Segmentation, Reconstruction, and Analysis of Blood Thrombus Formation in 3D 2-Photon Microscopy Images

We study the problem of segmenting, reconstructing, and analyzing the structure growth of thrombi (clots) in blood vessels in vivo based on 2-photon microscopic image data. First, we develop an algorithm for segmenting clots in 3D microscopic images based on density-based clustering and methods for dealing with imaging artifacts. Next, we apply the union-of-balls (or alpha-shape) algorithm to reconstruct the boundary of clots in 3D. Finally, we perform experimental studies and analysis on the reconstructed clots and obtain quantitative data of thrombus growth and structures. We conduct experiments on laser-induced injuries in vessels of two types of mice (the wild type and the type with low levels of coagulation factor VII) and analyze and compare the developing clot structures based on their reconstructed clots from image data. The results we obtain are of biomedical significance. Our quantitative analysis of the clot composition leads to better understanding of the thrombus development, and is valuable to the modeling and verification of computational simulation of thrombogenesis

Model predictions of deformation, embolization and permeability of partially obstructive blood clots under variable shear flow

Thromboembolism, one of the leading causes of morbidity and mortality worldwide, is characterized by formation of obstructive intravascular clots (thrombi) and their mechanical breakage (embolization). A novel two-dimensional multi-phase computational model is introduced that describes active interactions between the main components of the clot, including platelets and fibrin, to study the impact of various physiologically relevant blood shear flow conditions on deformation and embolization of a partially obstructive clot with variable permeability. Simulations provide new insights into mechanisms underlying clot stability and embolization that cannot be studied experimentally at this time. In particular, model simulations, calibrated using experimental intravital imaging of an established arteriolar clot, show that flow-induced changes in size, shape and internal structure of the clot are largely determined by two shear-dependent mechanisms: reversible attachment of platelets to the exterior of the clot and removal of large clot pieces. Model simulations predict that blood clots with higher permeability are more prone to embolization with enhanced disintegration under increasing shear rate. In contrast, less permeable clots are more resistant to rupture due to shear rate-dependent clot stiffening originating from enhanced platelet adhesion and aggregation. These results can be used in future to predict risk of thromboembolism based on the data about composition, permeability and deformability of a clot under specific local haemodynamic conditions

Influence of Milk Thistle Shot on Quality Parameters of the Sour-milk Beverage

Modern complicated ecological conditions cause a general necessity in improving the food structure of the population due to improving the quality, biological value and taste characteristics of products. Just that is why, the aim of the work was to study the influence of milk thistle shot on quality parameters of the sour-milk beverage – kefir. It was established, that it had the homogenous consistence with a broken clot and color from white to creamy with shot particles. The increase of the milk thistle shot dose to 3 % and 4 % results in the taste with the brightly expressed milk thistle smack and brown color with the expressed milk thistle content. The viscosity of kefir with fms 2,5 % changes during seven days of storage, although remains rather high at 8 day, namely 47 s. The increase of the viscosity of kefir with milk thistle shot is explained by its hygroscopic properties, in which result free moisture of the product is bound. The analysis of microbiological parameters in the process of storage of kefir with milk thistle shot allows to make a conclusion about the satisfactory sanitary condition of the new product and its harmlessness for consumer's health. In the kefir with milk thistle shot the general amount of amino acids grew by 11,6 %, including irreplaceable ones – by 10,1 %, replaceable – by 12,6 %, that indicates its biological value.Addition of milk thistle shot to kefir didn't cause changes of the biological value of the protein component of the combined product. It is testified by the mean value of amino acid scores of control (129,3 %) and experimental (127,9 %) samples of kefir. Some growth of the valine score (3,6 %) can be noted.So, the combined product is characterized by the balanced amino acid composition. Due to adding milk thistle shot, consumption of amino acids, necessary for synthesis of proteins and essential number of compounds, vitally important for the human organism, grew

Simulation of a mechanical thrombectomy device based in the use of self-expandable stents for the blood clots extraction

Recently, we have presented some studies concerning the analysis, design and optimization of one experimental device developed in the UK - GPTAD - which has been designed to remove blood clots without the need to make contact with the clot itself, thereby potentially reducing the risk of problems such as downstream embolisation. Based on the idea of a modification of the previous device, in this work, we present a model based in the use of stents like the SolitaireTM FR, which is in contact with the clot itself. In the case of such devices, the stent is self-expandable and the extraction of the blood clot is faciliatated by the stent, which must be inside the clot. Such stents are generally inserted in position by using the guidewire inserted into the catheter. This type of modeling could potentially be useful in showing how the blood clot is moved by the various different forces involved. The modelling has been undertaken by analyzing the resistances, compliances and inertances effects. We model an artery and blood clot for range of forces for the guidewire. In each case we determine the interaction between blood clot, stent and artery

Endovascular Embolization by Transcatheter Delivery of Particles: Past, Present, and Future.

Minimally invasive techniques to occlude flow within blood vessels, initially pioneered in the 1970s with autologous materials and subsequently advanced with increasingly sophisticated engineered biomaterials, are routinely performed for a variety of medical conditions. Contemporary interventional radiologists have at their disposal a wide armamentarium of occlusive agents to treat a range of disease processes through a small incision in the skin. In this review, we provide a historical perspective on endovascular embolization tools, summarize the current state-of-the-art, and highlight burgeoning technologies that promise to advance the field in the near future

Study of Influence of Calcium Content in Milk on Quality Indicators of Cottage Cheese

The analysis was realized, and the dependence between the calcium content and organoleptic and functional-technological properties of milk as a raw material for producing sour milk cheese was determined. It was demonstrated, that alongside with other factors, the important role in milk clotting belongs to calcium, which role is in binding of free OH-groups of phosphoric acid of casein micelles. As a result of the aforesaid, their negative charge and colloid stability decrease with further hydrophilicity decrease with further aggregation of casein molecules. It was established, that the excessive content of calcium in milk is negative that is manifested in formation of the dry and brittle consistence of sour milk cheese. There was offered the way of calcium content regulation in milk by its decalcification using the natural sorbent of sodium alginate. Regulation of the milk salt system, especially, the calcium content as an initial raw material for producing sour milk cheese by the change of the content and condition of calcium allowed to correct parameters of the process of sour milk cheese making and its functional-technological properties, especially, moisture-keeping ability, form stability and other. It was established, that the decrease of the calcium content in milk provides getting sour milk cheese with the soft, easily smearing consistence, without whey separation. The obtained data on the characteristic of organoleptic indicators fully correlate with studies of the microstructure and dispersity of sour milk cheese. It was determined, that milk decalcification results in raising dispersity of sour milk cheese at the synchronous increase of the percent content of protein particles with minimal size characteristics in the system. It was elucidated, that the microstructure of studied samples consists of protein grains of the same form, evenly distributed by the whole volume. Based on the obtained experimental data, there were corrected parameters of the technological process of sour milk cheese production. There were elaborated ways of formation of the culinary products assortment on the base of sour milk cheese, produced of decalcified milk

Behaviour of milk protein ingredients and emulsions stabilised by milk protein ingredients in the simulated gastrointestinal tract : a thesis presented in partial fulfilment of the requirements for the degree of Master of Food Technology, Massey University, Manawatu, New Zealand

Milk clotting behaviours in the stomach impact the digestion rates of protein and fat. A variety of milk protein products are applied as functional ingredients in many foods. This research was conducted to investigate the digestion behaviours of various commercial dairy ingredients and lipids in emulsions stabilised by these ingredients using a dynamic in vitro digestion model, i.e., a human gastric simulator (HGS), with a focus on the effect of different structures of clots formed in dairy ingredients during gastric digestion on hydrolysis of proteins and/or lipids. Skim milk powder (SMP), milk protein concentrate (MPC) 4851, MPC 4861, sodium caseinate, whey protein isolate (WPI) and heated (90°C, 20 min) WPI were used in the present study. Results showed that SMP and MPC 4851, which contained casein micelles, formed ball-like clots with a relatively dense network after 10 min of gastric digestion. These clots did not disintegrate after 220 min of digestion. MPC 4861 and sodium caseinate generated clots at around 40 min, and a loose, fragmented structure was observed at the end of the gastric digestion due to a lacking micellar structure of caseins. No clot was observed in WPI or heated WPI after 220 min gastric digestion, although aggregation occurred at around 40 min in heated WPI. These differences in coagulation behaviours apparently affected the rate of gastric emptying and protein hydrolysis by pepsin in the gastric system. In SMP and MPC 4851, the gastric emptying and hydrolysis of caseins was much slower than that observed in MPC 4861 and sodium caseinate. The most rapid gastric emptying of proteins was observed in the WPI samples both with and without heating. This is attributed to the formation of varied structured clots at different times under the gastric conditions. The effect of protein concentration on the gastric behaviour of these dairy ingredients in solution was then examined, with a particular emphasis on the structure of clots. SMP and MPC 4851 have been selected as model protein ingredients. Their gastric behaviours were investigated over a protein concentration range of 0.5-5.0% (w/w). The results showed that the digestion behaviour of SMP and MPC 4851 followed a similar pattern. The rate of pH changes in the emptied digesta during digestion was protein concentration dependent. With an increase in protein concentration, the decrease in pH slowed. The protein concentration had no apparent impact on the casein clotting time. Clots were formed in the first 10 min of digestion in all samples. However, in both SMP and MPC 4851, when protein concentration was lower than 2.0% (w/w) the clots consisted of small protein pieces with a loose, porous and open structure after a 220 min digestion. Whereas a cheese ball-like clot with a denser network was observed at the end of gastric digestion when the protein concentration varied from 2.0% to 5.0% (w/w). Such a difference in the structure apparently affected the rate of protein hydrolysis. A more rapid hydrolysis (P < 0.05) of the clotted protein was observed when protein concentration was lower than 2.0% (w/w) compared to the samples containing a higher proportion of protein (2.0%-5.0%, w/w). To study the effect of different coagulation behaviours on the digestion of oil droplets in oil-in-water emulsions, these dairy ingredients (with the exception of SMP) were used to prepare an oil-in-water emulsion (20.0% soy oil and 4.0% protein, w/w). They were digested under the dynamic gastric conditions using the HGS. The gastric digesta was emptied at 20 min intervals. Then all digesta were mixed to investigate the lipid digestion under the small intestinal conditions. Changes in physicochemical properties of emulsions, involving the particle size, the microstructure, the oil content of the emptied gastric digesta and the amount of free fatty acids (FFAs) released during the small intestine stage, were determined using an in vitro small intestinal digestion model. Aggregation of MPC 4851-stabilised emulsion took place after 5 min of digestion in the HGS with the largest size. The aggregates remained in the stomach and did not disappear during the whole gastric digestion. The hydrolysis of the aggregated network by pepsin was largely slowed by the reduced ability of the simulated gastric fluid (SGF, containing pepsin) to diffuse into the larger sized aggregates. MPC 4851-stabilised emulsion thus resulted in the slowest release of oil droplets into the small intestine. In comparison, MPC 4861 and sodium caseinate-stabilised emulsions aggregated in the stomach at approximately 40 min, forming smaller sized aggregates. These aggregates disintegrated at the mid and late-stages of digestion in these two emulsions. Therefore, MPC 4861 and sodium caseinate-stabilised emulsions had a more rapid delivery of oil droplets into the small intestine. In relation to the WPI-stabilised emulsions both with and without heating, the aggregations formed at a similar time to that which was observed in MPC 4861 and sodium caseinate-stabilised-emulsions; i.e., at approximately 40 min. However, they had the smallest sized aggregates amongst all samples and they disintegrated quickly with further digestion. WPI-stabilised emulsions both with and without heating had the fastest gastric emptying and hydrolysis by pepsin in the early and mid-stages of the gastric digestion process. Thus, the highest level of oil content contained in the emptied gastric digesta was produced from both WPI-stabilised emulsions. In the mixed gastric digesta, which were subjected to the small intestinal digestion, the oil contents contained in the different emulsion samples varied. This difference impacted the extent of lipid digestion by pancreatic lipase. The sample with a higher oil content released a greater amount of FFAs compared to the sample with a lower oil content. The extent of lipid digestion of different emulsion samples adhered to the following pattern: MPC 4851-stabilised emulsion < MPC 4861-stabilised emulsion < sodium caseinate-stabilised emulsion, WPI-stabilised emulsions both with and without heating. Overall, the gastric behaviours of dairy ingredients either in solutions or emulsions were affected by the formation of structured clots/aggregates. The differences in clotting/aggregation times and their structures were greatly dependent on the component and structure of protein, the processing prior to digestion and the susceptibility to proteases. These differences in protein coagulation/aggregation behaviour impacted the rates of protein hydrolysis and gastric emptying. The oil content and protein composition of the gastric digesta transferred into small intestine and the extent of lipid digestion in small intestine were also affected. These results are important in an application perspective. They provide useful information for the design and development of healthier food products by allowing greater control over the manipulation of protein bioavailability, which subsequently provides greater control over lipid metabolism

RGS10 shapes the hemostatic response to injury through its differential effects on intracellular signaling by platelet agonists.

Platelets express ≥2 members of the regulators of G protein signaling (RGS) family. Here, we have focused on the most abundant, RGS10, examining its impact on the hemostatic response in vivo and the mechanisms involved. We have previously shown that the hemostatic thrombi formed in response to penetrating injuries consist of a core of fully activated densely packed platelets overlaid by a shell of less-activated platelets responding to adenosine 5\u27-diphosphate (ADP) and thromboxane A2 (TxA2). Hemostatic thrombi formed in RGS10-/- mice were larger than in controls, with the increase due to expansion of the shell but not the core. Clot retraction was slower, and average packing density was reduced. Deleting RGS10 had agonist-specific effects on signaling. There was a leftward shift in the dose/response curve for the thrombin receptor (PAR4) agonist peptide AYPGKF but no increase in the maximum response. This contrasted with ADP and TxA2, both of which evoked considerably greater maximum responses in RGS10-/- platelets with enhanced Gq- and Gi-mediated signaling. Shape change, which is G13-mediated, was unaffected. Finally, we found that free RGS10 levels in platelets are actively regulated. In resting platelets, RGS10 was bound to 2 scaffold proteins: spinophilin and 14-3-3γ. Platelet activation caused an increase in free RGS10, as did the endothelium-derived platelet antagonist prostacyclin. Collectively, these observations show that RGS10 serves as an actively regulated node on the platelet signaling network, helping to produce smaller and more densely packed hemostatic thrombi with a greater proportion of fully activated platelets