Temporal sequence of the human RBCs' vesiculation observed in nano-scale with application of AFM and complementary techniques

Based on the multimodal characterization of human red blood cells (RBCs), the link between the storage-related sequence of the nanoscale changes in RBC membranes in the relation to their biochemical profile as well as mechanical and functional properties was presented. On the background of the accumulation of RBCs waste products, programmed cell death and impaired rheological properties, progressive alterations in the RBC membranes including changes in their height and diameter as well as the in situ characterization of RBC-derived microparticles (RMPs) on the RBCs surface were presented. The advantage of atomic force microscopy (AFM) in RMPs visualization, even at the very early stage of vesiculation, was shown based on the results revealed by other reference techniques. The nanoscale characterization of RMPs was correlated with a decrease in cholesterol and triglycerides levels in the RBC membranes, proving the link between the lipids leakage from RBCs and the process of vesiculation

Spectroscopic signature of red blood cells in a D-galactose-induced accelerated aging model

This work presents a semi-quantitative spectroscopic approach, including FTIR-ATR and Raman spectroscopies, for the biochemical analysis of red blood cells (RBCs) supported by the biochemical, morphological and rheological reference techniques. This multi-modal approach provided the description of the RBC alterations at the molecular level in a model of accelerated aging induced by administration of D-galactose (D-gal), in comparison to natural aging. Such an approach allowed to conclude that most age-related biochemical RBC membrane changes (a decrease in lipid unsaturation and the level of phospholipids, or an increase in acyl chain shortening) as well as alterations in the morphological parameters and RBC deformability are well reflected in the D-gal model of accelerated aging. Similarly, as in natural aging, a decrease in LDL level in blood plasma and no changes in the fraction of glucose, creatinine, total cholesterol, HDL, iron, or triglycerides were observed during the course of accelerated aging. Contrary to natural aging, the D-gal model led to an increase in cholesterol esters and the fraction of total esterified lipids in RBC membranes, and evoked significant changes in the secondary structure of the membrane proteins. Moreover, a significant decrease in the phosphorous level of blood plasma was specific for the D-gal model. On the other hand, natural aging induced stronger changes in the secondary structures of the proteins of the RBCs' interior. This work proves that research on the aging mechanism, especially in circulation-related diseases, should employ the D-gal model with caution. Nonetheless, the D-gal model enables to imitate age-related rheological alterations in RBCs, although they are partially derived from different changes observed in the RBC membrane at the molecular level

Trends in biomedical analysis of red blood cells – Raman spectroscopy against other spectroscopic, microscopic and classical techniques

Application of modern and innovative spectroscopic and microscopic approaches to biomedical analysis opens new horizons and sheds new light on many unexplored scientific territories. In this review, we critically summarize up-to-date Raman-based methodologies for red blood cells (RBCs) analysis used in biology and medicine, and compare them with both classical, as well as other spectroscopic and microscopic approaches. The main emphasis is placed on the advantages, disadvantages and capabilities of each technique for detection of RBC deteriorations and RBC-related diseases. Although currently used classical techniques of medical analysts serve as a gold standard for clinicians in diagnosis of erythropathies, they provide insufficient insight into RBC alterations at the molecular level. In addition, there is a demand for non-destructive and label-free analytical techniques for rapid detection and diagnosis of erythropathies. Their recognition often requires multimodal methodology comprising application of methods including sophisticated spectroscopy-based techniques, where Raman-based approaches play an important role

Temporal relationship between systemic endothelial dysfunction and alterations in erythrocyte function in a murine model of chronic heart failure

Endothelial dysfunction (ED) and red blood cell distribution width (RDW) are both prognostic factors in heart failure (HF), but the relationship between them is not clear. In this study, we used a unique mouse model of chronic HF driven by cardiomyocyte-specific overexpression of activated Gαq protein (Tgαq*44 mice) to characterise the relationship between the development of peripheral ED and the occurrence of structural nanomechanical and biochemical changes in red blood cells (RBCs).Systemic ED was detected in vivo in 8-month-old Tgαq*44 mice, as evidenced by impaired acetylcholine-induced vasodilation in the aorta and increased endothelial permeability in the brachiocephalic artery. ED in the aorta was associated with impaired nitric oxide (NO) production in the aorta and diminished systemic NO bioavailability. ED in the aorta was also characterised by increased superoxide and eicosanoid production. In 4- to 6-month-old Tgαq*44 mice, RBC size and membrane composition displayed alterations that did not result in significant changes in their nanomechanical and functional properties. However, 8-month-old Tgαq*44 mice presented greatly accentuated structural and size changes and increased RBC stiffness. In 12-month-old Tgαq*44 mice, the erythropathy was featured by severely altered RBC shape and elasticity, increased RDW, impaired RBC deformability, and increased oxidative stress (GSH/GSSH ratio). Moreover, RBCs taken from 12-month-old Tgαq*44 mice, but not from 12-month-old FVB mice, co-incubated with aortic rings from FVB mice, induced impaired endothelium-dependent vasodilation and this effect was partially reversed by an arginase inhibitor (ABH, 2(S)-amino-6-boronohexanoic acid).In the Tgαq*44 murine model of HF, systemic endothelial dysfunction accelerates erythropathy and, conversely, erythropathy may contribute to endothelial dysfunction. These results suggest that erythropathy may be regarded as a marker and a mediator of systemic endothelial dysfunction in HF. In particular, targeting RBC arginase may represent a novel treatment strategy for systemic endothelial dysfunction in HF. RBC arginase and possibly other RBC-mediated mechanisms may represent novel therapeutic targets for systemic endothelial dysfunction in HF.Endothelial dysfunction (ED) and red blood cell distribution width (RDW) both have prognostic value for heart failure (HF), but it is not known whether these pathologies are related. We comprehensively characterized endothelial and RBC functional status in a unique murine model of chronic heart failure with a prolonged time course of HF progression. Our results suggest that ED accelerates erythropathy and, conversely, erythropathy may contribute to ED. Accordingly, erythropathy in HF reflects ED and involves various changes (in functional, structural, nanomechanical, and biochemical levels) that could have diagnostic and therapeutic significance for HF

The development of a FTIR- based analysis of plasma from an animal model of breast cancer

Nowotwór piersi należy do grupy chorób, które przy odpowiednio wczesnej diagnostyce, są aktualnie łatwe do wyleczenia. Problem pojawia się wówczas, gdy guz pierwotny zaczyna się rozprzestrzeniać za pomocą naczyń krwionośnych lub limfatycznych do innych narządów w organizmie, takich jak płuca, wątroba czy mózg. Mechanizm przerzutu nie jest jeszcze do końca znany, co zdecydowanie utrudnia wczesne wykrycie przerzutowości jeszcze przed pojawieniem się nowotworu wtórnego. Obecnie poszukuje się nowych metod badawczych, które pozwoliłyby nie tylko na rozpoznanie choroby nowotworowej, ale także śledzenie jej postępu. Ogromne nadzieje wiązane są z mikroskopią FTIR, która daje możliwość zbadania całego profilu biochemicznego tkanek czy płynów ustrojowych także na poziomie mikronowym. W niniejszej pracy przy wykorzystaniu metody mikroskopii FTIR scharakteryzowano zmiany spektralne widm osocza modelu zwierzęcego zachodzące podczas rozwoju nowotworu sutka. Analiza otrzymanych wyników pokazała, że na poszczególnych etapach progresji choroby istnieją różnice w składzie biochemicznym krwi. Za pomocą otrzymanych widm FTIR możliwe było wyróżnienie szczególnych cech odróżniających konkretne stopnie zaawansowania choroby. Stwierdzono, że kluczowe znaczenie w tym przypadku mają zakresy spektralne związane z lipidami, białkami oraz kwasami nukleinowymi.Breast cancer belongs to the group of diseases which can be cured by an early diagnosis. However, an issue appears when the primary tumour starts to metastase to distant organs such as lungs, liver or brain through blood or lymph vessels. A full understanding of a mechanism of metastasis is unknown yet and this fact limits its early detection before emergence of secondary tumour. Nowadays, scientists search new tools of diagnosis which will enable not only recognise cancer disease but also detect its progression and metastasis. One of techniques which can be very useful for such an application is FTIR microscopy that easily provide an overall biochemical composition of tissues and bodily fluids on a micron level. In this study we characterized spectral changes of animal model plasma using microscopy FTIR. A murine model represents metastasis of breast cancer to lungs. A spectroscopic analysis of blood plasma showed that consecutive steps of metastasis progression exhibited significant differences in biochemical composition of blood plasma. Several IR features we were observed which discriminated precisely steps of cancer progression. A key spectral changes were found in regions attributed to vibrations of lipids, proteins and nucleic acids