Increase In Red Blood Cells Research Articles

Evaluation of Acute Toxicity of the Alkaloids of Peganum harmala L Seeds in Albino Wistar Female Rats

The Peganum harmala, commonly known as syrian street, belongs to the family Zygophyllaceae. Peganum harmala L is widely used in traditional medicine and pharmacology. The evaluation of toxic properties of Peganum harmala is crucial when considering public health protection because exposure to plant extracts can result in undesirable effects on consumers. Hence, in this study the acute IP toxicity of the total alkaloids of Peganum harmala L seeds was investigated in female rats. The oral dose (60 mg/kg) of tested plant extract was administered to two groups of animals (group I and II) in single dose and third group III is control. The first treated group is sacrificed after 24 hours and the second treated group and control group are sacrificed after 5 days. Throughout the treatment, changes in behavioral pattern, clinical sign and body weight of rats in treatment groups are observed. Also, a significant decrease in relative liver mass and serum biochemical parameters (AST, ALT, PAL, Bil D and Bil T) were observed. An increase in red blood cells, hemoglobin and hematocrit was recorded in group II. Histological examination shows that the seed extract of Peganum harmala L has no remarkable toxic effect on the liver of the treated animals of the second group. The result indicates that the IP administration of the total alkaloids of Peganum harmala L seeds plant did not produce any significant toxic effect in albino rats.

Antibodies against the erythroferrone N-terminal domain prevent hepcidin suppression and ameliorate murine thalassemia

AbstractErythroferrone (ERFE) is produced by erythroblasts in response to erythropoietin (EPO) and acts in the liver to prevent hepcidin stimulation by BMP6. Hepcidin suppression allows for the mobilization of iron to the bone marrow for the production of red blood cells. Aberrantly high circulating ERFE in conditions of stress erythropoiesis, such as in patients with β-thalassemia, promotes the tissue iron accumulation that substantially contributes to morbidity in these patients. Here we developed antibodies against ERFE to prevent hepcidin suppression and to correct the iron loading phenotype in a mouse model of β-thalassemia [Hbb(th3/+) mice] and used these antibodies as tools to further characterize ERFE's mechanism of action. We show that ERFE binds to BMP6 with nanomolar affinity and binds BMP2 and BMP4 with somewhat weaker affinities. We found that BMP6 binds the N-terminal domain of ERFE, and a polypeptide derived from the N terminus of ERFE was sufficient to cause hepcidin suppression in Huh7 hepatoma cells and in wild-type mice. Anti-ERFE antibodies targeting the N-terminal domain prevented hepcidin suppression in ERFE-treated Huh7 cells and in EPO-treated mice. Finally, we observed a decrease in splenomegaly and serum and liver iron in anti–ERFE-treated Hbb(th3/+) mice, accompanied by an increase in red blood cells and hemoglobin and a decrease in reticulocyte counts. In summary, we show that ERFE binds BMP6 directly and with high affinity, and that antibodies targeting the N-terminal domain of ERFE that prevent ERFE–BMP6 interactions constitute a potential therapeutic tool for iron loading anemias.

Hematological and biochemical blood indices of rainbow trout depending on the level of protein in feeds

Protein nutrition of fish is one of the main factors of ensuring their active growth. Finding the optimal concentration of protein is an actual direction of research. The purpose of the experiment was to establish the effect of different protein nutrition levels in commercial rainbow trout feeding on hematological and biochemical parameters of their blood. For this purpose, five experimental groups were formed by the method of analogs. The experiment lasted 210 days and was divided into two periods: equalization (10 days) and main (200 days). During the equalization period, the feeding ration was the same for fish in the control and experimental groups. In the main period, the level of protein in experimental feeds for different experimental trout groups ranged from 44 to 52% per 1 kg. It was found that different levels of rainbow trout’s protein nutrition did not cause significant changes in blood morphological composition. It is proved that rearing trout using compound feeds with a crude protein content of 50% and 52% promotes the increase of red blood cells, hemoglobin, white blood cells and phagocytic activity in blood. The use of 44% and 46% low crude protein diets in fish feeding reduces the morphological components in blood. As a result of studies on the phagocytic activity of neutrophils in fish blood, a slight increase was revealed under the influence of the studied factor. The analysis of the obtained results showed that with increasing levels of crude protein in the diets of rainbow trout, the concentration of total protein and its fractions in blood serum increased. Accordingly, the amount of total protein in blood of fish in the control group was 60.0 g/L, in the experimental groups – it was within the range of 59.6- 65.3 g/L. The increase in the level of rainbow trout’s protein nutrition in the experimental groups 4 and 5 caused an increase in the number of albumins, β-globulins and γ-globulins compared to control. As a result of experimental studies, an increase in the mineral status of rainbow trout blood during use of feeds with the crude protein content of 50-52 % was established. No differences were found during the assessment of trout white blood cell counts. The use of the obtained results during the production of compound feed will allow having a positive effect on the hematological parameters of commercial rainbow trout

The influence of "Stimix Zoostim" and "Normosil" probiotics on fecal microflora, hematologic indicators, nutrient digestibility, and growth of mother-bonded calves.

Aim:This paper presents the results of the studies on “Stimix Zoostim” and “Normosil” probiotics and their influence on fecal microflora, hematologic indicators, immunoresistance, nutrient digestibility, and growth intensity of mother-bonded calves.Materials and Methods:The calves of the control group were fed with their basic diet (BD) without “Stimix Zoostim” or “Normosil,” whereas the calves of the experimental group were fed with their BD supplemented with “Stimix Zoostim” and Normosil.” For 10-20-day-old calves, the daily dose was 10 mL per head, whereas 21-90-day-old calves received 15 mL of probiotics per head per day. The calves of the experimental group were administered probiotics every day. Calves aged 10 to 60 days received probiotics with milk, and then at the age of 61 to 90 days, received probiotics with water. (This is a sequential process. At 60 days old, calf rearing with milk stops, and after that, we use probiotics added to water to rear them). Both groups were administered probiotics twice a day, specifically 50% of the daily dose at each feeding time.Results:The results in this study proved that “Stimix Zoostim” and “Normosil” probiotics exhibit high probiotic activity and have a positive effect on the calves’ fecal microflora. Feeding the calves with probiotics resulted in a significant increase in the number of normal flora, such as lactic acid bacteria and bifidobacteria, and a decrease in the amount of Escherichia. It also resulted in an increase in red blood cells, hemoglobin, and γ-globulins within the physiological range. In our study, we found that the phagocytic reaction in the blood serum of the experimental group was slightly enhanced, suggesting a high response of the body to infectious agents.Conclusion:Thus, the group of calves receiving probiotic “Stimix Zoostim” exhibited an exceedance of phagocytic activity by 4.8% and the group receiving “Normosil” by 4.4%, in comparison with the control group. The daily dose of 10 mL of probiotics per head for 10-20-day-old calves and 15 mL per head for 21-90-day-old calves also had a positive effect on nutrient digestibility, growth, and forage consumption. The economic benefit per animal was 149.23 and 157.0 rubles, respectively.

Effect of Gonadotropin Releasing Hormone Injection on Physiological Changes and Reproductive Hormones in Clarias gariepinus

Evaluation of gonadotropin releasing hormone (GnRH) to physiological changes, spawning and reproductive parameters of African catfish, Clarias gariepinus in the current study has been conducted to determine the effectiveness of GnRH in stimulation the maturation and ovulation of African catfish. GnRH at 4, 8 and 12 μg/kg body weight dissolved in 0.6% NaCl under a natural temperature regime. The fish from Abbasa farm were fed with 30 % protein diet during experiment. Fish550 - 600 g body weight were injected with a doses of (4, 8 and 12 μg/kg body weight) three times a week. Result showed multiple ovulations and spawns within a period of approximately 30 days. Levels of 17β- estradiol (E2), testosterone (T) and follicular stimulating hormone (FSH) were elevated at 30 days’ post treatment, preceding the spawns with the highest fecundity. These results show that administration of GnRH induces multiple spawns in fishwhere, approximately 100% increase in red blood cells (RBCs), white blood cells (WBCs), hematocrit value (Hc) and hemoglobin content (Hb) respectively. Also, there are increased in plasma glucose level, total protein, aspartate and alanine amino transferase activities (AST and ALT) and increased in plasma creatinine and uric acid concentration after administration of GnRH. These data suggest that fish respond to administration of GnRH by increasing their hematology, enzymes of liver functions and kidneyfunctions. Injection of GnRH at 4, 8 and 12 µg/kg was found to be effective to induce ovulation in African catfish. 100% ovulation was also observed for the fish treated with GnRH. Also, plasma level of E2, FSH and T increased significantly with the association of ovulation. The present study indicates that administration of GnRH is effective for sexual maturation and ovulation in African catfish.

Дисфункция эндотелия как главный фактор кардиоваскулярного риска, лимитирующий работоспособность юных спортсменов: методы оценки и коррекция с помощью специализированных продуктов питания на основе апифитокомпонентов

Objective: to assess with exercise tests the endothelial dysfunction (ED) and performance before and after the treatment with specialized athlete’s nutrition product (SANP) with apian and herbal components. Materials and methods: 88 athletes aged 16-18 we tested. The main group included 58 athletes, who took SANP (candy bar «Gold F25 ApiSpeis Light») for 2 months. The control group-1 included 32 schoolchildren and students, non-athletes, with normal endothelium-dependent vasodilatation function of the brachial artery (EDVD BA) at the peak of the reactive hyperemia test (RHT). Control group-2, 30 athletes who did not take SANP, was formed for the comparison with the main group after taking SANP. For all individuals, we evaluated the level of EDVD, endothelium-dependent vasoconstriction (EDVC BA) at the peak of the hyperventilation test (HVT), the coefficients (C) of the endothelial sensitivity to shear stress and the blood flow velocity (Vps) of the BA at the peak of the RHT and HVT, the endothelial sensitivity index (ESI), and the maximum load (Wmax), the anaerobic threshold time (ATT). We also did general and biochemical blood tests. Results: the earliest manifestation of endothelial dysfunction is a decrease in sensitivity to shear stress during RHT and HVT resulting from endotheliocyte overstrain which can be assessed with ESI (sensitivity 80.9%, specificity 99.4 The EDVD diagnostic significance was 5.6 times lower than that of EIS, while 10.5% of athletes with over 10% EDVD actually had ED, with a predominant vasospasm and high cardiovascular risk. After the course of SANP in the main group, EDVD increased by 70%, EDVC decreased by 2.2 times, and the ESI increased by 2.3 times, against the background of improved performance. Total cholesterol decreased by 13%, triglycerides by 17%, cortisol by 14%, lactate by 25%. Red blood cell and mineral levels increased. A correlation was found between an increase in ATT a decrease in cortisol (r=-0.53; p<0.0001), an increase in red blood cells (r=0.62; p<0.0001), and an increase in Vps in BA during of hyperventilation with an increase in Wmax (r=0.52; p<0.0001). Conclusions: ED is the main factor of cardiovascular risk, limiting working capacity and forming a spastic type of regional hemodynamics of young athletes. The ESI criterion enables the detection of a decrease in the endothelial sensitivity to shear stress during a hyperventilation test, which causes the predominant vasospasm. Th e tested SANP can be used in the system of biomedical support for young athletes.

Transformer mineral oil ingestion induces systemic sub-acute toxicity in Wistar rats

We investigated the potential toxicities associated with the sub-acute ingestion of transformer mineral oil (TMO) at a heated low dose (HLD-50 mg/kg), heated high dose (HHD-500 mg/kg) and unheated high dose (UHD-500 mg/kg) in Wistar rats. There were increases in red blood cells and haemoglobin levels in HHD females and UHD males respectively versus control. The serum total proteins, albumin, and creatinine of the HHD females showed a significant increase versus control. The HHD males and UHD groups showed significant increase in liver malondialdehyde versus control. The livers of HHD groups had bile duct proliferation while those of HLD females and UHD groups showed focal areas of periportal chronic inflammation. HHD groups had kidneys with mild chronic inflammation and the HHD and UHD groups showed small intestines with chronic inflammation. In conclusion, sub-acute oral administration of TMO induced various degrees of dermal, haematological, hepatic, renal and small-intestinal toxicities in rats.

ACUTE AND SUB-ACUTE TOXICITY STUDIES OF ETHANOLIC STEM AND ROOT EXTRACTS OF ARISTOLOCHIA RINGENS AND SECURIDACA LONGEPEDUNCULATA IN WISTAR RATS

The aim of this study was to examine the acute and sub-acute toxicity of the ethanolic extract of the stem and root of Aristolochia ringens and Securidaca longepedunculata, respectively, in Wistar rats. The animals were divided into 5 per group and oral administration of extract at doses of 100, 250, 500 mg/kg were given daily for 28 days. Control group was given distilled water and animals were weighed at 7 day intervals. Animals were kept under close observation for 24 h and 28 days for acute and sub acute toxicity study, respectively. Body and relative organ weight, biochemical and haemotological analysis and phytochemical screening were carried out. The extract did not exhibit significant change in the percentage body weight when compared to the control group. The stem extract of A. rigens did not show any significant difference in the weight of the organs, weight while the root extract of S. longepedunculata exhibited a significant dose-dependent decrease in the weight of the liver and heart in all the treated groups (p&lt;0.05). Extract showed significant increase in red blood cell (RBC), haemoglobin (HGB) and haemotocrit (HCT) levels in A. ringens, while it decreased in S. longepedunculata extract in the study. There was no significant alteration in the haematological parameters (WBC, MCHC, MCV and MCH) of the treated rats when compared to the control rats. All blood serum biochemical parameters were not significantly (p&lt;0.05) different as compared with the control group, except for a significant (p&lt;0.05) non-dose-dependent increase of TP, HDL-c and LDL-c of A. ringens extract. The result indicate non toxicity of ethanolic stem and root extracts of A. ringens and S. longepedunculata, respectively, were found to be safe when orally administered.

The Haematological Effects of Oleanolic Acid in Streptozotocin-Induced Diabetic Rats: Effects on Selected Markers.

Background Sustained hyperglycaemia leads to the development of haematological alterations which, if left untreated, is associated with cardiovascular complications. Insulin is the mainstay drug in type 1 diabetes mellitus (T1D); however, the use of insulin is associated with haematological alterations that could further worsen cardiovascular complications. Therefore, the aim of the study was to investigate the haematological effects of oleanolic acid (OA) in streptozotocin- (STZ-) induced diabetic rats. Methods The animals were separated into five groups; the nondiabetic group (ND), the diabetic control group (DC), and the treatment groups of insulin (170 μg/kg, s.c), metformin (500 mg/kg, p.o), and OA (80 mg/kg, p.o). OA was administered orally twice a day. Thereafter, the animals were sacrificed, and blood and tissues were collected for haematological, hormonal, and oxidative status analysis. Results Untreated diabetic rats exhibited hyperglycaemia, elevated glycated haemoglobin (HbA1c), oxidative stress, and a reduced erythropoietin (EPO) concentration when compared to ND rats. However, administration of OA attenuated hyperglycaemia, HbA1c, and EPO concentrations compared to DC rats. The reduction of blood glucose concentration, HbA1c, and improved EPO concentrations was further associated with a notable increase in red blood cell (RBC) count and other RBC indices. We also observed an increase in the antioxidant status of the RBCs with a concomitant decrease in oxidative stress. Conclusion These findings suggest that OA improves diabetes-induced haematological changes caused by hyperglycaemia and attenuates the progression of cardiovascular complications in DM individuals.

Lobe specificity of iron binding to transferrin modulates murine erythropoiesis and iron homeostasis

AbstractTransferrin, the major plasma iron-binding molecule, interacts with cell-surface receptors to deliver iron, modulates hepcidin expression, and regulates erythropoiesis. Transferrin binds and releases iron via either or both of 2 homologous lobes (N and C). To test the hypothesis that the specificity of iron occupancy in the N vs C lobe influences transferrin function, we generated mice with mutations to abrogate iron binding in either lobe (TfN-bl or TfC-bl). Mice homozygous for either mutation had hepatocellular iron loading and decreased liver hepcidin expression (relative to iron concentration), although to different magnitudes. Both mouse models demonstrated some aspects of iron-restricted erythropoiesis, including increased zinc protoporphyrin levels, decreased hemoglobin levels, and microcytosis. Moreover, the TfN-bl/N-bl mice demonstrated the anticipated effect of iron restriction on red cell production (ie, no increase in red blood cell [RBC] count despite elevated erythropoietin levels), along with a poor response to exogenous erythropoietin. In contrast, the TfC-bl/C-bl mice had elevated RBC counts and an exaggerated response to exogenous erythropoietin sufficient to ameliorate the anemia. Observations in heterozygous mice further support a role for relative N vs C lobe iron occupancy in transferrin-mediated regulation of iron homeostasis and erythropoiesis.

Preoperative Anaemia in Primary Hip and Knee Arthroplasty.

Approximately one in three patients has untreated preoperative anaemia, which in turn is associated with an increased need for transfusion of allogenic red blood cell concentrates (RBC) and complications in the context of a surgical intervention. Here, the prevalence of preoperative and postoperative anaemia as well as their effects on transfusion rate, hospital length of stay and hospital mortality in primary hip and knee arthroplasty has been analysed. From January 2012 to September 2018, 378,069 adult inpatients from 13 German hospitals were analysed on the basis of an anonymized registry. Of these, n = 10,017 patients had a hip and knee joint primary arthroplasty. The primary endpoint was the incidence of preoperative anaemia, which was analysed by the first available preoperative haemoglobin value according to the WHO definition. Secondary endpoints included in-hospital length of stay, number of patients with red blood cell concentrate transfusion, incidence of hospital-acquired anaemia, number of deceased patients, and postoperative complications. The preoperative anaemia rate was 14.8% for elective knee joint arthroplasty, 22.9% for elective hip joint arthroplasty and 45.0% for duo-prosthesis implantation. Preoperative anaemia led to a significantly higher transfusion rate (knee: 8.3 vs. 1.8%; hip: 34.5 vs. 8.1%; duo-prosthesis: 42.3 vs. 17.4%), an increased red blood cell concentrate consumption (knee: 256 ± 107 vs. 29 ± 5 RBC/1000 patients; hip: 929 ± 60 vs. 190 ± 16 RBC/1000 patients; duo-prosthesis: 1411 ± 98 vs. 453 ± 42 RBC/1000 patients). Pre-operative anaemia was associated with prolonged hospital stay (12.0 [10.0; 17.0] d vs. 11.0 [9.0; 13.0] d; p < 0.001) and increased mortality (5.5% [4.6 - 6.5%] vs. 0.9% [0.7 - 1.2%]; Fisher p < 0.001) compared to non-anaemic patients. In patients aged 80 years and older, the incidence of preoperative anaemia and thus the transfusion rate was almost twice as high as in patients under 80 years of age. Preoperative anaemia is common in knee and hip primary arthroplasty and was associated with a relevant increase in red blood cell concentrate consumption. In the context of patient blood management, a relevant potential arises, especially in elective orthopaedic surgery, to better prepare elective patients, to avoid unnecessary transfusions and thus to conserve the valuable resource blood.

Stimulation of Bladder Acupoints by Cloprostenol for Treating Back Soreness in Athletic Horses

Twenty-five Thoroughbred jumper geldings suffered back soreness with poor performance, and 5 control horses were assessed by archived computer data, clinical examination, and laboratory analyses of complete blood picture, serum enzymes, and cortisol level, before and after cloprostenol-pharmacopuncture. The 25 diseased horses before therapy showed significant increases in aspartate aminotransferase and creatine phosphokinase with clinical pains scored mild in 15 horses, moderate in 9 horses, and severe in one horse, without changes in the hormonal and hematological data. After therapy, they responded by an increase of heart rate (57.8 ± 4.3 bpm), body temperature (38.5 ± 0.7°C), respiration rate (28.3 ± 2.1 bpm), and capillary refilling time (CRT) (1.0 ± 0.0). On the 2nd day, a significant decrease in the mean levels of aspartate aminotransferase and creatine phosphokinase (P = 0.001) was detected, while on the 4th day, they mimed the level of the 5 controls, and on the 6th day, they showed a significant decrease (P = 0.002). The serum cortisol level showed a significant increase on the 6th day of treatment (P = 0.013). The blood picture showed significant increases in red blood cells, mean corpuscular volume, platelets, white blood cells, hemoglobin, mean corpuscular hemoglobin, mean corpuscular hemoglobin concentration, mean platelet volume, platelet distribution width, lymphocytes, plateletcrit, and large platelet concentration ratio (P < 0.05) and nonsignificant changes in hematocrit, granulocytes, and midocytes. The improved blood parameters, enzymes, hormones, and performance progress after cloprostenol-pharmacopuncture proved its effectiveness in treating back soreness in athletic horses.

Determination of grapefruit (Citrus paradisi) peel extract bio‐active substances and its application in Caspian white fish (Rutilus frisii kutum) diet: Growth, haemato‐biochemical parameters and intestinal morphology

In this study, total phenolic and flavonoid contents of grapefruit peel extract (GPE) were equal to 117.3 ± 0.3 µg of gallic acid/mg and 39.30 ± 0.1 µg of quercetin/mg respectively. Caspian white fish (n = 180, 4 ± 0.9 g body weight) was fed with supplemented diets, including 0, 6.25, 12.5 and 25 mg of GPE/kg for 60 days at 25 ± 1°C. The growth performance was markedly improved in fish fed with 25 mg/kg of GPE compared to others (p < 0.05). Moreover, fish fed with 25 mg/kg of GPE showed a significant increase in red blood cell (2.65·106 cell/mm3), white blood cell (17.75·103 cell/mm3), packed cell volume (48%) and haemoglobin concentrations (8.75 g/dl) compared to the control (p < 0.05). However, the highest alanine aminotransferase (140 U/L), alanine transaminase (14.5 U/L), and alkaline phosphatase (18.5 U/L) were observed in control group. Morphological analysis of intestine revealed the highest amount of villus width (8.4 µM), height (32.86 µM) and surface area (342.7 µM2) in fish fed with 25 mg/kg of GPE (p < 0.05). In conclusion, supplementing feed with GPE at 25 g/kg can improve growth performance and haemato-biochemical parameters of Caspian white fish fry.

Preclinical Evaluation of the Haematinic Activity of an Oral Indiffusible Mixture of Tamarindus indica L. Leaf Extract.

Tamarindus indica L. is known to be a multipurpose traditional plant in India. It is used to treat some bacterial infections, parasitic infestations, constipation, and inflammation. It is also used as a blood tonic and for wound healing. This study was designed to substantiate the traditional claim of haematinic activity for T. indica. T. indica leaf extract was formulated into an oral indiffusible mixture (TIM) and evaluated for its haematinic activity in phenylhydrazine (single dose of 10 mg/kg per oral for 8 days) induced anaemia. Wistar rats were grouped into six (n=6). Groups I and II served as normal control and disease control groups, respectively. Group III received the standard drug (haematinic suspension 2 mL/kg). Groups IV, V, and VI received the formulated oral indiffusible mixture of T. indica at a dose of 100, 200, and 400 mg/kg, respectively. The TIM was formulated and pharmaceutically optimized. It produced significant increases in red blood cells, hemoglobin, and packed cell volume and a decrease in mean corpuscular volume. The results showed that the treatment with TIM reversed phenylhydrazine induced anemia. However, the short duration of the present study is regarded as a limitation, and therefore a longer duration is required for obtaining better responses.

Повышение эффективности дегельминтизации котят и щенков с помощью препарата гамавит

The purpose of the research is to evaluate therapeutic efficiency of Gamavit in the cases of dehelminthization of puppies and kittens. Materials and methods . Researches were conducted on 18 puppies and 20 kittens of different breeds at the age from 3 weeks to 3 months which had been divided based on analogues into two groups on each animal type. Precautionary dehelminthization was conducted for one part of animals, forced dehelminthization for the other part of animals (presence of helminths' ootids in fecal masses Toxocara canis, Toxascaris leonina, Uncinaria stenocephala, Ancylostoma caninum). Anthelmintic Dirofen-suspension was administered to animals of the first group orally as prescribed by instructions. Gamavit at a dose of 0.2 ml/kg was additionally administered as a single dose subdermally during dehelminthization of kittens and puppies of the second group. Results and discussion . Vomiting, decreased appetite were noted in animals of control group (without administration of Gamavit) after dehelminthization. Visible adverse changes were not noted in animals of experimental group (with administration of Gamavit). Weakening of leukocytosis and lowering of ERS, increase in red blood cells and hematocrit level, descreased activity of transferring enzymes and LDH level were noticed in animals of this group suggestive of weakening of intoxication and inflammatory reaction. In contrast, increased activity of transferring enzymes and LDH level were noticed in animals of control group which allows to infer about persisting inflammatory events. Consequently, high therapeutic efficiency of Gamavit in the cases of precautionary and forced dehelminthization were elucidated.

Lack of GDF11 Does Not Ameliorate Erythropoiesis in β-Thalassemia and Does Not Prevent the Activity of the Trap-Ligand RAP-536

Mutations in the HBB gene causes β-thalassemia (BT). Treatment for BT presents a major clinical challenge in the United States, as patients require chronic and expensive treatment for survival. A new drug in Phase III clinical trials, Luspatercept (ACE-536), has been shown to improve BT symptoms via an erythropoietin (EPO) -independent pathway. ACE-536 is a peptide drug identical to the extracellular domain of activin receptor IIB (ACVR2B). Upon administration, it competes with ACVR2B to bind members of the transforming growth factor (TGF) β superfamily. Growth differentiation factor 11 (GDF11) has been pinpointed as the primary target by which the trap ligand exerts its therapeutic efforts. Studies in murine models of BT using RAP-536 (the mouse analog of ACE-536), have suggested that Gdf11 is overexpressed in erythroblasts and that overexpression functions to inhibit erythroid differentiation. Interestingly, however, ACE-536 and RAP-536 have been shown to stimulate RBC synthesis in healthy humans and mice, where GDF11/Gdf11 overexpression has not been reported. Additionally, the expression data in mice has been questioned because of the unavailability of antibodies that can discriminate between Gdf11 and other TGF-β ligands.Due to the novelty of RAP-536 promoting erythropoiesis through an Epo-independent pathway and the lack of specific antibodies to distinguish between TGF-β ligands, we resorted to genetic tools to investigate the role of Gdf11 in erythropoiesis. For our study, we generated Hbb+/+Gdf11flox/flox and Hbbth3/+Gdf11flox/flox mice and crossed them with EpoRCre and VavCre transgenic lines, resulting in offspring harboring the Gdf11 deletion in erythroid cells and the complete hematopoietic compartment. If Gdf11 is secreted by erythroid cells, and it plays a role in inhibiting erythroid differentiation, then mice lacking Gdf11 in either erythroid cells or all hematopoietic cell lineages should show some increase in red blood cell (RBC) production, hemoglobin (Hb) and hematocrit (Hb). Furthermore, in Hbbth3/+mice, where Gdf11 has been proposed to be overexpressed, improvements in erythroid cell differentiation should be most apparent. Surprisingly, we did not detect any differences in RBC number, Hb or Hct levels of Gdf11 deficient Hbb+/+ or Hbbth3/+ mice compared to their Gdf11 containing controls. The discrepancy between our results and published data could be explained if Gdf11 is produced by non-hematopoietic tissues and indirectly influences erythropoiesis. Since Gdf11-/- are embryonic-lethal, we crossed Hbb+/+ Gdf11flox/floxand Hbbth3/+ Gdf11flox/flox mice with a tamoxifen (TAM) inducible Cre recombinase under the global Rosa26 promoter (RosaCre) to assess the effect of a pancellular deletion of Gdf11. No detectable differences were found in RBC, Hb or Hct levels of fl these animals after TAM treatment either acutely nor up to 5-6 months post deletion of Gdf11. Administration of RAP-536 significantly improved and increased hematopoietic parameters in the peripheral blood in all six models lacking Gdf11. In the RAP-536-treated Hbbth3/+ models, amelioration of anemia was noted by a decrease in spleen size and improved ineffective erythropoiesis indicated by an increased hematological parameters and increased ratio of mature to immature erythroblasts in spleen analyzed by FACS. Therefore, lack of Gdf11 at the erythroid, hematopoietic and pancellular level did not prevent a response to the drug.Next, we investigated the effects of RAP-536 directly on erythroid cells. Since the drug causes increases in RBC and Hb of normal patients, we challenged CD34+ cells with RAP-536 at various concentrations. Results showed no increases in cell numbers, erythroid viability, hemoglobin content nor differentiation. Currently we are investigating the mRNA expression of activin receptors IIA and IIb along with TGF-β ligands in healthy and BT CD34+ cells as well as in erythroid specific progenitors of the Hbbth3/+ mouse model.Our findings suggest that Gdf11 is not the sole target of RAP-536, nor that Gdf11 is required to promote improvement of erythropoiesis. Most importantly, we show that in the absence of Gdf11, RAP-536 is effective at increasing hematological parameters in both Hbb+/+ and Hbbth3/+ mice. The results of this study demonstrate potential alternative target(s) for the action of RAP-536. Future work will focus on identifying the unknown targets of RAP-536. DisclosuresCasu:Aevi Genomic Medicine, Inc: Research Funding; Ionis Pharmaceuticals, Inc.: Research Funding. Martinez:Acceleron Pharma: Employment. Suragani:Acceleron Pharma: Employment. Kumar:Acceleron Pharma: Employment. Rivella:Ionis Pharmaceuticals, Inc: Consultancy; Protagonist: Consultancy; Disc Medicine: Consultancy; MeiraGTx: Other: SAB.

Does the Use of an Infusion Pump for Red Blood Cells Increase Hemolysis?

Transfusions of red blood cells (RBCs) are frequently administered for a variety of clinical conditions. The use of an infusion pump has become common practice in nursing. Lack of evidence regarding hemolysis associated with transfusing older RBCs using an infusion pump led one 361-bed acute care suburban medical center to maintain its nursing policy to infuse all blood products by gravity. A combined simulated and clinical study was conducted with 25 units of RBCs. No significant hemolysis was demonstrated as measured by hemolysis index and serum potassium levels. As a result of this study, the nursing policy at the medical center was changed.

English

This study investigates the anti-diabetic and some hematological effects of aqueous and ethanol leaf extracts of Eriosema psoraleoides in alloxan-induced diabetic Wistar rats. Forty-eight albino mice were used for the lethal dose (LD50) study, while twenty-eight Wistar rats were used for the diabetic and haematological study. The result of the phytochemical analysis shows some important phytochemicals that could confer some health benefits. The extracts had lethal doses (LD50) of 4000 and 5000 mg/kg&shy;-1 body weight, which indicate that the extract was safe to a greater extent. The baseline blood glucose concentration presents non-diabetic glucose level 86.25&plusmn;8.26 mg dL&shy;&shy;-1. Diabetes was induced with alloxan monohydrate. A significant decrease (p&lt;0.05) in blood glucose levels in the test animals were observed when compared with positive control. The extracts had a significant effect on blood glucose and haematological levels of the treated rats closer to that of the standard drug (glibenclamide) with significant (p&lt;0.05) increase in red blood cell (RBC) and significant (p&lt;0.05) decrease in neutrophils and lymphocytes in all test groups relative to positive control. However, white blood cell (WBC) showed significant decrease (p&lt;0.05) in groups which received aqueous extracts when compared with positive control. The results of the study showed that the extracts of E. psoraleoides leaves exhibited significant anti-diabetic activities against alloxan-induced diabetes Wistar rats. It suggests the extracts possess bioactive compounds that when properly harnessed could help in improving the health state of diabetic patients. &nbsp; Key words: Eriosema psoraleoides, anti-diabetic, alloxan induced diabetic rats, haematological parameters.

ASSESSMENT OF BIOLOGICAL SAFETY OF FERMENTED PHYLLANTHUS EMBLICA FRUIT JUICE

Objective: The present study evaluated the subchronic toxicity of Lactobacillus mediated fermented Phyllanthus emblica fruit juice (FPJ) using a rat as a model system.Methods: FPJ was prepared, and estimated the changes in pH by pH meter, and microbial load by a plating method. Rats were fed with different dose of FPJ for 60 days. The changes in the body mass were noted. The blood and organs of the experimental rats were collected, after 60 days of intervention. Then, they were analyzed for the selected hematological and biochemical parameters by following standard hospital protocols.Results: The pH of FPJ after 30 days of fermentation was 3.16. FPJ was rich in probiotic Lactobacillus spp. (7.23 Log CFU per mL) without contamination. The supplementation of FPJ was not significantly affected the body weight of the experimental animals, except the female rats in posteffective dose (PED) group showed significant changes (20.83±8.49 g) compared to control (40±17.22 g). The internal organs of the rat were not affected by the FPJ supplementation. The changes observed in blood urea nitrogen, creatinine, cholesterol, triglyceride, aspartate aminotransferase; alanine aminotransferase, and alkaline phosphatase level of experimental rats, both male and female, were not significantly differed from the respective controls. The average of lymphocytes level was significantly increased in continuous dosing group of males and females. Interestingly, the increase in red blood cell and hemoglobin (HGB) were statistically significant for ED group and PED in both sexes, except for females with no effect on HGB content.Conclusion: The prepared FPJ was enriched with probiotic Lactobacillus spp. The supplementation of FPJ (up to 9 mL/kg/day) for 60 days was not significantly influenced the body weight, internal organs, biochemical and hematological parameters of experimental rats (both male and female). The results revealed that FHJ is suitable for the human consumption.

Exploring the relationship of peripheral total bilirubin, red blood cell, and hemoglobin with blood pressure during childhood and adolescence

ObjectiveTotal bilirubin is beneficial for protecting cardiovascular diseases in adults. The authors aimed to investigate the association of total bilirubin, red blood cell, and hemoglobin levels with the prevalence of high blood pressure in children and adolescents. MethodsA total of 3776 students (aged from 6 to 16 years old) were examined using cluster sampling. Pre‐high blood pressure and high blood pressure were respectively defined as the point of 90th and 95th percentiles based on the Fourth Report on the Diagnosis, Evaluation, and Treatment of High Blood Pressure in Children and Adolescents. Both systolic and diastolic blood pressure were standardized into z‐scores. ResultsPeripheral total bilirubin, red blood cell and hemoglobin levels were significantly correlated with age, and also varied with gender. Peripheral total bilirubin was negatively correlated with systolic blood pressure in 6‐ and 9‐year‐old boys, whilst positively correlated with diastolic blood pressure in the 12‐year‐old boys and 13‐ to 15‐year‐old girls (p<0.05). Higher levels of red blood cell and hemoglobin were observed in pre‐high blood pressure and high blood pressure students when compared with their normotensive peers (p<0.01). The increases in red blood cell and hemoglobin were significantly associated with high blood pressure after adjusting for confounding factors. The ORs (95% CI) of each of the increases were 2.44 (1.52–3.92) and 1.04 (1.03–1.06), respectively. No statistical association between total bilirubin and high blood pressure was observed (p>0.05). ConclusionTotal bilirubin could be weakly correlated with both systolic and diastolic blood pressure, as correlations varied with age and gender in children and adolescents; in turn, the increased levels of red blood cell and hemoglobin are proposed to be positively associated with the prevalence of high blood pressure.