Control of oral anticoagulant therapy with whole blood prothrombin time devices: the future has begun.
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Anticoagulants↗
Biomedical subjects
A Tripodi
Publications and source records attributed to A Tripodi.
A comparison of two sodium citrate concentrations in two evacuated blood collection systems for prothrombin time and ISI determination.
The prothrombin time is usually measured in citrated plasma. The W.H.O. recommended concentration of sodium citrate for blood collection for laboratory control of oral anticoagulant therapy is 0.109 M. Some evacuated blood collection systems include 0.105 M sodium citrate. The purpose of the present study was to establish the difference in ISI calibration between 0.109 and 0.105 M citrate, using 7 types of thromboplastin and various types of instrumentation. The two citrate concentrations were provided in both evacuated siliconised glass tubes and in evacuated polyethylene terephtalate (PET) tubes. The ISI difference between the two citrate concentrations was 5.4% for one system but not greater than 3% for all other systems when blood samples were collected with either siliconized glass or PET tubes. Most of the ISI differences between the two citrate concentrations were not significant at the 5% level. It is concluded that the ISI differences between 0.105 M and 0.109 M citrate are not of practical importance. In contrast, ISI differences between siliconised glass and PET tubes, using either 0.105 or 0.109 M citrate, were significant (p <0.05) for most thromboplastin systems and amounted to 7%. ISI interchange between these glass and PET tubes could induce INR differences amounting to 14%, which could affect clinical dosage of oral anticoagulants.
Blood Specimen Collection↗
Fetal echocardiography: a comparison of different techniques.
There is great availability on the market of echocardiographic instruments that have innovative technological systems which use second harmonic imaging. The true usefulness and applicability of these instruments in fetal echocardiography have yet to be verified. The objective of the present study was to verify the impact of this echographic technology in the diagnostic evaluation of the fetal cardiovascular system. Thus, an Acuson Sequoia echograph which uses native tissue harmonic imaging (NTHI) was used to examine 30 pregnant women between the 28th and 32nd week of gestation. The b-mode and m-mode images were acquired and recorded by the conventional method and by the second harmonic imaging method without contrast medium. Two experts in the field analyzed and compared the images recorded by the two methods and made diagnostic and qualitative judgements. The 30 cases examined were unanimously determined to have no visible echo pathologies. The morpho-biometric evaluation obtained by the two methods did not show any significant differences and the quality of the images obtained by the traditional method was found on average to be higher. In conclusion we believe that the use of second harmonic imaging for studies of the fetal heart does not seem to offer any advantages with respect to the conventional method.
Echocardiography↗
Familial overexpression of beta antithrombin caused by an Asn135Thr substitution.
We have investigated the basis of antithrombin deficiency in an asymptomatic individual (and family) with borderline levels (approximately 70% antigen and activity) of antithrombin. Direct sequencing of amplified DNA showed a mutation in codon 135, AAC to ACC, predicting a heterozygous Asn135Thr substitution. This substitution alters the predicted consensus sequence for glycosylation, Asn-X-Ser, adjacent to the heparin interaction site of antithrombin. The antithrombin isolated from plasma of the proband by heparin-Sepharose chromatography contained amounts of beta antithrombin (the very high affinity fraction) greatly increased (approximately 20% to 30% of total) above the trace levels found in normals. Expression of the residue 135 variant in both a cell-free system and COS-7 cells confirmed altered glycosylation arising as a consequence of the mutation. Wild-type and variant protein were translated and exported from COS-7 cells with apparently equal efficiency, in contrast to the reduced level of variant observed in plasma of the affected individual. This case represents a novel cause of antithrombin deficiency, removal of glycosylation concensus sequence, and highlights the potentially important role of beta antithrombin in regulating coagulation.
Adult↗
Is it only a mechanical matter? Histologic modifications of the aorta underlying external banding.
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Aged↗
Intimal-type primary sarcoma of the thoracic aorta presenting as a saccular false aneurysm: report of a case with evidence of rhabdomyosarcomatous differentiation.
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Aged↗
The effect of sample size on fresh plasma thromboplastin ISI determination.
The possibility of reduction of numbers of fresh coumarin and normal plasmas has been studied in a multicentre manual prothrombin (PT) calibration of high international sensitivity index (ISI) rabbit and low ISI human reference thromboplastins at 14 laboratories. The number of calibrant plasmas was reduced progressively by a computer program which generated random numbers to provide 1000 different selections for each reduced sample at each participant laboratory. Results were compared with those of the full set of 20 normal and 60 coumarin plasma calibrations. With the human reagent, 20 coumarins and seven normals still achieved the W.H.O. precision limit (3% CV of the slope), but with the rabbit reagent reduction < 50 coumarins with 17 normal plasmas led to unacceptable CV. Little reduction of numbers from the full set of 80 fresh plasmas appears advisable. For maximum confidence, when calibrating the ISI of a new reagent, it would always seem worthwhile to use the full number of patients' and normal plasmas at each centre for the initial calibration, i.e. 60 fresh coumarin and 20 fresh normal plasmas. Smaller numbers should achieve the required precision with the lower ISI reagent.
Animals↗
The risks of heparin-neutralizing substances.
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Blood Coagulation Tests↗
Risk of venous thromboembolism and clinical manifestations in carriers of antithrombin, protein C, protein S deficiency, or activated protein C resistance: a multicenter collaborative family study.
Deficiencies of antithrombin (AT), protein C (PC) or protein S (PS), and activated protein C resistance (APCR) are very well-established coagulation defects predisposing to venous thromboembolism (VTE). We performed a retrospective cohort family study to assess the risk for VTE in individuals with AT, PC, or PS deficiency, or APCR. Five hundred thirteen relatives from 9 Italian centers were selected from 233 families in which the proband had had at least 1 episode of VTE. We calculated the incidence of VTE in the whole cohort and in the subgroups after stratification by age, sex, and defect. The overall incidence of VTE (per 100 patient-years) in the group of relatives was 0.52. It was 1.07 for AT, 0.54 for PC, 0.50 for PS, 0.30 for APCR, and 0.67 in the group with a double defect. The incidence was associated with age, but not with sex. The mean age at onset was between 30 and 40 years for all the coagulation defects. Women had the peak of incidence in the age range of 21 to 40 years, earlier than men. The lifetime risk for VTE was 4.4 for AT versus APCR, 2.6 for AT versus PS, 2.2 for AT versus PC, 1.9 for PC versus APCR, and 1.6 for PS versus APCR. AT deficiency seems to have a higher risk for VTE than the other genetic defects. There is a relation between age and occurrence of thrombosis for both men and women. The latter had the peak of incidence earlier than the former.
Activated Protein C Resistance↗
Correction for lack of coincidence of normal and abnormal calibration slopes in ISI determination.
An underlying assumption of the WHO orthogonal regression model for prothrombin time standardisation is that a single line describes the relationship between PT of abnormal and normal plasmas. The aim was to evaluate Tomenson's correction for lack of coincidence using the model of the ECAA human reagent with lyophilized plasma calibrations. Local ISI calibrations using ECAA lyophilized normal and abnormal plasmas were performed on coagulometers with the ECAA human thromboplastin. They gave a high incidence of non-linearity of the two calibration slopes. The mean percentage deviation from assigned values of the target coumarin plasmas coagulometer INR at 45 centres before local ISI correction was 16.6% and 11.9% respectively in two studies. After local ISI calibration. the unsigned deviation was reduced to 5.54% and 6.61% respectively. Tomenson's correction for non-linearity further reduced the mean deviation to 3.08% and 2.96% respectively in the two studies demonstrating the value of the procedure.
Humans↗
Multicenter evaluation of lyophilized and deep-frozen plasmas for assignment of the International Normalized Ratio.
The interlaboratory variation of the International Normalized Ratio (INR) in various external quality assessment schemes is still relatively high. This is partly caused by inaccuracy of manufacturers' stated International Sensitivity Index (ISI) and/or local instrumentation effects. The interlaboratory variation and accuracy of INR determinations may be improved by a local calibration procedure based on lyophilized plasmas with assigned INRs. The purpose of the present study was to determine INR values for different types of lyophilized plasmas to be used for local calibration. A total of 13 lyophilized plasmas (one normal, six from coumarin-treated patients, six artificially depleted) were analyzed by 10 laboratories, each using five calibrated prothrombin time (PT) systems. INRs were calculated for each plasma using each laboratory's specific ISI and mean normal prothrombin time values. In the same way, five deep-frozen pooled plasmas from coumarin-treated patients were analyzed. There were significant INR differences for the lyophilized plasmas between the prothrombin time systems. The differences were relatively small for the deep-frozen coumarin plasmas (CV 2.6-3.3%) and three lyophilized coumarin plasmas from one manufacturer (CV 3.7-4.8%). Important INR differences were observed for three lyophilized coumarin plasmas from another manufacturer (CV 9.5-14.1%) and several artificially depleted plasmas (CV 5.3-12.8%). The citrate concentrations in the artificially depleted plasmas were lower than those in the normal and coumarin plasmas. These differences should be considered in the selection and certification of plasmas as calibrants for local calibration of PT systems. The lyophilized plasmas' INR values obtained in the present study will be used for a field study of local PT calibration to assess their efficacy.
Animals↗
Calibration of local systems with lyophilized calibrant plasmas improves the interlaboratory variability of the INR in the Italian external quality assessment scheme.
Calibration with lyophilized calibrant plasmas certified in terms of PT with International Reference Preparations for thromboplastin has been proposed to minimize the effect of coagulometers on the INR. Aim of this study was to test the ability of local calibration with lyophilized calibrant plasmas, combined with a modified statistical approach, to improve the interlaboratory variability of the INR measured on two test plasmas (one coumarin and one artificially-depleted) by participants in the External Quality Assessment Scheme (EQAS). Sets of lyophilized calibrant and test plasmas were sent to the participants in the EQAS, who were asked to determine PT with their own reagent/ instrument combination (local system). Results were returned as PT together with information on the type of local system, the stated International Sensitivity Index (ISI) and the geometric mean of PTs determined by testing with the local system fresh plasmas from 20 healthy subjects. Ninety-two participants using 9 and 11 brands of reagents and instruments returned results. The CV of the INR determined with the stated ISI for the coumarin (Mean INR = 4.39) and artificially-depleted (Mean INR = 4.23) test plasmas were 11.2% and 10.3% and were reduced on the average by 34% and 54%, respectively, when the INR was calculated with the local ISI. In conclusions, results from this field study involving laboratories and testing systems representative of the real situation in oral anticoagulant monitoring in our country, indicate that local calibration by artificially-depleted plasmas, combined with the proposed statistical approach, is suitable to improve the interlaboratory agreement on the INR.
Freeze Drying↗
Benefits of antibiotic prophylaxis in laparoscopic gynaecological surgery.
The prevention of infectious complications by antibiotic prophylaxis has made traditional or laparoscopic surgery much safer but at the same time has contributed to an uncontrolled and often irrational use of every kind of antibiotic. In this study we wanted to show that often mini-invasive surgery like laparoscopy can be practised without the use of antibiotics. Thus, postoperatively several patients undergoing laparoscopic, diagnostic and operative interventions were followed-up. The results showed that subjects without antibiotic therapy did not have any symptomatology that could be ascribed to bacterial infections. In conclusion this study has demonstrated that laparoscopic surgery, especially without any complications, should follow the elementary rules of surgical techniques and surgical asepsis and that antibiotic prophylaxis is not always necessary.
Adolescent↗
Influence of three types of automated coagulometers on the international sensitivity index (ISI) of rabbit, human, and recombinant human tissue factor preparations--a multicenter study.
Five tissue factor reagents and three types of automated instruments for prothrombin time (PT) determination were studied in an international multicenter collaborative exercise. The purpose of this work was to determine the international sensitivity index (ISI) for each combination of reagent and instrument against the international reference preparation RBT/90. Each type of instrument was used by 3 or 4 centers to assess the interlaboratory variation of the ISI. The interlaboratory variation of the ISI for each combination of reagent and instrument ranged between 0.4% and 7.8% coefficient of variation. For three reagents, the mean ISI values for ACL (nephelometric) and STA (mechanical) were practically identical, but the mean ISI values for MLA (photo-optical) were at least 10% higher. For two other reagents prepared from rabbit tissue, the mean ISI values increased in the order ACL, STA, MLA. The widest range of mean ISI values was noted with one rabbit tissue factor reagent: 1.68 for ACL and 2.00 for MLA. Exclusion of patient specimens with INR <1.5 and INR >4.5 determined by the international reference preparation resulted in a slight decrease of the mean ISI. The interlaboratory variation of the International Normalized Ratio (INR) was assessed from the results obtained with common lyophilized and deep-frozen plasmas. The use of instrument-specific ISI values resulted in reduced interlaboratory variation of the INR. It is recommended that thromboplastin manufacturers provide instrument-specific ISI values.
Animals↗
A comparison of artificially-depleted, lyophilized coumarin and fresh coumarin plasmas in thromboplastin calibration. European Concerted Action on Anticoagulation.
Artificially-depleted lyophilized plasmas and lyophilized coumarin plasmas were prepared and compared with fresh coumarin plasmas to assess their comparative reliability in local thromboplastin calibration using the manual prothrombin time (PT) technique. Their certified PT values were inserted in turn on the vertical axis in place of the PT obtained with fresh coumarin plasmas. PT results were obtained at eight ECAA national laboratories ('test centres') and inserted on the horizontal axis. The resulting thromboplastin calibration slopes were compared with conventional fresh coumarin plasma calibration slopes at the same 'test centres'. When 60 artificially-depleted plasmas were substituted for 60 fresh plasmas, the mean calibration slopes with the human plain International Reference Preparation (IRP) were 4.2% higher. For comparison with 20 lyophilized coumarins, three sets of 20 artificially-depleted plasmas were selected in sequential order from the 60. The lyophilized coumarin plasmas gave a mean deviation of 9.6% from the fresh plasma calibration slopes compared with values of 2.0%, 6.1% and 11.7% for the three sets of 20 depleted plasmas. Although both types of lyophilized plasma calibration slopes give measurable differences from conventional fresh plasmas, these may be regarded as acceptable in clinical terms.
Blood Coagulation Tests↗
A comparison of linear and orthogonal regression analysis for local INR determination in ECAA coagulometer studies. European Concerted Action on Anticoagulation.
International sensitivity index calibrations based on the W.H.O. recommended method depend on orthogonal regression analysis. As this is not readily available in statistical packages, comparison has been made with simple linear regression analysis in a study of coagulometer effects on the International Normalized Ratio (INR) at 155 European centres. Sets of seven lyophilized normal and 20 lyophilized artificially depleted abnormal plasmas were provided with five coumarin test plasmas and two European Concerted Action on Anticoagulation reference thromboplastins (low International Sensitivity Index (ISI) human and high ISI rabbit). Local ISI based on the artificially depleted lyophilized plasmas using conventional orthogonal regression gave good correction for local coagulometer effects on the human reagent and minimal correction with the rabbit reagent INR. Results were considerably worse after attempts at correction using calibration based on linear regression analysis with both reagents. The results indicate that calibration of coagulometer prothrombin time systems using simple linear regression is not appropriate.
Animals↗
Ultrastructural features of bovine cumulus-corona cells surrounding oocytes, zygotes and early embryos.
Integrated transmission and scanning electron microscopic (TEM and SEM) techniques have provided the first detailed description of the ultrastructural features of the bovine cumulus-corona (CC) cells surrounding oocytes at the time of final maturation, zygotes and early cleaving embryos (2/4 to 6/8 blastomeres). TEM revealed the presence of rough endoplasmic reticulum and Golgi complexes in the cytoplasm of CC cells surrounding immature, mature and fertilized eggs, and also revealed an increasing amount of smooth endoplasmic reticulation membranes, lipid droplets and mitochondria with villiform and/or tubular cristae in the cytoplasm of CC cells during maturation and fertilization of the oocyte. In addition, a loss of cell-to-cell junctions between CC cells was evident. TEM also demonstrated that a few residual CC cells were still associated with early embryos and that these cells showed rather degenerative or apoptotic patterns, the latter pattern also observed on cells associated with fertilized eggs. SEM revealed that the complex of CC cells of immature oocytes was compact with narrow intercellular spaces, which progressively enlarged in size around mature oocytes. This phenomenon is mostly due to the production of abundant extracellular matrix. Immature CC cell complexes possessed characteristic long and filiform microvilli whereas the surface of CC cells surrounding mature oocytes showed numerous blebs and occasional large cytoplasmic protrusions as well as microvilli. Zygotes and early embryos were covered with a few polyhedral CC cells possessing scarce and short microvilli and a large amount of pleomorphic blebs. This study demonstrated a precocious luteinization occurring in bovine CC cells at ovulation until zygote segmentation, and this process was associated with a progressive apoptotic mechanism that ended in the complete denudation of the zona pellucida covering the early embryo. The presence of CC cells around the maturing oocyte and fertilized egg could have important functions related to the microenvironmental requirements of ovum maturation as well as facilitating activities related to fertilization.
Animals↗
Minimum lyophilized plasma requirement for ISI calibration. European Concerted Action on Anticoagulation.
The minimum requirement of lyophilized plasma samples for a reliable International Sensitivity Index (ISI) was assessed by calibrations based on reducing numbers from a maximum of 60 depleted and 20 plasma samples from patients receiving coumarin using a manual technique and low ISI thromboplastin. The probability of achieving an international normalized ratio (INR) result within a clinically important range of 20% deviation has been assessed at European Concerted Action on Anticoagulation (ECAA) national laboratories in calibrations of ECAA reference thromboplastin against the certified prothrombin time (PT) with a reference thromboplastin. With conventional orthogonal regression using a certified PT and linear regression using certified INR, deviations and the coefficient of variation of the calibration slope increased with reduced numbers. The INR deviations became marked when the number of abnormal plasma samples was reduced to fewer than 20. Calibrations of 3 abnormal plasma samples and 1 lyophilized normal plasma sample gave a high incidence of deviations greater than 10% with an INR of 3.0. The study demonstrates that with both methods of analysis, an optimum minimum number of lyophilized plasma samples is needed for a reliable local ISI.
Calibration↗