Extracellular vesicles (EVs) as membrane-bound particles released by various cells are potential tools for diagnosis and treatment. Blood cells, particularly platelets, are the source of circulating ...EVs.
EVs were enriched with gradient ultracentrifugation and measured by nanoparticle tracking assay. A flow cytometric multiplex assay was used for cellular source determination. Activation of platelets was measured as a percentage of CD62p+/CD61+ platelets and correlated with the concentration and size of released EVs.
In general there was no statistically significant correlation between EVs` concentration and degree of platelet activation. EVs from different cellular sources were detected. Comparing different needle thicknesses, there was a decrease in the EVs concentration for the 16G needle versus the 21G needle, but no difference was observed for EVs` size and phenotype or platelets activation. During blood storage, platelet activation increased, but there was no effect on the EVs` concentration, size, or phenotype.
Preanalytical factors like needle thickness and storage time can affect the MVs' properties. Activation of platelets during blood collection or blood storage occurs; however, it is difficult to determine its effect on the physiological properties of EVs since the mechanisms of EVs` biogenesis and especially clearness are not precisely known.
This study examined the influence of platelet activation on the characteristics of EVs in blood•Two different needle sizes influenced EVs` concentration but not size and phenotype or platelet activation•Short blood storage infuenced platelet activation, but not EVs` concentration, size or phenotype•It appears that blood EVs` concentration is in an equilibrium between cellular release and uptake•We have detected EVs derived from platelets and WBC, bone marrow, epithelium, endothelium, and other body cells
Heat inactivation of a patient's sample is not systematically performed in the diagnostics of heparin-induced thrombocytopenia (HIT). Some authors recommend that the patient's sample is ...heat-inactivated to avoid the effect of thrombin on platelet activation in a functional assay. Others do not find this additional step essential or even advise against it.
An enzyme-linked immunosorbent assay (ELISA) and flow cytometry-based functional assay with CD62P as a marker of platelet activation were performed. Forty-seven patients with suspected HIT and three healthy controls were included in the study. Each serum sample was divided into two aliquots: one was heat-inactivated and the other was not. Both aliquots were tested in parallel using the same donor platelets from four randomly selected individuals. We designed an index of platelet activation for both protocols to assess platelet activation in the assay and to compare the results.
We observed a higher percentage of platelet activation in heat-inactivated compared to non-heat-inactivated sera. This phenomenon was seen in low and high heparin steps, although it did not occur for all samples. There were discrepant results in seven samples, which tested negative in the non-heat-inactivated protocol and positive in the heat-inactivated protocol. There was no case in which the result of a non-heat-inactivated aliquot was positive and the corresponding heat-inactivated aliquot was negative.
Due to the higher percentages of donor platelet activation, all seven non-compliant cases met our current criteria for a positive result. However, those results were probably false-positive based on ELISA optical density value and 4T score. Therefore, in the current settings, heat inactivation of serum is not suitable for our flow cytometric functional assay since it can cause an elevated risk of creating false-positive results.
The aim of the present research was to determine the haemato-biochemical profile and blood acid–base status of Croatian spotted goats in a traditional Mediterranean production system. The 60 ...non-gravid female Croatian spotted goats of different ages were included in the research. They were divided into four groups of 15 goats according to age: group I – ≤1 year old; group II – 2–3 years; group III – 3–6 years; and group IV – 7–10 years. Haematological parameters were determined in whole blood, biochemical parameters in serum and acid–base status in plasma by automatic analyser. Total leukocyte number (WBC), haemoglobin (HGB) and mean corpuscular volume (MCV) in the blood were the highest, while mean haemoglobin concentration in erythrocytes (MCHCs) was the lowest in yearlings compared to other groups. Concentrations of urea, Mg, Cl, non-esterified fatty acids (NEFAs) and lactate were the highest in yearlings. Concentrations of Ca, Na, total cholesterol, high-density lipoprotein (HDL), very low-density lipoprotein (VLDL) and beta hydroxybutyrate (BHB) as well as the activity of alanine aminotransferase (ALT) were higher in older goats compared to yearlings, while the opposite was determined for the activities of creatine kinase (CK) and alkaline phosphatase (ALP). Values of pH, the strong ion difference (SID), anion gap (AG) and z values as well as the content of HCO3 and total pressure of carbon dioxide (ctCO2) were higher in older goats compared to yearlings. The results obtained may help in monitoring the health and nutritional status and improve the management of Croatian spotted goats. Based on the results of the present study, the effect of age needs to be included in the model when preparing the reference values for the haemato-biochemical profile and acid–base status of goats.
The research was conducted on days 60, 90, 120 and 150 during the lactation with the aim to determine the effect of lactation stage on the concentration of crucial elements and chemical composition ...in Croatian spotted goats' milk. Milk yield significantly decreased (from 1.15 to 0.76 kg/day), while the concentration of Ca, Mg, P, Zn and Mo in the milk of the goats significantly increased during lactation. The concentrations of Cr, Cd and Pb in milk were very low, with no significant deviations during lactation. The quality of milk from Croatian spotted goats is adequate compared with other breeds in the surrounding area of Croatia.
The aim of the present study was to determine the lactation stage effect on the concentration of essential and selected toxic elements in the sheep’s milk of Dubrovačka ruda. The research was ...conducted with 23 sheep, average age of 4 years, of 3rd lactation, while the milk samples were taken during the early (60th day), middle (90th day) and late (120th day) lactation stage. The sheep were selected according to uniformed body development, adequate health status, body condition, equable age (4 years), parity (3rd lactation), stage of lactation (±7 days) and litter size (single). Sheep were reared on the extensive Mediterranean pastures, reared indoors afterwards, fed with hay ad libitum and feed mixtures in average 0.5 kg/day. Milk sample was collected during morning milking from each sheep. The digested samples were analyzed with continuous flow hydride generation technique using inductively coupled plasma for Ca, Mg, K, P, Na, Cu, Fe, Zn, Mn, Ni, Mo, Co, Cr, Cd and Pb concentrations. Significant increase of Mg, Na, Se, Mn, Mo and Cd concentrations were found in milk as well as decrease of K concentration during the lactation. Although the concentration of Ca, Cu, Cr and As in milk during the lactation is increased, the differences between the lactation stages were not observed. Concentrations of P, Fe, Ni, Pb and Hg in milk of Dubrovačka ruda did not differ during the lactation. The low concentrations of Cr, Cd, Pb, As, Hg in milk indicate the safety for consumers and preserved environment of Dubrovnik-Neretva County.
Background and Objectives: Most patients who are successfully resuscitated from cardiac arrest remain comatose, and only half regain consciousness 72 h after the arrest. Neuroprognostication methods ...can be complex and even inconclusive. As mitochondrial components have been identified as markers of post-cardiac-arrest injury and associated with survival, we aimed to investigate cytochrome c and mtDNA in comatose patients after cardiac arrest to compare neurological outcomes and to evaluate the markers’ neuroprognostic value. Materials and Methods: This prospective observational study included 86 comatose post-cardiac-arrest patients and 10 healthy controls. Cytochrome c and mtDNA were determined at admission. Neuron-specific enolase (NSE) was measured after 72 h. Additional neuroprognostication methods were performed when patients remained unconscious. Cerebral performance category (CPC) was determined. Results: Cytochrome c was elevated in patients compared to healthy controls (2.029 0.85–4.97 ng/mL vs. 0 0.0–0.16, p < 0.001) but not mtDNA (95,228 52,566–194,060 vs. 41,466 28,199–104,708 copies/μL, p = 0.074). Compared to patients with CPC 1–2, patients with CPC 3–5 had higher cytochrome c (1.735 0.717–3.40 vs. 4.109 1.149–8.457 ng/mL, p = 0.011), with no differences in mtDNA (87,855 47,598–172,464 vs. 126,452 69,447–260,334 copies/μL, p = 0.208). Patients with CPC 1–2 and CPC 3–5 differed in all neuroprognostication methods. In patients with good vs. poor neurological outcome, ROC AUC was 0.664 (p = 0.011) for cytochrome c, 0.582 (p = 0.208) for mtDNA, and 0.860 (p < 0.001) for NSE. The correlation between NSE and cytochrome c was moderate, with a coefficient of 0.576 (p < 0.001). Conclusions: Cytochrome c was higher in comatose patients after cardiac arrest compared to healthy controls and higher in post-cardiac-arrest patients with poor neurological outcomes. Although cytochrome c correlated with NSE, its neuroprognostic value was poor. We found no differences in mtDNA.
Respiratory transfusion-related reactions are not very frequent, partly also because recognition and reporting transfusion reactions is still underemphasized. Tis article describes the most important ...respiratory transfusion reactions, their pathophysiology, clinical picture and treatment strategies. Respiratory transfusion related reactions can be primary or secondary. The most important primary transfusion-related reactions are TRALI - transfusion-related acute lung injury, TACO – transfusion-associated circulatory overload, and TAD - transfusion-associated dyspnea. TRALI is immuneassociated injury of alveolar basal membrane, which becomes highly permeable and causes noncardiogenic pulmonary edema. Treatment of TRALI is mainly supportive with oxygen, fluids (in case of hypotension) and in cases of severe acute respiratory failure also mechanic ventilation. TACO is caused by volume overload in predisposed individuals, such as patients with heart failure, the elderly, infants, patients with anemia and patients with positive fluid balance. Clinical picture is that of a typical pulmonary cardiogenic edema, and the therapy is classical: oxygen and diuretics, and in severe cases also non-invasive or invasive mechanical ventilation. TAD is usually a mild reaction of unknown cause and cannot be classified as TACO or TRALI, nor can it be ascribed to patient’s preexisting diseases. Although the transfusion-related reactions are not very common, knowledge about them can prevent serious consequences. On the one hand preventive measures should be sought, and on the other early recognition is beneficial, so that proper treatment can take place.
Respiratory transfusion-related reactions are not very frequent, partly also because recognition and reporting transfusion reactions is still underemphasized. Tis article describes the most important ...respiratory transfusion reactions, their pathophysiology, clinical picture and treatment strategies. Respiratory transfusion related reactions can be primary or secondary. The most important primary transfusion-related reactions are TRALI - transfusion-related acute lung injury, TACO – transfusion-associated circulatory overload, and TAD - transfusion-associated dyspnea. TRALI is immuneassociated injury of alveolar basal membrane, which becomes highly permeable and causes noncardiogenic pulmonary edema. Treatment of TRALI is mainly supportive with oxygen, fluids (in case of hypotension) and in cases of severe acute respiratory failure also mechanic ventilation. TACO is caused by volume overload in predisposed individuals, such as patients with heart failure, the elderly, infants, patients with anemia and patients with positive fluid balance. Clinical picture is that of a typical pulmonary cardiogenic edema, and the therapy is classical: oxygen and diuretics, and in severe cases also non-invasive or invasive mechanical ventilation. TAD is usually a mild reaction of unknown cause and cannot be classified as TACO or TRALI, nor can it be ascribed to patient’s preexisting diseases. Although the transfusion-related reactions are not very common, knowledge about them can prevent serious consequences. On the one hand preventive measures should be sought, and on the other early recognition is beneficial, so that proper treatment can take place.
Transfuzijske reakcije niso pogoste, vendar kljub temu verjetno pogostejše, kot jih ocenjujemo na splošno, saj je prepoznavanje in poročanje o transfuzijskih reakcijah še vedno pomanjkljivo. Članek ...opisuje najpogostejše respiracijske transfuzijske reakcije, patofziologijo le-teh, klinično sliko in zdravljenje. Med primarnimi transfuzijskimi reakcijami so najpomembnejše: s transfuzijo povezana akutna okvara pljuč oz. TRALI, s transfuzijo povezana volumska preobremenitev oz. TACO ter s transfuzijo povezana dispneja oz. TAD. Pri TRALI pride do aktiviranja imunskega sistema, katerega posledica je povečana prepustnost bazalne membrane v pljučnih mešičkih, kar vodi v nekardiogeni pljučni edem. Zdravljenje je podporno z dodajanjem kisika, tekočin ob hipotenziji, v hujših primerih dihalne odpovedi tudi mehansko predihavanje. Klinična slika TACO je kardiogeni pljučni edem, do katerega pride zaradi volumske obremenitve pri predisponiranih osebah, ki so: dojenčki, starostniki, bolniki s srčnim popuščanjem, anemijo in bolniki s pozitivno tekočinsko bilanco. TACO zdravimo po načelih zdravljenja kardiogenega pljučnega edema z dodajanjem kisika, diuretiki, po potrebi pa tudi z neinvazivnim ali invazivnim mehanskim predihavanjem. Pri TAD pa gre za etiološko nepojasnjeno dispnejo, ki se pojavi po transfuziji in je ne moremo opredeliti kot TACO ali TRALI, prav tako pa je ne pojasnijo bolnikove kronične bolezni. Ponavadi je TAD blaga reakcija. Čeprav transfuzijske reakcije niso zelo pogoste, je znanje o prepoznavanju, zdravljenju in preprečevanju respiracijskih transfuzijskih reakcij nujno potrebno, da se preprečijo resne posledice pri bolnikih.
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