Proteomic and biomarker studies and neurological complications of pediatric sickle cell disease

Biomarker analysis and proteomic discovery in pediatric sickle cell disease has the potential to lead to important discoveries and improve care. The aim of this review article is to describe proteomic and biomarker articles involving neurological and developmental complications in this population. A systematic review was conducted to identify relevant research publications. Articles were selected for children under the age of 21 years with the most common subtypes of sickle cell disease. Included articles focused on growth factors (platelet-derived growth factor), intra and extracellular brain proteins (glial fibrillary acidic protein, brain-derived neurotrophic factor), and inflammatory and coagulation markers (interleukin-1β, l -selectin, thrombospondin-1, erythrocyte, and platelet-derived microparticles). Positive findings include increases in plasma brain-derived neurotrophic factor and platelet-derived growth factor with elevated transcranial Dopplers velocities, increases in platelet-derived growth factor isoform AA with overt stroke, and increases in glial fibrillary acidic protein with acute brain injury. These promising potential neuro-biomarkers provide insight into pathophysiologic processes and clinical events, but their clinical utility is yet to be established. Additional proteomics research is needed, including broad-based proteomic discovery of plasma constituents and blood cell proteins, as well as urine and cerebrospinal fluid components, before, during and after neurological and developmental complications.     

Biomarkers for the central nervous system complications of sickle cell disease: are we there yet?

Sickle cell disease (SCD, OMIM #603903), an autosomal recessively inherited β-hemoglobinopathy, was the first human disorder delineated at a molecular level. The putative single nucleotide mutation in the HBB gene generates an abnormal hemoglobin species, which polymerizes in deoxygenated conditions causing irreversible changes in erythrocyte shape and function. Sickling erythrocytes are in turn responsible for microvascular vaso-occlusion, hemolysis and a systemic vasculopathy in patients. SCD has represented an attractive field for proteomic investigation since its methodological infancy. Clinically actionable biomarkers, especially for the prevention of cerebrovascular complications in children with the condition, are urgently needed and their discovery remains a major challenge. In this issue, Lance and colleagues report of their unbiased proteomic studies on samples from the participants of the landmark prospective, randomized, single-blind SIT trial (NEJM 2014). Their results reveal numerous brain-enriched plasma proteins specific for SCD, and for silent cerebral infarcts in this disorder, and further analyses highlight novel cellular mechanisms behind the brain damage in SCD. Although the goal of identifying reliable biomarker candidates for cerebrovascular complications could not be met, the dataset produced by the authors constitutes a significant contribution to the field and opens new horizons for further clinical and laboratory investigation.     

Ergonomics in sickle operation

Sickle operation in harvesting has been analysed with reference to design features of nine different types of sickles, and field and laboratory based investigations on biomechanical stresses and physiological valuation on six farmers. It has been indicated that the blade geometry contributes significantly to human performance and there is ample scope for further design optimisation. The suggested modifications are: (i) sickle weight - 200 g; (ii) total length of sickle - 33 cm; (iii) handle length - 11 cm; (iv) handle diameter - 3 c cm; (v) radius of blade curvature - 15 cm; (vi) blade concavity - 5 cm; (vii) serrated sickle: tooth pitch - 0.20 cm and tooth angle - 60 degrees; (viii) ratio of the length of cutting surface to chord length - 1.20.     

Heat-shock protein-27, -70 and peroxiredoxin-II show molecular chaperone function in sickle red cells: Evidence from transgenic sickle cell mouse model

Sickle cell disease (SCD) is an autosomal recessive genetic red cell disorder characterized by the production of a defective form of hemoglobin, hemoglobin-S, that is worldwide-distributed. The acute clinical manifestations of SCD are related to hemoglobin cyclic-polymerization and to the generation of rigid, dense red blood cells (RBCs). We studied RBCs membrane proteome from human sickle RBCs, fractioned according to density compared to normal RBCs. 2-DE followed by MS analysis was carried out. We identified 65 proteins differently expressed, divided into five major clusters according to their functions: (i) membrane-cytoskeleton proteins; (ii) metabolic enzymes; (iii) ubiquitin-proteasome-system; (iv) flotillins; (v) chaperones. HSP27, HSP70 and peroxiredoxin-II (Prx-II) showed the most relevant changes. They were differently recruited to sickle RBCs membrane in response to in vitro hypoxia. Potential markers were then validated in a transgenic-mouse model for SCD, the SAD mice, exposed to hypoxia mimicking acute SCD vaso-occlusive-crisis (VOCs); we found that HSP70 and HSP27 bound to RBCs membrane respectively after 12?h and 48?h of hypoxia, while Prx-II membrane binding was modulated during hypoxia. Our data indicate that HSP27 and HSP70 play a novel role as RBCs membrane protein protectors and as possibly new markers of severity of RBCs membrane damage during acute VOCs.     

Biomarkers as disease definition: mantle cell lymphoma as an example

The introduction in pathology practice of a new biomarker that is contributing to define a disease requires a series of investigations. Now that new biomarkers are being discovered continuously it is important to learn from successful examples of markers that are presently widely used. In this historical account the steps are described that have led to the use of immunohistochemical staining of tissue samples with an antibody against cyclinD1 to diagnose mantle cell lymphoma. Furthermore, a short outlook is given on the introduction of proteomics as a tool in the diagnosis of lymphoma and the potential route to be taken for introducing this technology into clinical practice.     

Detection of anaemia disease in human red blood cells using cell signature,neural networks and SVM

Multimedia Tools and Applications - Anaemia disease attacks and deforms the circular red blood cells. Latterly, it has classified as a very dangerous disease. Many papers have been presented...     

A framework for diagnosing cervical cancer disease based on feedforward MLP neural network and ThinPrep histopathological cell image features

In this paper, Levenberg–Marquardt feedforward MLP neural network (LMFFNN) was proposed to classify cervical cell images obtained from 100 patients including healthy, low-grade intraepithelial squamous lesion and high-grade intraepithelial squamous lesion cases. This neural network along with extracted cell image features is a new model for cervical cell image classification. The semiautomated cervical cancer diagnosis system is composed of two phases: image preprocessing/processing and feedforward MLP neural network. In the first stage, image preprocessing is done to reduce the existing noises without lowering the resolution. After that, image processing algorithms were applied to manually cropped cell images to achieve a linear plot which includes real components, were used as LMFFNN inputs for classification of cervical cell images. Based on the results, cervical cell images were classified successfully with 100 % correct classification rate using the proposed method. Moreover, the rates of sensitivity and specificity were calculated as 100 % using LMFFNN method. It was shown there was a good agreement between the expert decision and values gained from the ANN model.     

Redox signalling in cardiovascular disease

Oxidative stress has almost universally and unequivocally been implicated in the pathogenesis of all major diseases, including those of the cardiovascular system. Oxidative stress in cells and cardiovascular biology was once considered only in terms of injury, disease and dysfunction. However, it is now appreciated that oxidants are also produced in healthy tissues, and they function as signalling molecules transmitting information throughout the cell. Conversely, when cells move to a more reduced state, as can occur when oxygen is limiting, this can also result in alterations in the function of biomolecules and subsequently cells. At the centre of this 'redox signalling' are oxidoreductive chemical reactions involving oxidants or reductants post translationally modifying proteins. These structural alterations allow changes in cellular redox state to be coupled to alterations in cell function. In this review, we consider aspects of redox signalling in the cardiovascular system, focusing on the molecular basis of redox sensing by proteins and the array of post-translational oxidative modifications that can occur. In addition, we discuss studies utilising proteomic methods to identify redox-sensitive cardiac proteins, as well as those using this technology more broadly to assess redox signalling in cardiovascular disease.     

Neuroproteomics in stem cell differentiation

The term "proteome" is used to describe the entire complement of proteins in a given organism or in a system at a given time. Proteome analysis in neuroscience, also called "neuroproteomics" or "neuromics" is in its initial stage, and shows a deficit of studies in the context of brain development. It is the main objective of this review to illustrate the potential of neuroproteomics as a tool to unravel the differentiation of neural stem or progenitor cells to terminally differentiated neurons. Experimental results regarding the rat striatal progenitor model cell line ST14A are presented to illustrate the large rearrangements of the proteome during the differentiation process of neural progenitor cells and their modification by neurotrophic factors like the glial cell line-derived neurotrophic factor (GDNF). Thereby native stem cells and cells transfected with GDNF gene were investigated at the proliferative state and at seven time points up to 72?h after induction of differentiation. In addition, the immortalized human fetal midbrain stem cell line ReNcell VM was analyzed in order to detect stem cell differentiation associated changes of the protein profile. This review gives also an outlook on technical improvements and perspectives of application of neural stem cell proteomics.     

Mechatronics in fuel cell systems

Power generation from fuel cells (FCs) requires the integration of chemical, fluid, mechanical, thermal, electrical, and electronic subsystems. This integration presents many challenges and opportunities in the mechatronics field. This paper highlights important design issues and poses problems that require mechatronics solutions. The paper begins by describing the process of designing a toy school bus powered by hydrogen for an undergraduate student project. The project was an effective and rewarding educational activity that revealed complex systems issues associated with FC technology.     

Autowave processes in cell structures

Autowave processes connected with activation of calcium ions depots in myocytes are investigated. Mechanisms of autoorganization (structurization) are proposed and dissipative structure reorganizations of calcium activity waves are considered.     

Proteomics of citrullination in cardiovascular disease

Citrullination is an enzymatic posttranslational modification (PTM), which has become a topic of recent research due to its involvement in various physiologic and pathologic processes. This review will focus primarily on the cardiovascular pathology associated to date with citrullination, including myocardial citrullination as well as the potential role of citrullination in atherosclerosis as a driver inflammation, especially in patients with rheumatoid arthritis (RA). There is extensive citrullination within normal and RA myocardium as well as the atherosclerotic plaque; and increased levels of antibodies to citrullinated proteins have been associated with increased cardiovascular burden in patients with RA. Given robust citrullination in both RA as well as non-RA patients, there also exists the potential for protein citrullination to contribute to cardiovascular pathology in the general population. However, to investigate this possibility will require development of improved biochemical and proteomic tools for the study of protein citrullination. The remainder of this review will discuss current and developing methodologies to study protein citrullination and discuss their applicability for the analysis of complex samples. The ability to identify and quantify citrullinated protein is a key to understanding the role of this PTM. Methodologies and limitations of current technology for the identification of citrullination are discussed.     

Manufacturing cell formation in the presence of flexible cell locations and material transporters

The effect of cell locations and material transporters in the formation of manufacturing cells is investigated in this paper. Automated guided vehicles (AGVs) using a tandem configuration are considered and a first-come-first-served (FCFS) principle is applied for transporting the material between machines or between the input/output (I/O) and a machine. Using the time taken to perform material transfers as a suitable measure, a polynomial programming model is developed for the problem. As the model can be shown strongly NP-hard, a higher-level heuristic algorithm based upon a concept known as ‘tabu search’ is presented. An example problem is solved to further demonstrate that cell locations indeed have a significant impact when material transfers are used in the design of manufacturing cells.     

The proteomic approach in Parkinson's disease

Parkinson's disease (PD) is a complex, multifactorial neurodegenerative disease affecting about 2% of the population over 65?years. Etiopathogenetic mechanisms of PD are not fully understood, although a number of factors contributing to the selective degeneration of substantia nigra neurons have been identified, including mitochondrial dysfunction, proteasomal impairment, oxidative stress, excitotoxicity, and inflammation. Although a global view of the disease at the molecular level can be obtained only from the biochemical analysis of the affected human tissue, difficulties in obtaining human specimens of the affected area have limited substantially the number of reports published to date. Therefore, cellular and animal models of the disease have been developed to investigate single factors contributing to disease pathogenesis, e.g., protein aggregation or altered dopamine homeostasis. In this review, we report how proteomic methodologies have been used so far to investigate cellular and animal models of PD, as well as to compare postmortem specimens of substantia nigra of affected patients to that of control subjects. Proteomic studies concur to highlight the role of a compromised antioxidant defense in PD pathogenesis. The proteomic approach in the investigation of etiopathogenetic mechanisms of PD is still at its beginning, however, the findings reviewed here should serve as a useful foundation to further work.     

Modelling immunomagnetic cell capture in CFD

The separation of cells from a complex sample by immunomagnetic capture has recently obtained increased attention for microfluidic applications. Here, we present a simulation approach for immunomagnetic separation in a flow-through microfluidic environment that for the first time takes binding kinetics of beads to target cells as well as binding of multiple beads per cell into account. The approach is implemented into a computational fluid dynamics code and facilitates the tailored design of microfluidic magnetophoretic devices with an optimised separation performance. Although the specific computational model under study is constrained to a 2D geometry, appropriate parameter sets that allow for a continuous separation of cell/bead complexes from non-magnetic particles could be derived. In addition, based on magnetophoretic mobilities, a critical threshold value of beads per cell is revealed, where further binding is considerably reduced or the reaction cascade ceases.     

Single cell trapping in larger microwells capable of supporting cell spreading and proliferation

Conventional cell trapping methods using microwells with small dimensions (10–20 μm) are useful for examining the instantaneous cell response to reagents; however, such wells have insufficient space for longer duration screening tests that require observation of cell attachment and division. Here we describe a flow method that enables single cell trapping in microwells with dimensions of 50 μm, a size sufficient to allow attachment and division of captured cells. Among various geometries tested, triangular microwells were found to be most efficient for single cell trapping while providing ample space for cells to grow and spread. An important trapping mechanism is the formation of fluid streamlines inside, rather than over, the microwells. A strong flow recirculation occurs in the triangular microwell so that it efficiently catches cells. Once a cell is captured, the cell presence in the microwell changes the flow pattern, thereby preventing trapping of other cells. About 62% of microwells were filled with single cells after a 20 min loading procedure. Human prostate cancer cells (PC3) were used for validation of our system.     

Developments in Cartesian cut cell methods

This paper describes the Cartesian cut cell method, which provides a flexible and efficient alternative to traditional boundary fitted grid methods. The Cartesian cut cell approach uses a background Cartesian grid for the majority of the flow domain with special treatments being applied to cells which are cut by solid bodies, thus retaining a boundary conforming grid. The development of the method is described with applications to problems involving both moving bodies and moving material interfaces.