Drawing Blood Samples Research Articles

Pain Assessment among African Neonates

Neonates who require treatment and venous drawing of blood samples in the newborn units are subjected to acute and painfully invasive procedures. Several tools to assess pain among newborns have been developed and are widely used in developed countries, but in Africa, there is limited experience in the assessment pain among newborns. This study assessed physiological and behavioral responses to pain among neonates during invasive procedures performed in a newborn unit in Rwanda. A total of 60 neonates born at gestational age of 28-42 weeks at the National University of Rwanda Teaching and Referral Hospital in the year 2005 were enrolled into this study. Blood pressures, heart and respiratory rates, oxygen saturation levels, the Neonatal Facing Coding System (NFCS) and Neonatal Acute Pain (APN) pain tools were and scores recorded before, immediately and 5, 10, 15 and 20 minutes after procedures were recorded. Physiological parameters were compared using the Wilcoxon Signed Ranks Test while the NFCS and APN were compared using the McNemar Test. All (100%) neonates experienced acutely peak pain in the first 5 minutes of the invasive procedures with peak responses recorded at 2.5 minutes and no pain (resolution) after 15 minutes among 81% of the neonates and only 6% experienced pain after 20 minutes. The increases in systolic blood pressures immediately after inflicting pain, 5, 10, 15 and 20 minutes were statistically significant (p<0.001, p<0.001, p<0.005 and p<0.046) respectively compared to the diastolic blood pressures whose significant increases were at 5 and 10 minutes, (p<0.001 and p<0.001) respectively. Respiratory rates were significantly high at the onset (p<0.001), 5 minutes (p<0.001) and 10 minutes later (p<0.002). Heart rates significantly increased at the onset of the procedures (p<0.000), 5 minutes (p<0.001) and after 10 minutes (p<0.033). Decrease in oxygen saturation immediately after the procedures was significant (p<0.001). Oxygen saturation immediately after the procedures significantly increased up to 5 minutes (p<0.001) and 10 minutes (p<0.001). Invasive procedures caused acute pain among neonates in the African settings but to date, neonatal practice had not been given its due consideration with the aim of reducing pain among African neonates.

Genetic counseling in monogenic diabetes GCK MODY.

Genetic testing in families with monogenic GCK MODY has predictive, diagnostic, and preventive utility. Predictive tests relate to people who have no features of the disorder themselves at the time of testing. Diagnostic tests relate to family members who have been previously diagnosed with diabetes mellitus or glucose metabolism disturbances. The preventive value of genetic testing for families is to raise awareness of the circumstances leading to glucose metabolism disorders. The detection of mutation carriers among family members of patients with GCK MODY and the determination of the clinical significance of the genetic test result. The study group included 27 families of adolescent patients with GCK MODY (39 (75%) of parents and 19 (73.08%) of siblings) monitored in the Department of Pediatrics, Endocrinology and Diabetes and in the Diabetes Clinic of John Paul II Upper Silesian Child Health Centre in Katowice in the years 2007-2012. Subjects underwent a blood sample drawing for genetic and biochemical testing. Through the genetic diagnostics we diagnosed GCK MODY in 14 (63.64%) mothers, 6 (35.29%) fathers and in 7 (36,84%) siblings. Genetic testing has contributed to the detection of 7 (26.92%) asymptomatic carriers of GCK gene mutation among parents and 3 (15,79%) asymptomatic carriers among siblings declaring no carbohydrate metabolism disturbances (before genetic testing there were no indications suggesting carbohydrate metabolism disturbances; OGTT were performed after positive genetic testing). Each case of mutation detection, which is the cause of monogenic diabetes in a patient, justifies the genetic testing in other members of his/her family. Awareness of the genetic status may allow sick family member to confirm the diagnosis, while asymptomatic mutation carriers could benefit from an early clinical observation. Consequently, in each case it gives an opportunity to take diagnostic and therapeutic measures in accordance with the current state of knowledge.

Humoral response markers in GCK MODY

The prevalence of antibodies to pancreatic islets in monogenic diabetes remains unknown and the incidence estimation is difficult as the occurrence of autoantibodies in patient is one of the well-known exclusion criteria for further genetic diagnostics. They has been found not only among patients with type 1 diabetes, but also in other types of diabetes: Type 2 diabetes, Latent Autoimmune Diabetes in Adults (LADA) (16) and monogenic diabetes (MD). Immunological characteristic of GCK MODY patients. The study group included families of 27 adolescent patients with GCK MODY (39 parents and 19 siblings) monitored in the Department of Pediatrics, Endocrinology and Diabetes and in the Diabetes Clinic of John Paul II Upper Silesian Child Health Centre in Katowice in the years 2007-2012. All patients and family members with GCK MODY underwent a blood sample drawing for immunological (classic humoral response markers: ICA, GAD, IA-2, IAA) and biochemical diagnostics. Pediatric, diabetes and family medical history was collected from the subjects and parents. Immunological diagnostics was performed in all patients except 1 (96.3%). Immunological diagnostics included 17 (89.5%) parents and 7 (87.5%) siblings with diagnosed GCK MODY. 8 (30.8%) adolescent patients with GCK MODY, 3 subjects (17.64%) among parents (with GCK MODY), as well as 2 subjects (28.57%) among siblings (with GCK MODY) showed a positive antibodies screen. The results of our study in children with GCK MODY and their family members suggest that the occurrence of classic antibodies directed against pancreatic islets antigens is fairly common in patients with GCK MODY. Despite various observations and many legitimate discussions, it is difficult to clarify the pathogenesis of the occurrence of autoantibodies in monogenic diabetes.

An improved micro-method for the measurement of steroid profiles by APPI-LC–MS/MS and its use in assessing diurnal effects on steroid concentrations and optimizing the diagnosis and treatment of adrenal insufficiency and CAH

Our goals were to (1) develop an improved micro-method usable for neonates for steroid profile measurements and a method to measure androsterone, a key steroid in the recently described androgen backdoor pathway together, with dehydroepiandrosterone and (2) to assess if dehydroepiandrosterone diurnal concentration fluctuations exist potentially necessitating strict adherence to time of blood sample draw and requirement of separate time-dependent reference intervals. Liquid chromatography–tandem mass spectrometry was performed with an atmospheric pressure photoionization source [1]. For each sample 50μL (100μL for the backdoor pathway) of serum was deproteinized by adding 75μL (150μL for the backdoor pathway) of acetonitrile containing the internal standards. After centrifugation, 75μL (150μL for the backdoor pathway) of supernatant was diluted with 250μL of water and injected onto a Poroshell 120 EC-C8 column (SB-C8 column for the backdoor pathway). Within-run coefficients of variation ranged from 2.4 to 10.4% and between-day coefficients of variation from 2.9 to 11.2%. Comparison studies yielded correlation coefficient between 0.97 and 1.00 with recoveries of 90% or greater. Our methods analyze a 9 steroid profile and an additional 2 steroid profile (backdoor pathway) with minimal sample volume (usable in neonates optimizing early diagnosis of endocrinopathies and genetic diseases). Low limits of quantitation make these methods ideal for steroid measurement in women and prepubertal children. As diurnal variations of dehydroepiandrosterone and other steroids [2] concentrations are clinically significant we recommend that separate reference intervals be developed for 8 am, 8 pm, and midnight sample draws. The use of this approach in improving the diagnosis of patients with adrenal insufficiency and congenital adrenal hyperplasia is discussed.

Volunteer experiences and perceptions of the informed consent process: Lessons from two HIV clinical trials in Uganda.

BackgroundInformed consent as stipulated in regulatory human research guidelines requires that a volunteer is well-informed about what will happen to them in a trial. However researchers are faced with a challenge of how to ensure that a volunteer agreeing to take part in a clinical trial is truly informed. We conducted a qualitative study among volunteers taking part in two HIV clinical trials in Uganda to find out how they defined informed consent and their perceptions of the trial procedures, study information and interactions with the research team.MethodsBetween January and December 2012, 23 volunteers who had been in the two trials for over 6 months, consented to be interviewed about their experience in the trial three times over a period of nine months. They also took part in focus group discussions. Themes informed by study research questions and emerging findings were used for content analysis.ResultsVolunteers defined the informed consent process in terms of their individual welfare. Only two of the volunteers reported having referred during the trial to the participant information sheets given at the start of the trial. Volunteers remembered the information they had been given at the start of the trial on procedures that involved drawing blood and urine samples but not information about study design and randomisation. Volunteers said that they had understood the purpose of the trial. They said that signing a consent form showed that they had consented to take part in the trial but they also described it as being done to protect the researcher in case a volunteer later experienced side effects.ConclusionVolunteers pay more attention during the consent process to procedures requiring biological tests than to study design issues. Trust built between volunteers and the research team could enhance the successful conduct of clinical trials by allowing for informal discussions to identify and review volunteers’ perceptions. These results point to the need for researchers to view informed consent as a process rather than an event.

Cyclophilin A in Ruptured Intracranial Aneurysm: A Prognostic Biomarker.

Cyclophilin A (CyPA), an oxidative stress-induced factor, was found to play an important role in the aneurysm formation. Our working hypothesis was that the plasma level of CyPA in ruptured intracranial aneurysm could predict the neurological outcome.From 2011 to 2013, a total of 36 patients with ruptured saccular intracranial aneurysm were recruited in our study. Before coil embolization, we draw blood samples at the orifice of a culprit aneurysm and in the remote peripheral vein for measurements of the CyPA levels. We utilized the modified Rankin scale 30 days after aneurysm rupture as the outcome measure. Generalized linear models were used to estimate the adjusted odds ratios of the poor neurological outcome given the presence of high plasma level of CyPA.The aneurysmal and venous CyPA levels were significantly associated with the initial clinical severity (P = 0.004 and 0.03, respectively) and 30-day outcome (P = 0.01 and 0.02, respectively). The aneurysmal CyPA levels modestly correlated with age and high Fisher grade (ρ = 0.39 and 0.41; P = 0.02 and 0.01, respectively). The aneurysmal CyPA levels strongly correlated with the venous counterpart (ρ = 0.89; P < 0.001). Patients with high levels of aneurysmal CyPA were 15.66 times (95% CI, 1.48–166.24; P = 0.02) more likely to have worse neurological outcome than those with the low levels after adjustment of the age, gender, and the documented confounding factors.High plasma level of CyPA is a significant prognostic biomarker for poor neurological outcome in patients with ruptured intracranial aneurysm.

Transfusion of Packed Red Cells

About four million units of packed red cells are transfused in Germany every year. The safety of blood transfusions is further improved by modern production methods and molecular diagnostic techniques. This review is based on selected publications,including the German guidelines and regulations and the German Transfusion Act. Packed red blood cells are transfused to prevent tissue hypoxia. As the clinical manifestations of anemia are nonspecific, the indication for transfusion is based on surrogate parameters, such as the hemoglobin (Hb) concentration, in addition to clinical criteria. For patients with unimpaired cardiopulmonary and vascular function, transfusion is generally indicated at hemoglobin values of 6 g/dL (3.7 mmol/L) or less. Randomized controlled trials have shown that a restrictive transfusion strategy (trigger: Hb 7-8 g/dL) in certain patient groups is as effective as a more liberal strategy (trigger: Hb about 10 g/dL). The most frequent causes for transfusion errors are lack of informed consent, lack of identity checking and/or AB0 identity testing, and the drawing of blood samples in unlabelled tubes. Overtransfusion, undertransfusion, as well as other transfusion errors can be markedly reduced by means of appropriate organizational measures and training.

Abstract 185: Using the Platelet Proteome to Identify Novel Biomarkers in Heart Failure With Preserved Ejection Fraction

Introduction: The platelet proteome is an untapped resource for identifying novel proteins that may reflect a disease process. The platelet proteome is understudied in heart failure. We hypothesize that the platelet proteome will change composition in patients with heart failure with ejection fraction (HFpEF). Using mass-spectral analysis, we examined the platelet proteome from subjects with heart failure with preserved ejection fraction (HFpEF) to identify proteins associated with the disease process. Methods/Results: We conducted a case-controlled prospective study in which we drew blood samples from hospitalized subjects with symptoms of HFpEF (n=9), the same subjects several weeks later without symptoms (n=7) and control subjects (n=8). The subjects with HFpEF were older and had a higher incidence of atrial fibrillation and smoking compared to the control group. There were no significant differences in body mass index, and history of hypertension, hyperlipidemia, coronary artery disease or diabetes. The HFpEF group had increased LV wall thickness, left atrial volume, and diastolic dysfunction. Mass spectrometry of the platelet rich abstract obtained from the blood samples of the 3 groups identified 6,102 proteins with 5 scans with peptide probabilities of ≥0.85. Of the 6,102 proteins, 165 were present only in symptomatic subjects, 78 were only found in asymptomatic subjects and 157 proteins were unique to the control group. S100A8 and its receptor, CD36 were two proteins identified consistently in HFpEF samples when compared with controls. Using ELISA, we further confirmed that plasma S100A8 levels were increased in subjects with HFpEF (728±358) vs. control (314±154). Conclusion: Platelets may harbor proteins associated with HFpEF. S100A8 is present in the platelets of subjects with HFpEF and increased in the plasma of the same subjects. S100A8 has been linked with other cardiovascular disease such as atherosclerosis and risk for myocardial infarction, stroke, or death. This is the first report on the association between S100A8 and HFpEF.

Performance of computer vision in vivo flow cytometry with low fluorescence contrast.

Detection and enumeration of circulating cells in the bloodstream of small animals are important in many areas of preclinical biomedical research, including cancer metastasis, immunology, and reproductive medicine. Optical in vivo flow cytometry (IVFC) represents a class of technologies that allow noninvasive and continuous enumeration of circulating cells without drawing blood samples. We recently developed a technique termed computer vision in vivo flow cytometry (CV-IVFC) that uses a high-sensitivity fluorescence camera and an automated computer vision algorithm to interrogate relatively large circulating blood volumes in the ear of a mouse. We detected circulating cells at concentrations as low as 20 cells/mL. In the present work, we characterized the performance of CV-IVFC with low-contrast imaging conditions with (1) weak cell fluorescent labeling using cell-simulating fluorescent microspheres with varying brightness and (2) high background tissue autofluorescence by varying autofluorescence properties of optical phantoms. Our analysis indicates that CV-IVFC can robustly track and enumerate circulating cells with at least 50% sensitivity even in conditions with two orders of magnitude degraded contrast than our previous in vivo work. These results support the significant potential utility of CV-IVFC in a wide range of in vivo biological models.

Effects of balloon inflation and cough trick methods on easing pain in children during the drawing of venous blood samples: a randomized controlled trial.

The purpose was to determine the effects of the balloon inflation and cough trick methods on easing pain in children during the drawing of venous blood samples. In this prospective randomized controlled study, 9- to 12-year-old children in the intervention groups were asked to cough or inflate balloons during the venipuncture procedure. The Faces Pain Scale-Revised was used to assess pain intensity. Pain intensity significantly differed between the control (n = 44) and intervention groups (balloon inflation [n = 44] and cough trick [n = 44], p < .001). Coughing and inflating balloons during venipuncture do not require preparation and are time saving, easy, accessible, and effective in reducing pain.

WinFood data from Kenya and Cambodia: constraints on field procedures.

Researchers face myriad challenges in the design and implementation of randomized, controlled trials. Apart from summaries on limitations, these challenges are rarely documented in detail to inform future research projects. To describe methodological challenges encountered during randomized, controlled trials (WinFood Study) designed to assess the efficacy of locally produced complementary foods based on traditional animal-source foods (edible termites and spiders) to support growth and nutritional status in Kenyan and Cambodian infants. In a randomized, controlled design, infants received WinFood or corn-soy blend (CSB) for 9 months from 6 to 15 months of age. Lean mass accrual and blood nutrition indicators (lipid profile, iron and zinc status) were measured cross-sectionally at 9 and 15 months of age, respectively. Lean mass was determined by measuring deuterium oxide enrichment in saliva samples following a standard dose of deuterium solution (0.5 g/kg body weight) to infants. Blood nutrition indicators were determined following the drawing of 3 mL of blood by venipuncture. Challenges included rapid depletion of food rations, high rate of loss to follow-up, delayed ethical approval, lack of local food-processing capacity, low capacity among staff to draw blood, and lack of laboratory capacity to perform both deuterium oxide and micronutrient status measurements. Spillage of deuterium oxide solution during dosing was a major challenge in the Kenya context. A high rate of morbidity among infants made some assessments very difficult, especially drawing of blood and saliva samples. The challenges were largely contextual. Improvement of local laboratory capacity, training of staff and sensitization of the communities and the Ethics Review Committee are highly recommended.

A nutrigenomic approach to detect nutritional stress from gene expression in blood samples drawn from Steller sea lions

Gene expression profiles are increasingly being used as biomarkers to detect the physiological responses of a number of species to disease, nutrition, and other stressors. However, little attention has been given to using gene expression to assess the stressors and physiological status of marine mammals. We sought to develop and validate a nutrigenomic approach to quantify nutritional stress in Steller sea lions (Eumetopias jubatus). We subjected 4 female Steller sea lions to 3 feeding regimes over 70-day trials (unrestricted food intake, acute nutritional stress, and chronic nutritional stress), and drew blood samples from each animal at the end of each feeding regime. We then extracted the RNA of white blood cells and measured the response of 8 genes known to react to diet restriction in terrestrial mammals. Overall, we found that the genomic response of Steller sea lions experiencing nutritional stress was consistent with how terrestrial mammals respond to dietary restrictions. Our nutritionally stressed sea lions down-regulated some cellular processes involved in immune response and oxidative stress, and up-regulated pro-inflammatory responses and metabolic processes. Nutrigenomics appears to be a promising means to monitor nutritional status and contribute to mitigation measures needed to assist in the recovery of Steller sea lions and other at-risk species of marine mammals.

Point and trend accuracy of continuous glucose monitoring using intravenous microdialysis in critically ill patients

Insulin infusion in critically ill patients mandates frequent measurements of the blood glucose level [1]. Microdialysis is a well-established technology that offers the opportunity to sample blood analytes with high accuracy, without the need for drawing blood samples [2,3]. We aimed to determine point and trend accuracy of microdialysis-based continuous glucose monitoring (CGM) (EIRUS®; Maquet Critical Care AB, Solna, Sweden).

Abstract 1816: Assessment of anti-tumor activity of the cathepsin L inhibitor, KGP94

Abstract Purpose: The cysteine cathepsin proteases are known to play an important role in tumor progression, invasion and angiogenesis. The present study evaluated the anti-tumor activity and pharmacokinetics of the small-molecule cathepsin L inhibitor KGP94. Materials and Methods: Male CDF1 or C3H/HeNHsd mice were inoculated on the right rear foot with a C3H mammary carcinoma or a SCCVII carcinoma, respectively. A solution of KGP94 was prepared each day by dissolving in a mixture of 10% Tween 80 and 90% HEPES-buffer. It was intraperitoneally injected at 0.01 ml/g mouse bodyweight. Various doses (1-20 mg/kg) were administered between 1-20 days, starting from the day of tumor inoculation. Anti-tumor activity was assessed by determining the tumor growth time, which was the time in days to reach a volume of 500 mm3 (TGT500). Pharmacokinetics of KGP94 were investigated by drawing blood samples before, and up to 24 hours after KGP94 injection. Results are listed as Mean (± Standard Error). One-way ANOVA comparison of group means was performed, and a P&amp;lt;0.05 was considered significant. Results: The TGT500 for control animals was 18.0 days (± 0.3) for the C3H mammary carcinoma and 13.6 days (± 0.7) for the SCCVII carcinoma. Treating tumors with KGP94 significantly increased the TGT500 for both tumor models to around 21 and 17 days respectively, when doses were at 10.0 mg/kg or higher. At a dose of 5.0 mg/kg the TGT500 of the C3H mammary carcinoma was significantly increased to 21.4 days (± 1.1), whereas the TGT500 of the SCCVII carcinoma was unchanged at 13.8 (± 0.2). At lower KGP94 doses neither tumor model showed significant growth delay. Additional experiments with the C3H mammary carcinoma model showed that treatment was most effective when started immediately after tumor inoculation, and that treatment for 5 days was just as effective as 20 days. Pharmacokinetic analysis of KGP94 levels in mouse plasma is ongoing. Conclusion: In the C3H mammary carcinoma KGP94 significantly delayed tumor growth, but the effect was primarily on the early phase following tumor cell inoculation. This suggests that KGP94 may have a more significant role in inhibiting the establishment of metastasis rather than influencing the growth of established tumors. Significant growth delay was also observed in the SCCVII tumor model. However, it occurred with higher doses of KGP94 compared to the C3H mammary carcinoma. The higher dose could be explained by the more aggressive nature of the SCCVII tumor model. Supported by The Danish Cancer Society, The Danish Council for Independent Research: Medical Sciences, and OXiGENE Inc. Citation Format: Thomas R. Wittenborn, Michael Stratford, Mary Lynn Trawick, Kevin G. Pinney, David J. Chaplin, Dietmar W. Siemann, Michael R. Horsman. Assessment of anti-tumor activity of the cathepsin L inhibitor, KGP94. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 1816. doi:10.1158/1538-7445.AM2014-1816

Improving validity of informed consent for biomedical research in Zambia using a laboratory exposure intervention.

BackgroundComplex biomedical research can lead to disquiet in communities with limited exposure to scientific discussions, leading to rumours or to high drop-out rates. We set out to test an intervention designed to address apprehensions commonly encountered in a community where literacy is uncommon, and where complex biomedical research has been conducted for over a decade. We aimed to determine if it could improve the validity of consent.MethodsData were collected using focus group discussions, key informant interviews and observations. We designed an intervention that exposed participants to a detailed demonstration of laboratory processes. Each group was interviewed twice in a day, before and after exposure to the intervention in order to assess changes in their views.ResultsFactors that motivated people to participate in invasive biomedical research included a desire to stay healthy because of the screening during the recruitment process, regular advice from doctors, free medical services, and trust in the researchers. Inhibiting factors were limited knowledge about samples taken from their bodies during endoscopic procedures, the impact of endoscopy on the function of internal organs, and concerns about the use of biomedical samples. The belief that blood can be used for Satanic practices also created insecurities about drawing of blood samples. Further inhibiting factors included a fear of being labelled as HIV positive if known to consult heath workers repeatedly, and gender inequality. Concerns about the use and storage of blood and tissue samples were overcome by a laboratory exposure intervention.ConclusionSelecting a group of members from target community and engaging them in a laboratory exposure intervention could be a useful tool for enhancing specific aspects of consent for biomedical research. Further work is needed to determine the extent to which improved understanding permeates beyond the immediate group participating in the intervention.

‘Short‐term treatment with methotrexate does not affect microvascular endothelial function in patients with psoriasis’

Psoriasis is associated with increased risk of cardiovascular disease (CVD), possibly due to chronic low-grade systemic inflammation. Systemic anti-inflammatory treatment might reduce the risk of CVD. Our aim was to investigate if short-term treatment with methotrexate influences microvascular endothelial function (MEF), an early surrogate marker of atherosclerosis, in patients with psoriasis. We prospectively studied a hospital cohort of patients with psoriasis. Measurements of MEF were performed with the Endo-PAT2000© device at baseline and after 8-10weeks of treatment with methotrexate. At the same time points, we recorded anamnestic information, measured body mass index (BMI), waist and hip circumferences and blood pressure, and drew blood samples (lipid profile, HbA1 and hs-CRP). Psoriasis severity was evaluated by psoriasis area and severity index (PASI) and the dermatology life quality index (DLQI). A total of 32 patients with psoriasis were included. Median age was 46 (range 18-82) years, and 50% were men. Twenty-seven patients completed the study. After 8-10weeks, median PASI had decreased significantly by 6.2 (from 9.8 to 3.6), and DLQI had decreased by 7 (from 9 to 2). No significant changes were observed in MEF, expressed by reactive hyperaemia index and augmentation index. Also, we saw no significant changes in BMI, waist-hip ratio, blood pressure and blood samples. Short-term treatment with methotrexate did not affect MEF in patients with psoriasis. Further studies are warranted.

EPA-0232 - Epigenetic alteration in ALDH2-metabolism in alcohol-dependent patients

Introduction Changes in DNA-methylation patterns in alcohol-dependent patients have been described in various studies. However, epigenetic regulation of genes responsible for the ethanol metabolism has not been in the focus of recent research. Objective Aim of our study was to evaluate changes in DNA-methylation of the aldehyde dehydrogenase 2 (ALDH2) in respect to genetic variants (rs886205 ALDH2-SNP) in blood samples of alcohol-dependent patients compared with healthy controls. Methods During an inpatients detoxification treatment we draw blood samples from 82 alcohol-dependent, male patients on day 1, day 7 and day 14. Bisulfite-treated DNA was subjected to methylation analysis via sequencing. Findings of the alcohol-dependent group were compared with results of a healthy control group (N=34) Results Our results show decreasing methylation levels of a repressive ALDH2 promoter fragment to control levels only in homozygous A-allele patients of the rs886205 SNP. Conclusion Findings of this study reveal a specific genetic-epigenetic interaction in ALDH2 regulation which may have crucial importance for the understanding of epigenetic regulation of ethanol metabolism in humans

Obtaining Blood Cultures by Venipuncture versus from Central Lines Impact on Blood Culture Contamination Rates and Potential Effect on Central Line–Associated Bloodstream Infection Reporting

Reduce the frequency of contaminated blood cultures that meet National Healthcare Safety Network definitions for a central line-associated bloodstream infection (CLABSI). An observational study. A 500-bed university-affiliated hospital. A new blood culture policy discouraged drawing blood samples from central lines. Phlebotomists were reeducated regarding aseptic technique when obtaining blood samples by venipuncture. The intravenous therapy team was taught how to draw blood samples by venipuncture and served as a backup when phlebotomists were unable to obtain blood samples. A 2-nurse protocol and a special supply kit for obtaining blood samples from catheters were developed. Rates of blood culture contamination were monitored by the microbiology laboratory. The proportion of blood samples obtained for culture from central lines decreased from 10.9% during January-June 2010 to 0.4% during July-December 2012 (P < .001). The proportion of blood cultures that were contaminated decreased from 84 (1.6%) of 5,274 during January-June 2010 to 21 (0.5%) of 4,245 during January-June 2012 (P < .001). Based on estimated excess hospital costs of $3,000 per contaminated blood culture, the reduction in blood culture contaminants yielded an estimated annualized savings of $378,000 in 2012 when compared to 2010. In mid-2010, 3 (30%) of 10 reported CLABSIs were suspected to represent blood culture contamination compared with none of 6 CLABSIs reported from mid-November 2010 through June 2012 (P = 0.25). Multiple interventions resulted in a reduction in blood culture contamination rates and substantial cost savings to the hospital, and they may have reduced the number of reportable CLABSIs.

A Low Cost Metal-Free Vascular Access Mini-Port for Artifact Free Imaging and Repeated Injections in Mice

PurposeSmall injection ports for mice are increasingly used for drug testing or when administering contrast agents. Commercially available mini-ports are expensive single-use items that cause imaging-artifacts. We developed and tested an artifact-free, low-cost, vascular access mini-port (VAMP) for mice.ProceduresLeakage testing of the VAMP was conducted with high speed bolus injections of different contrast agents. VAMP-induced artifacts were assessed using a micro-CT and a small animal MRI (9.4T) scanner ex vivo. Repeated contrast administration was performed in vivo.ResultsWith the VAMP there was no evidence of leakage with repeated punctures, high speed bolus contrast injections, and drawing of blood samples. In contrast to the tested commercially available ports, the VAMP did not cause artifacts with MRI or CT imaging.ConclusionsThe VAMP is an alternative to commercially available mini-ports and has useful applications in animal research involving imaging procedures and contrast agent testing.

Individual Platelet Adhesion Assay: Measuring Platelet Function and Antiplatelet Therapies in Whole Blood via Digital Quantification of Cell Adhesion

Widespread monitoring of platelet function and the effect of antiplatelet drugs will improve outcomes in cardiovascular patients, but platelet function testing is not routine in clinical practice. We report a rapid, accurate methodology to quantify platelet-protein interactions: a microarray of contact-printed 6-μm fibrinogen dots on a transparent substrate binds platelets from whole blood, one platelet per dot. The fractional occupancy of an array of fibrinogen dots after a predefined incubation time quantitatively assays platelet adhesion to the protein matrix. We demonstrate this technique by measurement of platelet adhesion to fibrinogen as a means to quantify the effect of the P2Y12 and αIIbβ3 receptor inhibitors cangrelor and abciximab, respectively, both in vitro--by incubating the drug with a freshly drawn blood sample--and in blood from patients treated with antiplatelet agents. The effects of single- and dual-antiplatelet therapy are also assessed. Results from this platelet-binding assay are well correlated with standard techniques including flow cytometry and light transmission aggregometry. This assay technology, readily integrated with microfluidic platforms, is generally applicable to the assay of cell-protein interactions and promises more effective, rapid assay of drug effects in cardiovascular disease patients.