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PubMedDate not supplied

A good practice guide to the administration of substances and removal of blood, including routes and volumes.

This article is the result of an initiative between the European Federation of Pharmaceutical Industries Associations (EFPIA) and the European Centre for the Validation of Alternative Methods (ECVAM). Its objectives are to provide the researcher in the safety evaluation laboratory with an up-to-date, easy-to-use set of data sheets to aid in the study design process whilst at the same time affording maximum welfare considerations to the experimental animals. Although this article is targeted at researchers in the European Pharmaceutical Industry, it is considered that the principles underpinning the data sets and refinement proposals are equally applicable to all those who use these techniques on animals in their research, whether in research institutes, universities or other sectors of industry. The implications of this article may lead to discussion with regulators, such as those responsible for pharmacopoeial testing. There are numerous publications dealing with the administration of test substances and the removal of blood samples, and many laboratories also have their own "in-house" guidelines that have been developed by custom and practice over many years. Within European Union Directive 86/609EEC1 we have an obligation to refine experiments to cause the minimum amount of stress. We hope that this article will provide background data useful to those responsible for protocol design and review. This guide is based on peer-reviewed publications whenever possible, but where this is not possible we have used "in-house" data and the experience of those on the working party (as well as helpful comments submitted by the industry) for a final opinion. The guide also addresses the continuing need to refine the techniques associated with the administration of substances and the withdrawal of blood, and suggests ways of doing so. Data-sharing between laboratories should be encouraged to avoid duplication of animal work, as well as sharing practical skills concerning animal welfare and scientific problems caused by "overdosing" in some way or another. The recommendations in this guide refer to the "normal" animal, and special consideration is needed, for instance, during pregnancy and lactation. Interpretation of studies may be confounded when large volumes are administered or excessive sampling employed, particularly if anaesthetics are used.

Animals↗
PubMed1995

Effects of recombinant insulin-like growth factor-I and growth hormone on bone turnover in elderly women.

We evaluated the effects of recombinant insulin-like growth factor-I (IGF-I) and growth hormone (GH) on calciotropic hormones and bone turnover markers in 16 healthy elderly women 71.9 +/- 1.3 years of age (mean +/- SEM). Subjects consumed a fixed diet providing 1000 mg of calcium and 0.9 g/kg of protein for 10 days before starting baseline 24-h urine and blood collections. Specimens were collected for 6 consecutive days before initiating subcutaneous injections of GH (25 micrograms/kg/day, n = 5) and IGF-I at 60 micrograms/kg b.i.d. (high-dose, n = 5) or at 15 micrograms/kg b.i.d. (low-dose, n = 6) for 28 days. Resorption markers included urine hydroxyproline (OHP), total pyridinolines (PYD), and N-telopeptide; formation markers include osteocalcin, skeletal alkaline phosphatase (sALP), and type I procollagen carboxy-terminal extension peptide (CICP). For each subject, baseline daily turnover markers varied substantially (DV = 16-22%). With GH and high-dose IGF-I, resorption and formation markers increased progressively to maximum levels at day 21. For GH, the increase in day 21 PYD, N-telopeptide, osteocalcin, and CICP was 143, 111, 53, and 81%, respectively (p < 0.96-0.02). For high-dose IGF-I, these increases were 108, 81, 77, and 111% (p < 0.02-0.002). However, with low-dose IGF-I no change was observed in resorption markers while osteocalcin and CICP increased progressively (day 21, % increases = 88 +/- 51, 36 +/- 14). Twenty-four hour urine collections during the last days of baseline and of study drug were taken as six 4 h aliquots. When deoxyPYD was measured on these samples in the low-dose IGF-I group, a significant increase was observed only on the 0800-1200 h aliquot. Serum phosphorus concentrations increased with GH (21.2 +/- 3.3%) and high-dose IGF-I (8.8 +/- 3.6%) by day 21 but actually decreased by day 28 (-9.7 +/- 2.7, p < 0.02) with low-dose IGF-I. Urinary phosphorus excretion decreased with high-dose IGF-I only. Twenty-four hour calcium excretion increased with all treatments. These results indicate that both GH and high-dose IGF-I activate remodeling osteons. By contrast, low-dose IGF-I may directly increase osteoblastic function with only a minimal increase in bone resorption and may therefore provide a useful means to increase bone mass. The results also suggest some of the GH action on renal phosphorus handling represents a direct action of GH on the nephron which does not involve the intermediacy of IGF-I. Finally, even under controlled conditions bone turnover markers exhibit substantial daily variation so that a very large treatment effect will be required for these markers to have clinical utility.

Aged↗
PubMed2006

Potential clinical impact of taking multiple blood samples for research studies in paediatric oncology: how much do we really know?

Pharmacological studies of anti-cancer agents in children are essential to determine their clinical safety and efficacy, both of which can differ considerably from that observed in adults. However, the potential clinical impact of taking blood samples, in addition to those required for standard clinical practice is commonly a concern for both medical and allied staff and parents. Frequently quoted 'safe limits' of 3%-5% of total blood volume taken on any one study day are not based on published data and may not be acceptable for all patients. This article reviews some of the reasons why clinical pharmacology data for anti-cancer drugs is often lacking in a paediatric patient population, summarises data from a retrospective study investigating the potential impact of repeated blood sampling for research purposes and discusses how this issue may be more systematically addressed in future studies. Research involving children with cancer should be limited to those studies addressing key scientific questions and should be designed to limit both the number and volume of blood samples required.

Adolescent↗
PubMed1988

The fetal intrahepatic umbilical vein as an alternative to cord needling for prenatal diagnosis and therapy.

Seventy-one fetal blood samplings (FBS) were attempted from the intrahepatic portion of the umbilical vein (IHV) at 18-34 weeks; 54 were attempted primarily and 17 secondarily after a failed attempt at the placental cord insertion. Fetal blood was obtained in 89 per cent of the cases. Intravascular transfusion (IVT) was attempted on 31 occasions and successful in 24 (77 per cent). In all cases of failed sampling or transfusion via the IHV, prenatal diagnosis and/or therapy was accomplished using alternative procedures. On only one occasion was the procedure postponed. There were no losses or neonatal morbidity attributable to the procedure. FBS from the IHV may be considered as an alternative approach to sampling the placental cord insertion. It is recommended in cases where the approach to the placental cord insertion is difficult or hazardous.

Blood Specimen Collection↗
PubMed1994

Plasmodium falciparum: selective growth of subpopulations from field samples following in vitro culture, as detected by the polymerase chain reaction.

Analysis of the Plasmodium falciparum parasites circulating in the blood of infected persons frequently reveals the presence of two or more genetically distinct parasite populations. P. falciparum parasites cultured in vitro, from blood specimens collected in the field, are often used for biological, immunological, and drug-resistance investigations relating to the epidemiology in the area concerned or on the assumption that the parasites which grow in vitro are in general representative of all P. falciparum parasites. By using the polymerase chain reaction to detect and characterize a number of parasite polymorphic genes with great sensitivity, the composition of P. falciparum populations from 51 isolates were compared on the day of collection and following 2 months of in vitro culture. It was found that substantial changes in the parasite population profile could be detected in ca. 70% of the samples analyzed. The implications of this observation for studies using parasite isolates cultured in vitro are discussed.

Animals↗
PubMed1988

Blood sampling in very low birth weight infants receiving different levels of intensive care.

Sixty very low birth weight infants (birth weight 560-1450 g) were studied during the first 28 days of life. The infants were classified as group A (n = 19 infants who never required ventilator support), group B (n = 20 infants mechanically ventilated for minor respiratory problems), and group C (n = 21 infants ventilated for respiratory distress syndrome). Diagnostic blood sampling was measured, infants were checked for clinical symptoms and laboratory signs of anaemia 24 h before and after the transfusion of packed red cells. A total of 7998 punctures (average: 4.8 per infant per day) were performed, the mean blood loss due to diagnostic sampling was 50.3 ml/kg per 28 days (range 7-142) for all infants. A high correlation (rs = +0.91) was found between the blood volumes sampled and transfused. In group A, the mean blood loss was 24 ml/kg, and a total of 29 blood transfusions were administered. The most frequent symptoms of anaemia were poor weight gain and apnoeic spells. In group B, the mean blood loss was 60 ml/kg and a total of 97 blood transfusions were administered. In group C, the mean blood loss was 67 ml/kg and a total of 116 blood transfusions were administered. In both groups B and C, poor weight gain, pallor and distended abdomen were the most frequent symptoms of anaemia. Following the blood transfusion, haematocrit rose and blood pressure remained unchanged. The symptoms that responded most favourably to the blood transfusion were: poor weight gain, oxygen requirement, and distended abdomen. The results emphasize the need for miniaturizing laboratory techniques and monitoring blood sampling.

Anemia↗
PubMed1993

Evaluation of lymphocyte immunity in breast cancer patients.

One hundred forty-two breast cancer patients were evaluated for three functional immunologic parameters: the ability of their lymphocytes to proliferate in response to general T-(phytohemagglutinin) and B-lymphocyte (pokeweed mitogen) stimulators and their ability to proliferate in response to specific autologous tumor antigens. Tests were performed on patient blood specimens collected approximately 2 hours prior to surgery or 2-4 weeks following chemotherapy. T-lymphocyte functional competence was impaired in 83/142 (58%) of the patients, while B-lymphocyte competence was impaired in 34/142 (24%) of these patients. A total of 21/52 (40%) of the patients had lymphocyte immunity against autologous tumor antigen. There were weak associations between the ability of patients' T- and B-lymphocytes to function normally, and their ability to respond to autologous tumor-antigen. There was no relationship of age, tumor burden (clinical or pathological tumor size), extension to skin and/or muscle, or metastasis to any of the three immunological parameters. A relationship (p = 0.0463) between T-lymphocyte competence and pathological nodal status was observed; individuals that were node positive for disease, tended to have impaired T-lymphocyte function. When evaluating T- and B-lymphocyte competence and lymphocytic immunity against tumor antigen in pre- and post-chemotherapy patients, an immunologic rebound (increase in immune parameters shortly after completion of chemotherapy) was observed in some patients. These results demonstrate the utility of measuring these immune parameters in breast cancer patients, their relevance to the natural biology of the disease, and the influence that chemotherapy may have on host immune function.

Antigens, Neoplasm↗
PubMed1993

Inactivation of viruses in fresh-frozen plasma.

Methylene blue (MB) or solvent/detergent (SD) treatment is used for the inactivation of lipid-enveloped viruses in plasma. One important characteristic of the SD treatment is the necessity to pool plasma from different donors, thus inducing the risk of spreading infectious particles. MB treatment can be applied to single-donor plasma, causing no greater infectious risk than conventional fresh-frozen plasma (FFP). However, the virucidal efficacy of the SD method regarding HIV, HBV and HCV has been significantly better examined and proven than the MB treatment. Most of the therapeutic constituents of both plasma products are well maintained; coagulation factors decrease by roughly 5-20%. SD treatment reduces protein S and alpha 2-antiplasmin by approximately 40%, whereas MB treatment leads to a significant photooxidative alteration of fibrinogen with a disturbance of fibrin polymerization. As current studies show, the use of either plasma product is obviously not limited by acute or chronic toxicity. Several studies are in progress to evaluate the relevance of alterations in FFP quality which may affect the clinical efficacy of virus-inactivated plasma.

Antiviral Agents↗
PubMed1986

[Determination of reference ranges of rheologic parameters: study of 653 randomly selected probands of the Aachen district].

To determine reference ranges for rheologic parameters (hematocrit, plasma viscosity, erythrocyte aggregation, erythrocyte rigidity) a randomized study involving 653 subjects was carried out. Conditions of sampling, transportation and storing of blood specimens were established prior to the survey. Only 283 subjects met the criteria for enrollment in the study; the others were rejected because of inconspicuous history, normal findings in physical and Doppler-sonographic examination and absence of the risk factors hypertension, diabetes mellitus, overweight, rheumatic diseases, and smoking. The reference range for hematocrit was determined by an impedance-measuring device to equal 39-52% for males and 34-50% for females. The reference range for plasma viscosity, measured by a capillary-tube-plasma viscometer, was found to vary from 1.14 mPas to 1.34 mPas. The reference values for the standardized erythrocyte aggregation index was determined with the mini erythrocyte aggregometer to range from 8 to 21. Erythrocytes measured with the selecting-erythrocyte rigidometer showed a rigidity reference range between 0.83 and 1.19. Analysis of the results revealed that the parameters were independent of age (except in young children) and sex (with the exception of hematocrit).

Age Factors↗
PubMed2005

Detection of needle puncture to blood vessel using puncture force measurement.

Although blood sampling is frequently performed, a system to take blood samples automatically has not yet been developed. In the paper, as a first step towards automatic blood sampling, an examination of an automatic method for puncturing blood vessels is described. The force waveforms produced by puncturing rabbit ear veins were measured. A characteristic peak, possibly associated with the needle piercing the vessel wall, was observed in each waveform of ten successful cases of 14 trials. An algorithm that allowed the detection of this peak was developed, and parameters of a filter to improve the accuracy of the algorithm were determined. Using this algorithm, automatic needle punctures were performed in a rabbit ear vein and then were simulated using the data derived from manual needle puncture on four other rabbits. The results gave 30 fully successful cases of 33 trials of needle puncture and showed that measurement of the puncture force may be important for automatic needle puncture of blood vessels.

Animals↗
PubMed2006

Flame-broiled food, NAT2 acetylator phenotype, and breast cancer risk among women with benign breast disease.

PURPOSE: The objective of this study was to examine the association between flame-broiled food consumption, a source of heterocyclic amine exposure, and the development of breast cancer among cohort of women with benign breast disease (BBD). The variation of the association by acetylation phenotype, as determined by the genotypes of selected N-acetyltransferase 2 (NAT2) enzymes, was also examined. METHODS: Among participants in an ongoing cohort study, 1187 women reported having a breast biopsy for BBD and completed a food frequency questionnaire. NAT2 G857A, NAT2 T341C, and NAT2 G590A genotypes were determined using DNA extracted from blood specimens collected in 1989. Incident cases of breast cancer were identified through linkage of the cohort participants with the Washington County Cancer Registry and the Maryland State Cancer Registry. Follow-up for the BBD cohort began at study entry in 1989 and ended on April 28, 2003. RESULTS: Of the women in this study, 77 subsequently developed breast cancer. Results showed that, among rapid acetylators, flame-broiled food intake was associated with a statistically significant increase in the risk of breast cancer (odds ratio (OR) 2.62; 95% confidence interval (CI) 1.06, 6.46). No association was observed between flame-broiled food intake and breast cancer among slow acetylators (OR 0.75; 95% CI 0.39, 1.43). CONCLUSIONS: These findings suggest that flame-broiled food may be a modifiable risk factor for the progression of BBD to invasive breast cancer among women who have genotypes consistent with rapid acetylation.

Acetylation↗
PubMed1975

Cesarean section prevents placenta-to-infant transfusion despite delayed cord clamping.

We have adapted and calibrated an accurate method for measuring residual placement blood volume (RPBV). We used this method in 33 normal vaginal deliveries and 18 cesarean section deliveries in which the umbilical cord was clamped from 5 to 118 seconds after delivery. Infants born vaginally received 10 to 20 ml. per kilogram of body weight of placental transfusion when umbilical cord was not clamped until 30 seconds or more after delivery. Equivalent delay of umbilical cord clamping in cesarean section infants produced no placental transfusion.

Birth Weight↗
PubMed1988

Donation of blood by the pregnant patient for autologous transfusion.

A study was conducted to determine the safety and utility of autologous blood donation in third trimester pregnancy. Thirty-seven obstetric patients, 32 with an obstetric risk factor, donated an average of 485 ml of blood. Twenty-one of the 37 patients were expected to undergo cesarean section. Nonstress testing was performed before and after phlebotomy. Continuous fetal heart rate monitoring was maintained throughout the donation, which lasted an average of 9 minutes. All nonstress test results were normal before and after the phlebotomy except in one case. All fetal heart rates remained stable during phlebotomy and premature labor was not precipitated. All fetal outcomes were normal. One patient delivered on the day of phlebotomy, 6 hours after the procedure. Only one of the autologous units was used, in a patient who had a pelvic infection and moderate anemia. The incidence of primary cesarean section was 35%. Phlebotomy of the mother appears to be safe for the fetus at term. Further investigation is needed to determine the safety of removal of more than 1 unit of blood and blood donation at earlier gestational ages.

Adult↗