Red Cell Pyruvate Kinase Deficiency With Hypertriglyceridemia: A Case Report.

Red cell pyruvate kinase (PK) deficiency is a genetic disorder affecting the enzyme PK in red blood cells. A deficiency in PK leads to hemolytic anemia. Hypertriglyceridemia means elevated levels of triglycerides in the blood. The hypertriglyceridemia disorder can be primary or secondary to an underlying disease. Hypertriglyceridemia with ß-thalassemia major is a known association and is called hypertriglyceridemia-thalassemia syndrome. A four-month-old male child was found to have milky serum. On investigation, there was severe anemia, with triglycerides at 1197 mg/dL and high lactate dehydrogenase (LDH). The child had severe pallor, mild icterus, a dysmorphic face, and splenohepatomegaly. Ophthalmic examination showed lipemia retinitis. The child was treated with medium-chain fatty acid formula feed. Regular blood transfusions, folic acid supplements, and avoidance of salicylate group drugs were advised. The child improved and is doing well. Thus, early diagnosis and treatment can change the prognosis and help maintain a near-normal life for affected infants.

Hyperglycemia induced cathepsin L maturation linked to diabetic comorbidities and COVID-19 mortality.

Diabetes, a prevalent chronic condition, significantly increases the risk of mortality from COVID-19, yet the underlying mechanisms remain elusive. Emerging evidence implicates Cathepsin L (CTSL) in diabetic complications, including nephropathy and retinopathy. Our previous research identified CTSL as a pivotal protease promoting SARS-CoV-2 infection. Here, we demonstrate elevated blood CTSL levels in individuals with diabetes, facilitating SARS-CoV-2 infection. Chronic hyperglycemia correlates positively with CTSL concentration and activity in diabetic patients, while acute hyperglycemia augments CTSL activity in healthy individuals. In vitro studies reveal high glucose, but not insulin, promotes SARS-CoV-2 infection in wild-type cells, with CTSL knockout cells displaying reduced susceptibility. Utilizing lung tissue samples from diabetic and non-diabetic patients, alongside Leprdb/dbmice and Leprdb/+mice, we illustrate increased CTSL activity in both humans and mice under diabetic conditions. Mechanistically, high glucose levels promote CTSL maturation and translocation from the endoplasmic reticulum (ER) to the lysosome via the ER-Golgi-lysosome axis. Our findings underscore the pivotal role of hyperglycemia-induced CTSL maturation in diabetic comorbidities and complications.

People with diabetes are at greater risk of developing severe COVID-19 and dying from the illness, which is caused by a virus known as SARS-CoV-2. The high blood sugar levels associated with diabetes appear to be a contributing factor to this heightened risk. However, diabetes is a complex condition encompassing a range of metabolic disorders, and it is therefore likely that other factors may contribute. Previous research identified a link between an enzyme called cathepsin L and more severe COVID-19 in people with diabetes. Elevated cathepsin L levels are known to contribute to diabetes complications, such as kidney damage and vision loss. It has also been shown that cathepsin L helps SARS-CoV-2 to enter and infect cells. This raised the question of whether elevated cathepsin L is responsible for the increased COVID-19 vulnerability in patients with diabetes. To investigate, He, Zhao et al. monitored disease severity and cathepsin L levels in patients with COVID-19. This confirmed that people with diabetes had more severe COVID-19 and that higher levels of cathepsin L are linked to more severe disease. Analysis also revealed that cathepsin L activity increases as blood glucose levels increase. In laboratory experiments, cells exposed to glucose or fluid from the blood of people with diabetes were more easily infected with SARS-CoV-2, with cells genetically modified to lack cathepsin L being more resistant to infection. Further experiments revealed this was due to glucose promoting maturation and migration of cathepsin L in the cells. The findings of He, Zhao et al. help to explain why people with diabetes are more likely to develop severe or fatal COVID-19. Therefore, controlling blood glucose levels in people with diabetes may help to prevent or reduce the severity of the disease. Additionally, therapies targeting cathepsin L could also potentially help to treat COVID-19, especially in patients with diabetes, although more research is needed to develop and test these treatments.

Transfusion sample mislabelling and wrong blood in tube in the UK: Insights from the national comparative audits of blood transfusion in 2012 and 2022.

BACKGROUND: Samples for transfusion rejected due to mislabelling can lead to harm when a patient has to be re-bled or has a transfusion or procedure delayed. Electronic labelling systems which scan the patient's identification band and generate a label at their side aim to reduce mislabelling and misidentification leading to wrong blood in tube (WBIT) errors. The 2022 National Comparative audit of sample collection aimed to compare national rates of sample mislabelling and WBIT to the 2012 audit and to examine the impact of electronic systems. METHOD: All UK hospitals were invited to provide data on rejected transfusion samples and WBIT incidents in 1 month (October 2022) and were asked if they had electronic labelling. RESULTS: Twenty-three thousand five hundred and eighty-four rejected samples were reported by 179 sites in 1 month. The rejection rate of 4.4% represents a 47% increase compared to 2012 (2.99%). There were 92 WBIT incidents, an incidence of 1 in 5882 samples-a 45% increase compared to 1 in 8547 in 2012. Twenty-three percent of sites can print a sample label at the patient's side, up by 224%. The six sites using only electronic sample labelling had a 46.9% lower rejection rate than sites using only hand-labelling but still reported WBIT. CONCLUSIONS: The increase in sample rejection and WBIT may reflect pressures facing clinical staff, zero tolerance policies and the two-sample rule. A human factors approach to understanding and tackling underlying reasons locally is recommended. Electronic systems are associated with fewer labelling errors, but careful implementation and training is needed to maximise their safety benefits.

Adapting the SMART tube technology for flow cytometry in feline full blood samples.

Flow cytometry of blood samples is a very valuable clinical and research tool to monitor the immune response in human patients. Furthermore, it has been successfully applied in cats, such as for infections with feline immune deficiency virus (FIV). However, if cells are not isolated and frozen, analysis of anticoagulated blood samples requires mostly prompt processing following blood collection, making later analysis of stored full blood samples obtained in clinical studies often impossible. The SMART Tube system (SMART TUBE Inc., California, United States; SMT) allows fixation and long-term preservation of whole blood samples at -80°C. However, this system has so far only been applied to human biological samples. In the present study, a new flow cytometry SMART Tube protocol adapted for feline whole blood samples was successfully established allowing quantification of T-helper cells, cytotoxic T-cells, B-cells, monocytes, and neutrophils up to 2 years post sampling. Results obtained from frozen stabilized and fresh blood samples were compared for validation purposes and correlated to differential blood counts from a conventional hematology analyzer. Clinical applicability of the new technique was verified by using samples from a treatment study for feline infectious peritonitis (FIP). Using the new SMT protocol on retained samples, it could be demonstrated that long-term storage of these SMT tubes is also possible. In summary, the newly adapted SMT protocol proved suitable for performing flow cytometry analysis on stored feline whole blood samples, thus opening up new avenues for veterinary research on a variety of aspects of clinical interest.

Biochemical Changes in Adult Male Gamers During Prolonged Gaming: Pilot Study.

BACKGROUND: Gaming has become an integrated part of life for children and adults worldwide. Previous studies on the impact of gaming on biochemical parameters have primarily addressed the acute effects of gaming. The literature is limited, and the study designs are very diverse. The parameters that have been investigated most thoroughly are blood glucose and cortisol. OBJECTIVE: This exploratory study is the first to investigate the effects of long gaming sessions on the biochemical parameters of healthy male adults. The extensive testing allowed us to observe short-term changes (within 6 hours), long-term changes during the duration of the gaming sessions, and follow-up after 1 week to determine whether any changes were longer lasting. METHODS: In total, 9 experienced gamers completed 2 back-to-back 18-hour gaming sessions interspersed with a 6-hour rest period. All participants adhered to a structured sleep pattern due to daytime employment or attending university. Blood, saliva, and urine samples were collected from the participants every 6 hours. Linear mixed-effect models were used to analyze the repeated-measures data accumulated during the study. A total of 51 biochemical parameters were investigated. RESULTS: In total, 12 of the 51 biochemical parameters significantly changed during the study: alkaline phosphatase, aspartate aminotransferase, bilirubin, chloride, creatinine, glucose, hemoglobin, immature reticulocyte fraction, lactate, methemoglobin, sodium, and thrombocytes. All changes were within the normal range. The mean glucose level of the participants was 4.39 (SD 0.07) mmol/L at baseline, which increased significantly by 0.24 (SD 0.07) mmol/L per 6 hours during the first period and by 0.38 (SD 0.07) mmol/L per 6 hours in the second period (P<.001). The glucose levels during the second session increased even though the participants had little energy intake. Cortisol levels did not change significantly, although the cortisol pattern deviated from the typical circadian rhythm. During both gaming sessions, we observed increasing cortisol levels from 6 AM until noon. The participants were relatively dehydrated at the start of the study. The patients were asked to fast before the first blood sampling. Within the first 6 hours of the study, the participants rehydrated, followed by relative dehydration during the remainder of the study. This pattern was identified using the following parameters: albumin, creatinine, hemoglobin, erythrocytes, potassium, and platelets. CONCLUSIONS: This study is the first of its kind, and many of the analyses in the study yielded novel results. The study was designed to emulate the behavior of gamers during the weekend and other long gaming sessions. At this point, we are not able to determine the difference between the effects of gaming and behavior during gaming. Regardless, the results of this study suggest that healthy gamers can partake in long gaming sessions, with ample amounts of unhealthy foods and little rest, without acute impacts on health.

Analysis of arterial blood gas values when discarding different volumes of blood samples in an arterial heparin blood collector during thoracoscopic surgery.

BACKGROUND: Arterial blood gas analysis (ABGA) plays a vital role in emergency and intensive care, which is affected by many factors, such as different instrumentation, temperature, and testing time. However, there are still no relevant reports on the difference in discarding different blood volumes on ABGA values. METHODS: We enrolled 54 patients who underwent thoracoscopic surgery and analysed differences in blood gas analysis results when different blood volumes were discarded from the front line of the arterial heparin blood collector. A paired t test was used to compare the results of the same patient with different volumes of blood discarded from the samples. The difference was corrected by Bonferroni correction. RESULTS: Our results demonstrated that the PaO2, PaCO2, and THbc were more stable in the 4th ml (PaO2 = 231.3600 ± 68.4878 mmHg, PaCO2 = 41.9232 ± 7.4490 mmHg) and 5th ml (PaO2 = 223.7600 ± 12.9895 mmHg, PaCO2 = 42.5679 ± 7.6410 mmHg) blood sample than in the 3rd ml (PaO2 = 234.1000 ± 99.7570 mmHg, PaCO2 = 40.6179 ± 7.2040 mmHg). CONCLUSION: It may be more appropriate to discard the first 3 ml of blood sample in the analysis of blood gas results without wasting blood samples.

Insights into the Age-Dependent Variation in Nutrition-Related Trace Elements in Diabetes Blood Using Total Reflection X-Ray Fluorescence.

The prevalence of diabetes has reached alarming levels in India, making it essential to understand the concentration of nutritional-trace elements (Fe, Cu, Zn, Cr. and Se) in blood samples from diabetic adults. In this study, 208 whole blood samples from diabetic (n = 104) and non-diabetic (n = 104) adults across various age groups were analyzed using total reflection X-ray fluorescence (TXRF) spectroscopy with a sample dilution method. Statistical analysis was performed to assess descriptive statistics and determine a significant correlation between elemental concentrations in the blood samples of diabetic and non-diabetic adults. The mean concentration of nutritional-related trace elements in diabetic blood was as follows: Fe (46 ± 5) > Zn (1.28 ± 0.14) > Cu (0.10 ± 0.01) > Cr (0.05 ± 0.004) > Se (0.013 ± 0.001) in mg/L, respectively. Additionally, this study investigated the influence of nutrition-related trace element concentrations across various age groups such as 25-40 years (young adults), 41-55 years (middle-aged adults), and 56-70 years (older adults). In this investigation, Zn (p < 0.001) and Cr (p < 0.05) concentrations differed significantly between diabetic and non-diabetic adults aged 56-70 years. These findings will help us to understand age-dependent changes in element concentrations, clarify their role in diabetes, and improve risk factor management associated with diabetes.

Challenges and Advances in Biomarker Detection for Rapid and Accurate Sepsis Diagnosis: An Electrochemical Approach.

Sepsis is a life-threatening condition with high mortality rates due to delayed treatment of patients. The conventional methodology for blood diagnosis takes several hours, which suspends treatment, limits early drug administration, and affects the patient's recovery. Thus, rapid, accurate, bedside (onsite), economical, and reliable sepsis biomarker reading of the clinical sample is an emergent need for patient lifesaving. Electrochemical label-free biosensors are specific and rapid devices that are able to perform analysis at the patient's bedside; thus, they are considered an attractive methodology in a clinical setting. To reveal their full diagnostic potential, electrode architecture strategies of fabrication are highly desirable, particularly those able to preserve specific antibody-antigen attraction, restrict non-specific adsorption, and exhibit high sensitivity with a low detection limit for a target biomarker. The aim of this review is to provide state-of-the-art methodologies allowing the fabrication of ultrasensitive and highly selective electrochemical sensors for sepsis biomarkers. This review focuses on different methods of label-free biomarker sensors and discusses their advantages and disadvantages. Then, it highlights effective ways of avoiding false results and the role of molecular labels and functionalization. Recent literature on electrode materials and antibody grafting strategies is discussed, and the most efficient methodology for overcoming the non-specific attraction issues is listed. Finally, we discuss the existing electrode architecture for specific biomarker readers and promising tactics for achieving quick and low detection limits for sepsis biomarkers.

Preanalytical Errors in Clinical Laboratory Testing at a Glance: Source and Control Measures.

The accuracy of diagnostic results in clinical laboratory testing is paramount for informed healthcare decisions and effective patient care. While the focus has traditionally been on the analytical phase, attention has shifted towards optimizing the preanalytical phase due to its significant contribution to total laboratory errors. This review highlights preanalytical errors, their sources, and control measures to improve the quality of laboratory testing. Blood sample quality is a critical concern, with factors such as hemolysis, lipemia, and icterus leading to erroneous results. Sources of preanalytical errors encompass inappropriate test requests, patient preparation lapses, and errors during sample collection, handling, and transportation. Mitigating these errors includes harmonization efforts, education and training programs, automated methods for sample quality assessment, and quality monitoring. Collaboration between laboratory personnel and healthcare professionals is crucial for implementing and sustaining these measures to enhance the accuracy and reliability of diagnostic results, ultimately improving patient care.

Possible alternative strategies to implement basophil activation testing in multicentric studies.

The Basophil Activation Test (BAT) enables flow cytometry characterization of basophil reactivity against specific allergenic molecules. The focus now revolves around democratizing this tool, but, as blood sample stability could be challenging, after having developed a simplified approach, herein, we aimed to characterize two strategies for implementing BAT in multicentric studies: store and ship blood before or after sample processing. Fresh heparin- and EDTA-anticoagulated whole blood samples followed both BAT workflows: "collect, store, process & analyze" or "collect, process, store & analyze". Storage temperatures of 18-25 °C or 2-8 °C and preservation times from 0 to 7 days were considered. Interleukin-3 was also evaluated. With the "collect, store, process & analyze" workflow, heparin-anticoagulated blood and 18-25 °C storage were better than other conditions. While remaining possible, basophil activation exhibited a possible reactivity decay after 24 h. Under the conditions tested, interleukin-3 had no role in enhancing basophil reactivity after storage. Conversely, the "collect, process, store & analyze" workflow demonstrated that either heparin- or EDTA-anticoagulated blood can be processed and kept up to 7 days at 18-25 °C or 2-8 °C before being analyzed. Various strategies can be implemented to integrate BAT in multicentric studies. The "collect, store, process & analyze" workflow remains a simplified logistical approach, but depending on time required to ship from the clinical centers to the reference laboratories, it might not be applicable, or should be used with caution. The "collect, process, store & analyze" workflow may constitute a workflow improvement to provide significant flexibility without impact on basophil reactivity.

Optimizing d-mannose and glyceraldehyde concentrations as glucose preservatives without clinically affecting biochemical test results.

Objectives: The objectives were to evaluate blood additives that combined lithium heparin (LH)-salt with glyceraldehyde (GLY) or d-mannose (MAN) for preserving glucose levels in plasma samples and to simultaneously determine the compatibility of these additives with 14 other biochemical tests. Methods: Blood samples from 40 subjects, equally divided into healthy and diabetic groups, were collected using five different additives. The three most effective additives, LH/GLY, LH/MAN, and LH/GLY/MAN, were selected for ensuring the best preservation of glucose levels and compatibility with 14 biochemical tests. One-way analysis of variance was used to analyze the mean paired differences of glucose level and biochemical tests. Simultaneously, the clinical criteria from Johns Hopkins Hospital were used to guide the interpretation and set acceptable thresholds for measurements that exceeded the standards. Results: The combination of 160 mmol/L GLY, 8.4 mmol/L MAN, and LH, maintained glucose levels at approximately 93.4-93.7 % for healthy subjects and 91.3-92.8% for subjects with diabetes mellitus over 8 h. The mean paired differences of glucose levels in preservation were statistically insignificant. The biases in 14 biochemical tests for LH/GLY/MAN and LH/MAN remained within the acceptable clinical criteria during the 8 h. Conclusions: Combining 160 mmol/L GLY, 8.4 mmol/L MAN, and LH, proved more effective in maintaining glucose levels than individual additives or the conventional sodium fluoride preservative. It did not yield clinical discrepancies in the 14 biochemical tests during 8 h at room temperature.

A novel fluorescent probe based imprinted polymer-coated magnetite for the detection of imatinib leukemia anti-cancer drug traces in human plasma samples.

Myeloid leukemia is a chronic cancer, which associated with abnormal BCR-ABL tyrosine kinase activity. Imatinib (IMB) acts as a tyrosine kinase inhibitor and averts tumor growth in cancer cells by controlling cell division, so it is urgent to develop an effective assay to detect and monitor its IMB concentration. Therefore, an innovative fluorescent biomimetic sensor is a promising sensing material that constructed for the efficient recognition of IMB and displays excellent selectivity and sensitivity stemming from molecularly imprinted polymer@Fe3O4 (MIP@Fe3O4). The detection strategy depends on the recognition of IMB molecules at the imprinted sites in the presence of coexisting molecules, which are then transferred to the fluorescence signal. The synthesized MIP@Fe3O4 was characterized using Fourier-transform infrared spectroscopy (FT-IR), scanning electron microscopy (SEM), and atomic force microscopy (AFM). Furthermore, computational studies of the band gap (EHOMO-ELUMO) of the monomers, IMB, and their complexes were performed. These results confirmed that the copolymer is the most appropriate and has high stability (Binding energy; 0.004 x 10-19 KJ) and low reactivity. A comprehensive linear response over IMB concentrations from 5 × 10-6 mol/L to 8 × 10-4 mol/L with a low detection limit of 9.3 × 10-7 mol/L was achieved. Furthermore, the proposed technique displayed long-term stability (over 2 months), high intermediate precision (RSD<2.1 %), good reproducibility (RSD <1.9 %), and outstanding selectivity toward IMB over analogous molecules with similar chemical and spatial structure (no interference by 100 to 150-fold of the competitors). Owing to these merits, the proposed fluorescence sensor was utilized to detect IMB in drug tablets and human plasma, and satisfactory results (99.3-100.4 %) were obtained. Thus, the synthesized fluorescence sensor is a promising platform for IMB sensing in various applications.

The Impact of Blood Sample Processing on Ribonucleic Acid (RNA) Sequencing.

In gene quantification and expression analysis, issues with sample selection and processing can be serious, as they can easily introduce irrelevant variables and lead to ambiguous results. This study aims to investigate the extent and mechanism of the impact of sample selection and processing on ribonucleic acid (RNA) sequencing. RNA from PBMCs and blood samples was investigated in this study. The integrity of this RNA was measured under different storage times. All the samples underwent high-throughput sequencing for comprehensive evaluation. The differentially expressed genes and their potential functions were analyzed after the samples were placed at room temperature for 0h, 4h and 8h, and different feature changes in these samples were also revealed. The sequencing results showed that the differences in gene expression were higher with an increased storage time, while the total number of genes detected did not change significantly. There were five genes showing gradient patterns over different storage times, all of which were protein-coding genes that had not been mentioned in previous studies. The effect of different storage times on seemingly the same samples was analyzed in this present study. This research, therefore, provides a theoretical basis for the long-term consideration of whether sample processing should be adequately addressed.

The Effect of Repeated Blood Harvesting from Pregnant Mares on Haematological Variables.

Studies have been carried out on the effect of large-volume blood harvesting from horses, but they were performed on nonpregnant horses of various breeds other than Icelandic horses. This study aimed to investigate the effect on the haematological variables indicating erythropoiesis of repeated blood harvesting from pregnant mares. To account for regional variation, two herds of mares were chosen, both kept under free-range conditions. Sequential EDTA blood samples were collected weekly from 160 mares and analysed for haematological variables in an automated analyser. Serum samples from 115 mares were analysed for total protein. In both herds, after three harvests, mares began to measure below the minimum value for erythrocyte numbers, and mild anaemia was present in up to 37% at one time. Mares in only one herd had moderate or marked anaemia, 14.3% of the herd. Both herds showed evidence of increased erythropoiesis, but there was a difference between the herds in the intensity of the response. In both herds, however, all mares had reached the minimum normal haematocrit value three weeks after the last harvest. It is important to investigate the causes for the differences between these herds, which might be addressed to reduce the risk of anaemia.

ALISTER - Application for lipid stability evaluation and research.

BACKGROUND AND AIMS: In lipidomic and metabolomic studies, pre-analytical pitfalls enhance the risk of misusing resources such as time and money, as samples that are analyzed may not yield accurate or reliable data due to poor sample handling. Guidance and pre-analytic know-how are necessary for translation of omics technologies into routine clinical testing. The present work aims to enable decision making regarding sample stability in every phase of lipidomics- and metabolomics-centered studies. MATERIALS AND METHODS: Data of multiple pre-analytic studies were aggregated into a database. Flexible approaches for evaluating these data were implemented in an RShiny-based web-application, tailored towards broad applicability in clinical and bioanalytic research. RESULTS: Our "Application for lipid stability evaluation & research" - ALISTER facilitates decision making on blood sample stability during lipidomic and metabolomic studies, such as biomarker research, analysis of biobank samples and clinical testing. The interactive tool gives sampling recommendations when planning sample collection or aids in the assessment of sample quality of experiments retrospectively. CONCLUSION: ALISTER is available for use under https://itmp.shinyapps.io/alister/. The application enables and simplifies data-driven decision making concerning pre-analytic blood sample handling and fits the needs of clinical investigations from multiple perspectives.

Highly electrochemically active Ti3C2Tx MXene/MWCNT nanocomposite for the simultaneous sensing of paracetamol, theophylline, and caffeine in human blood samples.

The facile fabrication is reported of highly electrochemically active Ti3C2Tx MXene/MWCNT (3D/1D)-modified screen-printed carbon electrode (SPE) for the efficient simultaneous electrochemical detection of paracetamol, theophylline, and caffeine in human blood samples. 3D/1D Ti3C2Tx MXene/MWCNT nanocomposite was synthesized using microwave irradiation and ultrasonication processes. Then, the Ti3C2Tx/MWCNT-modified SPE electrode was fabricated and thoroughly characterized towards its physicochemical and electrochemical properties using XPS, TEM, FESEM, XRD, electrochemical impedance spectroscopy, cyclic voltammetry, and differential pulse voltammetry techniques. As-constructed Ti3C2Tx-MWCNT/SPE offers excellent electrochemical sensing performance with good detection limits (0.23, 0.57, and 0.43 µM) and wide linear ranges (1.0 ~ 90.1, 2.0 ~ 62.0, and 2.0-90.9 µM) for paracetamol, caffeine, and theophylline, respectively,  in the human samples. Notably, the non-enzymatic electroactive nanocomposite-modified electrode has depicted a semicircle Nyquist plot with low charge transfer resistance (Rct∼95 Ω), leading to high ionic diffusion and facilitating an excellent electron transfer path. All the above results in efficient stability, reproducibility, repeatability, and sensitivity compared with other reported works, and thus, it claims its practical utilization in realistic clinical applications.

[Determination of Perchloroethylene in blood by headspace gas chromatography-mass spectrometry].

Objective: To establish a method for determination of Perchloroethylene (PCE) in blood by headspace gas chromatography-mass spectrometry (HS/GC-MS) . Methods: From Dctober to December 2021, A total of 3 mL blood samples were taken into a 10 mL headspace bottle, after heated at 60 ℃ for 30 mins, PCE in the top air was separated by VF-WAXms capillary column and detected by GC-MS. The retention time and external standard method were used for qualitative and quantitative analysis of PCE in samples, respectively. Results: There was good linear relationship in the range of 5.09-200.17 µg/L. The linear correlation coefficient was 0.9993.The detection limit was 0.21 µg/L and the lower limit of quantitation was 0.70 µg/L. The recovery rates of samples with different concentrations were 95.3%-103.8%. The intra-batch relative standard deviations (RSD) were 3.2%-4.6%, and inter-batch RSD was 4.0%-6.1%. The samples can be stored at 4 ℃ for three days and at -20 ℃ for seven days. Conclusion: This method is proved to be simple, practical and highly sensitive, which is suitable for the determination of PCE in blood.

An Automated Centrifugal Microfluidic Platform for Efficient Multistep Blood Sample Preparation and Clean-Up towards Small Ion-Molecule Analysis.

Sample preparation for mass spectroscopy typically involves several liquid and solid phase clean-ups, extractions, and other unit operations, which are labour-intensive and error-prone. We demonstrate a centrifugal microfluidic platform that automates the whole blood sample's preparation and clean-up by combining traditional liquid-phase and multiple solid-phase extractions for applications in mass spectroscopy (MS)-based small molecule detection. Liquid phase extraction was performed using methanol to precipitate proteins in plasma separated from a blood sample under centrifugal force. The preloaded solid phase composed of C18 beads then removed lipids with a combination of silica particles, which further cleaned up any remaining proteins. We further integrated the application of this sample prep disc with matrix-assisted laser desorption/ionization (MALDI) MS by using glancing angle deposition films, which further cleaned up the processed sample by segregating the electrolyte background from the sample salts. Additionally, hydrophilic interaction liquid chromatography (HILIC) MS was employed for detecting targeted free amino acids. Therefore, several representative ionic metabolites, including several amino acids and organic acids from blood samples, were analysed by both MALDI-MS and HILIC-MS to demonstrate the performance of this sample preparation disc. The fully automated blood sample preparation procedure only took 35 mins, with a throughput of three parallel units.

A novel mini-invasive finger-prick protocol for preparing blood micro-samples for morphological analysis of human erythrocytes by scanning electron microscopy.

Here we describe the development and optimization of a new protocol for the preparation and surface imaging by scanning electron microscope of human erythrocytes from blood micro-samples obtained by finger prick. By testing several key pre-analytical conditions for blood sampling, erythrocyte preservation, storage and imaging, we designed a rapid new minimally-invasive reproducible method for obtaining uniform deposition of an adequate number of erythrocytes with well-preserved morphology on a substrate. The possibility of obtaining reliable reproducible high resolution morphometric data on peripheral erythrocytes makes this protocol valuable for diagnostic and basic research purposes.

Complement Components in Peripheral Blood from Adult Patients with IDH Wild-Type Glioblastoma.

BACKGROUND: The complement system seems to influence cancer pathophysiology. The primary aim of this study was to explore complement components associated with the classical pathway (CP) of the complement system in peripheral blood from patients with IDH-wild-type (IDH-wt) glioblastoma. METHODS: Patients undergoing primary surgery due to glioblastoma in the years 2019-2021 were prospectively included in the present study. Blood samples were collected prior to surgery, and analyzed with regard to CP complement components, as well as standard coagulation tests. RESULTS: In total, 40 patients with IDH-wt glioblastomas were included. C1q was reduced in 44% of the cases compared to the reference interval. C1r was reduced in 61% of the analyzed samples. Both C1q and C1r are parts of the initial steps of the classical complement activation pathway, which, however, was not correspondingly altered. Activated pro-thromboplastin time (APTT) was shorter in 82% of the analyzed samples compared to the reference interval. APTT was shorter in those with reduced C1q and C1r levels. C1q is an important link between the innate and acquired immunity, and C1q and C1r also interact with the coagulation system. Patients who displayed reduced levels of both C1q and C1r preoperatively had a significantly shorter overall survival compared with the rest of the cohort. CONCLUSIONS: Our findings demonstrate that there are alterations in C1q and C1r concentrations in peripheral blood from patients with IDH1-wt glioblastoma compared with the normal population. Patients who displayed reduced C1q and C1r levels had a significantly shorter survival.