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•Single-cell MALDI TOF MS imaging of CD19+ lymphocytes was developed.•Single-cell MS provides CD19+ lymphocytes’ classification accuracy >90 %.•CD19+ lymphocytes’ mass spectra show ...numerous specific changes in CLL.•Metabolic pathways affected in CLL include single-carbon metabolism and apoptosis.
Modern mass spectrometry imaging (MSI) enables single cells’ metabolism exploration. Aims of this study were development of the single-cell MSI of human CD19+ lymphocytes and metabolic profiling of chronic lymphocytic leukemia (CLL).
Blood donor (BD) samples were used for the optimization of CD19+ lymphocyte isolation and single-cell matrix-assisted laser desorption/ionization time-of-flight (MALDI TOF) MSI. Independent set of 200 CD19+ lymphocytes coming from 5 CLL patients and 5 BD was used for the CD19+ lymphocytes classification assessment and the untargeted metabolic profiling. CLL vs BD lymphocyte classification was performed using partial least squares-discriminant analysis (PLS-DA) using normalized single-cell mass spectra recorded in 300–600 and 600–950 Da ranges was applied.
Accuracy assessed by 10-fold cross-validation of CD19+ lymphocyte PLS-DA classification reached >90.0 %. Volcano plots showed 106 significantly altered m/z signals in CLL of which 9 were tentatively annotated. Among tentatively annotated m/z signals formaldehyde and glutathione metabolites and tetrahydrofolate stand out.
A method for single-cell MALDI TOF MSI of CD19+ lymphocytes was successfully developed. The method confirmed the significance of oxidative stress and single-carbon metabolism, pyruvate and fatty acid metabolism and apoptosis in CLL and it provided metabolic candidates for diagnostic applications.
The aim of this study was to develop and evaluate matrix assisted LASER desorption ionization (MALDI) time-of-flight (TOF) mass spectrometry imaging (MSI) of blood smear. Integrated light microscope ...and MALDI IT-TOF mass spectrometer, together with a matrix sublimation device, were used for analysis of blood smears coming from healthy male donors. Different blood plasma removal, matrix deposition, and instrumental settings were evaluated using the negative and positive ionization modes while agreement between the light microscopy images and the lateral distributions of cellular marker compounds served as the MSI quality indicator. Red and white blood cells chemical composition was analyzed using the differential
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expression. Five seconds of exposure to ethanol followed by the 5 min of 9-aminoacridine or α-cyano-4-hydroxycinnamic acid deposition, together with two sets of instrumental settings, were selected for the MALDI TOF MSI experiments. Application of the thin and transparent matrix layers assured good correspondence between the LASER footprints and the preselected regions of interest. Cellular marker
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signals coincided well with the appropriate cells. A metabolite databases search using the differentially expressed
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produced hits which were consistent with the respective cell types. This study sets the foundations for application of blood smear MALDI TOF MSI in clinical diagnostics and research.
SAŽETAK
Radna skupina Hrvatskog društva za transfuzijsku medicinu pripremila je smjernice za određivanje Rh(D) krvne grupe i primjenu RhD genotipizacije. U smjernicama je opisan klinički značaj ...antigena D, povijest i ograničenja serološkog testiranja antigena D te mogućnosti RhD genotipizacije. Cilj smjernica bio je objava novih postupnika serološkog određivanja Rh(D) krvne grupe bolesnicima, trudnicama i novorođenčadi s uputama za specijaliste transfuzijske medicine u hitnom i redovnom radu te tumačenjima nalaza namijenjenim ginekolozima, pedijatrima, neonatolozima, anesteziolozima, internistima, liječnicima obiteljske medicine te svim liječnicima koji se u svom radu susreću s bolesnicima koji primaju krvne pripravke i donose odluku o primjeni RhIG imunoprofilakse. Kao rezultat provođenja smjernica predviđeno je praćenje i periodično izvještavanje u slučaju sumnje na RhD imunizaciju kod osoba nositelja D-varijante. Tijekom trudnoće postoji i mogućnost neinvazivnog određivanja prijenatalnog fetalnog RhD genotipa iz majčine plazme iza 16. tjedna gestacije, kao važnog alata u procjeni rizika razvoja hemolitičke bolesti fetusa i novorođenčeta. Radi lakšeg snalaženja navedene su vrste spremnika za uzorkovanje krvi i potrebna količina uzoraka. Navedena pretraga dostupna je u Hrvatskom zavodu za transfuzijsku medicinu na zahtjev ginekologa, a preporučuje se prvenstveno RhD imuniziranim trudnicama te u slučaju donošenja odluke o ranoj prijenatalnoj anti-D imunoprofilaksi i svim Rh(D) negativnim neimuniziranim trudnicama.
Heparin induced thrombocytopenia (HIT) is a life-threatening disorder which diagnosis depends on laboratory evaluation. The objective of this report is to present the impact of different laboratory ...methods for HIT detection on the diagnostic evaluation process. In this case, a 78-year old female patient previously diagnosed with monoclonal gammopathy of undetermined significance (MGUS) was administered with heparin for pulmonary embolism treatment. Patient's initial diagnostic work-up (determination of platelet count and prothrombin time measurement for monitoring of pharmacotherapy) was followed by the clinical estimation of HIT likelihood by "4Ts" score, two immunoassays (ID-PaGIA Heparin/PF4 Antibody Test and ELISA PF4 IgG assay) and one functional test called high-performance liquid chromatography serotonin release assay (HPLC-SRA). The result of "4Ts" score indicated a low likelihood of HIT but persistent thrombocytopenia that appeared days after discontinuation of heparin therapy suggested delayed-onset HIT. Both immunoassays were positive for presence of HIT-autoantibodies, while the functional HPLC-SRA was negative. Since different methods gave opposing results, their interpretation required great attention. In comparison to the HPLC-SRA, immunoassays are prone to the analytical interferences associated with the presence of non-specific antibodies, which may lead to false positive results. In this case, where the patient is known to produce antibodies of undetermined significance, HIT was ruled out as the possible cause of persistent thrombocytopenia primarily due to the negative result of HPLC-SRA, which is not prone to this type of interferences, but also due to the low "4Ts" clinical score.
Heparin induced thrombocytopenia (HIT) is a serious complication of heparin administration. In the last decade, this clinical syndrome has come into the focus of interest, primarily because of the ...severe thromboembolic complications that may lead to lethal outcome. In addition, great improvements have been made in the treatment with direct thrombin inhibitors and in laboratory diagnosis of HIT. As guidelines for diagnostic and management of HIT upgrade the quality of patient treatment, activities for their development have been launched in the Republic of Croatia. Based on British Committee for Standards in Haematology (BCSH) recommendations on diagnostic and treatment of HIT from 2006, activities for the introduction of new assays for anti-heparin antibodies were launched in 2008 and 2009, including algorithm of laboratory testing for HIT, sheet for clinical assessment of HIT (4T score), and education oftransfusiologists and clinicians. Upon evaluation of the results collected during one-year period, the Croatian Society of Haematology and Transfusion Medicine nominated a task force for the development of guidelines for HIT in January 2010. Following wide-ranging discussion, the guidelines were adopted in May 2011.
Trombocitopenija izazvana heparinom (HIT) teška je nuspojava heparinske terapije. U posljednjih desetak godina ovaj kliničkopatološki sindrom u središtu je interesa primarno zbog teških ...tromboembolijskih komplikacija, koje mogu imati i smrtni ishod. Znatno poboljšanje u liječenju HIT-a, postignuto je primjenom direktnih inhibitora trombina u zamjenu za heparin, a laboratorijsko ispitivanje antiheparinskih protutijela znatno je unaprijedilo dijagnostiku HIT-a. Uvođenje smjernica za dijagnostičko-terapijski postupak za HIT ima znatan učinak na kvalitetu liječenja bolesnika. Godine 2008. u Republici Hrvatskoj (RH) pokrenut je niz aktivnosti u cilju uspostavljanja smjernica za HIT, temeljenih na britanskim preporukama za dijagnostiku i liječenje trombocitopenije izazvane heparinom iz 2006. godine. Tijekom 2008/09. godine uvedeni su novi testovi za antiheparinska protutijela, algoritam laboratorijskog ispitivanja i obrazac za kliničku procjenu HIT-a te izobrazba transfuziologa i kliničara. U siječnju 2010. godine na stručnom sastanku Hrvatskog društva za hematologiju i transfuzijsku medicinu (HDHTM), nakon evaluacije rezultata jednogodišnje primjene preporuka osnovana je radna skupina za donošenje smjernica HDHTM-a za HIT. Nakon usuglašavanja i javne rasprave smjernice su prihvaćene u svibnju 2011. godine.
Nucleotides, glycosaminoglycans, and omega-3 essential fatty acids (O3s) could be used for improving skin health, although their modes of action, alone or in combination, are not yet fully ...understood. To gain some insight into these mechanisms, we performed two in vitro tests and one in vivo pilot trial. The effects on human dermal fibroblast proliferation and migration were evaluated with the following compounds and combinations: 0.156 mg/mL O3s, 0.0017 mg/mL hyaluronic acid (HA), 0.0004 mg/mL dermatan sulfate (DS), 0.0818 mg/mL nucleotides, and O3s + HA + DS and O3s + HA + DS + nucleotides at the same concentrations. In both in vitro assays, adding nucleotides to O3s + HA + DS provided significant improvements. The resulting combination O3s + HA + DS + nucleotides was then tested in vivo in dogs with atopic dermatitis by oral administration of a supplement providing a daily amount of 40 mg/kg nucleotides, 0.9 mg/kg HA, 0.18 mg/kg DS, 53.4 mg/kg EPA, and 7.6 mg/kg DHA. After 30 days, the pruritus visual analog scale (pVAS) score was significantly reduced, and no adverse effects were observed. In conclusion, the combination of nucleotides plus glycosaminoglycans and O3s could serve as a useful therapeutic alternative in skin health applications.
Relying on a recent re-conceptualization of psychosis proneness as a personality trait, its relations with the Big Five traits were investigated in a meta-analytic study. This re-conceptualized trait ...– named Disintegration – is articulated as a broad, hierarchically organized, nine-faceted behavioral disposition. Disintegration is postulated to be a basic personality trait distinct from the Big Five traits. In accordance with this conceptualization, all the articles considered for this meta-analysis carry information on the relationship between Disintegration-like phenomena (referring to various aspects of symptomatology with prefix ‘schizo-’, both at the clinical and the sub-clinical level), and at least one Big Five trait. The benchmark for assuming distinctness of the trait Disintegration was .40, based on the meta-analytically derived correlations found among the Big Five traits. By computing inverse sampling variance weighted mean correlation coefficients under a random-effects assumption, the following associations were found between Disintegration and N, E, O, A, and C, respectively: .24, −.27, 0, −.19, and −13. The differences in true correlations between the studies were substantial for each coefficient. Three variables were found to moderate Disintegration–personality correlations. The finding about the distinctness of Disintegration from other personality traits can have repercussions on the taxonomy of traits.
•Disintegration-like phenomena refer to various behaviors with prefix ‘schizo-’.•Meta-analysis was used to test if Disintegration is distinct from Big Five traits.•Disintegration phenomena and related clinical states are distinct from Big Five.•Disintegration–Big Five relations differ for positive compared to negative symptoms.•Disintegration–Big Five relations are the same in clinical and nonclinical samples.
This meta-analytic study investigates the relations
between Disintegration-like phenomena (i.e., various aspects of symptomatology
with the prefix "schizo-," both at the clinical and the
subclinical ...level) and the traits of the Psychobiological Model of Personality
(PBMP). The empirically based benchmark for assuming the distinctness of the
trait Disintegration was .30. The sample included 26 manuscripts with 30 studies
and 424 effect sizes. By computing inverse sampling variance weighted mean
correlation coefficients under a random-effects assumption, the following
associations were found between Disintegration and Harm Avoidance, Novelty
Seeking, Reward Dependence, Persistence, Self-Directedness, Cooperativeness, and
Self-Transcendence: .23, .04, −.15, −.02, −.23,
−.16, and .17, respectively. Two variables were found to moderate the
Disintegration-Self-Transcendence correlation. Despite the theoretical
expectation and some empirical evidence that Self-Transcendence (and other
character traits) should capture variations in Disintegration-like phenomena,
our results suggest that schizo-type phenomena are not adequately covered by the
PBMP.
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