Abstract—This paper employs the gradient theory to study the elastic properties and deformability of red blood cell (RBC) membrane using the first-order Cauchy-Born rule as an atomistic-continuum hyperelastic constitutive model that directly incorporates the microstructure of the spectrin network. The well–known Cauchy–Born rule is extended to account for a three-dimensional (3D) reference configuration. Using the strain energy density function and the deformation gradient tensor, the elastic properties of the RBC membrane were predicted by minimizing the potential energy in the representative cell. This extended formulation was then coupled with the meshfree method for numerical modeling of the finite deformation of the RBC membrane by simulating the optical tweezer experiment using a self–written MATLAB code. The results obtained provide new insight into the elastic properties and deformability of RBC membrane. In addition, the proposed method performs better when compared with those found in literature in terms of prediction accuracy and computation efficiency.
Abstract: This study was aimed at providing information on how Na sulphate, K sulphate, Ca sulphate and Mg sulphate supporting electrolytes affect the permeation of hypotonic Na chloride, K chloride, Ca chloride and Mg chloride ions through Red Blood Cell (RBC) membrane while the RBCs haemolyse using a UV/Vis spectrophotometer. The effects of the supporting electrolytes were deduced from the changes in their respective rates of permeation through RBC membranes during haemolysis. Na 2 SO 4 added to NaCl and MgSO 4 to MgCl 2 resulted in a decrease in permeation rate of the resultant solutions
The erythrocyte membranes refer to the lipid bilayer (phospholipids and cholesterol) containing the trans- membrane proteins of the cytoskeleton and an assembly of proteins regularly interconnected [1,2]. Multiple pro- teins make up the erythrocyte membrane cytoskeleton, which interacts with both the lipid bilayer and trans- membrane proteins to give the red blood cell its cha- racteristics strength and integrity of major intrinsic proteins. Ankyrin provides the primary linkage between α- and β-spectrin tetramers and the cytoplasmic domain of the major integral protein band 3, a relationship en- hanced by interactions with protein 4.2. The junctional complexes formed by spectrin tetramers and their inter- action with tropomyosin, adducin, actin and other pro- teins ensure the stability between the cytoskeleton and the lipid bilayer . In some abnormal conditions of the RBCs membrane, such as hereditary spherocytosis (HS), hereditary elliptocytosis (HE), hereditary pyropoikilocytosis (HPP) or the Southeast Asian Ovalocytosis (SAO), these membrane proteins are usually defective because of their specific inherited gene mutations . The high perme- ability of RBC membranes is due to the presence of water channel proteins called aquaporins (AQP) . An important characteristic of RBCs’ water diffusional per- meability is its inhibition by the PCMB . There are two basic strategies for measuring water exchange through the RBC membranes: the non-stationary me- thod via volume changes and the stationary method using NMR techniques .
the observation of Hu et al. . The mechanism behind the observed reduction in hypertensive subjects remains unclear. Potassium and sodium play important roles in the maintenance of cellular functions, and raised or lowered serum potassium level may be harmful to health . Sodium, the main extracellular cation has long been considered as the pivotal environmental factor for hypertension. Changes in serum urea, are associated with impairment of renal function . Reabsorption of filtered sodium by the renal tubules is increased in primary hypertension because of stimulation of several sodium transporters located at the luminal membrane, as well as the
We also investigated whether the SW technique caused any further photo-toxicity to the specimen compared to widefield epifluorescence imaging. It was observed that when using either imaging method, the red blood cell appeared to decay at a similar rate and in a manner that resembled the decay observed previously over 12 minutes under periodic focused laser illumination . In the study of Wong et. al , it was found that membrane damage occurred even at low laser powers as the result of reactive oxygen species being generated via photo-hemolysis. This process has been hypothesized to be due to interactions between the reactive oxygen species and the membrane proteins, band 3 and spectrin , which has also been reported in studies using widefield illumination [63,64]. It has also been found that when using multiphoton microscopy, red blood cell photodamage could be observed on timescales in the region of tens of seconds , however, when we carried out our video-rate experiments we observed no photo-toxicity effects over the 33 second duration.
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Phosphopeptide quantitation across all eight unique treatment groups indicate that the ERK1/2 pathway activa- tion in SS RBCs could be responsible for alteration of mul- tiple phenotypic and functional properties of the red cell, by affecting phosphorylation of thirty-six peptides from twenty-one phosphoproteins involved in adhesion, cAMP production, anion transport by band 3 and band 3 traffick- ing, RBC shape, flexibility, cell morphology maintenance, glucose and glutamate transport, degradation of misfolded proteins, and receptor ubiquitination, all of which play a significant role in the complex pathophysiology of the dis- ease. Interestingly, these data revealed that glycophorin A phosphorylation was highly differentiated between healthy and sickle RBCs, and its levels of phosphorylation were modulated by the presence of the MEK1/2 inhibitor U0126 and the presence of exogenously spiked ERK2. Glyco- phorin A is the major sialoglycoprotein, and increased SS RBC adhesion to vascular endothelial cells has been postu- lated to result from clustering of negatively charged glycophorin-linked sialic acid moieties at the RBC surface [6,39]. In addition, alteration in glycophorin A phosphoryl- ation could subsequently result in decreases in both anion transport by band 3 and band 3 trafficking. Thus, our stud- ies further confirm ERK1/2 as a potential therapeutic target to ameliorate multiple functions of the sickle red cell, in- cluding adhesion  and vaso-occlusion [10,18], chronic hemolysis and ischemic tissue damage , all of which are associated with the pathophysiology of SCD. Finally, follow-up validations will be addressed on additional physiologically relevant molecules presented in Table 2, such as cytoskeletal proteins, and the effect of their phos- phorylation by ERK1/2 on RBC function.
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Figure 2 depicts that the nature of the curves of os- motic pressure-volumetirc ratio relationship of RBC in- fected with plasmodium falciparum parasite from ana- lytical result and the spherical inflation of HRBC during gradual swelling in experimental observation . The curves are nonlinear and they show different mechanical behavior. Malaria IRBCs becomes stiffer and enhance cell rigidity and develop properties of cytoadherence [1, 8]. Due to the parasite matures and multiply within the cell, both the internal pressure and volume of the cell increases until a certain limit point is attained before hemolysis as shown from the curve of the IRBC in the figure. From the figure, at a fixed pressure, we analyze that the HRBC show better flexibility (deformability) than the IRBC. As the assumption of  that the red cell membrane cannot tolerate any stretch without he- molysis. Therefore, the curves in the figure shows loss of cell deformability regarding to the cell size (volume) increases before hemolysis. In order for a cell to be de- formable, it must have a large membrane area-to-volume ratio .
Lower RBC membrane AA levels are associated with de- creased niacin sensitivity, but this expected correlation has only been described in a healthy control group (Messamore et al., 2010). Although lower AA levels were found in patients who did not flush in response to oral niacin (Glen et al., 1996), no correlation between AA levels and niacin response was ob- served in two di ﬀ erent groups of patients with SZ (Maclean et al., 2003; Messamore et al., 2010). On the other hand, an unexpected correlation was observed between maximal niacin- induced blood flow and red blood cell membrane levels of adrenic acid (22:4 n-6), the elongation product of AA. Adrenic acid has vasodilatory actions in bovine coronary arteries and in arteries of the cortical layer of the adrenal gland (Kop et al., 2010). Lack of correlation between AA levels and niacin re- sponse may result from a homeostatic imbalance within the n-6 PUFAs pathway in SZ. The significance of adrenic acid levels as they relate to niacin-induced blood flow response de- serves further attention.
Methods: This case-control study (40 cases and 40 controls) was conducted with newly diagnosed BCC adults who were recruited from Razi Hospital. To measure fatty acids of red blood cell membrane, fatty acids were extracted and injected to gas chromatography. Case and control groups were matched based on sex, age, and body mass index (BMI). All subjects also completed two 24-hour dietary recalls by nutritionist help, which included two randomly selected days.
The therapeutic potential of nanoparticle-based drug carriers depends largely on their ability to evade the host immune system while delivering their cargo safely to the site of action. Of particular interest are simple strategies for the functionalization of nanoparticle surfaces that are both inherently safe and can also bestow immunoevasive properties, allowing for extended blood circulation times. Here, we evaluated a recently reported cell membrane-coated nanoparticle platform as a drug delivery vehicle for the treatment of a murine model of lymphoma. These biomimetic nanoparticles, consisting of a biodegradable polymeric material cloaked with natural red blood cell membrane, were shown to efficiently deliver a model chemotherapeutic, doxorubicin, to solid tumor sites for significantly increased tumor growth inhibition compared with conventional free drug treatment. Importantly, the nanoparticles also showed excellent immunocompatibility as well as an advantageous safety profile compared with the free drug, making them attractive for potential translation. This study demonstrates the promise of using a biomembrane-coating approach as the basis for the design of functional, safe, and immunocompatible nanocarriers for cancer drug delivery.
Anaemia is a condition that develops when your blood lacks enough healthy red blood cells or hemoglobin. Hemoglobin is a main part of red blood cells and binds oxygen which refers to a de- creased number of circulating red blood cells and is the most common blood disorder. Symptoms can include headaches, chest pains, and paleness of skin. Anemia currently affects more than 3 million Americans and an estimated 1.62 billion people, globally. There are three main types of anemia: that due to blood loss, that due to decreased red blood cell production and that due to increased red blood cell breakdown. Causes of blood loss include trauma and gastrointestinal bleeding, among others. Causes of decreased production include iron deficiency, a lack of vitamin B 12 , thalassemia, and a
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Hereditary spherocytosis (HS) is a heterogeneous disorder in which abnormalities of red blood cell structural proteins lead to loss of erythrocyte membrane surface area, resulting in spherical-shaped, hyperdense, poorly deformable red blood cells (Fig 1) with a shortened life span. 1–5 HS occurs worldwide and affects individuals from all racial and ethnic groups. Individual pediatricians encounter HS uncommonly, but hospitals and health care systems with large delivery services regularly deal with this condition, particularly in white neonates of northern European ancestry, in whom the condition can be as frequent as 1 in 1000 to 2000 births. Early suspicion and diagnosis of HS allow appropriate management, including provision of anticipatory guidance to parents, which can reduce the risk of adverse outcomes.
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 and heart failure . On the other hand, HbA1c is a marker of long term glucose homeostasis and is routinely used to assess the adequacy of glycemic control in diabetic patients. Many researches have been noted that HbA1c is associated with type 2 diabetes patients. As well as, HbA1c is a marker of long term glucose homeostasis and is routinely used to assess the adequacy of glycemic control in diabetic patients. [25-27]. HbA1c level increase with age, chronic subclinical in lammation and possibly oxidative stress and also in conditions that can adversely affect red blood cell survival. For example, hemolytic anemia, blood transfusion (BT0, chronic malaria or major blood loss) [25,28,29]. Platelets, another element of the complete blood count (CBC), play a key role in the development of atherothrombosis, a major role in intravascular thrombosis on top of atherosclerosis, resulting in cardiovascular diseases in diabetes with respect to altered platelet morphology and function [30,31]. Mean platelet volume (MPV) is and indiactor of the average size and was suggested as an indicator of platelet activity and the state of thrombogenesis [32,33]. Some studies antiplaletes have been demonstrated as to be very effective at decreasing myocardial infarction, stroke and death . In addition to, leukocytes are cognizant as to participate in the in lammatory process. Accompanying atherosclerosis. They are come round at the site of endothelial injury and form foam cells in the atherosmatosis plaque. WBC count is positively associated with increased cardiovascular mortality, mainly from coronary heart disease . Previously our researches have been shown the association of mean platelet volume and platelet parameter with HbA1c . In this study, we aimed to evaluate the relationship between RBC parameter such as RBC count, PCV, MCV, MCHC, RDW-CV and HbA1c value ≥7.5 of diabetics’ patients. So that RBC parameter could be used as prognicator of diabetes instead of HbA1c test.
Different petal extracts of Crocus sativus “Cashmerianus” were examined for anti-inflammatory activity by in-vitro human red blood cell (HRBC) membrane stabilization method and in-vivo carrangeenan- induced rat paw edema. The petroleum ether, chloroform, ethyl acetate, methanol and aqueous extracts of petals showed 49.36%, 59.39%, 48.10%, 70.37% and 68.89% protection at the dose of 400mg/ml respectively by in-vitro method. However, in-vitro most promising effective chloroform, methanol and aqueous extracts were selected for in-vivo anti-inflammatory method. The in-vivo anti- inflammatory activity was found as 50%, 63.16% and 57.89% inhibition of paw volume after 5h of chloroform, methanol and aqueous extracts treated at the dose of 400mg/kg groups respectively. The potency of the petals extracts were compared with Diclofenac (10 mg/ml) for in-vitro and 10mg/kg for in-vivo model. The methanol and aqueous extract showed the most promising anti-inflammatory activity in membrane stabilizing action on human red blood cell membrane and reduction of edema in carrageenan induced rat paw edema model. The results of present and first time reporting this study on petals extracts of Crocus sativus “Cashmerianus” demonstrate that extracts of petals possess significant (p<0.05) anti- inflammatory potential.
Twenty four male Wistar rats were picked according to body weight and placed into wooden cages labelled A to D. Six male Wistar rats each were placed per cage, with the group labelled A-serving as the control group, while B, C and D were the test groups. The animals in the control group (A) were administered daily with standard feed and clean tap water orally, using cannula and syringe. The animals in group B were administered orally with same volume (0.5 mL) corresponding to 200 mg/ kg body weight of the ethanol leaf extract, group C was administered pefloxacin (400 mg/5 mL) alone, while group D was co-administered pefloxacin and the extract (1:1) for 21 days. The animals in each group were sacrificed 24 h after the completion of their respective doses for blood samples collection by cardiac puncture and organs harvested for future evaluation . The animals were handled humanely in accordance with the guidelines of European convention for the protection of vertebrate animals and use for scientific purposes- ETS-123 .
Haemolytic assays have long been used to measure free radical damage and counteraction by antioxidants. It is useful for screening for oxidizing or antioxidizing molecules. Many primary or secondary plant metabolites have been found to protect cells from oxidative damage. These compounds have been evidenced to stabilize RBC membrane by scavenging free radicals and reducing lipid peroxidation. Human erythrocytes are the key target for the free radicals or reactive oxygen species because of polyunsaturated fatty acid membrane and also their oxygen transport linked redox activity. H 2 O 2 is a potent oxidizing
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From the results, compounds B, E and H show high activity compared with other compounds. The active compounds B, E, and H contain a nitro group at meta position. Replacing nitro group with carboxyl group will be minimize the activity. Therefore, the activity is the best when a nitro group located at meta position. In salicylic acid moiety, the activity of protection was found increased when methyl group located at para-position compared with the meta- methyl and meta-methoxy groups. Compound E contain meta nitro group and meta methoxy group, it is the compound with meta-methoxy group has high activity than other methoxy derivatives. The results showed compounds A, D, F and I have less activity. The action of the azo compounds could be related to the binding of compounds with erythrocyte membrane, especially phospholipids. Since the compounds has polar and nonpolar groups can bonded with same groups of phospholipids. This prevents membrane damage by physical interaction of osmotic pressure differences, which is causative to hemolysis of red blood cells 19 .
The human red blood cell (HRBC) membrane stabilization method was used as one of the methods for the determination of anti-inflammatory activity invitro).The blood was collected from healthy human volunteer who was not taken any NSAIDS for two weeks prior to the experiment and mixed with equal volume of Alsever solution and centrifuged at 3,000 rpm and the packed cells were separated. The packed cells were washed with isosaline and a 10% suspension was made. This HRBC suspension was used for the estimation of anti-inflammatory property. Different concentration of extract, reference sample and control were separately mixed with 1ml of phosphate buffer, 2ml hypo saline and 0.5 ml of HRBC suspension were added. All the assay mixtures were incubated at 37 0 C for 30 min in a BOD incubator and sufficiently centrifuged at 3,000-rpm. The supernatant liquid was decanted and the hemoglobin content was estimated spectrophotometrically at 560 nm. The percentage hemolysis was estimated by spectrometrically at 560 nm. The percentage hemolysis was estimated by assuming the hemolysis produced in the control as 100%.
Determination of Anti-inflammatory Activity of Human Red Blood Cell (HRBC) Membrane Stabilization Assay: Inflammation could be described as a defense response that is activated upon harmful stimuli to tissues. Uncontrolled inflammatory could also occur as a result of allergies, cardiovascular dysfunctions, metabolic syndrome, cancer and autoimmune diseases. During inflammation, lysosomal hydrolytic enzymes are released into cites which causes damages of the surrounding organelles and tissues with attendance variety of disorders. Several methods are employed to screen and study drugs or herbal preparations that exhibit anti-inflammatory
To evaluate the in-vitro anti inflammatory activity of Elaeocarpus ganitrus roxb of bark extracts by HRBC membrane stabilization. The plant Elaeocarpus ganitrus roxb (Elaeocarpaceae) commonly known as Rudraksha the king of herbal medicines. The plants are distributed throughout India, Southeast Asia, Japan, Malaysia, Southern China, Australia etc. The leaves, fruits, seeds and stems of the plant have been used widely in traditional folk medicine in many parts of India. In the information regarding the use of bark of Elaeocarpus ganitrus Roxb for several disorders systemic in vitro studies were carried out in our laboratory to establish the anti inflammatory potential by using HRBC membrane stabilization method. HRBC membrane is equals to lysosomal membrane which plays the role in the process of inflammation. The in-vitro HRBC membrane stabilization method showed significant anti inflammatory property of various Elaeocarpus ganitrus tested extracts. It was found that the chloroform extract shows significant anti inflammatory activity at the concentration of 500µg/ml which is comparable to the standard Diclofenac sodium.