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Biomedical subjects

G Lippi

Publications and source records attributed to G Lippi.

At least 73 records · Page 4Linked to original sources
PubMed1997

Clinical performance of leukocyte differential on the new Roche Cobas Vega haematological analyzer.

The Roche Cobas Vega is a five-part differential haematological analyzer evolving from the former Cobas Helios, Argos and Micros. As the leukocyte differential of Cobas Vega displays several interesting features, we analyzed its clinical performance and compared it either to three other commercial haematological analyzers (Technicon-Bayer H*2, Coulter STKS and Abbott CD-3500) or to the manual reference method, as described in the National Committee for Clinical Laboratory Standards (NCCLS) H20-A protocol. Within- and between-batch coefficients of variations (CVs) of the white blood cell differential were satisfactory and close, or even better than the ones reported for the other instruments: 0.81% (neutrophils), 1.87% (lymphocytes), 6.76% (monocytes), 7.73% (eosinophils) and 22.55% (basophils). The identification of abnormalities in the white blood cell differential was obtained either on the basis of the instrument-specific ranges or on the generation of one or more flags by comparison with results of the manual reference method. The Vega demonstrated remarkable performance in terms of specificity (90.0%), sensitivity (91.6%) and both positive (97.7%) and negative (69.1%) predictive values; likewise, the global efficiency was the best (90.3%) among the analyzers we evaluated. The flags generated in the presence of morphological abnormalities of the samples displayed excellent sensitivity (from 88.9% to 100%), specificity (from 93.5% to 98.3%) and satisfactory positive (from 51.1% to 66.7%) and negative (from 98.3% to 100%) predictive values. We conclude that the analytical performance of the new Cobas Vega haematological analyzer can significantly improve the identification of various haematological abnormalities as an important contribution to the accuracy is provided by the new staining for granulocytes.

Evaluation Studies as Topic↗
PubMed1997

Evaluation of five new liquid stable applications on the Roche Cobas Integra.

In the present study the analytical performances of five new liquid applications on the Roche Cobas Integra were evaluated: urea and high density lipoprotein (HDL) cholesterol in serum and glucose, creatinine and inorganic phosphorus in urine. The analytical evaluation consisted of imprecision, linearity and method comparison performed against either the actual Cobas Integra granulate applications or the corresponding methods on a Hitachi 704, according to the National Committee for Clinical Laboratory Standards protocols. Over 3700 results were obtained within 3 months. Average values of within-run and between-day coefficients of variation (CVs) were 1.15% and 1.48%, respectively, holding to a mean total CV of 2.17%. The linearity was excellent for all the five applications evaluated as the relative non-linearity was always within 1.53%, thus completely fulfilling the 2.5% upper limit. A strict correlation was observed by comparing results of 120 samples with either the corresponding granulate applications on Cobas Integra or the Hitachi reagents. Linear regression analysis of the results yielded correlation coefficients always above 0.987 and the slopes of the Passing & Bablok regression lines did not deviate by more than 7% from unity. No drift was observed over 4 hours of operations. In conclusion, the performance of these new Cobas Integra liquid applications, as demonstrated by the present study, proved them to be highly suitable for routine use in clinical laboratories.

Chemistry, Clinical↗
PubMed1996

Significant reduction of the bias among commercial immunoassays for lipoprotein(a) after use of a uniform calibrator.

Despite the increasing interest in the measurements of lipoprotein(a) (Lp(a) in serum or plasma, at present there is no effective standardization for Lp(a) assays; the main problems to solve are represented either by the lack of a suitable primary standard or by the absence of a reliable and widely available reference method. A first step is hence the uniformity of calibration of different immunoassays. We calibrated three commercial immunoassays for Lp(a) (enzyme linked immunosorbent assay (ELISA), latex-enhanced immunonephelometric assay (LINA), and immunonephelometric assay (INA) with either proprietary standards or purified Lp(a) material obtained with a rapid and simple procedure. Final results of purified Lp(a) calibration were reported in terms of protein Lp(a) mass whereas we were able to quantify the exact protein concentration of our purified lipoprotein. The uniformity of the calibration of the different assays led to a significant improvement of regression slopes (from 1.88 to 0.90 ELISA vs. LINA, from 1.45 to 0.95 ELISA vs. INA and from 1.27 to 0.96 INA vs. LINA) and correlation coefficients (from 0.990 to 0.994 ELISA vs. LINA, from 0.987 to 0.990 ELISA vs. INA and from 0.985 to 0.987 INA vs. LINA). Furthermore, the significant differences among Lp(a) values obtained after calibration with proprietary standards were minimized, becoming non-significant in two out of three cases. In conclusion, we demonstrated that a better agreement of Lp(a) values obtained with different commercial assays could be simply reached by uniformity of calibration and by employing standards with values accurately measured.

Bias↗
PubMed1996

Simple and rapid procedure for the purification of lipoprotein(a).

Lipoprotein(a) [Lp(a)] is a low-density lipoprotein-like particle displaying strong athero-thrombotic properties. Highly purified Lp(a) is increasingly requested for standardization of Lp(a) measurements and for biological studies. Several procedures have been described for Lp(a) separation and purification but none of them appear completely suitable. We present here a procedure for Lp(a) purification based on sequential elutions after lysine-Sepharose affinity chromatography. We were able to identify four distinct subspecies of Lp(a) showing different affinity to epsilon-amino groups of lysine-Sepharose, simply by modifying molarity and pH of the eluents; the fraction obtained in highly purified state represented the major form and could be eluted with 0.5 M sodium phosphate buffer (pH 4.4). Advantages of this procedure are represented by simplicity, rapidity and final yield.

Chromatography, Affinity↗
PubMed1996

Testosterone-induced suppression of lipoprotein(a) in normal men; relation to basal lipoprotein(a) level.

The concentration of lipoprotein(a) [Lp(a)] in human plasma is largely genetically determined and is inversely correlated to the size of apolipoprotein(a) [apo(a)]. Additionally, Lp(a) values are relatively stable within individuals and are only marginally susceptible to therapeutic treatment. The aim of our study was to evaluate the effect of exogenous testosterone on plasma Lp(a) concentration. The study was carried out on 19 healthy men who were receiving weekly intramuscular injections of 200 mg testosterone enanthate. Lp(a) values were determined at multiple time-points by a double monoclonal antibody-based enzyme immunoassay. This method is not sensitive to variation in Lp(a) size and the values are expressed in nmol/l. Apo(a) size isoforms were determined by agarose gel electrophoresis followed by immunoblotting. No correlation was found between the baseline Lp(a) values and the baseline values of testosterone or estradiol. The Lp(a) response to testosterone treatment varied widely among subjects and was dependent upon the pretreatment Lp(a) concentration. For 10 subjects with low Lp(a) values (< 25 nmol/l), no significant decrease in Lp(a) was observed while, for the nine individuals with Lp(a) values > 25 nmol/l, there was a significant and consistent reduction in Lp(a) ranging from 25 to 59%. Lp(a) levels returned to baseline values following cessation of testosterone administration. Apo(a) size polymorphism did not appear to play a role in the determination of Lp(a) response to testosterone.

Adult↗
PubMed1996

Effects of anticoagulants on lipoprotein(a) measurements with four commercial assays.

Lipoprotein(a) levels in plasma are considered an independent risk factor for atherosclerosis at different sites. Although Lp(a) measurements have recently gained interest in clinical laboratories, several problems are still unresolved. A potential source of pre-analytical variability lies in the treatment of the specimens, since it has been reported that values of several lipid quantities are lower when measured in plasma instead of serum. Lp(a) was measured in serum and in EDTA-treated, heparinised and citrated plasma from 15 healthy volunteers. Four analytical methods were used: two enzyme linked immunosorbent assays [ELISA] based on a polyclonal anti-apolipoprotein(a) antibody and a polyclonal anti-apolipoprotein B antibody, respectively; and two immunonephelo-metric assays [INA] based on a N antiserum to Lp(a) and on three monoclonal antibodies adsorbed on latex particles, respectively. Our measured Lp(a) values in plasma were lower than those found in serum, in particular for EDTA-treated (anti-apolipoprotein(a) ELISA: p < 0.01, anti-apolipoprotein B ELISA: p < 0.001 and Latex enhanced INA: p < 0.001) and citrated plasma (anti-apolipoprotein(a) ELISA: p < 0.05, anti-apolipoprotein B ELISA: p < 0.001 and INA: p < 0.001). Lp(a) values measured in heparinised plasma were also lower than those found in serum, but the difference was not statistically significant.

Antibody Specificity↗
PubMed1995

Effect of acute exercise on some haematological parameters and neutrophil functions in active and inactive subjects.

In this work we studied the possible effects of acute exercise on some haematological parameters and on some functions of neutrophils in seven active and six inactive subjects. Physical exercise (10 min on a cycle ergometer at a heart rate of 150 beats.min-1) induced a significant increase in total leucocyte, lymphocyte and neutrophil concentrations in active subjects; serum iron and ferritin concentrations were lower in active compared to inactive subjects. Cellular adhesion, bactericidal activity and superoxide anion production did not change after exercise, while we also observed some differences between active and inactive subjects before exercise. In particular, the neutrophils from active subjects showed a significantly higher percentage of adhesion, higher bactericidal activity and lower superoxide anion production. In conclusion, the training induced changes in some neutrophil functions, while acute exercise influenced, overall, leucocyte concentrations.

Adult↗
PubMed1995

Effect of pyridoxal 5'-phosphate on human neutrophil aggregation in vitro.

Pyridoxal 5'-phosphate inhibited polymorphonuclear leukocyte aggregation in vitro in a dose-dependent fashion at concentrations ranging from 0.5 to 0.001 mmol/l. At pyridoxal 5'-phosphate concentrations between 0.4 and 0.5 mmol/l mean inhibition of aggregation was about 50%-60%. In this range, and at lower pyridoxal 5'-phosphate concentrations, phorbol myristate acetate-activated polymorphonuclear leukocytes showed a normal increase in volume, whereas swelling was inhibited at higher concentrations of pyridoxal 5'-phosphate. Since pyridoxal 5'-phosphate can be administered as a vitamin to man without any relevant side-effects its role as a physiological anti-aggregant of polymorphonuclear leukocytes warrants further investigation.

Adult↗
PubMed1995

Lipoprotein(a) immunoassays: comparison of a semi-quantitative latex method and two monoclonal enzyme immunoassays.

Lipoprotein(a) is considered an independent risk factor for atherosclerosis. A variety of analytical methods have been proposed for lipoprotein(a) measurement, the majority of which require dedicated instruments and are costly to perform, particularly when the aim is to screen for high lipoprotein(a) concentrations in large populations. We evaluated the sensitivity and specificity of a newly developed semi-quantitative latex method to assess its suitability for identifying subjects with high lipoprotein(a) levels. Based on clinical data, a sensitivity limit of 20 mg/dl of total lipoprotein(a) particle was selected for the latex method. Results obtained by the latex method on 204 subjects were compared with two enzyme immunoassays using two anti-apo(a) monoclonal antibodies with different specificities. In one assay, the detecting monoclonal antibody (MAb a-5) is directed against an epitope present in a variable number depending on the apo(a) size isoforms in lipoprotein(a), while the other assay the detecting monoclonal antibody (MAb a-40) is directed against an epitope present only once in lipoprotein(a) particles, irrespective of their apo(a) size. Both the latex method and the MAb a-5 assay demonstrated a 100% sensitivity, in that no false-negative results were found using the MAb a-40 assay as the gold standard. Eleven subjects (5.4%) were misclassified as false positive by MAb a-5 assay and 23 (11.3%) were misclassified by the latex method. Based on its 100% sensitivity and 89% specificity, we conclude that the lipoprotein(a) latex method is a cost-effective rapid approach for screening large populations.

Adult↗
PubMed1995

Changes in several neutrophil functions in basketball players before, during and after the sports season.

Neutrophils play an important role in the immune system, forming the "first line of defence" against invading microorganisms and there are few data available concerning neutrophil functions in relation to exercise. We investigated in 7 basketball professional players possible changes before, during and after the sports season, in some haematological parameters and in several aspects of the phagocytic process of neutrophils, such as adhesion, superoxide anion release and bactericidal activity. Training and competitions produced a significant rise in the number of total leukocytes and differential counts, but the values returned to the pre-start levels 3 weeks after the end of the championship. The bactericidal activity and the superoxide anion released were significantly greater during the sports season, while the percentage of cellular adhesion significantly decreased during the championship; after the sports season the values returned to the control levels. As in the literature data concerning neutrophil functions in relation to exercise are non-convergent, it is important in our opinion, to understand whether the alterations induced by exercise can persist after repeated stimuli.

Adult↗
PubMed1995

"Non-dipper" hypertensive patients and progressive renal insufficiency: a 3-year longitudinal study.

Patients with a blunted or absent nocturnal blood pressure (BP) drop may be subject to increased risk for target organ damage. In this 3-year longitudinal case-control study we tested the hypothesis that an association exists between a reduced or absent night-time fall in BP and a future decline of kidney function in renal hypertensive patients. The case subjects were 48 hypertensives with renal insufficiency, divided into two groups according to the presence (dippers: n 20) or absence (non-dippers: n 28) of a nocturnal diastolic BP decline greater than 10% of daytime values, detected by ambulatory BP monitoring. At the baseline evaluation the two groups did not differ with respect to age, sex, body weight, office systolic and diastolic BP, mean daytime ambulatory BP, creatinine clearance, 24 h proteinuria. In the ambulatory BP profiles over a 3-year follow-up the nocturnal reductions of systolic and diastolic BP in the dippers were 14% and 15%, respectively, vs 7% and 5% in the non-dippers (p = 0.002/0.003). The non-dippers had a faster rate of creatinine clearance decline than the dippers (0.37 +/- 0.26 vs 0.27 +/- 0.09 ml/min/month; p = 0.002). Urinary protein excretion increase was higher in the non-dipper group than in the dipper group (993 +/- 438 vs 691 +/- 222 mg/24 h; p = 0.009). This longitudinal study suggests that the non-dipping pattern of ambulatory BP can be associated with a faster progression of renal insufficiency in renal hypertensives and that a proper nocturnal BP control is an additional aim of antihypertensive therapy.

Antihypertensive Agents↗
PubMed1994

Autosomal recessive hypermyelinating neuropathy.

We studied three patients from two kinships, affected by early onset hereditary motor and sensory neuropathy with probable autosomal recessive inheritance (HMSN type III). Morphological studies of sural nerve biopsies revealed an abnormal myelin proliferation. Two adult patients with long-term follow up, lost ability to walk at 28 and 22 years and showed severe involvement of the cranial nerves. Our observations suggest that "hypermyelination neuropathy" with early onset is a progressive disease with poor long-term prognosis. In one kinship the occurrence of the disease in two sibs of both sexes but not in parents, is consistent with an autosomal recessive inheritance. Familial cases of hypermyelination neuropathy have not been described in previous reports. Morphological aspects of this condition are compared with other forms of hypermyelination neuropathy.

Adult↗
PubMed1994

A simple assessment of human neutrophil adhesiveness.

The adhesiveness of human polymorphonuclear leukocytes was assessed in serum-coated polystyrene spectrophotometric cuvettes. Capped cuvettes, containing no more than 2 x 10(6) resting or concanavalin A-treated (100 micrograms/ml) polymorphonuclear leukocytes, were laid horizontally and subjected to three 90 degrees rotations on their major axis at fixed times. After incubation at room temperature, non-adherent cells remaining in suspension were counted on the Coulter counter STKS hematological analyzer. After a 16-min incubation (4 min each side of the cuvette) the adhesion of concanavalin A-activated neutrophils ranged from 98% to 100% and the adhesion of resting neutrophils from 30% to 35% (mean 32.4 +/- 2.2%, n = 10). An 8-min incubation (2 min each side) led to approximately 50% adhesion of concanavalin A-activated neutrophils (mean 49.9 +/- 2.2%, range 46%-54%, n = 16), whereas the adhesion of resting cells was about 21% (mean 21.4 +/- 1.6%, range 19%-24%, n = 16). The variation in percentage adhesion in repeated assays did not exceed 4% using concanavalin A-activated cells and 7.5% with resting neutrophils. The procedure is very rapid, easy to perform and precise, and no special apparatus or glassware is necessary. The method also allows microscopic evaluation of shape changes of adherent neutrophils through the clear sides of the cuvettes.

Animals↗
PubMed1994

Assessment of neutrophil aggregation by Coulter STKR and STKS haematological analysers.

We have studied an alternative method to aggregometry for the assessment of human polymorphonuclear (PMN) leucocyte aggregation. This simple, rapid and reliable procedure counts unaggregated cells on both Coulter STKS and STKR haematological analysers by the impedance principle. Aggregation of PMN was induced by 15 min incubation with fresh autologous serum (FAS) after a 10 min phorbol myristate acetate (PMA) activation of neutrophils in small aliquots (0.25 ml) of suspension containing about 4.0 x 10(9) PMN/1. Differences (x 100) between count of resting and PMA+FAS treated neutrophils/count of resting PMN reflect percent aggregation. By this procedure, PMN aggregation did not occur in autologous plasma from EDTA anticoagulated whole blood; it was partially inhibited by hydrocortisone, whereas inactivated or Zymosan activated sera gave values similar to those from FAS induced aggregation. PMA aggregation was dependent on Ca2+ + Mg2+ concentration. Intra-assay analytical variability did not exceed 4% on either instrument. Reference values (n = 20) of percent PMN aggregation were 50.7 +/- 4.7 on STKS and 47.1 +/- 4.8 on STKR. Most probably, the interindividual variance was due to the physiological variability of Mg2+ and/or Ca2+ concentrations in FAS. Thus, this procedure reflects the true PMN aggregability status in a given subject, and in a given electrolyte environment.

Adult↗
PubMed1994

High light scatter by neutrophils in the Bayer-Technicon H*2 analyzer: a screening test of morphologically defective responsiveness to in vitro chemotactic stimulation.

The Bayer-Technicon H*2 haematological analyser provides differential white blood cell count, including the assay of polymorphonuclear leukocytes by light scattering and the absorbance increase following the cytochemical reaction for myeloperoxidase. The mean value of polymorphonuclear leukocytes scatter, which reflects polymorphonuclear leukocytes volume, is printed in a separate report "for laboratory use only" as a ybar value in arbitrary units. In certain patients neutrophils displayed an unreported correlation between polymorphonuclear leukocytes high ybar basal values (> or = 37.00 arbitrary units) (determined on the H*2) and a defective response in vitro to the chemoattractant, formyl-methionyl-leucyl-phenylalanine (determined by microscopic evaluation of polymorphonuclear leukocytes shape change (polarization)). The patients showing no polymorphonuclear leukocyte response or a defective one to formyl-methionyl-leucyl-phenylalanine were all affected by "Systemic Inflammatory Response Syndrome (SIRS)". Therefore the predictive value of the positive test for SIRS is 100%. On the other hand 8.8% of SIRS patients had polymorphonuclear leukocytes < 37.00 arbitrary units of ybar basal value and a "normal" response to formyl-methionyl-leucyl-phenylalanine; the predictive value of the negative test being 90%. Since we demonstrated in vitro a dose-dependent deactivation of endotoxin or lipopolysaccharide-pretreated polymorphonuclear leukocytes, the "normal" response to formyl-methionyl-leucyl-phenylalanine of the "false negative" cases may occur because the endotoxaemia in these patients is too low to prevent it.(ABSTRACT TRUNCATED AT 250 WORDS)

Cell Size↗