Raman spectroscopy and Raman optical activity of blood plasma for differential diagnosis of gastrointestinal cancers.

Improving the early diagnosis of gastrointestinal cancers is a crucial step in reducing their mortality. Given the non-specificity of the initial symptoms, the ability of any diagnostic method to differentiate between various types of gastrointestinal cancers also needs to be addressed. To detect disease-specific alterations in biomolecular structure and composition of the blood plasma, we have implemented an approach combining Raman spectroscopy and its conformation-sensitive polarized version, Raman optical activity, to analyze blood plasma samples of patients suffering from three different types of gastrointestinal cancer - hepatocellular, colorectal and pancreatic. First, we aimed to discriminate any type of gastrointestinal cancer from healthy control individuals; inthenext step, the focus was on differentiating among the three cancer types studied. The more straightforward of the two statistical approaches tested, the combination of linear discriminant analysis and principal component analysis applied to the entire spectral dataset, allowed the discrimination of cancer and control samples with 87% accuracy. The three gastrointestinal cancers were classified with an overall accuracy of 76%. The second method, the linear discriminant analysis applied to a selection of spectral bands, yielded even higher values. Cancer and control samples were distinguished with 89% accuracy and hepatocellular, colorectal and pancreatic cancer with an overall accuracy of 87%. The results obtained in our study suggest that the proposed approach may become a disease-specific diagnostic tool in daily clinical practice.

The potential of multicylindrical dielectric barrier discharge plasma for diesel-contaminated soil remediation and biocompatibility assessment.

This study explored the use of multicylindrical dielectric barrier discharge (MC-DBD) plasma technology to eliminate diesel fuel contamination from the soil. This study also assessed the environmental impact of plasma-generated reactive species on soil properties, plant growth, and the safety of microbial and human skin cells using various analytical methods. MC-DBD plasma was characterized using the current-voltage analysis and optical emission spectroscopy (OES). Gas Fourier transform infrared spectroscopy was employed to detect reactive species, such as O3, NO, NO2, N2O, and HNO3, in the plasma-treated air. The diesel fuel concentration in the soil was measured before and after plasma treatment using a gas chromatography-flame ionization detector. The efficacy of the MC-DBD plasma treatment was evaluated based on soil characteristics (pH and moisture), discharge parameters (power), and reactive species (O3 and NOx). Using only power of 30 W, the MC-DBD achieved a 94.19% removal of diesel fuel from the soil and yielded an energy efficiency of 1.78 × 10-2 m3/kWh within a 60-min treatment period. Neutral soil with a moisture content of 2% proved more effective in diesel fuel removal compared with acidic or alkaline soil with higher moisture content. O3 was the most efficient plasma-generated reactive species for diesel fuel removal and is involved in oxidation-induced fragmentation and volatilization. Overall, the potential of the MC-DBD plasma technology for remediating diesel fuel-contaminated soils is highlighted, and valuable insights for future applications are provided.

Development of a biodegradable microfluidic paper-based device for blood-plasma separation integrated with non-enzymatic electrochemical detection of ascorbic acid.

In the present article, we developed an electrochemical microfluidic paper-based device (EµPAD) for the non-enzymatic detection of Ascorbic Acid (AA) concentration in plasma using whole human blood. We combined LF1 blood plasma separation membrane and Whatman grade 1 filter paper to separate plasma from whole blood through wax printing. A screen-printed electrode (SPE) was modified with spherical-shaped MgFe2O4 nanomaterial (n-MgF) to improve the catalytic properties of SPE. The n-MgF was prepared via hydrothermal method, and its material phase and morphology were confirmed via XRD, FTIR, TEM, SEM, and AFM analysis. The fabricated n-MgF/SPE/EµPAD exhibited detection of AA ranging from 0 to 80 µM. The obtained value of the detection limit, limit of quantification, sensitivity, and response time are 2.44 µM, 8.135 µM, 5.71 × 10-3 mA µM-1 cm-2, and 10 s, respectively. Our developed n-MgF/SPE/EµPAD shows marginal interference with the common analytes present in plasma, such as uric acid, glutamic acid, glucose, urea, lactic acid, and their mixtures. Overall, our low-cost, portable device with its user-friendly design and efficient plasma separation capability offers a practical and effective solution for estimating AA concentration from whole human blood in a single step.

From "wet" matrices to "dry" blood spot sampling strategy: a versatile LC-MS/MS assay for simultaneous monitoring caffeine and its three primary metabolites in preterm infants.

OBJECTIVES: To update traditional "wet" matrices to dried blood spot (DBS) sampling, based on the liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS) technique, and develop a method for simultaneous analyzing caffeine and its three primary metabolites (theobromine, paraxanthine, and theophylline), supporting routine therapeutic drug monitoring (TDM) for preterm infants. METHODS: DBS samples were prepared by a two-step quantitative sampling method, i.e., volumetric sampling of a quantitative 10 µL volume of peripheral blood and an 8 mm diameter whole punch extraction by a methanol/water (80/20, v/v) mixture containing 125 mM formic acid. Four paired stable isotope labeled internal standards and a collision energy defect strategy were applied for the method optimization. The method was fully validated following international guidelines and industrial recommendations on DBS analysis. Cross validation with previously developed plasma method was also proceeded. The validated method was then implemented on the TDM for preterm infants. RESULTS: The two-step quantitative sampling strategy and a high recovery extraction method were developed and optimized. The method validation results were all within the acceptable criteria. Satisfactory parallelism, concordance, and correlation were observed between DBS and plasma concentrations of the four analytes. The method was applied to provide routine TDM services to 20 preterm infants. CONCLUSIONS: A versatile LC-MS/MS platform for simultaneous monitoring caffeine and its three primary metabolites was developed, fully validated, and successfully applied into the routine clinical TDM practices. Sampling method switching from "wet" matrices to "dry" DBS will facilitate and support the precision dosing of caffeine for preterm infants.

A sensitive LC-MS/MS methotrexate assay capable of assessing adherence to methotrexate therapy in rheumatoid arthritis.

OBJECTIVES: To develop a sensitive liquid chromatography-tandem mass spectrometry method capable of measuring serum methotrexate in patients with rheumatoid arthritis to assess adherence to drug treatment. METHODS: Isotopically labelled internal standard and deionised water were added to sample prior to solid phase extraction using a Waters Oasis Max ion-exchange 96-well plate. Following extraction, samples were analysed by LC-MS/MS on a TQS-micro mass spectrometer. RESULTS: Mean recovery was 107 % for four different concentrations of methotrexate spiked into seven patient samples, whilst post extraction spiking gave a mean recovery of 100 %. Between-batch and within-batch CVs were ≤6 % at three different concentrations of methotrexate in fresh frozen plasma. Mean bias was <5 % for between-batch and within batch analysis at three different weighed in concentrations of methotrexate certified reference material. The lower limit of quantification of the assay was 0.1 nmol/L with linearity up to approximately 100 nmol/L. Dilution linearity studies were used to validate the dilution of patient samples prior to analysis. There was no significant interference in the method from lipaemia, haemolysis or icterus. CONCLUSIONS: A sensitive LC-MS/MS assay for methotrexate has been developed and validated. The method has been used to measure methotrexate adherence in patient samples from clinical trials and could be used in future research to assess the ability of the assay as a biofeedback intervention to improve adherence to methotrexate therapy.

Implementation and validation of a UHPLC-MS/MS method for quantification of the endocannabinoids AEA and 2-AG in cerebral interstitial fluid and plasma.

Endogenous endocannabinoids such as N-arachidonoylethanolamine (AEA) and 2-arachidonoylglycerol (2-AG) are involved in the patho-biochemistry of several neurological diseases and have been associated with mood-enhancing phenomena. Although they have been intensively studied in recent years, accurate and reliable quantification of these analytes in cerebral interstitial fluid (cISF) to elucidate their neuro-modulatory role is still challenging. Moreover, there is a need for an analytical method that can analyze plasma in addition to cISF and is thus able to address research questions in both preclinical and clinical studies. Aim was to implement a method for simultaneous quantification of AEA and 2-AG in cISF and plasma, to validate it by taking the requirements of the U.S. Food and Drug Administration into account, and to test its usability in three different case studies. A UHPLC-MS/MS method with preceding liquid-liquid extraction to determine AEA and 2-AG in cISF and plasma was successfully implemented, and the parameters selectivity, specificity, linearity, accuracy, precision, sensitivity, carry-over and stability met the validation criteria. The usability of the analytical method was demonstrated in an in vitro study with cerebral open flow microperfusion (cOFM), an in vivo cOFM study in rats, and a clinical study in human plasma. The developed method allowed quantification of AEA and 2-AG in the biologically relevant concentration ranges in cISF and plasma. The availability of a reliable, complementary, time-resolved dataset of endocannabinoid concentrations in both matrices can be of considerable future importance for the evaluation of drug efficacy.

An atypical presentation of anticoagulant rodenticide toxicosis in a dog.

A 5-month-old intact female Australian shepherd dog was referred to our clinic for neurologic signs including ataxia, a head tilt, and altered mentation following consumption of an unidentified rodenticide several days prior to developing clinical signs. A provisional diagnosis of bromethalin toxicosis had been made, given the neurologic signs seen and the general increased use of bromethalin-containing rodenticide products. However, on physical examination, the dog was noted to have scleral hemorrhage and bleeding at the venipuncture sites, which was inconsistent with bromethalin toxicosis. Coagulation testing was supportive of anticoagulant rodenticide toxicosis and the rodenticide was later identified as the first-generation anticoagulant rodenticide diphacinone. The neurologic signs seen were attributed to a coagulopathy causing multifocal hemorrhage into the central nervous system. Neurologic signs rapidly resolved following treatment with a frozen plasma transfusion and vitamin K1. This atypical presentation of an anticoagulant rodenticide toxicosis highlights the need for accurate product identification, if available, and thorough patient examination and laboratory testing. An atypical presentation of anticoagulant rodenticide toxicosis should be considered when neurologic signs are present with clinical bleeding, especially if the type of rodenticide is unknown, or even if it was not thought to have an anticoagulant as the active ingredient. Key clinical message: Given the change in commercially available rodenticide products, this case highlights the need for accurate product identification in cases of suspected toxicosis, and the variable clinical signs that can be seen following anticoagulant rodenticide toxicosis.

Présentation atypique d'une toxicose aux rodenticides anticoagulants chez un chien. Une chienne berger australien intacte âgée de 5 mois a été référée à notre clinique pour des signes neurologiques, notamment de l'ataxie, une inclinaison de la tête et une altération de l'état mental à la suite de la consommation d'un rodenticide non identifié plusieurs jours avant l'apparition des signes cliniques. Un diagnostic provisoire de toxicose à la brométhaline avait été posé, compte tenu des signes neurologiques observés et d'une utilisation historique accrue de produits rodenticides contenant de la brométhaline. Cependant, lors de l'examen physique, il a été constaté que le chien présentait une hémorragie sclérale et des saignements au niveau des sites de ponction veineuse, ce qui n'était pas cohérent avec une toxicose à la brométhaline. Les tests de coagulation ont confirmé la toxicose au rodenticide anticoagulant et le rodenticide a ensuite été identifié comme étant le rodenticide anticoagulant de première génération diphacinone. Les signes neurologiques observés ont été attribués à une coagulopathie provoquant une hémorragie multifocale du système nerveux central. Les signes neurologiques ont rapidement disparu après un traitement par transfusion de plasma congelé et de vitamine K1. Cette présentation atypique d'une toxicose aux rodenticides anticoagulants met en évidence la nécessité d'une identification précise du produit, si disponible, ainsi que d'un examen approfondi du patient et de tests de laboratoire. Une présentation atypique de toxicose des rodenticides anticoagulants doit être envisagée lorsque des signes neurologiques sont présents avec saignement clinique, en particulier si le type de rodenticide est inconnu, ou même si l'on ne pense pas qu'un anticoagulant soit l'ingrédient actif.Message clinique clé :Compte tenu de l'évolution des produits rodenticides disponibles dans le commerce, ce cas met en évidence la nécessité d'une identification précise du produit en cas de suspicion de toxicose et les signes cliniques variables qui peuvent être observés à la suite d'une toxicose au rodenticide anticoagulant.(Traduit par Dr Serge Messier).

Association of body mass index with perioperative blood transfusion and short-term clinical outcomes in patients undergoing isolated coronary artery bypass grafting.

BACKGROUND: Few studies have considered outcomes among low body mass index (BMI) cohorts undergoing coronary artery bypass grafting (CABG). This study aims to investigate the effects of low body weight on blood transfusion and perioperative outcomes in patients undergoing isolated CABG. METHODS: This retrospective study enrolled consecutive cases from a single-center between January 2008 and December 2018. Low body weight/underweight was defined as a BMI < 18.5 kg/m², while normal BMI was defined as 18.5 ≤ BMI < 24.0 kg/m². The primary endpoint was the perioperative red blood cell (RBC) transfusion rate. Secondary endpoints include platelet and plasma transfusion rates, transfusion volume for all blood components, hospital length of stay, and the occurrence of adverse events including prolonged mechanical ventilation, re-intubation, re-operation, acute kidney injury, and 30-day all-cause mortality. RESULTS: A total of 7,620 patients were included in this study. After 1:1 propensity score matching, 130 pairs were formed, with 61 pairs in the on-pump group and 69 pairs in the off-pump group. Baseline characteristics were comparable between the matched groups. Low body weight independently increased the risk of RBC transfusion (on-pump: OR = 3.837, 95% CI = 1.213-12.144, p = 0.022; off-pump: OR = 3.630, 95% CI = 1.875-5.313, p < 0.001). Moreover, within the on-pump group of the original cohort, BMI of < 18.5 kg/m² was independently correlated with increased risk of re-intubation (OR = 5.365, 95% CI = 1.159 to 24.833, p = 0.032), re-operation (OR = 4.650, 95% CI = 1.019 to 21.210, p = 0.047), and 30-day all-cause mortality (OR = 10.325, 95% CI = 2.011 to 53.020, p = 0.005). CONCLUSION: BMI < 18.5 kg/m² was identified as an independent risk factor for increased perioperative RBC transfusion rate in patient underwent isolated CABG with or without CPB. Only on-pump underweight patients in the original cohort exhibited an increased risk for re-intubation, re-operation, and 30-day all-cause mortality. Physicians and healthcare systems should consider these findings to improve management for this population.

[One case of bromadiolone poisoning leading to intestinal necrosis and severe coagulopathy].

Bromadiolone is still often used in life as a poisonous rodent agent. Bromadiolone poisoning is often manifested as coagulation dysfunction, resulting in organ bleeding, including cerebral hemorrhage, intestinal bleeding, abdominal hemorrhage, etc. At present, no case of intestinal necrosis caused by bromadiolone poisoning have been reported. This article reviewed one case of intestinal necrosis and severe coagulation dysfunction, and finally confirmed bromadiolone poisoning by poison detection. The patient recovered and was discharged after surgery, vitamin K injection, plasma transfusion and other treatment methods.

Prevalence, Risk Factors and Clinical Outcomes of Acute Kidney Injury after Paediatric Liver Transplantation.

BACKGROUND: Knowledge about acute kidney injury (AKI) in paediatric patients after liver transplantation is limited. This study focused on the prevalence and contributing factors of AKI and its impact on the postoperative outcomes of paediatric recipients. METHODS: The data of 211 paediatric patients (<12 years old) who, from December 2018 to November 2020, received first-time liver transplantation for end-stage liver disease or advanced hepatic failure in our hospital were processed for retrospective analysis. According to the criteria of the Kidney Disease Improving Global Outcomes, all patients were dichotomised into AKI and non-AKI groups. The preoperative characteristics of the patients, laboratory test results, donor type and intraoperative parameters between the two groups were compared. The effects of AKI on the postoperative outcomes of paediatric recipients were analysed. The risk factors for AKI after liver transplantation were analysed by multivariate logistic regression. RESULTS: The incidence of AKI within the first seven days after paediatric liver transplantation was 34.1%. AKI at stages I, II and III accounted for 69.4%, 22.2% and 8.3%, respectively. Compared with non-AKI patients, AKI patients had a higher proportion of hepatic cirrhosis (p = 0.039) and ascites (p = 0.003); Worse hepatic function (prolonged prothrombin time, activated partial thromboplastin time, decreased level of serum albumin and increased international normalised ratio and total bilirubin level); Higher paediatric end-stage liver disease (PELD) score (p = 0.008); And larger amount of intraoperative blood loss (p < 0.001), fluid positive balance (p = 0.035), red blood cells (RBCs) (p < 0.001) and fresh frozen plasma transfusion (p < 0.001). The multivariate logistic regression analysis demonstrated that ascites (odds ratio (OR): 2.273, p = 0.040), PELD (OR: 1.027, p = 0.013) and RBCs transfusion (OR: 1.033, p < 0.001) were independent contributing factors to AKI in paediatric patients who received liver transplantation. AKI contributed to prolonged hospital stays but did not increase hospital mortality. CONCLUSIONS: The onset of AKI can markedly prolong the hospital stay, and is common in paediatric patients undergoing liver transplantation. Poor hepatic function and large RBC transfusion contribute to AKI after liver transplantation.

High-Throughput Blood Plasma Extraction in a Dimension-Confined Double-Spiral Channel.

Microfluidic technologies enabling the control of secondary flow are essential for the successful separation of blood cells, a process that is beneficial for a wide range of medical research and clinical diagnostics. Herein, we introduce a dimension-confined microfluidic device featuring a double-spiral channel designed to regulate secondary flows, thereby enabling high-throughput isolation of blood for plasma extraction. By integrating a sequence of micro-obstacles within the double-spiral microchannels, the stable and enhanced Dean-like secondary flow across each loop can be generated. This setup consequently prompts particles of varying diameters (3, 7, 10, and 15 µm) to form different focusing states. Crucially, this system is capable of effectively separating blood cells of different sizes with a cell throughput of (2.63-3.36) × 108 cells/min. The concentration of blood cells in outlet 2 increased 3-fold, from 1.46 × 108 to 4.37 × 108, while the number of cells, including platelets, exported from outlets 1 and 3 decreased by a factor of 608. The engineering approach manipulating secondary flow for plasma extraction points to simplicity in fabrication, ease of operation, insensitivity to cell size, high throughput, and separation efficiency, which has potential utility in propelling the development of miniaturized diagnostic devices in the field of biomedical science.

Optimization for the analysis of 42 per- and polyfluorinated substances in human plasma: A high-throughput method for epidemiological studies.

There is an increasing awareness about the presence of per- and polyfluoroalkyl substances (PFAS) in many environmental and biological compartments, including human biofluids and tissues. However, the increase of PFAS replacements, including alternatives with shorter chain or less bioaccumulative potential, has broaden the exposure and the need for wider identification procedures. Moreover, the low volumes available for human blood or plasma, and the high number of samples needed to assess adequately epidemiologic studies, require particularly fast, reproducible and, if possible, miniaturized protocols. Therefore, accurate and robust analytical methods are still needed to quantify the PFAS's burden in humans and to understand potential health risks. In this study, we have developed and validated the analysis of 42 PFAS in human plasma by means of a Captiva 96-well micro extraction plate and a LC-q-Orbitrap. For the optimization of the analytical workflow, three extraction/clean-up methods were tested, and the selected one was validated using spiked artificial and bovine plasma at four concentration levels. The final method showed high absolute recoveries for the 42 PFAS, ranging from 52% to 130%, instrumental detection limits between 0.001-0.6 ng mL-1, overall good precision (CV < 20% for most of the PFAS) and a low uncertainty (< 30% of relative expanded deviation, k = 2). The method was further validated both with the NIST plasma Standard Reference Material 1950, showing that the accuracy of the provided results was between 63%-101%, and by the proficiency test arranged by the Arctic Monitoring Assessment Program (AMAP, 2022) obtaining satisfactory results within 95% confidence interval of the assigned value.

Blood plasma metagenomic next-generation sequencing for identifying pathogens of febrile neutropenia in acute leukemia patients.

To investigate the value of metagenomic next-generation sequencing (mNGS) in acute leukemia (AL) patients with febrile neutropenia (FN). We retrospectively reviewed 37 AL patients with FN and compared the results of mNGS with blood culture (BC) and the clinical features of the mNGS-positive group and the mNGS-negative group. A total of 14 detected pathogens were the final clinical diagnosis, of which 9 strains were detected only by mNGS and 5 strains were detected by both mNGS and BC. The top pathogens were Klebsiella pneumoniae, Pseudomonas aeruginosa and Stenotrophomonas maltophilia. A total of 67.57% (25/37) were bacterial infections, and 2.7% (1/37) were fungal or viral infections. The diagnostic positivity rate of mNGS (25/37, 67.6%) was significantly higher than that of BC (7/37, 18.9%), and the difference was statistically significant (p < 0.05). Then, we explored the clinical distinction between the mNGS-positive group and the mNGS-negative group, and 3 features were filtered, including lymphocyte count (LY), creatinine levels (Cr), and white blood cell count (WBC). Our study demonstrated that early implementation of mNGS can effectively improve the efficacy of pathogen detection in AL patients with FN. The higher diagnostic positivity rate and the ability to detect additional pathogens compared to BC made mNGS a valuable tool in the management of infectious complications in this patient population. Furthermore, the identified clinical features associated with mNGS results provided additional insights for the clinical indication of infection in AL patients with FN.

Rapid serum tubes reduce transport hemolysis and false positive rates for high-sensitivity troponin T.

INTRODUCTION: Hemolysis in the emergency department (ED) can significantly delay results and appropriate action. We evaluated the main sources of hemolysis during sample collection, and to evaluate the use of rapid serum tubes (RST) as a transport hemolysis-mitigating measure for high-sensitivity troponin T (hs-cTnT) testing. METHODS: We examined the effect of tube type, tube fill, types of sample draw and collection methods on hemolysis and hs-cTnT in samples (n = 158) from ED patients. We also compared hs-cTnT values in paired RST and plasma separate tube (PST) samples that were hemolysis-free. RESULTS: The primary source of hemolysis in samples collected in the ED was underfilling tubes. In both tube types, PST and RST, filled tubes showed a median reduction in hemolysis of 69.1 % (p < 0.0001). Blood collected in RST also experienced less hemolysis compared to PST. In hemolysis-free samples, false positive results in PST were noted in patients with hs-cTnT values < 50 ng/l. CONCLUSION: We suggest that proper tube filling during sample collection and use of RST tubes can significantly reduce the effects of hemolysis. In addition, laboratories should be aware that PST tubes have a non-trivial rate of false positives when hs-cTnT < 50 ng/l.

Identification of Sanguinarine Metabolites in Rats Using UPLC-Q-TOF-MS/MS.

Sanguinarine (SAN), as the main active component of a traditional Chinese veterinary medicine, has been widely used in the animal husbandry and breeding industry. However, the metabolites of SA are still uncertain. Therefore, this research aimed to investigate the metabolites of SA based on rats in vivo. The blood, feces, and urine of rats were collected after the oral administration of 40 mg/kg SAN. Ultra-high-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF-MS/MS) was employed to identify the metabolites of SAN. The elemental composition of sanguinarine metabolites was inferred by analyzing their exact molecular weight, and the structures of the metabolites were predicted based on their fragment ions and cleavage pathways. A total of 12 metabolites were identified, including three metabolites in the plasma, four in the urine, and nine in the feces. According to the possible metabolic pathways deduced in this study, SAN was mainly metabolized through reduction, oxidation, demethylation, hydroxylation, and glucuronidation. This present research has summarized the metabolism of SAN in rats, which is helpful for further studying the metabolic mechanism of SAN in vivo and in vitro.

Anti-Clogging Effect of Continuous Hemodiafiltration With Blood Recirculation.

Pediatric dialysis requires low flow from the body, but greater flow is needed to prevent clogging. As a solution, we developed a new continuous hemodiafiltration system with blood recirculation (CHDF-R), which enables separate settings for blood flow from the body and to the hemofilter. We compared CHDF-R with conventional continuous hemodiafiltration (CHDF) of bovine plasma and blood by monitoring the transmembrane pressure (TMP) and observing the hemofilter membrane surface. When using bovine plasma, the postdialysis TMP with CHDF-R was significantly lower than with CHDF (median CHDF, 23.7; median CHDF-R, 18.1; p = 0.029). Likewise, when using bovine blood, the postdialysis TMP was also significantly lower with CHDF-R than with CHDF (median CHDF, 150; median CHDF-R, 100; p = 0.029). Moreover, the area of clogged membrane was significantly smaller with CHDF-R than with CHDF, and the inner membrane surface showed less material deposition with CHDF-R than CHDF. CHDF-R thus appears to suppress accumulation of clogging substances by producing higher shear stress within hollow fiber membranes.

Phytocorrection of Age-Related Changes in the Composition of Blood Plasma, Lymph, and Interstitial Fluid.

We studied the effect of a new complex phytocomposition on age-related changes in the composition of biological fluids (blood, lymph, and interstitial fluid) in old (22-24-month-old) white Sprague-Dawley rats. A 3-month course of phytocomposition induced an increase in the volumes of interstitial fluid (IF) and plasma and stimulated lymph outflow and diuresis. In the blood and lymph, the clotting time increased, while the viscosity and all lipid indicators decreased. The phytocomposition increased the number of erythrocytes and leukocytes, the levels of immunoglobulins (except IgG) and lymphocyte subpopulations, which contributed to correction and improvement of the immune properties of the blood and lymph. Increased hydration of the tissues of the body and strengthening of the anti-atherogenic and immune properties of the lymph and blood led to recovery of the drainage and detoxification function of the lymphatic system. Due to the presence of bioflavonoids, microelements, and vitamins, the new complex phytocomposition produces a lymphotropic effect by changing the composition of the blood, lymph, and IF and stimulates fluid transport from IF into the vascular bed, thereby promoting natural lymph detoxification and increasing the immune properties of the blood and lymph.

Long-term follow-up of patients with congenital thrombotic thrombocytopenia purpura receiving a plasma-derived factor VIII (Koate) that contains ADAMTS13.

BACKGROUND: Hereditary thrombotic thrombocytopenia purpura (hTTP) is an ultra-rare disorder resulting from an inherited deficiency of ADAMTS13, a von Willebrand factor (VWF)-cleaving metalloprotease. The plasma-derived factor VIII/VWF Koate (FVIII/VWFKoate ) has been shown to contain ADAMTS13, allowing for its use to treat hTTP at home by the patient/caregiver. AIM: Based on prior demonstration of safe and effective use of FVIII/VWFKoate in eight patients with hTTP, we conducted a retrospective study to gather additional data regarding the use of FVIII/VWFKoate for hTTP. METHODS: This was a multicentre, retrospective, noninterventional chart review of patients who had received FVIII/VWFKoate for the management of hTTP. Data collected included demographics, medical history, relevant family history, past use and tolerability of fresh frozen plasma, and details regarding FVIII/VWFKoate therapy. RESULTS: The cohort included 11 patients (seven males, four females) with hTTP, ranging in age at study entry from 2 to 28 years. The average duration of FVIII/VWFKoate therapy was 4.8 years (range, 0.5-6.5 years). Among nine patients using FVIII/VWFKoate as prophylaxis, the normalized annual rate of breakthrough TTP episodes ranged from 0.2 to 1.1 episodes/year. All nine patients who received FVIII/VWFKoate prophylaxis had thrombocytopenia recorded at baseline, while eight (88.9%) did not have thrombocytopenia after using FVIII/VWFKoate . There was one AE (unspecified) attributed to FVIII/VWFKoate . CONCLUSION: These data suggest that FVIII/VWFKoate is a safe and well-tolerated source of the missing ADAMTS13 enzyme in patients with hTTP, producing a marked reduction in thrombocytopenia prevalence, low frequency of TTP episodes, and with the added benefit of self- or caregiver-administration.

Pronounced effect of lamination on plasma separation from whole blood by microfluidic paper-based analytical devices.

Many diseases are detected through blood tests. Currently, most blood tests are done on plasma instead of whole blood because of the interference of blood cells on detection results. Here, we developed a laminated microfluidic paper-based analytical device (L-µPAD) for the separation of plasma from whole blood without using plasma separation membrane (PSM). A lateral flow design consisting of a circular sampling zone and rectangular detection zone was patterned on the paper substrate using laser printing technology. The µPAD was then laminated after impregnation with KCl solution. Lamination and electrolyte addition represented synergistic effects on the separation by controlling the pore size of the paper. In addition, by preventing evaporation on one hand and squeezing paper pores on the other hand, lamination caused longer movement of the separated plasma, the longest plasma path reported so far. The separation process was monitored using colorimetric reagent bromocresol green and scanning electron microscopy. The process of separation was completed in less than 90s without significant hemolysis and the separated plasma was far from the interfering effect of red blood cells. We used the device for the determination of serum albumin. However, it represents the potential for point-of-care testing in multi-assay experiments too.

A large-scale, targeted metabolomics method for the analysis and quantification of metabolites in human plasma via liquid chromatography-mass spectrometry.

Metabolomics is the study of small molecules, primarily metabolites, that are produced during metabolic processes. Analysis of the composition of an organism's metabolome can yield useful information about an individual's health status at any given time. In recent years, the development of large-scale, targeted metabolomic methods has allowed for the analysis of biological samples using analytical techniques such as LC-MS/MS. This paper presents a large-scale metabolomics method for analysis of biological samples, with a focus on quantification of metabolites found in blood plasma. The method comprises a 10-min chromatographic separation using HILIC and RP stationary phases combined with positive and negative electrospray ionization in order to maximize metabolome coverage. Complete analysis of a single sample can be achieved in as little as 40 min using the two columns and dual modes of ionization. With 540 metabolites and the inclusion of over 200 analytical standards, this method is comprehensive and quantitatively robust when compared to current targeted metabolomics methods. This study uses a large-scale evaluation of metabolite recovery from plasma that enables absolute quantification of metabolites by correcting for analyte loss throughout processes such as extraction, handling, or storage. In addition, the method was applied to plasma collected from adjuvant breast cancer patients to confirm the suitability of the method to clinical samples.