Gel Tubes Research Articles

A Protocol to Validate Specimen Collection Tubes Without Requiring IRB Approval.

CLIA regulations require the validation of specimen collection tubes before reporting patient results (1, 2). Our laboratory recently encountered a problem with a new formulation of Vitamin D reagent (Abbott Diagnostics) when utilizing plasma processed from lithium heparin gel vacutainers (mint green top) (Becton, Dickinson and Company). Cells and particulates that remained above the gel after centrifugation were mixed into plasma samples during courier transportation from outpatient clinics. Analysis of these samples generated falsely increased values of vitamin D with the new formulation of reagent but not with the previous formulation. Removal of plasma from the gel tube followed by recentrifugation gave clinically sensible results that matched results obtained using the previous reagent formulation. This issue was not discovered during …

Rapid identification of pathogenic organisms positive for blood culture by MALDI-TOF mass spectrometry in combination with Sepsityper Kit or serum-separating gel tubes

Objective To evaluate the feasibility of using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) in combination with Sepsityper Kit or serum-separating gel tubes for identification of pathogenic organisms positive for blood culture. Methods A total of 153 clinical specimens with a positive result in blood culture were collected and tested with MALDI-TOF MS in combination with Sepsityper Kit or serum-separating gel tubes. Test results were compared with those by using culturing. Results The 153 positive blood culture specimens included 143 of single bacterial infection and 10 of multiple bacterial infections. The consistency rates at species and genus levels were 76.2% (109/153) and 7.8% (12/153) between Vitek 2 Compact method and MALDI-TOF MS combined with Sepsityper Kit, and 75.1% (101/153) and 8.5% (13/153) between Vitek 2 Compact method and MALDI-TOF MS combined with serum-separating gel tubes. In the identification of single bacterial infection specimens, the consistency rates of MALDI-TOF MS combined with Sepsityper Kit at species and genus levels were 95.2% (79/83) and 0% (0/83) for gram-negative bacteria, 57.5% (27/47) and 23.4% (11/47) for gram-positive bacteria and 33.3% (3/9) and 11.1% (1/9) for Candida, the consistency rates of MALDI-TOF MS combined with serum-separating gel tubes at species and genus levels were 90.4% (75/83) and 4.8% (4/83) for gram-negative bacteria, 55.3% (26/47) and 14.9% (7/47) for gram-positive bacteria and 0% (0/9) and 22.2% (2/9) for Candida. MALDI-TOF MS combined with Sepsityper Kit was only consistent with Vitek 2 Compact method in the identification of one specimen of multiple bacterial infections. Conclusion MALDI-TOF MS in combination with Sepsityper Kit or serum-separating gel tubes can identify the pathogens positive for blood culture within one hour and is of high consistency with culturing. In comparison with the traditional identification method for positive blood cultures, MALDI-TOF MS combined with Sepsityper Kit or serum-separating gel tubes has the advantages of rapidity and easy operation, which meet the clinical needs of rapid diagnosis and timely and effective antibacterial treatment, and is of great value in clinical application. Key words: Sepsityper Kit; Serum-separating gel tube; MALDI-TOF MS; Blood culture

Elevated Expression of miR302-367 in Endothelial Cells Inhibits Developmental Angiogenesis via CDC42/CCND1 Mediated Signaling Pathways

Rationale: Angiogenesis is critical for embryonic development and microRNAs fine-tune this process, but the underlying mechanisms remain incompletely understood.Methods: Endothelial cell (EC) specific miR302-367 line was used as gain-of-function and anti-miRs as loss-of-function models to investigate the effects of miR302-367 on developmental angiogenesis with embryonic hindbrain vasculature as an in vivo model and fibrin gel beads and tube formation assay as in vitro models. Cell migration was evaluated by Boyden chamber and scratch wound healing assay and cell proliferation by cell count, MTT assay, Ki67 immunostaining and PI cell cycle analysis. RNA high-throughput sequencing identified miR-target genes confirmed by chromatin immunoprecipitation and 3'-UTR luciferase reporter assay, and finally target site blocker determined the pathway contributing significantly to the phenotype observed upon microRNA expression.Results: Elevated EC miR302-367 expression reduced developmental angiogenesis, whereas it was enhanced by inhibition of miR302-367, possibly due to the intrinsic inhibitory effects on EC migration and proliferation. We identified Cdc42 as a direct target gene and elevated EC miR302-367 decreased total and active Cdc42, and further inhibited F-actin formation via the WASP and Klf2/Grb2/Pak1/LIM-kinase/Cofilin pathways. MiR302-367-mediated-Klf2 regulation of Grb2 for fine-tuning Pak1 activation contributing to the inhibited F-actin formation, and then the attenuation of EC migration. Moreover, miR302-367 directly down-regulated EC Ccnd1 and impaired cell proliferation via the Rb/E2F pathway.Conclusion: miR302-367 regulation of endothelial Cdc42 and Ccnd1 signal pathways for EC migration and proliferation advances our understanding of developmental angiogenesis, and meanwhile provides a rationale for future interventions of pathological angiogenesis that shares many common features of physiological angiogenesis.

The Effects of Glucomannan Hydrolysates and BV Gel on Nugent Score, Treg Cell Percentage, and TGF-? level in Bacterial Vaginosis

Background: Bacterial vaginosis is commonly experienced by women of reproductive age. The Nugent score is the gold standard for diagnosing bacterial vaginosis. Prebiotic Glucomannan hydrolysates (GMH) as a therapy in the treatment of bacterial vaginosis and has an immunomodulatory effect on the immune system and provides cellular immunity. The purpose of this study was to examine the effects of GMH and BV Gel on Nugent scores, Treg cell presentation, and TGF ? levels in bacterial vaginosis of women of childbearing age.Subjects and Method: This was an experimental study. A sample of 24 women aged 20 to 45 years old with bacterial vaginosis (Nugent score ?7) was divided into 4 groups: oral antibiotic group metronidazole (500mg), combination of GMH (300mg) and metronidazole, 5 ml BV Gel tube, and combination of GMH and BV Gel scores. The dependent variables were GMH and BV Gel administrations. The independent variables Nugent score, Treg presentation, and TGF-? level. Nugent score, Treg cell presentation, and TGF ? level were measured on day-0, day-11, and day-22. The data were analyzed by one way Anova.Results: The results of the analysis after treatment on day 22 showed that the GMH and BV gel were able to reduce Nugent scores, increase Treg cell presentation and TGF ? levels in bacterial vaginosis of women of childbearing age.Conclusion: GMH as an alternative therapy for bacterial vaginosis compared with antibiotic treatment can improve normal vaginal flora and stimulate the immune system in vitro and in vivo significantly.Keywords: Glucomannan Hydrolysates, BV Gel, Nugent sel Treg score, TGF ?Correspondence:Ambar Dwi Retnoningrum. Masters Program of Midwifery, Universitas Brawijaya, Malang, East Java. Email: adreambar@gmail.com. Mobile: +6281335743696.Indonesian Journal of Medicine (2018), 3(1): 33-43https://doi.org/10.26911/theijmed.2018.03.01.05

Co-culturing in multilayer polymer gel tube device fabricated by repeated molding

Co-culturing in multilayer polymer gel tube device fabricated by repeated molding

Determination of the Reference Values of the Tumor Marker HE4 in Female Population of Canton Sarajevo.

Introduction:Ovarian cancer has the highest mortality rate of all gynecological tumors and represent fifth leading cause of cancer death in women, due to the absence of early symptoms of the disease and the lack of screening tests. A new HE4 ovarian cancer biomarker was found in various studies, with different populations having quite different sensitivity and specificity values as well as reference values. The tumor marker HE4 is a useful marker for ovarian cancer detection with only minimal expression in normal ovarian tissue. HE4 is particularly promising as a marker for early detection, showing its potential in differentiating women with ovarian cancer than those with benign (nonmalignant) ovarian conditions.Material and methods:Samples were taken in serum gel tubes, centrifuged and stored until analysis. A total of 300 respondents was included, of whom 188 were premenopausal women and 112 postmenopausal women. Respondents were divided into two groups by age: women aged 50-75 years and Control group of women aged 20-50 years. The HE4 assay was performed by the Immunological Test (ECLIA) for the quantitative determination of HE4 in human serum or plasma. Solid phase test, two-step reaction using immune 2HP and 3HD8 monoclonal antibodies („sandwich principle“).Results:Tumor markers HE4 analysis by groups indicate that in the postmenopausal group there was 92 (82.14%) of subjects with normal tumor markers HE4 and 170 (90.43%) in the premenopausal group. The elevated values of HE4 tumor markers in the postmenopausal group were in 20 (17.86%) cases and in the premenopausal group 18 (9.57%). Statistical analysis (nonparametric method) established the upper limit of the reference value of HE4 tumor markers in premenopausal and postmenopausal women.Conclusion:The established reference value for the women population in Canton Sarajevo for premenopausal women is <78.6 pmol/l, and for postmenopausal women <122.5 pmol/l. We found that the values of HE4 tumor markers differ significantly in premenopausal and postmenopausal women (p=0.0391). Postmenopausal women have a higher incidence of high-value tumor markers HE4. By comparison of the reference interval of the tumor marker HE4 for the population of women from Canton Sarajevo, determined by our research, with a reference interval for the German and Asian populations, we found a significant difference between the upper limit of the reference value in postmenopausal women (p<0.05), while in premenopausal women the difference was not statistically significant (p=0.4314).

Dynamic Phantom for Flow Model in Magnetic Resonance Angiography

Purpose. To develop phantom for flow modeling in magnetic resonance angiography (MRA): relative contrast assessment, accuracy of the linear velocity and volumetric flow, what improve accuracy of diagnostic in cardiac and neurosurgical clinics (quality assessment of blood and cerebrospinal fluid motion). To compare scanners of different manufactures in points of the MRA efficiency using the developed phantom.Materials and methods. The main part of dynamic phantom consists of a disc filled with agarose gel (for linear and volumetric velocity control) and silicone tubes for fluid flow modelling. MR study was performed at MRI units of two manufactures for comparing quantitative assessments of MRA sequences: 2DTOF, 3DTOF, and at three MRI units of one firm for estimated accuracy calibration curve calculating and linear velocity and volumetric flow determination for PC MRA. Phantom study well correlate with clinical MRA results.Results. Obtained phantom scanning results in 2DTOF, 3DTOF sequences allow for objective comparing two MRI units of different manufactures. For 2DTOF mode was showed more effective signal enhancement affected by TOF effect for scanner of manufacture 2, then manufacture 1: 8.86 ± 0.88 и 6.07 ± 0.03 corresponding. For 3DTOF was observed rather more inflow relative contrast affected by TOF effect for scanner of manufacture 1: 6.06 ± 0.47 and 3.17 ± 0.83 corresponding. However, for manufacture 1 was showed more significant signal suppression for fat tissue, which improve vasculature visualization. Accuracy linear velocity fluid flow measurement in 2DPC is equal to ±2σ = ±0,4 by five pixels for three scanners of one manufacture. Using developed phantom was modelled MRA effects in 3DPC and Time-SLIP modes.Conclusions. The developed dynamic phantom can be used for calibration tests in MRA. The case of MRI units of two manufactures were compared quantitative assessments of MRA sequences and analyzed methods of enhancement fluid flow signal.

Good correlation between the Afinion AS100 analyser and the ABX Pentra 400 analyser for the measurement of glycosylated haemoglobin and lipid levels in older adults in Durban, South Africa.

The Afinion AS100 analyser is a small bench-top, multi-assay, point-of-care (POC) analyser that is able to measure glycated haemoglobin (HbA1c) and lipid levels. To assess performance of the Afinion analyser compared with a reference laboratory test for the measurement of HbA1c and lipid levels. The study involved men and women enrolled in a cross-sectional study, Sexual health, HIV infection and comorbidity with non-communicable diseases among Older Persons (SHIOP), which was conducted from February to May 2016. Whole blood was drawn aseptically by a trained study nurse into a serum separator gel tube and an ethylenediaminetetra-acetic acid (EDTA) tube. The EDTA whole blood was used to measure HbA1c levels, and serum to measure total cholesterol (TC), high-density lipoprotein cholesterol (HDL-C), low-density lipoprotein cholesterol (LDL-C) and triglyceride levels. Lin's correlation coefficient was used to assess the agreement between the Afinion and ABX Pentra 400 analysers for each marker. A total of 435 older individuals were included in the study. The proportion of HbA1c results that were correctly classified by the Afinion analyser was 92.2%. Bland-Altman analysis and linear regression analysis showed a very good agreement (correlation concordance 0.89) between the two analysers for the measurement of HbA1c. The two-way scatter plot for TC showed a substantial correlation (0.80). However, a total of 69 cholesterol results that were within the normal range on the Pentra were misclassified as abnormal on the Afinion. The readings obtained for HDL-C levels with the Afinion were shown to be slightly overestimated when compared with the Pentra. However, correlation for HDL-C on the two analysers was 0.93, indicating an almost perfect agreement. Seventy-four LDL-C results were erroneously classified as abnormal on the Afinion but were within the normal range on the Pentra, resulting in a substantial correlation of 0.75. An excellent agreement was observed between triglyceride measurements (0.99). This study supports the use of the Afinion AS100 analyser in POC testing for the measurement of HbA1c, triglycerides and HDL-C in a South African setting.

Elevated Systemic IL-6 Levels in Patients with Aneurysmal Subarachnoid Hemorrhage Is an Unspecific Marker for Post-SAH Complications.

Background: Aneurysmal subarachnoid hemorrhage (aSAH) is still a fatal and morbid disease, although bleeding aneurysms can be secured in almost all cases. Occurrence of post-SAH complications including cerebral vasospasm, delayed cerebral ischemia, hydrocephalus, epilepsy, and infections are the main determinants of clinical outcome. Hence, it is important to search for early predictors for specific post-SAH complications to treat these complications properly. Both cellular and molecular (cytokines) inflammation play a key role after aSAH during the phase of occurrence of post-SAH complications. Interleukin-6 (IL-6) is a well-known cytokine that has been extensively analyzed in cerebrospinal fluid (CSF) of patients after aSAH, but detailed studies exploring the role of systemic IL-6 in aSAH associated complications and its impact on early clinical outcome prediction are lacking. The current study aims to analyze the systemic IL-6 levels over two weeks after bleeding and its role in post-SAH complications. Methods: We recruited 80 aSAH patients prospectively who underwent peripheral venous blood withdrawal in serum gel tubes. The blood was centrifuged to harvest the serum, which was immediately frozen at −80 °C until analysis. Serum IL-6 levels were quantified using Immulite immunoassay system. Patient records including age, gender, post-SAH complications, aneurysm treatment, and clinical outcome (modified Rankin scale and Glasgow outcome scale) were retrieved to allow different subgroup analysis. Results: Serum IL-6 levels were significantly raised after aSAH compared to healthy controls over the first two weeks after hemorrhage. Serum IL-6 levels were found to be significantly elevated in aSAH patients presenting with higher Hunt and Hess grades, increasing age, and both intraventricular and intracerebral hemorrhage. Interestingly, serum IL-6 was also significantly raised in aSAH patients who developed seizures, cerebral vasospasm (CVS), and chronic hydrocephalus. IL-6 levels were sensitive to the development of infections and showed an increase in patients who developed pneumoniae. Intriguingly, we found a delayed increase in serum IL-6 in patients developing cerebral infarction. Finally, IL-6 levels were significantly higher in patients presenting with poor clinical outcome in comparison to good clinical outcome at discharge from hospital. Conclusion: Serum IL-6 levels were elevated early after aSAH and remained high over the two weeks after initial bleeding. Serum IL-6 was elevated in different aSAH associated complications, acting as a non-specific marker for post-SAH complications and an important biomarker for clinical outcome at discharge.

Post-analytical stability of 23 common chemistry and immunochemistry analytes in incurred samples

BackgroundStorage of blood samples after centrifugation, decapping and initial sampling allows ordering of additional blood tests. The pre-analytic stability of biochemistry and immunochemistry analytes has been studied in detail, but little is known about the post-analytical stability in incurred samples. MethodsWe examined the stability of 23 routine analytes on the Dimension Vista® (Siemens Healthineers, Denmark): 42–60 routine samples in lithium-heparin gel tubes (Vacutainer, BD, USA) were centrifuged at 3000×g for 10min. Immediately after centrifugation, initial concentration of analytes were measured in duplicate (t=0). The tubes were stored decapped at room temperature and re-analyzed after 2, 4, 6, 8 and 10h in singletons. The concentration from reanalysis were normalized to initial concentration (t=0). Internal acceptance criteria for bias and total error were used to determine stability of each analyte. Additionally, evaporation from the decapped blood collection tubes and the residual platelet count in the plasma after centrifugation were quantified. Results and conclusionWe report a post-analytical stability of most routine analytes of ≥8h and do therefore – with few exceptions – suggest a standard 8hour-time limit for reordering and reanalysis of analytes in incurred samples.

Comprehensive Lipoprotein Characterization Using Lipidomics Analysis of Human Plasma.

Lipoproteins are responsible for the transport of lipids and other nutrients in the circulation and therefore play an important role in lipid metabolism and dyslipidemia. They have also been linked to multiple metabolic disorders including cardiovascular disease, and thus understanding their lipid composition is of crucial importance. Characterization of lipoproteins is a challenging task due to their heterogeneity. In particular, their fractionation is often laborious and time-consuming, making large sets of clinical samples difficult to analyze. We developed and validated lipidomics analysis of lipoproteins including chylomicrons, very low-density, low-density, and high-density lipoproteins. Lipoproteins were first fractionated by polyacrylamide tube gel electrophoresis, and, after liquid-liquid extraction, lipids were analyzed by direct-infusion mass spectrometry. About 100 unique lipid species were detected with good reproducibility and reliability. In addition to their lipid composition, valuable information on the fatty acid composition of lipoproteins and lipids was obtained. The presented method offers in-depth analysis of the lipid as well as fatty acid composition of lipoproteins while allowing a good sample throughput. It is thus especially suited for studying lipid associated diseases in clinical cohorts.

Automated Sample Preparation Enables LC-MS/MS as a Routine Diagnostic Analysis for Serum Testosterone.

The advantage of LC-MS/MS for analyzing serum testosterone in female, pediatric, and hypogonadal male patient samples is well accepted (J Clin Endocrinol Metab 2010;95:4542-8). However, many clinical laboratories still use testosterone immunoassays because of the technical challenges of LC-MS/MS (Clin Chem 2010;56:1234-44). Although LC-MS/MS has been shown to have better accuracy and specificity than immunoassay, better reproducibility has been more elusive because of the complexities of sample preparation (Clin Chem 2008;54:1290-7; Rev Endocr Metab Disord 2013;14:185-205). We evaluated replacing automated immunoassay with LC-MS/MS for all testosterone analyses at the University of California San Diego Health System Center for Advanced Laboratory Medicine. We used a novel extraction media, AC Extraction Plate™ (AC Plate), from Tecan Schweiz with automated liquid handling for LC-MS/MS sample preparation. We modified the existing vendor application and validated the method for matrix effect, recovery, precision, trueness [accuracy relative to certified reference material (CRM)], specificity, reportable range, sample stability, and correlation with other methods. Method performance was excellent, with a reportable range of 4-1560 ng/dL (0.14-54.13 nmol/L), between-day CV <6%, mean accuracy for CRM of <4.0% bias, no interference from hemolysis, icterus, lipemia, serum separator tube gel, or common steroids/metabolites, and mean bias of 1.3% vs 4 other LC-MS/MS testosterone methods. A retrospective analysis of calibration stability suggests that sparse and/or historical calibration is feasible for routine use. We conclude that automated extraction with the AC Plate and a focus on robustness during method development delivered ease of use and method performance consistent with adopting LC-MS/MS for all testosterone testing.

Fabrication of micro-alginate gel tubes utilizing micro-gelatin fibers

Tissues engineered utilizing biofabrication techniques have recently been the focus of much attention, because these bioengineered tissues have great potential to improve the quality of life of patients with various hard-to-treat diseases. Most tissues contain micro-tubular structures including blood vessels, lymphatic vessels, and bile canaliculus. Therefore, we bioengineered a micro diameter tube using alginate gel to coat the core gelatin gel. Micro-gelatin fibers were fabricated by the coacervation method and then coated with a very thin alginate gel layer by dipping. A micro diameter alginate tube was produced by dissolving the core gelatin gel. Consequently, these procedures led to the formation of micro-alginate gel tubes of various shapes and sizes. This biofabrication technique should contribute to tissue engineering research fields.

Strategies to identify candidates for D variant genotyping.

RhD variants have altered D epitopes and/or decreased antigen copies per red cell. Individuals carrying these variants may test antigen negative, weakly positive, or positive by serology, and may or may not be at risk of alloimmunisation after exposure. There have been recommendations to perform RHD genotyping of patients, pregnant women and females of childbearing potential with serological weak D phenotype, to guide prophylactic use of Rh immune globulin (RhIG), and better conserve D-negative blood products. The purpose of this study was to evaluate the performance of a set of empirical criteria to identify such patients. A two-method strategy of gel testing (GT) and tube testing (TT) was used for Rh typing of patients with no historical blood type in the present institution. A monoclonal-polyclonal blend anti-D was used for Rh typing by TT at immediate spin. Three empirical criteria were used to identify candidates for genotyping: C1: discrepancy between the two test methods and a GT reaction strength >2+ stronger than TT; C2: weak serological reaction, defined as reaction strength ≤2+ regardless of testing method if both GT and TT were performed or reaction strength ≤2+ if only GT was performed, or reaction strength ≤1+ if only TT was performed; C3: presence of anti-D in D-positive patients with no history of RhIG use in the preceding 3 months and in whom alloanti-D is suspected. Overall, 50 patients, ranging from newly born to 93 years old, were identified. Genomic testing confirmed D variants in 49/50 cases with a positive predictive value of 98%. This two-method strategy is a powerful screening tool for identifying candidates for RHD genotyping. This strategy meets the current requirements of two blood type determinations/two specimens in pre-transfusion testing while simultaneously identifying candidates for RHD genotyping with a minimal increase in work load and cost.

Antibodies to biotinylated red blood cells in adults and infants: improved detection, partial characterization, and dependence on red blood cell-biotin dose.

Biotin-labeled red blood cells (BioRBCs) are used for in vivo kinetic studies. Because BioRBC dosing occasionally induces antibodies, a sensitive and specific anti-BioRBC detection assay is needed. Aims were to 1) develop a gel card assay to evaluate existing, naturally occurring and BioRBC-induced plasma antibodies, 2) compare gel card and tube agglutination detection results, and 3) test for a relationship of antibody induction and BioRBC dose. Reagent BioRBCs were prepared using sulfo-NHS biotin ranging from densities 18 (BioRBC-18) to 1458 (BioRBC-1458)µg/mL RBCs. Among BioRBC-exposed subjects, gel card and tube agglutination results were concordant in 21 of 22 adults and all 19 infant plasma samples. Gel card antibody detection sensitivity wasmore than 10-fold greater than tube agglutination. Twelve to 16 weeks after BioRBC exposure, induced anti-antibodies were detected by gel card in three of 26 adults (12%) at reagent densities BioRBC-256 or less, but in none of 41 infants. Importantly, induced anti-BioRBC antibodies were associated with higher BioRBC dose (p=0.008); no antibodies were detected in 18 subjects who received BioRBC doses less than or equal to BioRBC-18. For noninduced BioRBC antibodies, six of 1125 naïve adults (0.3%) and none of 46 naïve infants demonstrated existing anti-BioRBC antibodies using reagent BioRBC-140 or -162. Existing anti-BioRBCs were all neutralized by biotin compounds, while induced antibodies were not. The gel card assay is more sensitive than the tube agglutination assay. We recommend reagent BioRBC-256 for identifying anti-BioRBCs. Use of a low total RBC biotin label dose (≤ BioRBC-18) may minimize antibody induction.

CHARACTERISTICS OF CROSSMATCH TYPES IN COMPATIBILITY TESTING ON DIAGNOSIS AND BLOOD TYPES USING GEL METHOD

Cross matching is a method to investigate compatibility between recipients and donor blood. Furthermore, this method is applied for investigation of compatibility between donor’s erythrocyte antigen and recipient’ serum antibody (major test) and for investigation of compatibility between donor serum antibody and recipient’s erythrocyte antigen (minor test). Cross matching may be performed using tube method (convensional) and gel method. To determine the charateristic of blood group and diagnosis in crossmatch incompatibility using Gel method. An observational study with retrospective approach has been done in blood bank Dr. Wahidin Sudirohusodo Hospital using data from Hospital information system and medical record from June 2013 until June 2014. Total 213 samples consisted of 105 males (49.3%) and 108 females (50.7%). Age groups >17 years old had a higher number for incompatibility (83.6%). Incompatibility was found also higher in infection group as the main diagnosis (77; 36.2%), B blood group (70; 32.9%), positive rhesus (100%) and minor incompatibility (81.2%). Samples from patients without transfusion history had a higher number of incompatibility (97.7%) than with transfusion history (2.3%). A further research by taking direct medical history from patient was suggested to avoid bias so the result of incompatibility would be more valid. A study for antibodies screening and identification for any crossmatch incompatibility was also suggestion.

Comparison of antibody titers using conventional tube technique versus column agglutination technique in ABO blood group incompatible renal transplant.

INTRODUCTION:Measurement of alloantibody titer to a red cell antigen (ABO titers) is an integral part of management of ABO incompatible kidney transplants (ABOiKT).MATERIAL AND METHODS:There are different methods of titer estimation. Alloantibody detection by tube titration and Gel agglutination columns are accepted methodologies. It is essential to find the difference in titers between the two methods so as to set the 'cut-off' titer accordingly, depending upon the method used.RESULTS:We did a prospective observational study to compare and correlate the ABO titers using these two different techniques – conventional tube technique (CTT) and the newer column agglutination technique (CAT). A total of 67 samples were processed in parallel for anti-A/B antibodies by both tube dilution and column agglutination methods. The mean titer by conventional tube method was 38.5 + 96.6 and by the column agglutination test was 96.4 + 225. The samples correlated well with Spearman rho correlation coefficient of 0.94 (P = 0.01).CONCLUSION:The column agglutination method for anti A/B titer estimation in an ABO incompatible kidney transplant is more sensitive, with the column agglutination results being approximately two and half fold higher (one more dilution) than that of tube method.

Benchmarking sample preparation/digestion protocols reveals tube-gel being a fast and repeatable method for quantitative proteomics.

Sample preparation, typically by in-solution or in-gel approaches, has a strong influence on the accuracy and robustness of quantitative proteomics workflows. The major benefit of in-gel procedures is their compatibility with detergents (such as SDS) for protein solubilization. However, SDS-PAGE is a time-consuming approach. Tube-gel (TG) preparation circumvents this drawback as it involves directly trapping the sample in a polyacrylamide gel matrix without electrophoresis. We report here the first global label-free quantitative comparison between TG, stacking gel (SG), and basic liquid digestion (LD). A series of UPS1 standard mixtures (at 0.5, 1, 2.5, 5, 10, and 25 fmol) were spiked in a complex yeast lysate background. TG preparation allowed more yeast proteins to be identified than did the SG and LD approaches, with mean numbers of 1979, 1788, and 1323 proteins identified, respectively. Furthermore, the TG method proved equivalent to SG and superior to LD in terms of the repeatability of the subsequent experiments, with mean CV for yeast protein label-free quantifications of 7, 9, and 10%. Finally, known variant UPS1 proteins were successfully detected in the TG-prepared sample within a complex background with high sensitivity. All the data from this study are accessible on ProteomeXchange (PXD003841).

Erratum.

Veterinary Clinical PathologyVolume 45, Issue 4 p. 733-733 ErratumFree Access Erratum This article corrects the following: Preliminary evaluation of a gel tube agglutination major cross-match method in dogs Dania Villarnovo, Shelley A. Burton, Barbara S. Horney, Allan L. MacKenzie, Raphaël Vanderstichel, Volume 45Issue 3Veterinary Clinical Pathology pages: 411-416 First Published online: July 5, 2016 First published: 21 November 2016 https://doi.org/10.1111/vcp.12432Citations: 1AboutSectionsPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Share a linkShare onFacebookTwitterLinkedInRedditWechat When this article1 was published in the journal, an incorrect figure was included for Figure 2. The correct figure is: Figure 2Open in figure viewerPowerPoint Gel column images of control and sample pairings with atypical patterns using the RapidVet-H major cross-match gel tube method (Study 1). (A) Positive control tube. (B) Negative control tube. (C) Negative control tube with erythrocytes present at the top of the gel column. (D–G) Reaction tubes: (D, E, and G) Erythrocyte spreading throughout the gel columns. (F) Erythrocytes at both bottom and top of gel column. Pairings E–G were negative on the standard tube method and on the RapidVet-H method. The pairing D was positive on the standard tube method but negative on the Rapid-Vet-H method. References 1Villarnovo D, Burton SA, Horney BS, MacKenzie AL, Vanderstichel R. Preliminary evaluation of a gel tube agglutination major cross-match method in dogs. Vet Clin Pathol. 2016; 45: 411– 416. Citing Literature Volume45, Issue4December 2016Pages 733-733 FiguresReferencesRelatedInformation

A novel approach to evaluate spatial resolution of MRI clinical images for optimization and standardization of breast screening protocols.

Stringent quality assurance is required in MRI breast screening to ensure that different scanners and imaging protocols reach similar diagnostic performance. The authors propose a methodology, based on power spectrum analysis (PSA), to evaluate spatial resolution in clinical images. To demonstrate this approach, the authors have retrospectively compared two MRI sequences commonly employed in breast screening. In a novel approach to PSA, spatial frequency response curves (SFRCs) were extracted from the images. The SFRC characterizes spatial resolution describing the spatial frequency content of an image over a range of frequencies. Verification of the SFRCs was performed on MRI images of Eurospin agarose gel tubes acquired with different resolution settings. SFRCs of volunteer and patient images obtained with two clinical MRI sequences were then compared. The two sequences differed primarily in k-space coverage pattern, which was either radial (RAD) or linear (LIN). The computed SFRCs were able to demonstrate the differences between RAD and LIN sequences in relatively small groups of subjects. The curves showed a similar pattern of decay in both volunteer and patient images, indicating that the spatial frequency response is mainly determined by the imaging protocol and not by intersubject anatomical differences. The LIN protocol produced images with increased sharpness; this was reflected in the corresponding SFRCs, which showed a higher content of spatial frequencies associated with image details. The SFRC can provide an objective assessment of the presence of spatial details in the image and represent a useful quality assurance tool in the evaluation of different breast screening protocols. With a reference image, a comparative analysis of the SFRCs could ensure that equivalent image quality is achieved across different scanners and sites.