Rat Red Blood Cells Research Articles

Centaurium erythraea methanol extract protects red blood cells from oxidative damage in streptozotocin-induced diabetic rats

Ethnopharmacological relevanceCentaurium erythraea Rafn (CE) is a traditional medicinal herb in Serbia with antidiabetic, digestive, antipyretic and antiflatulent effects Aim of the studyTo investigate the potential protective effects of the methanol extract of the aerial parts of CE against glyco-oxidative stress in red blood cells (RBCs) in rats with experimentally induced diabetes. Material and methodsDiabetes was induced in Wistar rats by intraperitoneal (i.p.) injection of multiple low-dose streptozotocin (STZ) (40mg/kg, for five consecutive days), with the 1st day after the last STZ injection taken as the day of diabetes onset. The methanol extract of CE (100mg/kg) was administered orally and daily, two weeks before the first STZ injection, during the 5-day treatment with STZ, and for four weeks after the STZ injections (pre-treated group) or for four weeks after diabetes onset (post-treated group). The effect of CE extract administration on the redox status of RBCs was evaluated by assessing lipid peroxidation, the ratio of reduced/oxidized glutathione (GSH/GSSG), the level of S-glutathionylated proteins (GSSP) and the enzymatic activities of superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx) and glutathione reductase (GR) in RBCs four weeks after diabetes onset. The major biochemical parameters of diabetes, protein glycation/glycosylation of erythrocytes and parameters which correlate with their aggregation and deformability were also evaluated. ResultsDaily application of CE extract to STZ-induced diabetic rats provided important antidiabetic effects, observed in both pre-treated and post-treated groups of diabetic rats as elevated serum insulin concentration, reduction of blood glucose and glycated hemoglobin concentrations and an improved lipid profile. Antioxidant effects of CE extract were detected in RBCs of diabetic rats and observed as decreased lipid peroxidation and ameliorated oxidative damage as a result of increased SOD, CAT and GR activities, an improved GSH/GSSG ratio and reduced GSSP levels. Moreover, the CE extract protected RBC proteins from hyperglycemia-induced damage by reducing non-enzymatic glycation and enzymatic glycosylation processes. CE extract was more effective when applied before diabetes induction (pre-treated group). ConclusionsThe results of this study show that the Centaurium erythraea methanol extract protects RBCs in diabetic animals from oxidative damage. They provide additional support for the application of this traditionally used plant in diabetes management.

Anti-inflammatory and membrane stabilizing properties of methyl jasmonate in rats

The present investigation was carried out to evaluate anti-inflammatory and membrane stabilizing properties of methyl jasmonate (MJ) in experimental rat models of acute and chronic inflammation. The effects of MJ on acute inflammation were assessed using carrageenan-induced rat's paw edema model. The granuloma air pouch model was employed to evaluate the effects of MJ on chronic inflammation produced by carrageenan in rats. The number of white blood cells (WBC) in pouch exudates was estimated using light microscopy. The levels of biomarkers of oxidative stress, such as malondialdehyde (MDA), glutathione (GSH) and activity of antioxidant enzymes in the exudates, were determined using spectrophotometry. The membrane stabilizing property of MJ was assessed based on inhibition of hemolysis of rat red blood cells (RBC) exposed to hypotonic medium. Our results indicated that MJ (25–100 mg·kg−1, i.p.) produced significant anti-inflammatory activity in carrageenan-induced paw edema in rats (P < 0.05). MJ reduced the volume of pouch exudates and the number of WBC in carrageenan-induced granulomatous inflammation. It also exhibited potent antioxidant and membrane stabilizing activities. In conclusion, these findings suggest the therapeutic potentials of methyl jasmonate in disease conditions associated with inflammation and its anti-inflammatory activity may be related to its antioxidant and membrane stabilizing activities.

Young and old rats have different strategies of metabolic adaptation to Cu-induced liver fibrosis

We investigated the role of pro- and antioxidant and immune systems in age-related adaptive reactions to chronic exposure to toxicants (copper ions). For this purpose, we administered multiple doses of copper sulfate to younger and older rats in order to induce liver fibrosis and then measured the parameters of proand antioxidant and immune systems and physiological indices. It was found that Cu-induced liver fibrosis was accompanied by oxidative stress and a significant reduction in cell-mediated immune activity. If oxidative stress was eliminated by the administration of exogenous antioxidants (mix factor), the physical capacity, body weight, and content of red blood cells in rats with fibrosis was recovered. The parameters of cell-mediated immune response recovered more effectively in older animals than in younger ones. The metabolic strategies of adaptation to toxic exposure were different in younger and older animals.

Toxicological Evaluation of the Aqueous Extract of Caralluma europaea and Its Immunomodulatory and Inflammatory Activities.

Background:Caralluma europaea (CE) has been studied for its chemical constituents, and no information is available on its toxicity or its pharmacological activities.Objective:To determine the toxicity of an aqueous extract of CE stems in vitro and in vivo after acute and subchronic oral gavages in Swiss albino's mice and its immunomodulatory and inflammatory activities.Materials and Methods:The extract was administrated in single oral dose at 5 g/kg body weight for the acute toxicity test and by gavages daily at doses of 1, 2.5, or 5 g/kg for 30 consecutive days for the subchronic toxicity test. The immunomodulatory activities and inflammatory activities were tested by the evaluation of hemagglutination antibodies (HAs) titers and delayed-type hypersensitivity (DTH) response.Results:For the dose of 1 g/kg, no visible toxic effects were observed. However, for the higher doses, clinical observations of toxicity were noted after 1 week of treatment. This was confirmed by the biochemical parameters values and the histology analyses of the spleen, liver, and kidney tissues. The high cellular mortality rate in vitro when treated with CE extract confirmed their toxicity potential. There was also increase of “HA titer” and “DTH” response in mice treated with nontoxic dose of CE (1 g/kg) compared to control group. This immune activity was confirmed by the high number of lymphocytes infiltrates noted in the different organs.Conclusion:We conclude that CE at the dose up of 1 g/kg produced toxic effect in mice that induced an immune inflammatory reaction.SUMMARY Caralluma europaea (CE) has been studied for its chemical constituents, and no information is available on its toxicity or its pharmacological activities. The objective is to determine the toxicity of an aqueous extract of CE stems in vitro and in vivo after acute and subchronic oral gavages in Swiss albino's mice and its immunomodulatory and inflammatory activities. For the dose of 1 g/kg, no visible toxic effects were observed. However, for the higher doses, clinical observations of toxicity were noted after 1 week of treatment. This was confirmed by the biochemical parameters values and the histology analyses of the spleen, liver, and kidney tissues. The high cellular mortality rate in vitro confirmed their toxicity potential. There was also increase of “hemagglutination antibody titer” and “delayed-type hypersensitivity” response in mice treated with nontoxic dose of CE (1 g/kg) compared to control group. This immune activity was confirmed by the high number of lymphocytes infiltrates noted in the different organs. We conclude that CE at the dose up of 1 g/kg produced toxic effect in mice that induced an immune inflammatory reaction. Abbreviations Used: CE: Caralluma europaea, ALT: Alanine aminotransferase, AST: Aspartate aminotransferase, RRBCs: Rat red blood cells, DTH: Delayed-type hypersensitivity response, PBS: Phosphate buffer solution.

Relationship between Partition Coefficients of Carboxylic Acids and Changes in Osmotic Resistance in Rat Erythrocytes In vitro

We examined monocarboxylic and dicarboxylic acids and their partition coefficients for osmotic fragility (OF) in isolated red blood cells (RBCs) in rats. The dense packed RBC was incubated in a phosphate–NaCl buffer solution containing each carboxylic acid at 0 to 100 mM at 37oC for 1 h. The RBC suspensions were transferred into the OF test tubes containing NaCl from 0.2 to 0.9%. The hemoglobin concentration was determined and NaCl concentration inducing 50% hemolysis was calculated as OF value. The OF in RBCs was dose-dependently increased by exposure to some of monocarboxylic acids possessing certain length of hydrocarbons with more than 4 carbons. A positive and statistically significant correlation was obtained between the partition coefficients and the degree of change in OF for monocarboxylic acids. Dicarboxylic acids corresponded with the monocarboxylic acids had either no effect or rather decreased OF, and there was no correlation between the partition coefficients and change in OF for these acids. The partition coefficients of the monocarboxylic acids were higher than those for the corresponding dicarboxylic acids. Whereas monocarboxylic acids are thought to act on the hydrophobic acyl-chain of phospholipids, which exists in a deeper region, dicarboxylic acids act on the interface region, which is hydrophilic and in a shallower area of the RBC membrane. Both carboxylic acids are speculated to cause physicochemical changes in the deep or swallow portion of phospholipid layer through different mechanisms and result in changes in the resistance to osmotic pressure in the rat RBC membrane.

Inhibitors of Bruton's Tyrosine Kinase Reduce Anti-Red Blood Cell Response in a Murine Model of Autoimmune Hemolytic Anemia

Autoimmune hemolytic anemia (AIHA) is an antibody-mediated condition where the mainstays of treatment are corticosteroids and immunosuppressive therapies. This leads to substantial toxicity in patients with refractory or chronic disease and thus there is a need for improved therapies. BTK is a critical kinase in the B-cell receptor signaling cascade and we hypothesized that inhibition would decrease autoantibody responses. Supporting this, CLL patients taking ibrutinib, a first generation BTK inhibitor, experience very low rates of AIHA (Leukemia 2016). Based on this rationale we evaluated two BTK inhibitors, ibrutinib (ibr) and acalabrutinib (acal), in mouse models of AIHA to validate BTK as a target in this disease.Young C3H/B6 F1 mice were treated with acal, ibr, or vehicle (veh) via drinking water for 2 weeks then injected intraperitoneally with washed rat red blood cells (RBCs) weekly for 4 weeks using the method of Playfair and Marshall-Clarke (Nature New Biology 1973) to induce a response against autologous RBCs starting 14 days after initial injection and peaking days 28-35. Drug treatment was continued throughout the study and using this delivery method BTK occupancy was 99.1% for acal, 96.1% for ibr, and 0% for veh. Quantitation of immunoglobulin on mouse RBCs was performed by a direct enzyme-linked anti-globulin test (DELAT) and by flow cytometry for surface immunoglobulin as a confirmatory method. Results from DELAT are shown in figure 1. A repeated measures model was applied to log-transformed data. On day 7, prior to autoimmune response, there were no statistically significant differences between treatment groups. However, on day 21 significantly less autoantibody was detected on RBCs in the acal group vs. both veh (p=0.001) and ibr (p=0.008); the ibr group showed a trend toward less autoantibody compared to veh but this did not reach statistical significance (p=0.542). This relationship between treatments held true at days 42 and 56 (acal vs. vehicle, p=<0.001 and p=0.001; acal vs. ibr, p=0.008 and p=0.004; ibr vs. veh, p=0.542 and p=0.542). These results were confirmed by flow cytometry. Hematocrit tests were performed by capillary tube centrifugation with no mice developing anemia and no differences between treatment groups excluding rapid RBC turnover as a cause of reduced immunoglobulin on RBCs.As ibrutinib has been shown to alter macrophage function, we used a passive model to determine if BTK inhibitors changed clearance of antibody coated RBCs from circulation. After two weeks of treatment with acal (n=13), ibr (n=12), or veh (n=12) mice were injected with a single dose of anti-RBC antibody (anti-ter-119) and hematocrit was measured daily for 7 days. Median hematocrit values at baseline for the three groups were 53.7 (SD=1.3), 52.8 (SD=1.4), and 52.8 (SD=0.9) for acal, ibr, and veh respectively and fell to a low of 18.4 (SD=2.7), 18.0 (SD=3.2), and 15.8 (SD=1.9) on day 3 before recovering. A repeated measures model was applied to the data and no significant differences between drug treatment groups were found (p=0.177), indicating no effect of BTK inhibition on RBC clearance.To examine anti-rat RBC antibody response washed rat RBCs were incubated with mouse plasma and analyzed by flow cytometry. Treatment groups were compared at day 42 after initial rat RBC injection. An analysis of variance (ANOVA) model was applied to log-transformed data. There was significantly less anti-rat RBC antibody in the acal group vs veh (p=0.008). In addition, there was decreased, albeit not statistically significant, antibody production with ibr compared to veh and acal compared to ibr (p=0.193 and p=0.102, respectively). This is the same trend as for autoantibody production and argues that BTK inhibition may reduce antibody responses.In summary, treatment with acalabrutinib significantly reduced anti-RBC autoantibody response in our model compared to vehicle and ibrutinib, and autoantibody was consistently decreased in ibrutinib treated mice compared to vehicle although this did not reach statistical significance. The differences between inhibitors may be due to potency of BTK inhibition or effects on other targets of ibrutinib which is a less specific drug. This mechanism is under active investigation. These results support BTK as a potential therapeutic target in AIHA and the further development of acalabrutinib for this indication. [Display omitted] DisclosuresWoyach:Karyopharm: Research Funding; Morphosys: Research Funding; Acerta: Research Funding.

Patients with Sickle Cell Disease in Sicily Have Lower Rates of End Organ Damage, Allo-Immunization and Opioid Prescription Compared to a US Cohort

Background: In developed countries, childhood mortality in sickle cell disease (SCD) is less than 5%, where the SCD morbidity and mortality burden has shifted to adults. As age increases, so do end organ complications like chronic pain. Environmental factors like health economics and genetic factors may influence such outcomes. Recent data from UK, showed higher than expected survival (ASH 2015) in the setting of a national health care system. The Hospital of Gela, Sicily has a well characterized Caucasian SCD population, which also has consistent access to health care via a nationalized system. We hypothesized that traditional disease severity markers including end organ damage, the rate of red blood cell (RBC) alloimmunization and opioid use would be lower in Sicily, compared to a modern US population of African ancestry with inconsistent access to healthcare.Methods: 90 SCD patients in Sicily were followed for over 9 years (2006-2014) to identify clinical complications, alloimmunization rates and survival. Demographics and sickle and alpha globin genotypes were documented. In the US, 632 SCD patients (excluding those with SC) had the same parameters collected as part of the Bethesda Sickle Cell Cohort Study. Laboratory and clinical complications were compared between populations according to phenotype groups (SS/SB0 or SB+ thalassemia). Clinical manifestations included hospitalizations for pain per year, RBC alloimmunization, hydroxyurea prescription rate. Statistical analysis utilized univariate comparisons (t and Chi square tests).Results: In Sicily, 51 patients had SS/SB0 and 28 SB+. All were of self-described Caucasian ancestry. No SC patients were identified in Sicily, therefore all comparisons were limited to SS/SB0 and SB+ thalassemia. Sicilian SS/SB0 patients were older than the US (p<0.0001), had lower AST (p=0.04) and creatinine (p=0.005), and higher HbF (p<0.0001). SB+ patients had similar characteristics (Table 1). Sixteen Sicilian patients (18%) died at a mean age of 53 years (range 31-77). As suggested by these survival data, SCD complications were less frequent for both SS/SB0 and SB+ groups in Sicily compared to the US (Table 2). More Sicilians were prescribed hydroxyurea, especially among those with SB+, although this was not a significant difference. The pain crisis hospitalizations per year was also higher in the US (p=0.008). Consistent with less frequent hospitalizations, only 4 of the Sicilian patients (3 SS/SB0, 1 SB+) were taking long acting opioids, compared to more frequent opioid use in the US. Finally, RBC alloimmunization, another surrogate measure for disease severity, was twice as common in SS/SB0 patients from the US (30%) as in Sicily (14%, p=0.01).Conclusions: These data suggest that end organ complications, like chronic pain and alloimmunization, in self-described Caucasians with uniform access to health care in Sicily, are less common than in a US cohort of adults. These differences could be attributable to access to specialized care, genetic differences in the proportion of African ancestry or environmental factors like more frequent use of preventative therapies like hydroxyurea. RBC alloimmunization in Sicily is significantly lower than reported in the US , but is still higher than observed in thalassemics in Italy. Perhaps this could be explained a matched genetic background between blood donors and recipients in Sicily, as has been suggested as a strategy to reduce RBC alloimmunization in the US. Further studies comparing unique SCD populations from different geographic regions may be helpful to elucidate genetic or environmental risk factors for end organ complications and disease severity. [Display omitted] [Display omitted] DisclosuresNo relevant conflicts of interest to declare.

Physical Properties of Blood Are Altered in Young and Lean Women with Polycystic Ovary Syndrome.

Classic features of polycystic ovary syndrome (PCOS) include derangement of metabolic and cardiovascular health, and vascular dysfunction is commonly reported. These comorbidities indicate impaired blood flow; however, other than limited reports of increased plasma viscosity, surprisingly little is known regarding the physical properties of blood in PCOS. We aimed to investigate whether haemorheology was impaired in women with PCOS. We thus measured a comprehensive haemorheological profile, in a case-control design, of lean women with PCOS and age-matched healthy controls. A clinical examination determined similar cardiovascular risk for the two groups. Whole blood and plasma viscosity was measured using a cone-plate viscometer. The magnitude and rate of red blood cell (RBC) aggregation was determined using a light-transmission aggregometer, and the degree of RBC deformability was measured via laser-diffraction ektacytometry. Plasma viscosity was significantly increased in women with PCOS. Blood viscosity was also increased for PCOS at lower-to-moderate shear rates in both native and standardised haematocrit samples. The magnitude of RBC aggregation–a primary determinant of low-shear blood viscosity–was significantly increased in PCOS at native and 0.4 L·L-1 haematocrit. No difference was detected between PCOS and CON groups for RBC deformability measurements. A novel measure indicating the effectiveness of oxygen transport by RBC (i.e., the haematocrit-to-viscosity ratio; HVR) was decreased at all shear rates in women with PCOS. In a group of young and lean women with PCOS with an unremarkable cardiovascular risk profile based on clinical data, significant haemorheological impairment was observed. The degree of haemorheological derangement observed in the present study reflects that of overt chronic disease, and provides an avenue for future therapeutic intervention in PCOS.

Effect of yeast biomass with high content of carotenoids on erythrocyte deformability, NO production and Na,K-ATPase activity in healthy and LPS treated rats.

Measurements of red blood cell (RBC) deformability together with estimation of NO-synthase activity and Na,K-ATPase activity were used for characterization of RBC functionality in rats subjected to single dose of Escherichia coli lipopolysaccharides (LPS) at a dose of 1 mg/kg. We hypothesized that LPS might initiate a malfunction of RBC. We also investigated the potential effect of carotenoids (10 mg/kg/day) produced in red yeast biomass of Rhodotorula glutinis on RBC in LPS-challenged rats. LPS significantly reduced the deformability of RBC (by 14%) together with decrease of NO-synthase activity by 20%. Daily supplementation of carotenoids for 10 days attenuated the LPS-induced injury, as observed by 22% increase of RBC deformability and 23% increase of NO-synthase activity. The activity of Na,K-ATPase was also improved probably due to increased number of active enzyme molecules as indicated by 66% enhancement of Vmax value, hence maintaining the activity of erythrocyte Na,K-ATPase to the level even higher as compared with healthy control animals. It may be concluded that administration of yeast biomass with high content of carotenoids resulted in advanced function of erythrocytes as concerns their ability to squeeze through narrow capillaries of the circulation, better intrinsic production of NO and improvement of intracellular homeostasis of sodium.

БІОХІМІЧНІ ТА ФІЗІОЛОГІЧНІ ЗМІНИ В КРОВІ ПРИ ЕКСПЕРИМЕНТАЛЬНІЙ ЕМФІЗЕМІ

&lt;p&gt;РЕЗЮМЕ. У роботі показано, що під час розвитку експериментальної емфіземи в мембрані еритроцитів тварин спостерігаються суттєві біохімічні та структурні зміни.&lt;/p&gt;&lt;p&gt; &lt;/p&gt;

Study of Diseases Associated ESR above 100 among the Patients who Visited Shahid Sadoughi Hospital in the Second half of 2013

Introduction: ESR (Erythrocyte Sedimentation Rate) is the sedimentation rate of red blood cells per unit of time that is expressed in millimeters. ESR test is simple and inexpensive and fast and can be somewhat diagnostic manual to confirm the diagnosis, rule out the diagnosis, follow-up course of the disease and so on, so it is of interest to practitioners.Methods: Descriptive and cross-sectional study on 124 patients in all wards of Shahid Sadooghi hospital in the second 6 months of 1393, that had raised ESR of 100 at least one time. All information including gender, age, diagnosis of the presence or absence of fever and indigenous of them cases in the archive hospital patients were taken.Results: 56% female and 44% were male. 16% of patients in the age range 0-19 years, 56% aged 20-64 years and 28% were aged over 64 years. 41% of patients were admitted with fever. 59% were native and 41 % were non-native. Infectious causes (45.9%), malignancy (14.5%), collagen vascular disease (16.9%), internal disease and hematology (16.9%) and other causes (5.6%) were formed cases. Diabetic foot (7%), pneumonia (7%), RA (6%), ESRD (5.6%), metastatic lung cancer (4%), TB (3%) formed the majority of cases.In feverish cases pneumonia (11.5%), diabetic foot (7.6%), tuberculosis (5.7%), pyelonephritis (5.7%) and RA (5.7%) were the most frequent form. Infectious causes of fever (65.3%), collagen vascular (13.4%), malignancies (5.7%) and other causes (15.6%).conclusion: In patients with extremely high sediment in terms of gender segregation is almost the same of hospital commiunity.

Repeated administration of benzophenone induced oxidative stress in alterations of morphometry and cytomembrane morphology of rat red blood cell

Repeated administration of benzophenone induced oxidative stress in alterations of morphometry and cytomembrane morphology of rat red blood cell

Synthesis, Characterization, In Vivo Imaging, Hemolysis, and Toxicity of Hydrophilic Ag2S Near-Infrared Quantum Dots

Ag2S quantum dots (Qds) are a new type of near-infrared Qds and hold great potential for in vivo imaging. There is great demand for versatile, practical, and environmentally-friendly processes for formation of hydrophilic Ag2S Qds. The present study prepared hydrophilic Ag2S near-infrared Qds using a simple, easy, one-step method. The Qds were characterized by TEM, XRD, FT-IR, UV–Visible and PL spectroscopy and the effect of the reaction variables of time, temperature, precipitation agent feeding rate, and S:Ag molar ratio on the size and PL spectrum of the Ag2S Qds was investigated. The Qds exhibited near-infrared photoluminescence and good photostability, which was evaluated under different conditions. Cytotoxicity tests were carried out on human A549 and Hep G2 cell lines at concentrations of 6.25–200 µg/ml. MTT assay revealed that the Qds showed no significant toxicity. The effect of plasma and corona proteins on the hemolytic activity of Ag2S Qds in four concentrations was evaluated for rat and human red blood cells. In vivo imaging showed that the Ag2S Qds penetrated the body of mice with excellent brightness. The hydrophilic Ag2S Qds can be safely applied as near-infrared optical imaging probes for nanodiagnostics or in vivo imaging.

EFFECT OF L-ARGININE - NO ON PROOXIDANT-ANTIOXIDANT BALANCE IN ERYTHROCYTES OF RATS UNDER ALCOHOL INTOXICATION.

It was shown changes in the activity of antioxidant enzymes (superoxide dismutase, catalase) and NO-synthase (NOS), in the content. of stable metabolic products of nitric oxide and levels of lipid peroxidation products in erythrocytes of rats under alcoholic intoxication. It was shown that animals with alcohol intoxication under of the admission of the main substrate NOS - L-arginine activity of antioxidant protection enzymes was increased in twice on the fond of TBA-positive products decrease contents. Established in hemolisate red blood cells in rats with alcohol inrotoxication value of total NOS activity decreases by 65% compared to control. Not selective inhibitor Nto-nitro-L-arginine methyl ester (L-NAME), which is a structural analog of L-arginine, reduced output level of total NOS activity by 23.4% in the control and 25% under conditions of pathology. The consumption of rats L-arginine NOS total activity increased in the two study groups. The results testify that L-arginine has antioxidant properties, whereas L-NAME exerts a slightly stabilizing influence.

Effects of GlyT1 inhibition on erythropoiesis and iron homeostasis in rats.

Glycine is a key rate-limiting component of heme biosynthesis in erythropoietic cells, where the high intracellular glycine demand is primarily supplied by the glycine transporter 1 (GlyT1). The impact of intracellular glycine restriction after GlyT1 inhibition on hematopoiesis and iron regulation is not well established. We investigated the effects of a potent and selective inhibitor of GlyT1, bitopertin, on erythropoiesis and iron homeostasis in rats. GlyT1 inhibition significantly affected erythroid heme biosynthesis, manifesting as microcytic hypochromic regenerative anemia with a 20% steady-state reduction in hemoglobin. Reduced erythropoietic iron utilization was characterized by down-regulation of the transferrin receptor 1 (TfR1) on reticulocytes and modest increased iron storage in the spleen. Hepatic hepcidin expression was not affected. However, under the condition of reduced heme biosynthesis with reduced iron reutilization and increased storage iron, hepcidin at the lower and higher range of normal showed a striking role in tissue distribution of iron. Rapid formation of iron-positive inclusion bodies (IBs) was observed in circulating reticulocytes, with an ultrastructure of iron-containing polymorphic mitochondrial remnants. IB or mitochondrial iron accumulation was absent in bone marrow erythroblasts. In conclusion, GlyT1 inhibition in rats induced a steady-state microcytic hypochromic regenerative anemia and a species-specific accumulation of uncommitted mitochondrial iron in reticulocytes. Importantly, this glycine-restricted anemia provides no feedback signal for increased systemic iron acquisition and the effects reported are pathogenetically distinct from systemic iron-overload anemias and erythropoietic disorders such as acquired sideroblastic anemia.

The effect of aluminum chloride and acetic acid in some blood picture of male rats

The current study was conducted to investigate the poisonous effects of Aluminum Chlorid dissolved in water alone and its effects of dissolved in acetic acid 25% in blood variables for male rats of Mus musculus . fifteen rats of the species Mus musculus ages (3-4) weeks and weights of (100±150gm) were used in this study. The study included tow experiments, (25) rat used for each experiment, which included (5) groups with (5) rat each. The first group of each experiment groups was considered the control group ,while the remaining four groups were considered as treatment groups. The first group were treated with (70 mg/kg of bodyweight) of Aluminum Chloride for different intervals (2, 5, 10, and 20 days); the second group given (70 mg/kg bodyweight) of Aluminum Chloride dissolved in acetic acid 25%. the blood variables for all rats were calculated after the different injection intervals was finished and dissected and blood was taken directly from the heart and put in experimental tube containing anticoagulant EDTA , The results showed low corporate rate of red blood cells ( RBC) and other blood variables and high moral in the rate of white blood cells ( WBC) compared to the control group, these changes were gradual as the dosage period.

Peptide inhibitor of complement C1 modulates acute intravascular hemolysis of mismatched red blood cells in rats.

Acute hemolytic transfusion reactions have a broad clinical presentation from mild and transitory signs and symptoms to shock, disseminated intravascular coagulation, renal failure, and death. We have recently developed a rat model of acute intravascular hemolysis showing that the classical complement pathway mediates antibody-dependent hemolysis. The objective of this study was to evaluate the role of the classical pathway inhibitor peptide inhibitor of complement C1 (PIC1) in this animal model. Male Wistar rats received a 15% transfusion of human red blood cells (RBCs) and blood was isolated from the animals up to 120 minutes. Animals received PIC1 either 2 minutes before or 0.5 minutes after transfusion. Sham-, vehicle-, and cobra venom factor (CVF)-treated animals were used as control groups with a subset of rats also receiving an equivalent dose of intravenous immunoglobulin (IVIG) before transfusion. Blood was analyzed for transfused RBC survival by flow cytometry and free hemoglobin (Hb) in isolated plasma by spectrophotometry. Vehicle-treated rats showed decreased human RBC survival and increased free Hb as expected. Rats receiving PIC1 before transfusion showed increased human RBC survival and decreased Hb similar to CVF-treated rats. Notably, rats receiving PIC1 after initiation of transfusion showed similar decreases in hemolysis as animals receiving PIC1 before transfusion. Compared to IVIG and saline controls, PIC1-treated animals demonstrated decreased hemolysis and protection from acute kidney injury. These results demonstrate that PIC1 has efficacy in an animal model of acute intravascular hemolysis in both prevention and rescue scenarios.

Alanine-162 of Bacillus thuringiensis Cyt2Aa2 toxin is essential for membrane binding and oligomerisation

ABSTRACTCyt2Aa2 is a cytolytic toxin produced by Bacillus thuringiensis subsp. darmstadiensis. It is specifically toxic to dipteran larvae in vivo and is also active against several cell types, such as erythrocytes. The active toxin is proposed to bind to the cell membrane, and membrane pore formation by toxin oligomerisation leads to cell lysis. This study aimed to characterise the role of residues (I139, S159, L160, S161, A162, D209 and V215) potentially involved in the membrane binding of Cyt2Aa2. All mutants, except I139A and V215A, showed similar characteristics to the wild-type toxin after proteinase K cleavage. Three mutants, S159A, L160A and S161A, showed high haemolytic activity but low toxicity against Aedes aegypti. Membrane interaction assays showed that these mutants could bind to rat red blood cells (rRBCs) and oligomerise. The mutant D209N had no haemolytic activity but was still mildly toxic to A. aegypti. The mutant A162V could not lyse rRBCs, even at high concentrations, and showed no toxicity against A. aegypti. Our data suggest that alanine 162 of the Cyt2Aa2 toxin is involved in membrane binding and oligomerisation. Substitution of this amino acid altered the conformation of the toxin and affected its biological activity.

Effect of liposome-treated red blood cells in an anemic rat model

Context: Liposomes have been shown to improve human red blood cell (RBC) in vitro quality by minimizing membrane damage occurring during 42-d hypothermic storage. Small animal models are necessary to evaluate novel blood products and guide future clinical studies.Objectives: The aim of this study was to assess the effect of liposome treatments on rat RBC hypothermic storage lesion (HSL) and to examine in vivo outcomes of transfusing liposome treated RBCs in a rat model.Materials and methods: Unilamellar liposomes were synthesized which contained saturated (DPPC:CHOL, 7:3 mol%), unsaturated (DOPC:CHOL, 7:3 mol%), saturated charged (DPPC:CHOL:PS, 6:3:1 mol%), and unsaturated charged (DOPC:CHOL:PS, 6:3:1 mol%) phospholipids. After liposome treatment, rat RBC quality was assessed by percent hemolysis, deformability, aggregation, hematological indices, microvesiculation, and cholesterol/phospholipid concentrations. An anemic rat model of myocardial ischemia and reperfusion (I/R) was used to evaluate the outcomes of transfusing liposome-treated RBCs.Results: All four liposome treatments resulted in significant decreases in hemolysis, with the most prominent effect seen with DOPC-liposomes (DOPC: 1.6 ± 0.1% versus control: 3.1 ± 0.2%, p = 0.015). RBCs treated with uncharged liposomes had lower hemolysis compared with charged liposomes (3.4 ± 0.2% versus 3.9 ± 0.4%, p = 0.010). The in vivo study showed no significant difference in the hemoglobin levels and infarct size (53.3 ± 13.1% versus 45.3 ± 8.4%, p = 0.223) between liposome and control groups.Discussion and conclusion: Liposome treatment improved in vitro quality of stored rat RBCs. However, the changes observed in vitro were not sufficient to improve the in vivo outcomes of myocardial I/R in anemic rats transfused with liposome-treated RBCs.

The effect of centrifugation at various g force levels on rheological properties of rat, dog, pig and human red blood cells.

Laboratory investigations often require centrifugation of blood samples for various erythrocyte tests. Although there is a lack of data about the effect of centrifugation at various g force levels on erythrocyte rheological properties. We aimed to investigate the effect of a 10-minute centrifugation at 500, 1000 or 1500 g at 15°C of rat, dog, pig and human venous (K3-EDTA, 1.5 mg/ml) blood samples. Hematological parameters, erythrocyte deformability, cell membrane stability, osmotic gradient ektacytometry (osmoscan) and erythrocyte aggregation were determined. Hematological and erythrocyte deformability parameters showed interspecies differences, centrifugation caused no significant alterations. Cell membrane stability for human erythrocytes centrifuged at higher g level showed less decrease in deformability. Osmoscan O min parameter showed slight elevation in dog centrifuged aliquots. Erythrocyte aggregation parameters changed unexpectedly. Rat and dog erythrocyte aggregation indices significantly dropped in centrifuged aliquots. Pig erythrocyte aggregation indices increased significantly after centrifugation. Human erythrocyte aggregation was the most stable one among the investigated species. The used centrifugation protocols caused the largest alterations in erythrocyte aggregation in a controversial way among the investigated species. On the other hand, erythrocyte deformability parameters were stable, cell membrane stability and osmoscan data show minor shifts.