bluuebunny/crossref_metadata_2025_split huggingface dataset:
Multiplex In-cell Reverse Transcription-Polymerase Chain Reaction for the Simultaneous Detection of p210 and p190 BCR-ABL mRNAs in Chronic Myeloid Leukemia and Philadelphia-Positive Acute Lymphoblastic Leukemia Cell Lines
Authors: Suh Soon Pal , Kee Seung Jung , Lim Woo Hyun , Song Jeong Won , Lee Sang Khoo , Kim Jong Phil , Shin Jong Hee , Ryang Dong Wook 
Published: October 2004

Abstract
We designed a novel multiplex in-cell reverse transcription-
polymerase chain reaction method for the simultaneous
detection and differentiation of p190 and
p210 BCR-ABL mRNAs within single cells from the human
chronic myeloid leukemia and Philadelphia positive
acute lymphoblastic leukemia. Human K562
chronic myeloid leukemia and SUP B-15 Ph+ acute
lymphoblastic leukemia cell lines were used as positive
controls for p210 and p190 BCR-ABL mRNAs, respectively.
HL60 cell line was used as a negative control.
After the leukemia cells were fixed and
permeabilized, without extracting nucleic acids, the
mRNAs were reverse transcribed to cDNAs, and the
cDNAs were amplified by multiplex polymerase chain
reaction with fluorescent primers specific for p190 and
p210 BCR-ABL mRNAs. After transfer onto glass slides
by cytospin, the amplified cells were detected by fluorescence
microscopy. Fluorescence microscopy after
propidium iodide or 4′,6-diamidino-2-phenylindone
counterstaining showed that the positive K562 cells
exhibited a yellow-green fluorescent cytoplasm
around a red nucleus, and that the positive SUP B-15
cells exhibited an orange cytoplasm around a blue nucleus.
Only the red or blue nucleus was visible in respective
negative HL60 cells. The specificity of amplification
was confirmed by the absence of a signal when
control experiments were performed either with
RNase digestion of mRNA or without reverse transcriptase/Taq polymerase. We conclude that the multiplex
in-cell reverse transcription-polymerase chain
reaction method is capable of simultaneously detecting
and differentiating the p210 and p190 BCR-ABL
mRNAs of chronic myeloid leukemia and Philadelphia-positive
acute lymphoblastic leukemia cells, and that it
may be useful in quantitatively monitoring the minimal
residual disease during therapy.
Genetic Polymorphisms and Activity of Cholesterol Ester Transfer Protein (CETP): Should We Be Measuring Them?
Authors: Ordovas Jose M. 
Published: October 2004

Abstract
Cholesteryl ester transfer protein (CETP) is a plasma
glycoprotein that mediates the transfer of cholesteryl
ester from high density lipoproteins (HDL) to triglyceride-rich lipoproteins in exchange for triglycerides.
Several approaches are currently being used in research
laboratories to measure its activity and/or
mass. However, these assays are not standardized and
it is not possible to compare data from different laboratories.
Also, we lack enough information to assess
the value of this variable as a coronary heart disease
(CHD) predictor. Several genetic variants at CETP locus
have been identified and they have been generally
associated with increased HDL-cholesterol concentrations.
However, there is no consensus about the association
of this CETP-related increase in HDL-cholesterol
and protection against CHD. Nevertheless, the
most recent evidence from the common CETP-TaqI-B
polymorphism shows that the lower CETP activity associated
with the presence of this polymorphism decreases
CHD risk in men. Based on this and previous
evidence, there has been an interest in the development
of CETP inhibitors as a tool to increase HDL-cholesterol,
thus reducing CHD risk. However, it should be
noted that the evidence about the cardioprotective
role of these drugs is not yet available.
Detection of Haemoglobins with Abnormal Oxygen Affinity by Single Blood Gas Analysis and 2,3-Diphosphoglycerate Measurement
Authors: Guerrini Giovanni , Morabito Alberto , Samaja Michele 
Published: October 2004

Abstract
The aim is to determine if a single measurement of
blood 2,3-diphosphoglycerate combined with gas
analysis (pH, PCO2, PO2 and saturation) can identify
the cause of an altered blood-oxygen affinity: the presence
of an abnormal haemoglobin or a red cell disorder.
The population (n=94) was divided into healthy
controls (A, n=14), carriers of red cell disorders (B,
n=72) and carriers of high oxygen affinity haemoglobins
(C, n=8). Those variables were measured both in
samples equilibrated at selected PCO2 and PO2 and in
venous blood. In the univariable approach applied to
equilibrated samples, we correctly identified C subjects
in 93.6% or 96.8% of the cases depending on the
selected variable, the standard P50 (PO2 at which 50%
of haemoglobin is oxygenated) or a composite variable
calculated from the above measurements. After
introducing the haemoglobin concentration as a further
discriminating variable, the A and B subjects were
correctly identified in 91.9% or 94.2% of the cases, respectively.
These figures become 93.0% or 86.1%, and
93.7% or 94.9% of the cases when using direct readings
from venous blood, thereby avoiding the blood
equilibration step. This test is feasible also in blood
samples stored at 4°C for 48 h, or at room temperature
for 8 h.
Effect of Acute Phase Response on Cumulative Troponin T Release
Authors: Diris Jart H.C. , Kragten Johannes A. , Kleine Appie H. , Hermens Wim Th. , Dieijen-Visser Marja P. van 
Published: October 2004

Abstract
We studied a possible effect of the extent of the acute
phase response after acute myocardial infarction on
the cumulative release of troponin T. The height of the
acute phase response might influence the cumulative
release of troponin T, bound to the myofibrillar structures
of the heart, in a different way compared to the
free cytoplasmic cardiac marker hydroxybutyrate dehydrogenase
(EC 1.1.1.27). To investigate this, the cumulative
amount of C-reactive protein in plasma, i.e.
the quantified acute phase response, was related to
the cumulative plasma release of hydroxybutyrate dehydrogenase
(an established method for infarct sizing)
on the one hand and to that of troponin T on the other
hand.
The study was performed in patients receiving
(n=16) and in patients not receiving (n=6) thrombolytic
therapy. Cumulative protein release was calculated using
a two-compartment model for circulating proteins.
Conclusions: The cumulative amount of plasma C-reactive
protein is significantly higher in the patients
not receiving thrombolytic therapy, as is in accordance
with earlier studies. The cumulative amount of troponin
T released is significantly related to the cumulated
concentration of C-reactive protein, especially in
patients not receiving thrombolytic therapy. The intensity
of the acute phase response, estimated from cumulative
plasma C-reactive protein response, has no
effect on the relative proportions of troponin T and hydroxybutyrate
dehydrogenase released into plasma.
Personal Experience with the Application of Carbohydrate-Deficient Transferrin (CDT) Assays to the Detection of Congenital Disorders of Glycosylation
Authors: Colomé Catrina , Ferrer Imma , Artuch Rafael , Vilaseca M. Antònia , Pineda Mercè , Briones Paz 
Published: October 2004

Abstract
Congenital disorders of glycosylation (CDG) are genetic
multisystemic diseases due to various defects in
the biosynthesis or processing of glycoproteins. Our
aim is to present our experience in the selective
screening of CDG syndrome in a paediatric population
(421 patients) with clinical suspicion of the disease,
analysing serum carbohydrate-deficient transferrin
(CDT) by radioimmunoassay and/or immunoturbidimetry.
We established the normal values for our
paediatric population. The abnormal results were confirmed
and classified by isoelectric focusing of serum
sialotransferrins, and by enzymatic and molecular
studies. We found 14 patients (3.3%) with abnormal
serum CDT; 11 of them were classified as CDG type Ia
(CDG-Ia) and the other three showed altered isoelectrofocusing
patterns but remain untyped and are under
investigation. In conclusion, both CDT assays
proved to be useful tools for CDG screening. Isoelectric
focusing is a simple procedure but it requires specific
instruments that are not always available. Since the
immunoturbidimetric procedure is commonly used to
monitor for recent excessive alcohol consumption in
clinical laboratories and does not require special
equipment, it may also be reliably used to screen for
CDG in children under clinical suspicion.
Screening for Proteinuria in Japanese Schoolchildren: a New Approach
Authors: Pugia Michael J. , Murakami Mutsumi , Lott John A. , Ohta Yoshihide , Kitagawa Teruo , Yamauchi Kuniaki , Suhara Yutaka , Kasjima Junko 
Published: October 2004

AbstractBy governmental mandate, Japanese school children are screened annually for proteinuria, hematuria, and glucosuria to identify children with possible renal disorders. We added urine dipstick tests for albumin and creatinine to the Japanese screening protocol, and used their dipstick results for blood, glucose and protein. The sulfosalicylic acid precipitation test was used to confirm “trace” positive protein dipsticks. The Japanese and our screening protocol have in common the same data for glucosuria and proteinuria. Their scheme has an algorithm for repeat testing of children with abnormal results, and further testing and medical evaluation for those showing persistently abnormal values. Out of the 23,121 students, we found seven with likely nephritis, one with confirmed nephritis, one with nephrotic syndrome, 170 with persistent unexplained hematuria, 19 with persistent unexplained proteinuria, 14 cases of urinary tract infection, and 20 cases of likely diabetes mellitus. We conclude that dipstick testing for albumin, protein, creatinine, glucose and occult blood has significant value in a multilevel testing scheme for identifying children with urinary tract abnormalities or diabetes. The assay of albumin increases the sensitivity of the screening, and dividing the albumin by the creatinine concentration reduces the potential errors arising from concentrated or dilute urines.
Determination of a Redox Compensation Index and Its Relationships to Glycaemic Control in Type 2 Diabetes Mellitus
Authors: Sugherini Lidia , Valentini Melissa , Cambiaggi Caterina , Tanganelli Italo , Gragnoli Giorgio , Borgogni Patrizia , Comporti Mario , Pompella Alfonso 
Published: October 2004

Abstract
The measurement of single parameters of oxidative
stress in biological fluids can often give results difficult
to interpret as to the real involvement of oxidative
processes in a given disease condition. In the present
study we propose a novel integrated parameter, called
“redox compensation index”, obtained by combining
the results of two established and convenient procedures,
i.e. the Fox-2 assay for plasma lipid hydroperoxides
and the ferric reducing/antioxidant power (FRAP)
assay for total antioxidant potential of plasma. These
procedures were employed for the evaluation of oxidative
stress in a group of patients with type 2 diabetes
mellitus, a condition in which oxidative processes are
implicated in the development of complications. In type
2 diabetic patients, plasma lipid hydroperoxides were
directly correlated with levels of glycated hemoglobin.
On the other hand, a significant inverse correlation was
o b s e rved between levels of glycated hemoglobin and
redox compensation values. The data reported suggest
that the redox compensation index could represent a
convenient parameter for the direct appraisal of oxidative
status in clinical subjects, and are in support of the
proposed role of protein glycation in production of oxidative
alterations during type 2 diabetes mellitus.
Clinical Experiences with a New Semi-Quantitative Solid Phase Immunoassay for Rapid Measurement of Procalcitonin
Authors: Meisner Michael , Brunkhorst Frank-Martin , Reith Hans-Bernd , Schmidt Joachim , Lestin Heiko-Gundmar , Reinhart Konrad 
Published: October 2004

AbstractA self-developing solid-phase immunoassay (B.R.A.H.M.S. PCT-Q, B.R.A.H.M.S.-Diagnostica GmbH, Hennigsdorf, Germany) has recently become available for the semi-quantitative and rapid measurement of procalcitonin (PCT). In this study we examined the validity of this assay at daily clinical routine conditions at five different hospitals in a prospective study.After development of the assay (200 μl plasma, 30 minutes incubation), PCT levels were categorized into four groups (< 0.5 μg/l; ≥ 0.5−< 2 μg/l; ≥ 2−< 10 μg/l; ≥ 10 μg/l) according to the provided reference scale. Samples from patients with suspected elevation of PCT of different etiology (n=237) were read by various analyzers and compared with the results of the Lumitest®PCT (B.R.A.H.M.S.-Diagnostica GmbH, Hennigsdorf, Germany).A total of 74.7% of measurements were categorized according to the results of the Lumitest®PCT, 24.5% were read within the next lower or higher category. Using a ± 10% range at the reference concentrations (20% at 0.5 μg/l), 82.7% of samples were correctly categorized and 16.4% within the next categories. Using a cut-off value of 2.0 μg/l, 92.0% (94.1% for ± 10%) of the results were correctly categorized.The semi-quantitative solid phase immunoassay allows a rapid, simple and semi-quantitative measurement of plasma PCT. The validity of the test results and its ease of use are sufficient to support acute diagnostic decisions. However, for the follow-up of PCT concentrations and routine daily measurements, the quantitative luminometric assay should be preferred, when available.
Inter-Method Differences and Commutability of Control Materials for HbA2 Measurement
Authors: Mosca Andrea , Paleari Renata , Scimè-Degani Valeria , Leone Leonello , Leone Daniela , Ivaldi Giovanni 
Published: October 2004

Abstract
The intermethod variability of control materials and
patient blood samples for the measurement of hemoglobin
A2 (HbA2) were compared. A set of 54 blood
samples and 10 control materials were analyzed in duplicate
by HPLC and microcolumn methods. For each
set of methods the distances of the materials from the
regression line of patient blood results (expressed as
normalized residuals) were calculated. Four out of ten
controls had normalized residuals exceeding three
standard deviations from the regression line. Moreover,
total Hb and Hb derivatives analysis proved that
only a minority of the controls could be considered
similar to patients' blood samples. Intermethod calibration
performed “a posteriori” by the two best performing
control materials improved intermethod variability
among all the five tested methods. We conclude
that the use of high resolution HPLC methods together
with appropriate commutable control materials allows
for better harmonization of results in the field of
diagnosis of hemoglobin disorders in research and
clinical practice.
High Sensitivity C-Reactive Protein: Biological Variations and Reference Limits
Authors: Chenillot Olivier , Henny Joseph , Steinmetz Josiane , Herbeth Bernard , Wagner Carola , Siest Gérard 
Published: October 2004

Abstract
Serum C-reactive protein (CRP) concentration was determined
for 3605 subjects using an immunonephelometric
assay improved to provide greater sensitivity.
Subjects were from 5 to 75 years old and belonging to
1003 nuclear families recruited from the Stanislas Coh
o rt Study between January 1994 and August 1995.
Sample values for CRP ranged from 0.17 mg/l to 100
mg/l. Geometric means (mean − SD; mean + SD) were in
the 5–14 years old group 0.37 (0.17–1.07) mg/l, in the
15–28 years old group 0.47 (0.17–1.38) mg/l and in the
29–75 years old group 0.98 (0.34–2.85) mg/l. For
women, the geometric means were 0.38 (0.17–1.10)
mg/l, 0.62 (0.20–1.90) mg/l and 0.98 mg/l (0.31–3.13)
mg/l respectively. The interindividual variability ranged
from 138% to 759% among different age classes. Biological
factors associated with CRP concentration variations
were examined and accounted for 25% of the CRP
variability in men and 40% in women. The main biological
factors statistically associated with CRP concentration
variations in men were: drugs, leukocyte count,
body mass index, tobacco consumption, age, and in
women: drugs, leukocyte count, age, body mass index
and hemoglobin concentration. These factors were
used to define the exclusion and partition criteria when
obtaining the reference samples. Medians for reference
values ranged from 0.20 to 0.68 mg/l in males and from
0.20 to 0.78 mg/l in women.
A Model for a Multicentre Approach to the Derivation of Reference Intervals for Thyroid Hormones and Testosterone for Laboratories Using Identical Analysers
Authors: Andrew Charles E. , Hanning Ian , M. McBain Alasdair , Moody Denise , Price Alun 
Published: October 2004

Abstract
Ideally, every laboratory should derive their own reference
intervals for all analytes, but this is difficult in
practice. A survey, by questionnaire, of UK laboratories
using the Chiron Diagnostics ACS:180 (Chiron Diagnostics
Limited, Halstead, Essex, UK), for thyroid
function tests, showed that 10% of laboratories derived
their own reference intervals, 60% quoted values
“adapted” from intervals for previous methods, whilst
the remaining 40% quoted (often incorrectly) reference
intervals supplied by the manufacturer. In addition
only 13% of respondent laboratories derived their
own reference intervals for testosterone. As a result of
this survey, a study was devised to enable the users of
the Chiron Diagnostics ACS:180 immunoassay system
to develop and use within-method, between-laboratory
reference intervals for thyroid hormones and
testosterone. Laboratory collaboration provided the
recommended minimum number of data points by establishing
a reference sample group. This sample
group was used for the calculation of appropriate reference
intervals for each hormone according to the
guidelines published by the IFCC. We propose this approach
as a model for laboratories using identical instrumentation
to produce, through collaboration,
within-method, between laboratory reference intervals.
Comparison of Two Methods for the Detection of Urinary Iodine Used in Epidemiological Studies
Authors: Wuethrich Christian , Jaeggi-Groisman Silvia E. , Gerber Hans 
Published: October 2004

Abstract
Two methods for the determination of iodine in urine
or serum based on the Sandell-Kolthoff reaction were
compared. The first was an autoanalyser method
(AAII, Technicon) where the mineralisation takes place
in a continuous flow manner. This procedure was used
at the Department of Clinical Chemistry of the University
Hospital of Berne (Switzerland) from 1968 until
1993. The second method was evaluated and adapted
in our own laboratory. Each sample and the iodate-standards
are mineralised in a Pyrex glass tube and the
decolorisation reaction takes place in a 96-well-microtiter
plate which was read by a PC-controlled photometer
at 405 nm. This method, with a detection limit
of 0.1 μmol/l showed good analytical recovery (90 to
110%) and a low imprecision (intra-assay coefficient of
variation (CV) of 5% and an inter-assay CV of 7.5%). In
contrast to the autoanalyser method, the microtiter
plate-method is suitable both for series up to 24 samples
(3-fold) and for single samples. A comparison of
87 samples in the range of 0.1 to 60 μmol/l which were
measured with both Sandell-Kolthoff based methods
showed no obvious discrepancy.
These two methods showed a good agreement for
the determination of urinary iodine. This guarantees
that the results of earlier epidemiological studies can
be compared with recent studies performed in our and
many other laboratories.
Evaluation of the Clinical Chemistry Analyser Olympus AU400
Authors: Lasnier Elisabeth , Mario Nathalie , Boque Marie-Chantal , You Sok-Nay , Vaubourdolle Michel 
Published: October 2004

Abstract
The Olympus AU400 analyser (Olympus, Tokyo,
Japan) is an automated chemistry instrument for turbidimetric,
spectrophotometric and ion selective electrode
measurements. Overall analytical performances
of the AU400 and the reagents provided by Olympus
were evaluated according to the French Society of
Clinical Biology guidelines. Twenty parameters including
specific proteins, substrates, enzyme activities and
electrolytes were tested. The linearity exceeded the
specifications given by the manufacturer. Within- and
between-run imprecision (CV%), evaluated at two levels,
was below 1.5% for ion selective electrode parameters
and 3% for other analytes, except for CO2, alkaline
phosphatase at low levels and magnesium.
Results compared well with those obtained with the
analysers routinely used in our laboratory (Behring
BNII, Olympus AU800 and Beckman CX3 Delta). The
usual positive interferences from lipaemia and haemoglobin
on total protein measurement were observed.
Creatine kinase and alkaline phosphatase assays were
the subject of positive and negative interference by
haemoglobin, respectively. There was a negative interference
by bilirubin in the uric acid, aspartate-amino
transferase, creatine kinase and lactate dehydrogenase
assays and a positive interference in the calcium
assay. The system was found to be very easy to use
and the workstation is user-friendly.
Comparison of Proteolytic Enzymes and Glutathione S-Transferase
Levels in Non-Heart-Beating Donors' (NHBD) Kidney Perfusates
Authors: Balupuri Shlokarth , Talbot David , El-Sheikh Mohamed , Snowden Chris , Manas Derek M. , Kirby John , Mantle David 
Published: October 2004

Abstract
In order to identify biochemical markers of kidney
damage prior to transplantation, we determined the
levels of activity of a range of proteolytic enzymes in
kidney perfusate samples from non-heart-beating
donor (NHBD) cases. Urinary protease activities have
been described as indices of kidney damage in renal
disease; their potential as markers of tissue damage in
kidneys before transplantation has not been assessed.
In an attempt to identify additional/ improved biochemical
markers, the present study compared the
levels of total glutathione S-transferase (GST) with
corresponding levels of several proteolytic enzymes in
perfusate fluid from machine perfused NHBD kidneys.
Proteases were selected to represent factors that may
influence enzyme efflux, such as intracellular localization
or molecular size.
Methods: Perfusate samples were obtained over an
8-hour period from machine-preserved NHBD kidneys.
Protease activities in these samples were determined
by fluorometric assays and comparison made with total
GST activity. Individual proteases were analysed in
the transplanted and non-transplanted kidney groups
(discarded on the basis of other viability parameters).
Results: A correlation between protease activity and
total GST was obtained for only leucyl- and pyroglutamyl
aminopeptidase. Furthermore, in the transplanted
group, it was possible to set nominal upper
limits of activity for alanyl-arginyl- and dipeptidyl IV-aminopeptidase
(AP). In the non-transplanted kidney
group protease levels were increased above “normal”
upper limits for the same enzyme types. By the use of
alanyl AP it was possible to discriminate 75% of unsuitable
kidneys discarded by the use of other criteria.
Conclusion: The lack of correlation between total
GST and protease activity for most of the enzymes investigated
and alanyl AP levels in perfusate samples
could be related to differences in cellular localisation,
suggesting that assays of alanyl AP may give complimentary
biochemical information relating to kidney
tissue damage. Quantification of alanyl AP in machine
perfusate samples may be a valuable additional independent
biomarker of tissue damage.