Automated Blood Collection Research Articles

The Causes of Platelet Aggregation in Version 6.4 Trima Accel Automated Blood Collection System and the Comparison of Two Intervention Measures

To explore the causes of platelet aggregation in version 6.4 Trima Accel automated blood collection system and the effect of 2 intervention measures. The data on platelet aggregation (n=61) and non-aggregation (n=323) of 61 donors in 2020 were collected and the causes of aggregation were analyzed. Then the 72 donors with platelet aggregation in 2021 were randomized into intervention group A (increasing the anticoagulant-to-blood ratio) and intervention group B (wrapping the donor's arm with an electric blanket to keep warm and improve the blood flow speed). The collection time, average blood flow speed, number of machine alarms, anticoagulant usage, deaggregation and citrate reaction of the two groups were compared. Platelet aggregation was negatively correlated with the average blood flow speed (r =-0.394) and positively correlated with the collection time (r =0.458). The equations for predicting aggregation and non-aggregation were constructed based on Bayesian and Fisher discriminant analysis, and the predicted accuracy was 77.1%. The comparison of the effects of two intervention measures showed that the average blood flow speed in group B was higher than that in group A; the collection time, number of machine alarms, anticoagulant usage and proportion of citrate reaction in blood donors in group B were all lower than those in Group A, all these differences were significant (P < 0.05). In the entire cohort in 2021, 90.28% of the products were immediately deaggregated after collection, and 9.72% of the products were deaggregated within 4 hours. There was no statistically significant difference in deaggregation between the two intervention groups (P >0.05). During apheresis platelet collection, the predictive equations for aggregation and non-aggregation can be used to predict the occurrence probability of aggregation, and the intervention can be made in advance. Both intervention measures are effective in reducing platelet aggregation, however, measure B has the advantages of improving the speed of blood collection, shortening the collection time, reducing the alarm frequency and the anticoagulant usage, and reducing the incidence of citrate reaction in blood donors.

Design and development of robotic system for precise vascular access and autonomous venipuncture

AbstractThe medical robot systems have the effect of reducing the work load and preventing accidents by supporting task that are difficult for inexperienced medical practitioners. Recently, research on automated blood collection and injection robotic system has been increasing, but work is still necessary such as replacing needle tips, setting tourniquet, and fixing the patient's arm. This letter proposes an intelligent intravenous injection robot without having to see the medical workers face‐to‐face. A revolving system is designed for automatic needle replacement and an arm holder for vessel fixation. Finally, positioning the needle at the target point is experimented by position control based on mathematical modelling and velocity kinematics.

Clinical application of a fully automated blood collection robot and its assessment of blood collection quality of anticoagulant specimens.

To investigate the application of intelligent puncture blood collection robots in anticoagulated blood specimens, the satisfaction of subjects with the two blood collection methods, and the feasibility of intelligent blood collection devices to replace manual blood collection methods in clinical work. A total of 154 volunteers from Zhongshan Hospital Fudan University were recruited to compare the test results of anticoagulant blood samples between blood collection robot and manual blood collection, a questionnaire was used to inquire about the volunteers' feelings about the two blood collection methods; the blood collection data of 6,255 patients willing to use the robot for blood collection were collected to analyze the success rate of blood collection. The blood collection robot is superior to manual specimen collection in terms of volume and pain of specimen collection, and the puncture success rate is 94.3%. The anticoagulated blood specimens collected by the robot had 11 indexes statistically different from the results of manual blood collection, but the differences did not affect the clinical diagnosis and prognosis. The intelligent robotic blood collection is less painful and has better acceptance by patients, which can be used for clinical anticoagulated blood specimen collection.

Remote Neuronal Activation Coupled with Automated Blood Sampling to Induce and Measure Circulating Luteinizing Hormone in Mice.

Circulating luteinizing hormone (LH) levels are an essential index of the functioning of the hypothalamic-pituitary control of reproduction. The role of numerous inputs and neuronal populations in the modulation of LH release is still unknown. Measuring changes in LH levels in mice is often a challenge since they are easily disrupted by environmental stress. Current techniques to measure LH release and pulsatility require long-term training for mice to adapt to manipulation stress, certain restraint, the presence of the investigator, and working on individual animals, reducing its usefulness for many research questions. This paper presents a technique to remotely activate specific neuronal populations using Designer Receptor Exclusively Activated by Designer Drugs (DREADDs) technology coupled with automated sequential blood sampling in conscious, freely moving, and undisturbed mice. We first describe the stereotaxic surgery protocol to deliver adeno-associated virus (AAV) vectors expressing DREADDs to specific neuronal populations. Next, we describe the protocol for carotid artery and jugular vein cannulation and postsurgical connection to the CULEX automated blood sampling system. Finally, we describe the protocol for clozapine-N-oxide intravenous injection for remote neuronal activation and automated blood collection. This technique allows for programmed automated sampling every 5 min or longer for a given period, coupled with intravenous substance injection at a desired time point or duration. Overall, we found this technique to be a powerful approach for research on neuroendocrine control.

Factors Affecting Platelet Yield in a Single Donor Plateletpheresis

Background: This procedure, known as plateletpheresis, involves the removal of platelets from a donor's blood. This method produces eight times as many platelets as would be obtained by using whole blood. Aim: To evaluate the effect of different donor related factors on platelet yield. Method: This research was carried out at the Children's Hospital &amp; Institute of Child Health Science in Lahore, Pakistan, specifically in the Immunohematology &amp; Blood Transfusion Unit. Forty plateletpheresis donors meeting the criteria were enlisted for the study. The COM.TEC cell separator apparatus and the TRIMA automated blood collection system were used to perform the plateletpheresis. Before and after a plateletpheresis, the patient's hematocrit, haemoglobin, platelet count, white blood cell count, processed volume, and processing time were recorded. Results: Platelet yield demonstrated a substantial positive link with the pre-platelet count of the donors, as well as their age, weight, and the volume processed. Platelet production was not correlated in any way with blood group, total leukocyte count, high-content testing, or processing time. Conclusion: A patient's pre-platelet count, age, weight, and processed volume were positively correlated with their platelet production. Keywords: Plateletpheresis, Hemoglobin, Total leukocyte count, Platelet count, Platelet yield

Using automation to manage donor engagement and fine-tune supply and demand during the first year of the COVID-19 pandemic

BackgroundCOVID-19 disrupted blood center operations starting March 2020 and continues to affect donor presentation and blood availability today. The industry mobilized significant resources to collect COVID-19 convalescent plasma (CCP) to treat COVID-19 patients. At the same time, blood centers continued to collect platelets, plasma, and red blood cells (RBCs) to meet the needs of non-COVID-19 patients. The purpose of this study was to quantify how automation was used to fine-tune supply and demand and increase donor engagement during the first year of the pandemic. MethodsThis was a single-center retrospective study of blood collection and donor presentation at a mid-sized US blood center. Data was evaluated from January 1, 2020 through March 31, 2021. Parameters evaluated included donor presentation, platelets per procedure, concurrent RBC and plasma collections per procedure, operator compliance, total donor appointment count, and donor frequency. ResultsWith the cancelation of mobile blood drives, fixed sites increased total apheresis procedures by 37% and increased turns per bed by 46% whereas less products were collected per donor. By collecting only what was needed, platelet expiration rate decreased from 6.8% (pre-pandemic) to less than 4%. Donor engagement as measured by donor frequency increased from 1.6 in January 2020 to 1.8 in March 2021. ConclusionsUsing technological advances such as automated blood collection and information systems, the blood center improved donor engagement and avoided collecting a surplus of any one type of blood product over the course of the pandemic

Platelets Feeding Bacteria with Lactate during Room Temperature Storage: Mitigated By Refrigeration

Introduction: Platelet transfusion related sepsis is a serious concern limiting platelet storage time to 5 days. Room temperature (RT) storage of platelets increases the risk of bacterial growth and necessitates implementation of bacterial contamination monitoring. Cold storage of platelets represents an attractive alternative for improving platelet safety. In this study, we assessed bacterial growth in platelets stored either at room temperature (22oC) or refrigerated (4oC).Methods: Apheresis platelets in plasma (PLT) were obtained from healthy donors using the Terumo Trima Accel Automated Blood Collection System (Terumo BCT). Platelet poor plasma (PPP) was obtained from PLT aliquots centrifuged twice at 2,500 x g for 5 min. In some experiments aliquots of PPP were supplemented with 5 to 20 mM lactic acid. Aliquots of PLT or PPP were transferred to pH SAFE minibags (Blood Cell Storage, Inc) and inoculated with Acinetobacter baumannii strains Ci77 or Ci79, Escherichia coli, Pseudomonas aeruginosa, or PBS (uninfected control). Minibag aliquots stored at RT were agitated using an orbital shaker set to 60 rpm while refrigerated aliquots were stored under static conditions. Bacterial growth was monitored daily through dilution plating. Lactate levels in PLT aliquots were assessed by iSTAT (Abbott) using CG4+ test cartridges. Platelet activation and aggregation were assessed on days 0, 1, 3, and 5 by flow cytometry and Multiplate platelet aggregometry respectively.Results: Growth of A. baumannii progressed exponentially over the first 3 days post-collection in PLT aliquots stored at RT. However, growth was significantly (p < 0.05) reduced in PPP units. Growth of E. coli and P. aeruginosa, both of which are capable of utilizing glucose present in the anticoagulant used, grew at significantly faster rates and did not seem to be dependent on metabolically active PLTs. Lactate levels were assessed in PLT units and found to mirror growth of A. baumannii . Furthermore, addition of ≥ 15 mM lactic acid to RT stored PPP restored A. baumannii growth. Although lactate levels did not appear to contribute to growth of E. coli, lactate levels began to decrease in PLT units contaminated with the bacterium at a rapid rate after day 3. Lactate levels in units contaminated with P. aeruginosa increased throughout the observation window. Bacterial growth remained static throughout under all treatment conditions stored refrigerated and lactate levels were reduced. PLT also appeared to retain function throughout cold storage irrespective of bacterial contamination while RT storage resulted in a decrease in aggregation over time which was exacerbated by bacterial contamination.Conclusions: Bacterial growth remained static throughout under cold storage. Additionally, PLT lactate production was increased at RT and correlated with increased growth of A. baumannii . Growth of E. coli and P. aerugionsa appeared to proceed independently of lactate production or metabolically active PLTs. However, E. coli did appear to utilize lactate as a carbon source at later time points. These data demonstrate that bacterial growth can be controlled through refrigeration without loss of function and RT stored platelets may potentiate growth of certain bacterial strains through accelerated metabolism relative to cold storage. DisclosuresNo relevant conflicts of interest to declare.

Severe CD4+ T-cell lymphopenia is not observed in frequent plateletpheresis donors collected on the Fenwal Amicus.

In a recent study, we determined that 30% of frequent plateletpheresis donors collected using the Trima Accel Automated Blood Collection System (Terumo BCT) had a CD4+ T-cell count below 200 cells/μL. Whether CD4+ T-cell lymphopenia is associated with donation using other plateletpheresis instruments is unknown. We obtained blood samples from 30 current frequent Fenwal Amicus plateletpheresis donors. All participants had made 20 to 24 plateletpheresis donations in the most recent 365-day period, and all had previously donated over 50 times on the Fenwal Amicus instrument. Blood samples were analyzed to determine blood counts, including CD4+ and CD8+ counts. Of 30 study participants, none had a CD4+ count below 200 cells/μL. There was one participant with a CD4+ count between 200 and 300 cells/μL. This individual was over the age of 55 and had a history of more than 300 lifetime plateletpheresis sessions. One participant had a CD8+ count below the lower limit of normal (125 cells/μL) and a normal CD4+ count. We did not detect severe CD4+ lymphopenia in frequent platelet donors undergoing plateletpheresis with the Fenwal Amicus. Since the Fenwal Amicus does not incorporate a leukoreduction system chamber, this finding supports the hypothesis that such chambers-found in the Trima Accel instrument-contribute to CD4+ lymphopenia in frequent platelet donors.

Platelet Metabolism during Room Temperature Storage Contributes to Bacterial Growth: Effect Mitigated By Refrigeration

Introduction: Transfusion related sepsis is a serious concern limiting platelet storage time to 5 days at room temperature. While most units are screened for bacterial contamination when collected, most bacterial monitoring methods can take up to 7 days to detect potential contamination. Thus, cold storage of platelets represents an attractive alternative for improving platelet safety. In this study, we assessed bacterial growth in platelets stored either at room temperature (22oC) or refrigerated (4oC).Methods: Apheresis platelets in plasma (PLT) were obtained from healthy donors using the Terumo Trima Accel Automated Blood Collection System (Terumo BCT). Fresh plasma (FP) was collected similarly. Aliquots of PLT or FP were transferred to pH SAFE minibags (Blood Cell Storage, Inc) and inoculated with Acinetobacter baumannii, Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus, Staphylococcus epidermidis, or PBS (uninfected control). Minibag aliquots stored at RT were agitated using an orbital shaker set to 60 rpm while refrigerated aliquots were stored under static conditions. Bacterial growth was monitored daily through dilution plating. Lactate levels in PLT aliquots were assessed by iSTAT (Abbott) using CG4+ test cartridges while plasma glucose levels were assessed using blood glucose testing strips (Germaine Laboratories). Platelet activation and aggregation were assessed on days 0, 1, 3, and 5 by flow cytometry and Multiplate platelet aggregometry, respectively.Results: Bacterial growth progressed rapidly over the first 3-4 days post-collection in all PLT aliquots stored at RT except those challenged with S. epidermidis. Significant growth of S. epidermidis was not detected until day 4. No change in bacterial numbers were detected in any refrigerated aliquots through day 5. While refrigeration appeared to preserve PLT function throughout with low levels of activation irrespective of bacterial contamination, RT storage resulted in significantly (p < 0.05) decreased platelet aggregation over time which was exacerbated by bacterial contamination. In the absence of metabolically active PLTs, bacterial growth was significantly reduced, or at least delayed, in all test groups. FP aliquots challenged with Gram-negative pathogens exhibited a significant (p < 0.05) delay in bacterial growth at day 1. While growth of E. coli and P. aeruginosa recovered by day 2, growth was significantly (p < 0.05) inhibited in aliquots challenged with A. baumannii throughout the observation period. Conversely, no differences in bacteria growth were observed in aliquots challenged with Gram-positive pathogens until day 3, at which point growth appeared to be significantly (p< 0.05) stunted in FP relative to PLT aliquots. Bacterial growth appeared to correlate with PLT lactate production. However, only E. coli showed clear signs of lactate utilization as lactate levels diminished significantly after day 3. Despite this, A. baumannii, E. coli, and S. epidermidis, exhibited increased bacterial growth in FP aliquots supplemented with concentrations of lactic acid in excess of 15 mM.Conclusions: Bacterial growth, platelet activation and platelet lactate production appeared largely static throughout in refrigerated aliquots. Conversely, bacterial growth was significantly increased in all RT stored aliquots, as was lactate production suggesting platelet metabolism may contribute to bacterial growth. Illustrating this, lactic acid concentrations in excess of 15 mM modulated growth of A. baumannii, E. coli, S. epidermidis in FP. These data demonstrate that bacterial growth can be controlled through refrigeration without loss of function and RT storage may potentiate growth of certain bacterial strains through accelerated PLT metabolism. DisclosuresNo relevant conflicts of interest to declare.

CD4+ T-Cell Lymphopenia in Frequent Platelet Donors Who Have Ceased Platelet Donation for at Least 1 Year

IntroductionWe recently discovered that a number of frequent apheresis platelet donors at our donor center had CD4+ T-lymphocyte counts below 200 cells/µL. The cause appears to be repeated extraction of these cells by the Trima Accel Automated Blood Collection System's leukoreduction system chamber. Plateletpheresis at our donor center has been conducted exclusively with this instrument since 2006. How long CD4+ T-cell lymphopenia persists after stopping plateletpheresis is unknown. Whether there are infectious or other complications that could relate to CD4+ T-cell lymphopenia in former donors is also unknown. We therefore investigated the blood counts (including CD4+ T-lymphocyte counts) and medical histories of former platelet donors who had a history of frequent platelet donation but had stopped donating platelets for at least 12 months.MethodsWe mailed a questionnaire to former frequent apheresis platelet donors who had not donated platelets at our donor center for at least 12 months. Donors who replied to the questionnaire were contacted by phone to schedule a study visit. Frequent platelet donation was defined as 20-24 plateletpheresis sessions in at least one 365-day period starting in 2011. Donors who consented to participate in the study confirmed that they had not donated platelets in the prior 12 months, provided a blood sample for analysis, and completed a health questionnaire that included questions about opportunistic infections and malignancies. Medical records of these donors were also reviewed, where available. Approval for the study was obtained from the Partners HealthCare Institutional Review Board (2017P002880) and all participants provided written informed consent.ResultsOf 52 former frequent platelet donors identified as eligible to receive a questionnaire, one was known to have died of a myocardial infarction and another had requested not to receive communications from the donor center. Therefore, 50 potential study candidates were mailed a questionnaire. Twelve questionnaires were returned as undeliverable. Five potential study candidates returned the questionnaire but either declined to participate in the study or did not respond when contacted to schedule a study visit. Fourteen former frequent platelet donors elected to participate in the study after receiving the first mailing of the questionnaire. Of these 14 study participants, 6 were female and all were Caucasian. The median age was 65. The most common reason for ceasing platelet donation was a positive HLA antibody test (5 participants) followed by “low white blood cells,” “heart trouble,” and “inconvenience” (2 participants each). All former donors had tested negative for HIV while donating platelets. There were 2 participants with CD4+ T-lymphocyte counts below 200 cells/µL (Figure 1); one of these participants had prior CD4+ T-lymphocyte counts available for review. These showed that the CD4+ T-lymphocyte count in this former donor had improved slightly since ceasing donation one year earlier (Figure 2). Three study participants had a CD4+ T-lymphocyte count between 200 and 300 cells/µL. A review of prescription medications and medical problems did not identify an etiology for the low CD4+ T-lymphocyte counts. Responses to the health questionnaire showed that no study participant had ever had a severe infection, an infection with an unusual pathogen (“bug”), or thrush. Two former frequent platelet donors had a history of shingles, both of whom had CD4+ T-lymphocyte counts slightly below the normal range (441-2156 cells/µL) at the time of participation in the study. Five former donors had a history of cancer: Two episodes of squamous cell skin cancer (1 participant), papillary thyroid carcinoma (1 participant), glioblastoma (1 participant), basal cell skin cancer (1 participant), and squamous cell skin cancer as well as melanoma in situ (1 participant).ConclusionIn our small cohort, there is no evidence that CD4+ T-cell lymphopenia predisposes to opportunistic infections or to malignancies classically associated with immune dysregulation. Plateletpheresis-associated CD4+ T-cell lymphopenia has improved slightly in one former frequent platelet donor a year after ceasing plateletpheresis but the count remains below 200 cells/µL. Frequent plateletpheresis involving a leukoreduction system chamber should be considered in the differential diagnosis of idiopathic CD4+ lymphopenia. [Display omitted] DisclosuresGansner:Novimmune SA: Research Funding; UpToDate: Patents & Royalties.

Platelet Lactate Production during Room Temperature Storage Promotes Bacterial Growth

Introduction: Transfusion related sepsis is a serious concern for both military and civilian trauma centers. The primary source of bacterial contamination associated with such events is room temperature (RT) stored platelets. Due to high risk of bacterial contamination, RT stored platelets may only be administered up to 5 days post-collection. Although approved by the FDA, cold stored platelets may only be administered 3 days post-collection under current regulations. In this study, we utilized the opportunistic pathogen Acinetobacter baumannii - a reported cause of platelet contamination and important hospital acquired infection (HAI) - as a model organism to assess bacterial growth in platelets stored either at room temperature (22oC) or refrigerated (4oC), as well as determine contributors to bacterial growth.Methods: Apheresis platelets in plasma (PLT) were obtained from healthy donors using the Terumo Trima Accel Automated Blood Collection System (Terumo BCT). Platelet poor plasma (PPP) was obtained from PLT aliquots centrifuged twice at 2,500 x g for 5 min. Other PLT aliquots were stimulated with Phorbol 12-myristate 13-acetate (PMA) to generate activated platelets (aPLT). Following activation, aPLTs were pelleted through centrifugation at 2,500 x g for 5 min and the releasate (REL) was transferred to a separate container. Pelleted aPLTs were washed 3 times in sterile PBS and suspended in PPP. In some experiments aliquots of PPP were supplemented with 40 mM lactic acid. Aliquots of PLT, PPP, aPLT, and REL were transferred to pH SAFE minibags (Blood Cell Storage, Inc) and inoculated (a.k.a., contaminated) with A. baumannii clinical isolate Ci79. Minibag aliquots stored at RT were agitated using an orbital shaker set to 60 rpm while refrigerated aliquots were stored under static conditions. Bacterial growth was monitored daily through dilution plating. In some experiments, lactate levels in PLT aliquots were assessed by iSTAT (Abbott) using CG4+ test cartridges.Results: Bacterial growth progressed exponentially over the first 3 days post-collection in PLT aliquots stored at RT. However, growth was significantly (p < 0.05) reduced in PPP units (Fig. 1A). Neither platelet activation status nor released factors appeared to have any effect as no significant differences were observed in bacterial growth between contaminated PLT and aPLT units, nor between contaminated PPP and REL units (Fig. 1A). Growth progressed at a much faster rate and to a greater magnitude in the presence of live platelets (PLT and aPLT), suggesting the contribution of platelet metabolism. Thus, lactate levels were assessed in PLT units and found to mirror bacterial growth (Fig. 1B). Furthermore, addition of lactic acid to RT stored PPP restored bacterial growth (Fig. 1A). Growth remained static throughout under all treatment conditions stored refrigerated.Conclusions: Bacterial growth remained static over the 5 day post-collection observation period during cold storage. Additionally, bacterial growth at RT appeared to be related to increased production of lactate from pyruvate via NAD-dependent lactate dehydrogenase (nLDH) following glycolysis (Fig. 1C). To that end, many bacteria, including A. baumannii,as well as various Staphylococcus spp, and Streptococcus spp, possess genes encoding for NAD-independent lactate dehydrogenases (iLDH) which enable utilization of lactate as a carbon source (Fig. 1D). These data demonstrate that not only can bacterial growth be controlled through refrigeration, but RT stored platelets potentiate bacterial growth through their accelerated metabolism relative to cold storage. [Display omitted] DisclosuresNo relevant conflicts of interest to declare.

Platelet corrected count increments by apheresis platform

Automated blood collection platforms use different technological systems to isolate and collect individual blood components. These unique systems could potentially result in differences in platelet in vivo viability, as measured by the corrected count increment (CCI). This retrospective study evaluated CCI data of platelet transfusions among oncology patients who received multiple unmanipulated apheresis platelets between January 1, 2006 and December 31, 2009. Apheresis platelets were collected from our community blood center by standard procedures using two different collection systems and were transfused to patients in a blinded manner. The CCI of the platelet recipient was significantly higher at 0-2 hours post-transfusion among the individuals who received platelets collected on Trima Accel (Terumo BCT) (mean = 6281, standard deviation = 3650) compared to the platelets collected by the Amicus system (Fresenius Kabi) (mean = 5251, standard deviation = 3311, p = 0.004). These hypothesis-generating data suggesting improved recovery and survival of Trima Accel platelets demonstrate the need for the investigation and implementation of the best collection methods to provide better platelet transfusion support.

Assessment of factors affecting the platelet yield using continuous flow cell separator

Background: The quality of single donor platelets (SDP) in terms of yield influences the platelet recovery in the recipient. Various donor factors such as predonation platelet count and hemoglobin concentration may affect platelet yield. The aim of present study was to assess the effect of various donor factors on platelet yield and effect of plateletpheresis on hematological values of donors. Materials and methods : A total of 171 plateletpheresis procedures were performed using continuous flow cell separator (Amicus Automated Blood Collection System version 2.52/3.1) were evaluated for platelet yield. The pre and post donation hematological values such as platelet count, hemoglobin (Hb) concentration, mean platelet volume (MPV) and platelet distribution width (PDW) were measured in all blood donors. Results : The mean platelet yield was 3.8 + 1.65 X 10 11 /unit. The platelet yield correlated positively (r = 0.446, p < 0.001) with donor pre platelet count and negatively (r =- 0.004, p = 0.954) with donor pre-Hb. There was significant increase (p < 0.001) in Hb concentration and platelet count decreased significantly (p<0.001) after each procedure. While, there was no significant decease in MPV and PDW after each procedure. Conclusion: All transfusion centers should have their own plateletpheresis donor data based on demographic and hematological characteristics of the donor population. This will ensure donor safety, product quality and an effective donor registry.

Application of Automated Dried Blood Spot Sampling and LC–MS/MS for Pharmacokinetic Studies of AMG 517 in Rats

The use of dried blood spot (DBS) sampling technique is of particular interest for drug discovery pharmacokinetic studies due to the small blood volume requirement. In addition, automated blood sampling is an attractive approach for rat pharmacokinetic studies as animal handling work is minimized. The goal of this study was to use an automated DBS sampler for automated blood collection and spotting onto DBS paper for pharmacokinetic studies in rats. AMG 517, a potent and selective vanilloid receptor antagonist, was dosed to rats (n = 3) intravenously and blood samples were collected at nine time points over a 24 h period using the automated DBS sampler. After drying, storage and shipment, the DBS samples were extracted and analyzed by LC-MS/MS. The developed bioanalytical method for the analysis of DBS samples had good accuracy and precision within the context of a discovery, non-GLP analysis. The concentration-time data and pharmacokinetic parameters generated from automated spotted samples were very similar to those derived from manually spotted DBS samples. The manual DBS data were also comparable to plasma data after correction for blood-to-plasma ratio. The automated DBS sampling is a promising technique for rodent pharmacokinetic studies and will improve the efficiency and quality of DBS sampling.

Cesium cessation? An advantage of pathogen reduction treatments

Cesium cessation? An advantage of pathogen reduction treatments

Tracking deposition of a 14C-radiolabeled kudzu hairy root-derived isoflavone-rich fraction into bone

Hairy roots were induced in four genotypes from three kudzu species (Pueraria montana var. lobata, P. lobata and P. phaseoloides) in vitro using Agrobacterium rhizogenes to stimulate rapid secondary metabolite synthesis. Hairy roots from P. montana var. lobata (United States Department of Agriculture no. PI 434246) yielded the highest puerarin and total isoflavone content and the greatest new biomass per growth cycle among the genotypes evaluated. Hairy roots from this genotype were selected for radiolabeling using (14)C-sucrose as a carbon source. Isoflavones from radiolabeled kudzu hairy root cultures were extracted with 80% methanol, partitioned by solvent extraction, and then subfractionated by Sephadex LH-20 gel filtration. Radiolabeled isoflavones were isolated in a highly enriched fraction, which contained predominantly puerarin, daidzin and malonyl-daidzin and had an average radioactivity of 8.614 MBq/g (232.8 μCi/g) dry fraction. The (14)C-radiolabeled, isoflavone-rich fraction was orally administered at a dose of 60 mg/kg body weight to male Sprague-Dawley rats implanted with a jugular catheter, a subcutaneous ultrafiltrate probe and a brain microdialysate probe. Serum, interstitial fluid, brain microdialysate, urine and feces were collected using a Culex(®) Automated Blood Collection System for 24 h. At the end of this period, rats were sacrificed and major tissues were collected. Analysis by a scintillation counter confirmed that a bolus dose of (14)C-radiolabeled, isoflavone-rich kudzu fraction reached bone tissues, which accumulated 0.011%, 0.09% and 0.003% of the administered dose in femur, tibia and vertebrae, respectively. Femurs extracted with 80% methanol were analyzed by high-performance liquid chromatography with electrospray ionization-mass spectrometry and were found to contain trace quantities of puerarin, daidzein and puerarin glucuronide. This study demonstrates that kudzu isoflavones and metabolites are capable of reaching bone tissues, where they may contribute to the prevention of osteoporosis and the promotion of bone health.

Brief note about plasma catecholamines kinetics and submaximal exercise in untrained standardbreds

Four untrained standardbred horses performed a standardized exercise test on the treadmill and an automated blood collection system programmed to obtain blood samples every 15 s was used for blood collection in order to evaluate the kinetics of adrenaline and noradrenaline. The highest average values obtained for adrenaline and noradrenaline were 15.0 +/- 3.0 and 15.8 +/- 2.8 nmol/l respectively, with exponential accumulation of adrenaline (r = 0.977) and noradrenaline (r = 0.976) during the test. Analysis of the correlation between noradrenaline and adrenaline for each phase of the test shows that correlation coefficient decreases as the intensity of exercise increases (from r = 0.909 to r = 0.788). This suggests that during submaximal exercise, the process for release, distribution and clearance of adrenaline into blood circulation differs from that of noradrenaline.

In vivo metabolic tracking of 14C-radiolabelled isoflavones in kudzu (Pueraria lobata) and red clover (Trifolium pratense) extracts

Absorption, distribution and elimination of 14C-labelled isoflavone-containing extracts from kudzu (Pueraria lobata) root culture and red clover (Trifolium pratense) cell culture were investigated in an in vivo rat model. The predominant isoflavones in the kudzu extract were the glycosides puerarin, daidzin and malonyl daidzin, while in the red clover extract, the major isoflavones were formononetin and its derivatives, genistein and biochanin A, with radioactivities of 3.770 and 7.256 MBq/g, respectively. Male Sprague-Dawley rats, implanted with a jugular catheter and a subcutaneous ultrafiltrate probe, were orally administered with 14C-labelled isoflavone extracts from either kudzu or clover cell cultures. Serum, interstitial fluid (ISF), urine and faeces were collected using a Culex Automated Blood Collection System for 24 h. Analysis of bone tissues revealed that radiolabel accumulated in the femur, tibia and vertebrae at 0.04, 0.03 and 0.01 % of the administered dose, respectively, in both kudzu and red clover treatments. The liver accumulated the greatest concentration of radiolabel among the tissues tested, at 1.99 and 1.54 % of the administered kudzu and red clover extracts, respectively. Serum and ISF analysis showed that both extracts were rapidly absorbed, distributed in various tissues, and largely eliminated in the urine and faeces. Urine and faeces contained 8.53 and 9.06 % of the kudzu dose, respectively, and 3.60 and 5.64 % of the red clover dose, respectively. Serum pharmacokinetics suggest that extracts from kudzu may undergo enterohepatic circulation.

Automated collection of double red blood cell units with a variable‐volume separation chamber

Automated collection of blood components offers multiple advantages and has prompted development of portable devices. This study sought to document the biochemical and hematologic properties and in vivo recovery of red cells (RBCs) collected via a new device that employed a variable-volume centrifugal separation chamber. Normal subjects (n = 153) donated 2 units of RBCs via an automated blood collection system (Cymbal, Haemonetics). Procedures were conducted with wall outlet power (n = 49) or the device's battery source (n = 104). Units were collected with or without leukoreduction filtration and were stored in AS-3 for 42 days. The units were assessed via standard biochemical and hematologic tests before and after storage, and 24 leukoreduced (LR) and 24 non-LR RBCs were radiolabeled on Day 42 with Na(2)(51)CrO(4) for autologous return to determine recovery at 24 hours with concomitant determination of RBC volume via infusion of (99m)Tc-labeled fresh RBCs. Two standard RBC units (targeted to contain 180 mL of RBCs plus 100 mL of AS-3) could be collected in 35.7 +/- 2.0 minutes (n = 30) or 40.3 +/- 2.7 minutes for LR RBCs (n = 92). An additional 31 collections were conducted successfully with intentional filter bypassing. RBC units contained 104 +/- 4.1 percent of their targeted volumes (170-204 mL of RBCs), and LR RBCs contained 92 percent of non-LR RBCs' hemoglobin. All LR RBCs contained less than 1 x 10(6) white blood cells. Mean hemolysis was below 0.8 percent (Day 42) for all configurations. Adenosine triphosphate was well preserved. Mean recovery was 82 +/- 4.9 percent for RBCs and 84 +/- 7.0 percent for LR RBCs. The Cymbal device provided quick and efficient collection of 2 RBC units with properties meeting regulatory requirements and consistent with good clinical utility.

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