vivo bone repair. To investigate osteogenic potential, UMR-106 osteoblastic cells were cultured on BioS-2P and Bioglass 45S5 discs in osteogenic medium. The osteoinductive potential was evaluated using mesenchymal stem cells (MSCs) cultured on BioS-2P, Bioglass 45S5 and polystyrene in non-osteogenic medium. Rat bone calvarial defects were implanted with BioS-2P scaffolds alone or seeded with MSCs. UMR-106 proliferation was similar for both materials, while alkaline phosphatase (ALP) activity and mineralization were higher for BioS-2P. Bone sialoprotein (BSP), RUNX2 and osteopontin (OPN) gene expression and BSP, OPN, ALP and RUNX2 protein expression were higher on BioS-2P. For MSCs, ALP activity was higher on Bioglass 45S5 than on BioS-2P and was lower on polystyrene. All genes were highly expressed on bioactive glasses compared to polystyrene. BioS-2P scaffolds promote...

We studied the changes in blood glucose concentration in blood samples collected in heparinized specimen tubes containing no other preservative, or containing NaF, D-mannose, or a combination of NaF and D-mannose. Blood concentration in samples taken into NaF decreased by 0.40 mmol/L in the first 2 h; thereafter, there was no change. In samples collected into mannose there was a small but significant decrease in blood glucose concentration with time. When samples containing mannose were analyzed immediately after collection, the concentration of glucose was higher than in later analyses, probably because of an exchange of intracellular glucose for extracellular mannose. When a combination of NaF and mannose was used, the blood glucose concentration was relatively stable but slightly higher than nonpreserved samples for the next 24 h. However, samples containing mannose were unsuitable for electrolyte analysis. We conclude that a combination of D-mannose and NaF may be a better prese...

Malignant pleural mesothelioma (MPM) is a fatal disease with a median survival of less than 14 months. For the first time, a genetically engineered vaccinia virus is shown to produce efficient infection, replication, and oncolytic effect against MPM. GLV-1h68 is a replication-competent engineered vaccinia virus carrying transgenes encoding Renilla luciferase, green fluorescent protein (both inserted at the F14.5L locus), ␤-galactosidase (inserted at the J2R locus, which encodes thymidine kinase), and ␤-glucuronidase (at the A56R locus, which encodes hemagglutinin). This virus was tested in six human MPM cell lines (MSTO-211H, VAMT, JMN, H-2373, H-2452, and H-2052. GLV-1h68 successfully infected all cell lines. For the most sensitive line, MSTO-211H, expression of green fluorescent protein (GFP) started within 4 hr with increasing intensity over time until nearly 100% of cells expressed GFP at 24 hr. All cell lines were sensitive to killing by GLV-1h68, with the degree of sensitivity predictable by infectivity assay. Even the most resistant cell line exhibited 44 Ϯ 3.8% cell survival by day 7 when infected at a multiplicity of infection of 1.0. Viral proliferation assays demonstrated 2to 4-fold logarithmic replication of GLV-1h68 in the cell lines tested. In an orthotopic model, GLV-1h68 effectively prevented development of cachexia and tumor-related morbidity, reduced tumor burden, and cured MPM in both early and late treatment groups. GLV-1h68 was successfully used to treat MPM in vitro and in an orthotopic model (in vivo). These promising results warrant clinical investigation of GLV-1h68 as a novel agent in the treatment of MPM.

relevance of the observed effects. We conclude that the sequential measurement of OC and possibly also of bone ALP during the first 2 months after fracture might be used to predict delayed fracture healing. Previous studies have been very limited, and because of methodologic problems, they are only partly comparable with our study. Further studies are needed.

A series of 25 sera were assayed with the proposed system, in duplicate, each day for five days; the standard deviation of the results was ± 2.5 mg/dl. Correlation results for 33 sera assayed with the proposed system and with an automated o-toluidine one

Cancer biomarkers are commonly used in pediatrics to monitor cancer progression, recurrence, and prognosis, but pediatric reference value distributions have not been well established for these markers. The Canadian Laboratory Initiative on Pediatric Reference Intervals (CALIPER) sought to develop a pediatric database of covariate-stratified reference value distributions for 11 key circulating tumor markers, including those used in assessment of patients with childhood or adult cancers. Healthy community children from birth to 18 years of age were recruited to participate in the CALIPER project with informed parental consent. We analyzed serum samples from 400-700 children (depending on the analyte in question) on the Abbott Architect ci4100 and established reference intervals for α-fetoprotein (AFP), antithyroglobulin (anti-Tg), human epididymis protein 4 (HE4), cancer antigen 125 (CA125), CA15-3, CA19-9, progastrin-releasing peptide (proGRP), carcinoembryonic antigen (CEA), squamou...

The ABCA1 gene encodes for a member of subfamily A of the ATP-binding cassette transporters that plays an important role in cellular export of cholesterol and phospholipids; therefore, quantification of the ABCA1 mRNA is critical in many studies related to its expression and regulation by metabolic factors, nutritional status, and new antiatherogenic drug candidates. We developed a rapid, sensitive, specific, and reproducible real-time reverse transcription-PCR (RT-PCR) method for detection and quantification of ABCA1 transcripts in total RNA isolated from cultured human cells and tissues. To quantify ABCA1 mRNA, we generated a calibration curve from serial dilutions of in vitro-transcribed RNA corresponding to an amplified ABCA1 cDNA 205-bp fragment (homologous calibrator). Two pairs of fluorescent hybridization probes were used to detect the ABCA1 and porphobilinogen deaminase (PBGD) mRNAs; the latter served as an internal control. PCR was performed as real-time amplification of A...