Gel Tubes Research Articles

A novel method to detect intracellular metabolite alterations in MCF-7 cells by doxorubicin induced cell death.

Metabolic reprogramming within cancer cells has been recognized as a potential barrier to chemotherapy. Additionally, metabolic tumor heterogeneity is the one of factors behind discernible hallmarks such as drug resistance, relapse of the tumor and the formation of secondary tumors. In this paper, cell-based assays including PI/annexin V staining and immunoblot assay were performed to show the apoptotic cell death in MCF-7 cells treated with DOX. Further, MCF-7 cells were lysed in a hypotonic buffer and the whole cell lysate was purified by a novel and specifically designed metabolite (~ 100 to 1000Da) fractionation system called vertical tube gel electrophoresis (VTGE). Further, purified intracellular metabolites were subjected to identification by LC-HRMS technique. Cleaved PARP 1 in MCF-7 cells treated with DOX was observed in the present study. Concomitantly, data showed the absence of active caspase 3 in MCF-7 cells. Novel findings are to identify key intracellular metabolites assisted by VTGE system that include lipid (CDP-DG, phytosphingosine, dodecanamide), non-lipid (N-acetyl-D-glucosamine, N1-acetylspermidine and gamma-L-glutamyl-L-cysteine) and tripeptide metabolites in MCF-7 cells treated by DOX. Interestingly, we reported the first evidence of doxorubicinone, an aglycone form of DOX in MCF-7 cells that are potentially linked to the mechanism of cell death in MCF-7 cells. This paper reported novel methods and processes that involve VTGE system based purification of hypotonically lysed novel intracellular metabolites of MCF-7 cells treated by DOX. Here, these identified intracellular metabolites corroborate to caspase 3 independent and mitochondria induced apoptotic cell death in MCF-7 cells. Finally, these findings validate a proof of concept on the applications of novel VTGE assisted purification and analysis of intracellular metabolites from various cell culture models.

Determination of acrylic acid in workplaceair by on-line methylationgas chromatography

A method to determine acrylic acid in workplace air was developed by on-line methylation gas chromatography. Methods: the samples were absorbed by solvent desorption silica gel tube and desorbed by methanol. Desorption fluid in the injector at 350 ℃ in the presence of an organic alkali tetramethylamine hydroxide (TMAH, 25% methanol) , allowed quantitative analysis of acrylic acid as its corresponding methyl derivative by gas chromatography. Results: calibration curve of the method was linear within the range 0-258.4 mg/L and showed good linearity with r=0.999 4. The determination limit of the method was 0.9 mg/L, and the minimum detection concentration was 0.06 mg/m(3) (collect 15 L air) . The relative standard deviation (RSD) was 2.2%-2.7% (n=5) . Recoveries were between 96.9-101.6%. Conclusion: the results prove on-line methylation gas chromatography is an accurate, simple and high sensitive method for determining acrylic acid in workplace air.

A study on the assessment of stability of glucose concentrations in serum separator gel tubes

A study on the assessment of stability of glucose concentrations in serum separator gel tubes - IJCBR- Print ISSN No: - 2394-6369 Online ISSN No:- 2394-6377 Article DOI No:- 10.18231/2394-6377.2018.0069, International Journal of Clinical Biochemistry and Research-Int J Clin Biochem Res

3D printed portable gel electrophoresis device for rapid detection of proteins

The growing demand for rapid, portable, and economical detection methods for environmental analysis has resulted in increasing demands on the portability and miniaturization of analytical instruments. The miniaturization of scientific instruments facilitates analysis in the field of medicine, food, and environment, especially for the under-resourced areas. The gel electrophoresis devices currently available for protein separation are primarily used in laboratories. Miniaturized instruments that can be used for on-site and rapid separation of protein have not yet been reported. In this study, a portable gel electrophoresis device for rapid separation and detection of proteins was developed and manufactured by 3D printing in a laboratory, which was economical, convenient, and quick. First, four kinds of portable gel electrophoresis devices that included three kinds of columnar gel and one slab gel electrophoresis device were designed with computer-aided design software SolidWorks 2017 (Dassault Systemes SE, France); the components including gel tubes, gel plates, and gel electrophoresis tanks were then printed using a 3D printer after optimization of the printing parameters. Then, the performance of the four kinds of gel electrophoresis devices was investigated using prestained protein molecular weight standards. The results showed that the single-channel slab gel electrophoresis design can quickly separate proteins with the best separation efficiency. Moreover, the effect of different separation gel lengths (5, 10, 15, and 20 mm) on protein separation was studied and it was found that 10% separation gels with a length of 5 mm could effectively separate prestained protein molecular weight standards (in the range of 15-250 kD) in 20 minutes. Next, the battery was optimized for the portable GE device and a 25 V lithium battery (70 mm×60 mm×40 mm) was used as the power supply, which could provide a constant voltage of 25 V for 100 hours during gel electrophoresis. Then, the One-Step BlueTM reagent (Biotium, USA) was used to color the separation results of the five standard proteins (carbonic anhydrase, ovalbumin, bovine serum albumin, conalbumin, ribonuclease A), and the results were recorded by mobile phone. Finally, the proposed gel electrophoresis device was compared with the commercial device. The results showed that the two devices are comparable; however, the slab gel electrophoresis was faster, portable, and economical. In summary, this research designed and manufactured a portable gel electrophoresis device using 3D printing technique, which can be used for on-site analysis and detection of proteins. The device presents the following advantages compared with the commercial devices:1) small and portable:the size of the electrophoresis tank of the device is only 15 mm×20 mm×17 mm and the 25 V lithium battery has a working time of approximately 100 hours; 2) low cost:it can be processed in 5 hours using 3D printing technology, with 10 mL of printing material while the total cost is less than 400 RMB; 3) fast separation:this device can quickly achieve protein separation compared with commercial devices and can further use multiple electrophoresis tanks in parallel to analyze more samples at the same time. Besides, this research also highlights the advantages of 3D printing for the development of miniaturized analytical equipment. Though this study has achieved preliminary results for rapid separation of proteins using gel electrophoresis devices, the quantitative analysis of proteins following protein detection and the application of more samples need further research. Meanwhile, the continued application of 3D printing technology will promote the development of miniaturized and portable experimental equipment.

Correlation between levels of serum ferritin and lipid profile status.

Objectives: To analyse the correlation between ferritin level in serum and lipid profile in patients presenting to a Tertiary Care Centre. Study Design: Cross Sectional Descriptive study. Setting: Khyber Teaching Hospital Peshawar. Period: 1st January 2019 to 31st July 2019. Material & Methods: About 60 patients presenting to the laboratory referred for serum lipid profile or serum ferritin were included in the study. Sampling was done by purposive non probability technique. Blood was taken from patients in gel tube. Serum ferritin levels and lipid profile were done in all cases. Lipid profile included serum cholesterol, triglyceride level, low density lipoprotein and high density lipoproteins. The levels were recorded in a proforma. The serum ferritin level was correlated with the lipid profile status in all patient and the results were drawn accordingly. Results: About 60 patients were analysed. The mean age was 20.8± 5.2 years (range: 2 years to 60 years). There were 39 (65%) males and 21(35%) females. Out of 60 cases, serum ferritin level was normal, increased and lower in 15(25%), 40(66.6%) and 5 (8.3%) cases respectively. The cases where serum ferritin was abnormally raised, the significant changes in lipid profile were raised cholesterol and raised triglyceride (in 62.5% and 82.5% cases respectively). Conclusion: There is an association between raised serum ferritin and raised serum cholesterol level and serum triglyceride levels. This supports the link between serum ferritin regulation and lipid metabolism in the body. As dyslipidemia is a major cardiovascular risk factor. Therefore, a raised ferritin level must alert the physician about lipid profile of the patient and should prompt the physician to think about preventive measures against cardiovascular events in order to decrease the morbidity and mortality in patients at risk.

Investigation on the Behavior of κ -Carrageenan Hydrogels for Compressive Intra-Vessel Disintegration.

Gel disintegration via compression is a possible approach for the reversal of the occlusion of male vasa deferentia (VD) by hydrogels. κ -carrageenan (KC) hydrogels can be used for such an application. To determine the required forces for in-vessel compressive disintegration, a gel-tube model, preparing KC gels in different tubes, is studied. These gels are of alternating biopolymer (1-3% by mass) and potassium (100-300mM) concentration. Gel-filled tubes are uniaxially compressed at two different compression speeds (1 and 0.3mm s-1 ). Breakage compression strains are cross studied by shear breaking gel measurements using dynamic mechanical analysis. The measurements showed good agreement. Gel structure disintegration occurred below (62±8) % strain. During compression, three stages of gel disintegration are present. Gel-tube wall detachment, gel rupture, and gel expulsion. The force required for gel disintegration and tube deformation can be added arithmetically. From the modulus of a human aortae model, it is estimated that average human pinch forces are insufficient to disintegrate 2% and 3% by mass KC hydrogels in VD by massage. The compressive disintegration would require a compression device while evading tissue damage.

Evaluation of Some Clinical, Hematological and Biochemical Parameters in Healthy Gurcu Bucks and Does

The aim of the study is to evaluate some hematological parameters in healthy Gurcu bucks and female Gurcu goats. The number of studies in this area is very low. In the study, healthy, 2-4 years old, 15 male and 15 female animals were used. From the animals in the study, 10 mL blood samples were taken with the help of holder from Vena Jugularis and transferred to K3EDTA and gel tubes. Complete blood count was measured in a complete blood count device within 30 minutes. Total leukocyte (WBC) was determined as 11.95 x103/µL in females and 8.18 x103/µL in males. Hemoglobin was measured as an average of 7.32 g/dL in females and 8.75 (g/dL) in males. The total protein concentration was determined as an average of 7.40 g/dL in females, 7.26 g/dL in males and a total average of 7.33 g/dL. In addition, the total average concentrations of creatinine, urea nitrogen, albumin, calcium, phosphorus, magnesium and glucose, respectively; 0.66 mg/dL, 25.92 mg/dL, 2.56 g/dL, 8.62 mg/dL, 4.37 mg/dL, 2.77 mEq/L and 47.04 mg/dL. In this study, some blood parameters of healthy Gurcu bucks and does, where traditional pasture feeding was made, were determined. Hematological parameters have been determined according to gender and it is important in terms of total average levels.

Comparison between conventional tube method and gel card technique for blood crossmatching: A study in a tertiary care center of north India

The cross-matching of blood can be performed by various methods like saline tube method and gel card technique. Saline tube method is most commonly used in blood banks, whereas the latest method used is Gel card technique for cross-matching. Both techniques have their advantages and disadvantages. The main aim of this study was the comparison of gel card and conventional tube test for sensitivity and specificity, time and efficacy. This prospective study was done at the Department of Blood Transfusion, BPS GMC Khanpur Kalan, Sonipat, Haryana. A total of 500 samples collected at the Department of Blood Transfusion, were cross-matched using conventional tube method (spin tube method) with and without using AHG and Gel card Method. 497 (99.4%) samples were compatible, and 3 (0.6%) samples were incompatible with Gel card method, but by test tube method 492 (98.4%) samples were consistent, and 05(1.6%) were false positive (FP). Incompatibility of those 05 samples (FP) disappeared after incubation with AHG reagent at 37°C saline tube method. The sensitivity and specificity of both gel card method and saline tube method with AHG methods is 100% whereas the specificity of saline tube method without AHG is 98.9%. The average time taken by Gel card method was 20-30 minutes for a single compatibility test whereas in conventional spin tube method with the use of AHG (IAT) average time required was 90 minutes and without AHG it was 45-60minutes. The sensitivity and specificity of the spin tube method and the gel card method are comparable to each other. But the gel card method is easy to perform, rapid, reliable procedure and results can be recorded. In contrast, spin saline tube method is more time consuming, and results cannot be preserved. Thus gel card technique can be preferred over the spin tube method.

Stability of Glucose Concentrations in Frozen Plasma.

Conflicting information is available regarding the stability of glucose concentrations in frozen plasma samples. Clinical trials could benefit from such long-term storage because it would allow usage of a central laboratory with higher-quality laboratory analyzers in contrast to mobile analyzers in a decentralized setting. In this study, venous blood samples were collected in lithium-heparin gel tubes. Plasma was separated immediately after blood was drawn, and from each of the 21 plasma samples, 6 aliquots were prepared for measurement at 6 time points: immediately and after 2, 4, 6, 8, and 12 weeks. Between sampling and measurement, aliquots were stored at less than -20°C. Transport on dry ice was simulated by placing aliquots in a -80°C freezer for 5 days between weeks 8 and 12. Measurements were performed on a hexokinase-based laboratory analyzer.Average relative differences and corresponding 99% confidence intervals (CIs) were calculated between the stored aliquots' and the immediately measured aliquots' glucose concentrations. Glucose concentrations were deemed stable as long as average relative differences were ≤±2.5%. Over the whole 12-weeks duration, the largest average relative difference was -1.82% (99% CI: -2.25% to -1.39%). Shorter storage durations tended to lead to less bias. In this study, the stability of glucose concentrations in frozen plasma samples obtained with lithium-heparin gel tubes could be shown for up to 12 weeks. Future studies should be performed to assess whether this is independent of the glucose analyzer and the type of sampling tube used.

5-Years Prevalence Study of HBV and HCV infections in Babylon Province: Forensic Helping Data

HBV and HCV infections are serious global health care problem because they may end with chronichepatitis, cirrhosis, and hepatocellular carcinoma. HBV infection results in approximately 2 billion humaninfections while HCV approximately 160 million individuals. Raw database were collected from Generalhealth department at Babylon Health directorate for 715505 investigated person during a period from Jan.2015 to Dec. 2019. Blood samples was drawn in Gel tube from suspects and centrifuged to separate theserum. Serum submitted for investigation of HbsAg and HCV Ab by both rapid immunochromatographictest and ELISA test. Actually the database taken from Babylon Health directorate and the results analysisdepends upon dividing of data records from four health sectors, blood bank record lab, health care checkinglab, pre-surgery checking, Marriage lab, thalassemia lab, Hemodialysis lab Prisoner and foreigners lab. Theresults revealed that , total positive cases were 4284 and 2260 for “HBV and HCV” respectively. The highestno. of cases for both “HBV and HCV” were recorded in 2018 (976 and 467) and (961 and 557) for “HBVand HCV” respectively. The incidence rate of “HBV and HCV” were varies worldwide. The results revealedthat the occurrence rate (Mean ± SD) of “HBV” was (0.627±0.134) while for HCV was (0.327±0.084). Theresults revealed that, there is no defined month or semester for high rate infections. The high prevalencepercentage were showed for prisoner-foreigner category (48.53 and 45.95 for HBV and HCV respectively)followed by surgery (22.66 and 19.03 for HBV and HCV respectively), blood bank (14.03 and 13.95 forHBV and HCV respectively) and in married (7.79 and 3.92 for HBV and HCV respectively. The current studyconclude the high prevalence percentage of HBV (duplicate) than HCV with no seasonality of occurrenceand the Prisoners and foreigners, Surgery, blood donor and hemodialysis were mostly involved with HBVand HCV infection.

Improved Sample Quality and Decreased Turnaround Time When Using Plasma Blood Collection Tubes with a Mechanical Separator in a Large University Hospital.

Serum is commonly used for clinical chemistry testing but many conditions can affect the clotting process, leading to poor sample quality and impaired workflow. With serum gel tubes, we found a high proportion of sample probe aspiration errors on our Beckman AU5800 analyzers. We decided to implement the BD Barricor™ plasma tubes, and we validated an off-specification centrifugation scheme and verified that results obtained for 65 chemistry and immunochemistry tests were comparable to those obtained in serum gel tubes. Finally, we evaluated the impact of this new tube on sample error rate and laboratory turnaround time. To validate centrifugation settings, 50 paired samples were collected in Barricor tubes and centrifuged at 1912 × g for 10 min or 5 min (off-specification). To compare serum gel tubes with Barricor plasma tubes, 119 paired samples were collected from volunteers and results were analyzed using weighed Deming regression. Finally, the proportion of aspiration errors and laboratory TAT for potassium were measured before and after implementing Barricor tubes. Barricor tubes showed clinically acceptable equivalence to serum gel tubes for the studied analytes, and the off-specification centrifugation scheme did not affect the results. Implementing Barricor tubes improved the laboratory workflow by decreasing the aspiration error rates (2.01% to 0.77%, P < 0.001) and lowering hemolysis (P < 0.001). The laboratory TAT for potassium were also significantly lowered (P < 0.001). Use of Barricor tubes instead of serum gel tubes leads to better sample quality, shorter more reproducible laboratory TAT, and decreases costs associated with error management.

Exposures and Emissions in Coffee Roasting Facilities and Cafés: Diacetyl, 2,3-Pentanedione, and Other Volatile Organic Compounds

Roasted coffee and many coffee flavorings emit volatile organic compounds (VOCs) including diacetyl and 2,3-pentanedione. Exposures to VOCs during roasting, packaging, grinding, and flavoring coffee can negatively impact the respiratory health of workers. Inhalational exposures to diacetyl and 2,3-pentanedione can cause obliterative bronchiolitis. This study summarizes exposures to and emissions of VOCs in 17 coffee roasting and packaging facilities that included 10 cafés. We collected 415 personal and 760 area full-shift, and 606 personal task-based air samples for diacetyl, 2,3-pentanedione, 2,3-hexanedione, and acetoin using silica gel tubes. We also collected 296 instantaneous activity and 312 instantaneous source air measurements for 18 VOCs using evacuated canisters. The highest personal full-shift exposure in part per billion (ppb) to diacetyl [geometric mean (GM) 21 ppb; 95th percentile (P95) 79 ppb] and 2,3-pentanedione (GM 15 ppb; P95 52 ppb) were measured for production workers in flavored coffee production areas. These workers also had the highest percentage of measurements above the NIOSH Recommended Exposure Limit (REL) for diacetyl (95%) and 2,3-pentanedione (77%). Personal exposures to diacetyl (GM 0.9 ppb; P95 6.0 ppb) and 2,3-pentanedione (GM 0.7 ppb; P95 4.4 ppb) were the lowest for non-production workers of facilities that did not flavor coffee. Job groups with the highest personal full-shift exposures to diacetyl and 2,3-pentanedione were flavoring workers (GM 34 and 38 ppb), packaging workers (GM 27 and 19 ppb) and grinder operator (GM 26 and 22 ppb), respectively, in flavored coffee facilities, and packaging workers (GM 8.0 and 4.4 ppb) and production workers (GM 6.3 and 4.6 ppb) in non-flavored coffee facilities. Baristas in cafés had mean full-shift exposures below the RELs (GM 4.1 ppb diacetyl; GM 4.6 ppb 2,3-pentanedione). The tasks, activities, and sources associated with flavoring in flavored coffee facilities and grinding in non-flavored coffee facilities, had some of the highest GM and P95 estimates for both diacetyl and 2,3-pentanedione. Controlling emissions at grinding machines and flavoring areas and isolating higher exposure areas (e.g., flavoring, grinding, and packaging areas) from the main production space and from administrative or non-production spaces is essential for maintaining exposure control.

Association of MYH9-rs3752462 polymorphisms with chronic kidney disease among clinically diagnosed hypertensive patients: a case-control study in a Ghanaian population

BackgroundChronic kidney disease (CKD) is a significant comorbidity among hypertensive patients. Polymorphisms in the non-muscle myosin heavy chain 9 gene (MYH9) have been demonstrated to be significantly associated with CKD, among African- and European-derived populations. We investigated the spectrum of MYH9-associated CKD among Ghanaian hypertensive patients.MethodsThe study constituted a total of 264 hypertensive patients. Hypertensive patients with glomerular filtration rate (eGFR) < 60 ml/min/1.73m2 (CKD-EPI formula) or clinically diagnosed were defined as case subjects (n = 132) while those with eGFR ≥60 ml/min/1.73m2 were classified as control subjects (n = 132). Demographic data were obtained with a questionnaire and anthropometric measurements were taken. Five (5) millilitres (ml) of venous blood was drawn from study subjects into gel and EDTA vacutainer tubes. Two (2) mL of EDTA anticoagulated blood was used for genomic DNA extraction while three (3) mL of blood was processed to obtain serum for biochemical measurements. Genotyping of MYH9 polymorphisms (rs3752462) was done employing Tetra primer Amplification Refractory Mutation System (T-ARMS) polymerase chain reaction (PCR). Spot urine samples were also collected for urinalysis. Hardy-Weinberg population was assessed. Logistic regression models were used to assess the associations between single nucleotide polymorphisms and CKD.ResultsThe cases and control participants differed in terms of age, sex, family history, and duration of CKD (p-value < 0.001). The minor allele frequencies of rs3752462 SNP were 0.820 and 0.567 respectively among the control and case subjects. Patients with the heterozygote genotype of rs3752462 (CT) were more likely to develop CKD [aOR = 7.82 (3.81–16.04)] whereas those with homozygote recessive variant (TT) were protective [aOR = 0.12 (0.06–0.25)]. Single nucleotide polymorphism of rs3752462 (CT genotype) was associated with increased proteinuria, albuminuria, and reduced eGFR.ConclusionsWe have demonstrated that MYH9 polymorphisms exist among Ghanaian hypertensive patients and rs3752462 polymorphism of MYH9 is associated with CKD. This baseline indicates that further longitudinal and multi-institutional studies in larger cohorts in Ghana are warranted to evaluate MYH9 SNP as an independent predictor of CKD among hypertensive patients in Ghana.

Effectiveness of Flow Volume Measurement Training Using a Custom-Made Doppler Flow Simulator.

This study was conducted to analyze the effectiveness of a custom-made Doppler ultrasound (DUS) flow simulator, vascular phantom, and Doppler test fluid in the training of dialysis staff in flow volume (FV) measurements for arteriovenous (AV) access in hemodialysis (HD) patients. A DUS flow simulator was constructed using a continuous renal replacement therapy machine. Vascular phantoms were constructed using a rubber enema tube and keyboard cleaning gel. The Doppler test fluid consisted of freeze-dried instant coffee granules and 0.9% saline. This easy and affordable simulator was used to train 12 dialysis staff members, with no previous experience in DUS FV measurement. After a 3-day lecture course, the staff members performed DUS analyses on the AV access of HD patients. Thereafter, they underwent a 3-day training course using the simulator and then performed the DUS analyses of the AV access again. Each staff member assessed the FV 3 times, and the mean values of the measurements pretraining and posttraining were analyzed using paired t tests. The difference in the mean value of FV measurements and the reference value decreased from 131.6 mL/min to 62.5 mL/min (95% confidence interval = 30.0-108.0, P = 0.002), and the standard deviation of the FV measurements decreased from 96.9 mL/min to 47.0 mL/min (95% confidence interval = 7.9-91.8, P = 0.023) after DUS training with the simulator. The accuracy and reproducibility of FV measurements markedly improved after training with the simulator; it may be helpful for medical practitioners involved in AV access for HD treatment.

Comparison of test results obtained from lithium heparin gel tubes and serum gel tubes

Abstract Objectives There is currently trend that plasma might be alternative to serum due to some of its advantages. This study aimed to compare test results from heparinized plasma and serum. Methods Blood samples from total of 40 participants (20 healthy, 20 hemodialysis patients) were drawn into serum gel tubes with clot activator and lithium heparin gel tubes. Twenty-eight clinical chemistry analytes were measured in serum and plasma samples. To determine whether difference between test results is clinically significant, total error (TE) was calculated and compared total allowable error (TEa) limits. Results TE of below 5% was calculated for amylase, AST, calcium, total cholesterol, chloride, CK, glucose, HDL-cholesterol, iron, LDH, LDL-cholesterol, magnesium, sodium, total bilirubin, uric acid and urea. Albumin, ALT, creatinine, CRP, lipase, phosphorus, potassium, total protein, and triglyceride had TE of 5–7%. TE of 7–10% were determined for ALP, direct bilirubin, and GGT. TE values were within TEa limits for all analytes. Conclusions It was concluded that results of 28 analytes measured in lithium heparin gel tubes are comparable to those of serum gel tubes. It is thought that several advantages including reduced turnaround time might be provided by using plasma instead of serum for these tests.

Undetectable Free Aromatic Amino Acids in Nails of Breast Carcinoma: Biomarker Discovery by a Novel Metabolite Purification VTGE System.

Background: Metabolic reprogramming in breast cancer is depicted as a crucial change in the tumor microenvironment. Besides the molecular understanding of metabolic heterogeneity, appreciable attention is drawn to characterizing metabolite profiles in tumor tissue and derived biological fluids and tissue materials. Several findings reported on the metabolic alterations of free aromatic amino acids (FAAAs) and other metabolites in biological fluids. Furthermore, there is a significant gap in the development of a suitable method for the purification and analysis of metabolite biomarkers in nails of cancer patients.Methods: To address the metabolite alterations, specifically FAAA levels in nails, fingernail clippings of breast cancer patients (N = 10) and healthy subjects (N-12) were used for extraction and purification of metabolites. Here, we reported a novel and specifically designed vertical tube gel electrophoresis (VTGE) system that helped in the purification of metabolites in the range of 100–1,000 Da from nail materials. Here, the VTGE system uses 15% polyacrylamide under non-denaturing and non-reducing conditions, which makes eluted metabolites directly compatible with LC-HRMS and other analytical techniques. Qualitative and quantitative determination of FAAAs in nail lysates was done in positive ESI mode of the Agilent LC-HRMS platform.Results: The analysis on collected data of nail metabolites clearly suggested that FAAAs including tryptophan, tyrosine, phenylalanine, and histidine were undetectable in nail lysates of breast cancer over healthy subjects. This is a first report that showed highly reduced levels of FAAAs in nails of breast cancer patients. Furthermore, the present observation is in consonance with previous findings that showed cancer cachexia and high amino acid catabolism in breast cancer patients that drive metabolite-led cancer growth and proliferation.Conclusion: This paper provides a proof of concept for a novel and specifically developed VTGE process that showed first evidence on the undetectable level of FAAAs in nails of breast cancer patients as metabolite biomarkers. Here, the authors propose the potential use of a VTGE-assisted process to achieve metabolomic discovery in nails of breast cancer and other tumor types.

Development of preanalytical criteria assessment of quality of vacuum blood collection systems affecting clinical examination of patients

This paper presents comparative studies of preanalytical quality indices of vacuum gel systems (VGS) from four manufacturers, i.e., GL 795 (China), Vacuette (Austria), BD Vacutainer SST (USA) and Zdravmedtech (Russia). The analysis of the number of qualitative defects of the preanalytic phase of blood collection in vacuum gel tubes at the inpatient departments and in Department of Clinical Chemistry (preanalytic laboratory), in accordance with adopted criteria based on Russian industry standards, which are implemented with European indices of pre-analytic stage. The obtained results allowed us to identify a number of qualitative defects at pre-analytic stage for the vacuum gel systems from each manufacturer, including to summarize the data on number of these defects. Based on these results, the most common defects in each of the VGS presented by manufacturers are formulated. The results obtained can be used by manufacturers to improve the quality (structures) of the mentioned vacuum gel tubes.The results of the comparative characteristics of VGS of different manufacturers were evaluated by their influence on the indexes of hemolysis (IH), ictericity, lipemia. The resulting indices of ictericity and lipemia do not allow to use them as criteria for assessing quality of the gel systems. The frequency of IH occurrence in serum at &gt; 50 conventional units in VGS of different manufacturers was studied. It was revealed, that IH in the range from 10 to 50 conventional units and more, can be used as a criterion of VGS quality.Comparison of routine biochemical parameters in VGS of different manufacturers, i.e., alanine aminotransferase, aspartate aminotransferase, direct bilirubin, lactate dehydrogenase (LDH), serum K+, was also carried out. It was found that the largest deviations from the average value were obtained for aspartate aminotransferase indices of more than 12.47% in Russian test tubes, and for direct bilirubin of more than 12.25% in Chinese test tubes and more than 9.15% in Russian test tubes. The study showed influence of IH upon quantitative values of lactate dehydrogenase for VGS from different manufacturers. It is proposed to use lactate dehydrogenase as an analyte. On this basis, it is possible to recommend a comparative quality assessment of vacuum gel systems, with IH cutoff level of more than 19 conventional units. It remains relevant to search for other analytes that can be used to assess the quality of VGS from different manufacturers. Some areas of research in biochemical testing are outlined. Preliminary data are obtained on usage of vacuum systems from different manufacturers for quality assessment, as well as conditions for performing these tests, along with the level of the hemolysis index of more than 10 conventional units.

The Prevalence of Blood Groups in Domestic Cats in the Saskatoon and Calgary Areas of Saskatchewan and Alberta, Canada.

The purpose of this study was to evaluate blood types of domestic cats in two cities in Western Canada (Saskatoon, Saskatchewan and Calgary, Alberta), as well as to determine the risk of mismatched transfusion and neonatal isoerythrolysis. Several cat studies around the world have shown variability in the prevalence of blood types in domestic and pedigree cats. Canadian data based on feline blood types is based out of Montreal. In this study the cohort of cats revealed a higher than anticipated prevalence; of 5% type B and 0.6% AB blood types. In our study, blood typing was performed in 400 domestic cats; 200 in Saskatoon and 200 in Calgary. Blood typing was performed using the gel tube method and the risk of transfusion mismatch (MT) was estimated by adding the risk of a major transfusion reaction and the risk of a minor transfusion reaction. The risk of neonatal isoerythrolysis (NI) was estimated according to the equation (p2)(q2) + 2pq(q2), with q being the b allele frequency and p = 1 – q. There was an identical frequency for feline blood types in both Saskatoon and Calgary cats, with 96% type A, 4% type B, and 0% AB. Based on these percentages, the risks of MT and NI in domestic cats were 7.6 and 4 % respectively. The frequency of type B cats in the population was similar to that in the previous Canadian study. These results demonstrate regional differences in prevalence of type B blood in domestic shorthairs across the world and serve to reinforce recommendations to blood type prior to transfusion or mating.

Stability of 21 Antihypertensive Drugs in Serum Collected in Standard (Nongel) Serum Tubes Versus Tubes Containing a Gel Separator.

Background:Therapeutic drug monitoring of antihypertensive drugs is being increasingly used to optimize treatment and to assess nonadherence. Separator gels are often used in blood collection tubes to facilitate serum or plasma separation from other blood constituents before analyses. Drug adsorption into the separator gel presents a possible pre-analytical cause of falsely low concentrations or false negative results.Methods:Drug-free blood from blood donors was spiked with therapeutic concentrations of 21 antihypertensive drugs, transferred to serum tubes with and without separator gel (Vacuette gel plastic tubes and plain serum plastic tubes, respectively), and centrifuged. Serum was collected immediately after centrifugation and after 24 and 72 hours of room temperature storage, samples were analyzed in triplicates using liquid chromatography–mass spectrometry.Results:Serum samples collected immediately after centrifugation or 24 hours later, had the same drug concentrations in the gel and nongel tubes. After 72 hours of room temperature storage, verapamil and lercanidipine serum concentrations were 43% and 29%, respectively, lower in gel tubes than nongel tubes. Canrenone, diltiazem, and bendroflumethiazide showed between 10% and 20% concentration loss in gel tubes, compared with nongel tubes, with the 2 latter observed as unstable also in nongel tubes.Conclusions:Except for verapamil, lercanidipine, and canrenone, which showed substantial concentration loss in gel tubes, gel tubes may be used for therapeutic drug monitoring purposes for the most commonly used antihypertensive drugs. Transferring serum to gel-free containers immediately after centrifugation minimizes concentration loss; however, bendroflumethiazide and diltiazem are generally unstable at room temperature.

A Multicenter Evaluation of a Nongel Mechanical Separator Plasma Blood Collection Tube for Testing of Selected Therapeutic Drugs.

Some therapeutic drugs are unstable during sample storage in gel tubes. BD Vacutainer® Barricor™ Plasma Blood Collection Tube with nongel separator was compared with plasma gel tubes, BD Vacutainer PST™, PST II, and BD Vacutainer Serum Tube for acetaminophen, salicylate, digoxin, carbamazepine, phenytoin, valproic acid, and vancomycin during sample storage for up to 7 days. Seven hospital sites enrolled 705 participants who were taking at least one selected drug. The study tubes were collected and tested at initial time (0 h), after 48 h of storage at room temperature and on day 7 (after additional 5 days of refrigerated storage). The performance of BD Barricor tube was evaluated for each drug by comparing BD Barricor samples with samples from the other tubes at 0 h from the same participant; stability was evaluated by comparing test results from the same tube at 0 h, 48 h, and 7 days. At 0 h, BD Barricor showed clinically equivalent results for selected therapeutic drugs compared with the other tubes, except phenytoin in BD PST. Phenytoin samples ≥20 µg/mL in BD PST had 10-12% lower values than samples in BD Barricor. During sample storage, all selected drugs remained stable for 7 days in BD Barricor and in serum aliquots. In BD PST, all drugs remained stable except phenytoin and carbamazepine and in BD PST II except for phenytoin. The BD Barricor Tube is effective for the collection and storage of plasma blood samples for therapeutic drug monitoring without sample aliquoting.