Increase In Red Blood Cell Count Research Articles

Effect of different hypoxic and hypobaric interventions on blood gas and erythrocyte-related indicators in rats.

To explore the effects of hypoxic and hypobaric conditions on blood gas and erythrocyte-related indicators in rats. SD male rats were exposed to low-pressure hypoxic conditions simulating an altitude of 6500 m in a small or a large experimental cabin. Abdominal aortic blood samples were collected and blood gas indicators, red blood cells (RBCs) count, and hemoglobin (Hb) content were measured. The effects of exposure to different hypoxia times, different hypoxia modes, normal oxygen recovery after hypoxia, and re-hypoxia after hypoxia preconditioning on blood gas indicators, RBCs count and Hb content were investigated. The effect of blood gas indicators was correlated with the length of exposure time of hypoxia and the reoxygenation after leaving the cabin. Hypoxia caused acid-base imbalance and its severity was associated with the duration of hypoxia; hypoxia also led to an increase in RBCs count and Hb content, and the increase was also related to the time exposed to hypoxia. The effects of reoxygenation on acid-base imbalance in rats caged in a small animal cabin were more severe that those in a large experimental cabin. Acetazolamide alleviated the effects of reoxygenation after leaving the cabin. Different hypoxia modes and administration of acetazolamide had little effect on RBCs count and Hb content. Normal oxygen recovery can alleviate the reoxygenation and acid-base imbalance of hypoxic rats after leaving the cabin and improve the increase in red blood cell and hemoglobin content caused by hypoxia. The improvement of hypoxia preconditioning on post hypoxia reoxygenation is not significant, but it can alleviate the acid-base imbalance caused by hypoxia in rats and to some extent improve the increase in red blood cell and hemoglobin content caused by hypoxia. Due to excessive ventilation and elevated RBCs count and Hb content after hypoxia reoxygenation, oxygen partial pressure and other oxygenation indicators in hypoxic rats are prone to become abnormal, while blood gas acid-base balance indicators are relatively stable, which are more suitable for evaluating the degree of hypoxia injury and related pharmacological effects in rats.

Iron Restriction Alleviates β-Thalassemia By Stimulating ULK1-Mediated Autophagy of Free α-Globin

In β-thalassemia, mutations in the HBB gene cause reduced β-globin synthesis with accumulation of toxic free α-globin, leading to ineffective erythropoiesis and hemolysis. We showed previously that β-thalassemic erythroblasts can eliminate free α-globin by ULK1-mediated autophagy and that this process is stimulated by rapamycin inhibition of mTORC1, which phosphorylates ULK1 to inhibit its activity. Several studies have shown that iron restriction can reduce the accumulation of free α-globin precipitates to alleviate the pathophysiology of β-thalassemia. Considering that iron stimulates mTORC1 activity in numerous cell types including erythroblasts, we tested the hypothesis that iron restriction alleviates β-thalassemia by inhibiting mTORC1, thereby stimulating ULK1-mediated autophagy of free α-globin. Hbb Th3/+ β-thalassemic mice or Hbb Th3/+ Ulk1 -/- mice were treated with the clinical stage oral ferroportin inhibitor vamifeport (known also as VIT-2763) for 45 days to cause systemic iron restriction. In Hbb Th3/+ mice, vamifeport administration led to 66% and 40% reductions in the levels of serum iron (p<0.001) and ferritin (p=0.001), respectively, as expected. Iron restriction by vamifeport alleviated the pathophysiology of β-thalassemia compared to vehicle-treatment in Hbb Th3/+, as evidenced by a 21% increase in RBC count (p<0.001), 32% reduction in insoluble α-globin in red blood cells (RBC) (p=0.002), 41% reduction in reticulocyte count (p<0.001), 27% reduction in spleen mass (p=0.034) and enhanced maturation of spleen and bone marrow erythroid precursors, indicating reduced ineffective erythropoiesis. Multivariate analysis showed that beneficial effects of iron restriction in β-thalassemic mice were attenuated by concomitant disruption of both Ulk1 alleles. For example, vamifeport treatment of Hbb Th3/+ Ulk1 -/- mice caused only a 12% increase in RBC counts (p=0.015 compared to Hbb Th3/+). Additionally, iron restriction improved erythroid maturation in Hbb Th3/+ Ulk1 -/- mice, but to a significantly lesser extent than in thalassemic mice with intact Ulk1alleles. We obtained similar results by inducing iron deprivation with PR73, a peptide hepcidin mimetic. Mechanistically, long-term administration of vamifeport was associated with inhibition of mTORC1, as evidenced by 32% reductions in the phosphorylation of its substrate, ribosomal protein S6 kinase (p=0.004), and activation of the heme-regulated eIF2α kinase (HRI), indicated by increased phosphorylation of eIF2α and/or accumulation of the HRI effector transcription factor ATF4. Similar results were obtained with PR73. Administration of a single dose of PR73 to Hbb Th3/+ mice led to activation of HRI in Ter119 + erythroblasts after 8 hours, which preceded the downregulation of mTORC1 activity, detected at 24 hours. This is consistent with prior studies showing that HRI inhibits mTORC1. Loss of ULK1 had no effect on suppression of mTORC1 with either drug, consistent with ULK1 being the substrate of mTORC1; we are currently investigating whether loss of ULK1 alters the HRI pathway. Together, our data support a mechanism through which iron restriction alleviates the pathophysiology of β-thalassemia in part by stimulating ULK1-mediated autophagy of free α-globin. This process is likely mediated by iron restriction that activates HRI which in turn inhibits mTORC1 to block its negative effects on ULK1. Additional mechanisms for iron regulation of mTORC1 activity are also possible. Our findings provide new insight into the mechanisms by which iron restriction alleviates β-thalassemia and illustrate how metabolic regulators of erythropoiesis impact the severity of β-thalassemia by eliminating free α-globin through a key protein quality control pathway.

Changes in routine blood parameters of patients with generalized tonic clonic seizure: a retrospective study

To analyze the alterations in routine blood parameters in these patients. Generalized tonic clonic seizure (GTCS) is one main type of generalized epilepsy. Clinicians have recently found changes in routine blood parameters in GTCS patients. A retrospective study of 45 GTCS patients and 200 healthy controls was conducted. Blood routine tests in emergency ward and on the second day of admission were retrieved. Changes in routine blood parameters, including red blood cell (RBC) count, white blood cell (WBC) count, platelet count, lymphocyte count, neutrophil count, and neutrophil-to-lymphocyte ratio (NLR) on the seizure episode day and the second day after seizure were analyzed. The GTCS patients had increased platelet count, RBC count, WBC count, neutrophil count, lymphocyte count, and NLR, especially at onset of GTCS episode, compared to the controls. These parameters had decreased on the second day after seizure onset. The neutrophil count and WBC count in GTCS cases with seizure duration <5min were significantly lower than those with seizure duration >5 min. However, this phenomenon was not observed in other indices. The GTCS may induce a transient increase in RBC count, WBC count, and platelet count, and obvious elevation in neutrophil count. There is a certain correlation between WBC count and the duration of seizures. Changes in routine blood parameters observed in GTCS patients may provide useful information clinically.

Effect of Smoking on the Red Blood Cell Count, Hemoglobin Concentration, Hematocrit and Red Cell Indices in Adult Male Smokers

Background- Cigarette smoking is one of the common addictions. The effects of cigarette smoking on health are serious and, in many cases, deadly.&#x0D; Objectives: The present study was carried to observe the effect of smoking on Red Blood Cells (RBC) count, Hemoglobin (Hb) concentration, Hematocrit (HCT) and Red cell indices in adult male smokers and compare it with non-smokers to create awareness.&#x0D; Materials and Methods: In this study 50 adult male smokers and 50 adult male non-smokers aged from 20-45 years were taken. The smokers were regularly smoking for at least one year. Fresh peripheral blood sample from subjects were collected and analyzed for Red blood cell count, Hemoglobin concentration, Hematocrit and Red cell indices by using Mythmic-22 automated 5-part Hematology Analyzer.&#x0D; Result: In this study the values of RBCs count, Hb concentration and Hematocrit showed significant increase (p&lt;0.05) in smokers as compared to non-smokers. The values of Red cell indices (MCV, MCH, MCHC) were statistically not significant between two groups.&#x0D; Conclusion: This study concluded that smoking causes persistent state of hypoxia in the body due to smoke contents which cause increase in RBCs count, Hb concentration and Hematocrit.&#x0D; EMCJ. January 2022; 7(1): 1-5

Red Blood Cell Count: An Unrecognized Risk Factor for Nonalcoholic Fatty Liver Disease.

ObjectiveNonalcoholic fatty liver disease (NAFLD) is becoming a global public health challenge. A convenient NAFLD indicator will greatly facilitate risk appraisal and prevention. As a readily available and inexpensive hematological index in routine clinical examinations, red blood cells (RBCs) are gaining increasing attention in many diseases, such as metabolic syndrome, but their association with NAFLD is unknown.MethodsThis health management cohort study included 27,112 subjects (17,383 non-NAFLD and 9,729 NAFLD) with up to 5 years of follow-up (median 2.8 years). NAFLD was diagnosed by ultrasonography. NAFLD severity was categorized as mild, moderate, or severe. The generalized estimation equation (GEE), an extension of generalized linear models that allows for analysis of repeated measurements, was used to analyze the association between RBC count and NAFLD.ResultsOverall, 4,332 of 17,383 (24.9%) subjects without NAFLD at baseline developed NAFLD. Incident NAFLD risk was positively associated with RBC count. After adjustment for hemoglobin and other confounders, the risk of incident NAFLD was 21%, 32%, and 51% higher in the second, third, and fourth RBC count quartiles, respectively, than in the lowest quartile. In 1,798 of 9,476 (19.0%) subjects with NAFLD at baseline, the severity of NAFLD increased. NAFLD progression risk increased progressively as RBC count increased (P for trend < 0.001). Every one-unit (1012 cells/L) increase in RBC count was associated with a 53% [OR 1.53 (95% CI 1.32-1.77)] increased risk for NAFLD progression.ConclusionsElevated RBC count was independently associated with a high risk of NAFLD incidence and progression. This finding revealed a convenient NAFLD risk indicator.

Spleen Contraction During Step‐Transition Supine Cycling Exercise: Preliminary findings

The current literature suggests that the spleen plays a significant role during physiological stress like exercise or apnea, primarily due to its capacity to contract and release additional red blood cells (RBC) into the systemic circulation. Besides the RBC release, human spleen emptying is also associated with circulating platelets release. However, data on the circulating platelets release following acute exercise have not yet been examined thoroughly. This is especially important since the spleen platelet volume is ~20% larger compared to the volume of circulating platelets, and some authors suggest that increased spleen platelet concentration is associated with the greater risk of a cardiovascular or cerebrovascular event during exercise. The present study aims to explore the magnitude of spleen contraction during on-transient supine cycling exercise in healthy humans. We hypothesized that the spleen is capable of rapid contractions during on-transient cycling, thereby releasing an additional amount of both RBC and platelets into the systemic circulation. Such an increase in RBC count in particular may facilitate O2 transport, and result in faster oxygen kinetics during on transient supine cycling. Healthy, active volunteers (age= 22±3, V̇O2 max.= 48±5 mL·kg·min-1), completed three laboratory visits, including a detailed medical examination, resting ultrasound imaging of the spleen, V̇O2 max. test until voluntary exhaustion, and a supine step-transition test from 20 W to a moderate-intensity cycling power output (PO), corresponding to V̇O2 at 90% gas exchange threshold (GET). During the step-transitions breath-by-breath pulmonary oxygen uptake (V̇O2p), near-infrared spectroscopy derived muscle oxygenation, and beat-by-beat cardiovascular response (via Finapres) were continuously monitored. In parallel, minute by minute ultrasonic measurements of the spleen function were performed in successive, 3 sec. time frames. Blood samples were drawn from the antecubital vein before and immediately after the acute exercise intervention.Currently, six participants completed all study procedures. Compared to baseline readings, the largest spleen volume reduction (~40%) was observed during the 6th minute of cycling at 90% GET. Spleen volume recovered to baseline values after 20 minutes. Immediately after exercise cessation the mean RBC and platelet count increased significantly by 3.2% and 6.5%, respectively. In parallel, the V̇O2p was 46±10 sec, while the mean V̇O2 uptake was ~2.5 L·min-1. Secondary to these findings, transient rises in mean arterial pressure (MAP) and heart rate (HR), were observed during the step-transition exercise, with MAP and HR reaching the highest value of 110±4 mmHg and 149±2 bmp, respectively. In conclusion, these preliminary findings indicate that splenic contraction evoked by a series of on-transient cycling bouts at 90% GET results in a rapid increase in RBC and platelet count into the systemic circulation of healthy active individuals.

Effects of garlic (Allium sativum ) and vitamin E on blood profile, growth performance and internal organ characteristics of rabbit bucks

Studies have shown that garlic has medicinal, antimicrobial properties, speeds up digestion and widely used as preservatives, spice and condimentin many homes. Vitamin E on the other hand is a fat soluble vitamin with potent antioxidant properties essential for the stabilization of biological membranes, protecting cells from oxidative stress and inhibits angiogenesis. Thus, a study using thirty-six (36) pre-pubertal New Zealand White rabbit bucks was conducted to investigate the response of garlic and vitamin E supplementation on haematological indices, serum biochemistry and growth performance. The bucks were randomly assigned to four dietary treatments and replicated three times in a completely randomized design (CRD). Each dietary treatment consisted of rabbits per treatment with 3 rabbits per replicate. Four dietary treatments were formulated to meet the nutrient requirements of rabbits supplemented at 0% garlic and vitamin E (T ), 3% garlic (T ), Vitamin E (T ), and 3% garlic and Vitamin E (T ) respectively. Data collected on the growth performance, haematology and serum biochemistry parameters and on the internal organ characteristics were subjected to analysis of variance (ANOVA). Results showed that significantincrease (P&lt;0.05) were observed in final body weight, ear length, forelimb, hind- limb and body length for rabbits fed supplementation of garlic, vitamin E and its combination, while tail length, head to shoulder significantly decreases. Hematological parameters showed significant increases in red blood cellcount for vitaminE group while T was higher on MCHC. White blood cell and its differentials increased significantly on T, T and T Serum biochemical characteristics were significantly higher on total protein, globulin and creatinine; while cholesterol, glucose and ALP reduced significantly on T , T and T It could be concluded that combination of garlic and vitamin E improved performance and blood profile of rabbit bucks compared to when they are used individually.

Proof-of-concept study on improved efficacy of rHuEPO administered as a long-term infusion in rats

BackgroundHuman recombinant erythropoietin (rHuEPO) is often used in the treatment of diseases associated with a decreased production of red blood cells (RBC), such as chronic renal failure. rHuEPO is typically administered as an intravenous or subcutaneous (SC) injection every few days. The low minimum effective concentration (MEC) of rHuEPO, compared to the concentrations observed after standard doses, suggests that a low dose of the drug administered as a long-term infusion should be efficacious. This study aimed to compare the efficacy observed after a single subcutaneous administration of rHuEPO with that observed after a long-term infusion of rHuEPO via implanted osmotic pumps at a similar or lower dose.Materials and methodsIn this study three rats received rHuEPO as a single SC injection at a dose of 1350 IU/kg, nine via osmotic pumps at a rate of 0.25, 0.5 and 1 IU/kg and at a total dose of 333 IU/kg, 667 IU/kg, 1333 IU/kg. Three rats served as a control group. The erythropoietin concentrations, RBC count and hemoglobin were measured.ResultsAn increase in RBC count and hemoglobin was observed after SC infusion of rHuEPO. The baseline corrected area under the effect curve for hemoglobin and RBC count was more than 10-times higher for the SC infusion than for a single SC administration with a comparable dose.ConclusionsThis study demonstrates that administration of rHuEPO as a long-term infusion at a rate ensuring MEC allows to achieve a high efficacy of therapy using relatively small doses of the drug.

Effects of recreational scuba diving on erythropoiesis-"normobaric oxygen paradox" or "plasma volume regulation" as a trigger for erythropoietin?

Previous studies have shown an increase in erythrocyte lipid peroxidation and a decrease in red blood cell (RBC) count, hemoglobin, and hematocrit after only one recreational scuba diving session. The aim of this study was to examine the effect of repetitive scuba diving on RBC parameters and erythropoiesis. Divers (N = 14) conducted one dive per week over 5 weeks at a depth of 20-30m for 30min. For measuring RBC parameters, erythropoietin, iron, and ferritin, blood samples were collected before and after the first, third, and fifth dive. Between pre- and post-dive results, a statistically significant increase in RBC count, hemoglobin, hematocrit, mean corpuscular volume (MCV), RBC distribution width (RDW), iron, and ferritin was observed. Analysis of the results between the first, third, and fifth dive showed that the erythropoietin increase at the third (pre-dive p = 0.009; post-dive p = 0.004) and fifth dive (pre-dive p < 0.001; post-dive p = 0.003) was not accompanied by changes in RBC count, hemoglobin, iron, and ferritin. In parallel, a continuous increase in hematocrit, MCV, and RDW was observed, whereas mean corpuscular hemoglobin (MCH) and mean corpuscular hemoglobin concentration (MCHC) decreased. Changes in RBC indices and EPO elevation indicate that the occasional switch from hyperoxia to normoxia or mechanisms for plasma volume regulation may be a step in the maintenance of erythropoiesis.

S148 TFR2-HAPLOINSUFFICIENCY ENHANCES THE BENEFICIAL EFFECT OF TMPRSS6-ANTISENSE OLIGONUCLEOTIDE TREATMENT IN BETA-THALASSEMIA MICE

Background: Transferrin receptor 2 (TFR2) is a type II transmembrane protein mainly expressed in the liver and in the erythroid compartment. In the liver it acts as an activator of the iron hormone hepcidin and, when mutated, is responsible for type 3 hereditary hemochromatosis. In erythroid cells TFR2 binds the erythropoietin (EPO) receptor and serves as a brake of the EPO-EPOR signaling pathway. We have recently demonstrated that the complete loss of bone marrow (BM) Tfr2 increases erythroblast EPO sensitivity and improves anemia, ineffective erythropoiesis and iron-overload in a β-thalassemia intermedia murine model (Hbbth3/+), without reducing splenomegaly. On the contrary, approaches based on the induction of iron restriction were proven to be effective in ameliorating splenomegaly and anemia in thalassemic mice. Among these, targeting the hepcidin inhibitor TMPRSS6 is one of the most promising strategies. Aims: The main aim of the present work is to investigate whether the combination of Tfr2-haploinsufficiency in the BM and TMPRSS6 antisense oligonucleotide (ASOs) administration will reduce splenomegaly and provide a synergistic benefit on thalassemic erythropoiesis and anemia. Methods: We generated thalassemic mice with both or a single Tfr2 allele in the BM by transplanting BM cells from Hbbth3/+ or Tfr2+/-/Hbbth3/+ mutants into lethally irradiated wild-type mice. Mice were treated with TMPRSS6-ASO (or control ASO) twice a week for 6 weeks starting 9 weeks after BM transplantation. Complete blood count was determined at the beginning of the treatment and after 3 and 6 weeks. At sacrifice, blood and organs were collected for full phenotypic analysis. Results: Deletion of a single Tfr2 allele in Hbbth3/+ mice BM mildly increased Red Blood Cells (RBC) count and Hemoglobin (Hb) levels and improved erythropoiesis, with a concomitant decrease in EPO levels, but no reduction of spleen size. A similar phenotype was induced by TMPRSS6-ASO treatment of Hbbth3/+ animals, with additional partial correction of splenomegaly. The combination of BM Tfr2 haploinsufficiency and TMPRSS6-ASO had a striking effect on the thalassemic phenotype: indeed, an impressive increase in RBC count (up to 11.2 ± 1.2∗106/μl) and Hb (∼3 g/L) compared with Hbbth3/+ was attained, accompanied by the normalization of reticulocyte and EPO levels, improved erythropoiesis and spleen size reduction. Mean corpuscular volume and hemoglobin were strongly reduced. This indicates a disproportionate erythropoietic expansion relative to the available iron, whose circulating levels are decreased relative to untreated thalassemic controls. Summary/Conclusion: While Tfr2 haploinsufficiency does not impair systemic iron homeostasis, here we demonstrate that deletion of a single Tfr2 allele in the BM is sufficient to improve the hematological parameters of thalassemic mice. This finding paves the way toward a potential targeted therapy based on partial Tfr2 inhibition that might be beneficial for thalassemia (and potentially for other forms of anemia) without inducing defective hepcidin activation and iron overload. In addition, our results prove that targeting Tfr2 in combination with an anti-TMPRSS6 therapy may be a valuable therapeutic approach that strongly improves the thalassemic phenotype. However, Tmprss6 inhibition must be tightly controlled to avoid excessive iron restriction. Further experiments will allow setting up the optimal conditions.

Potential of plant stress hormone methyl Jasmonate against lipopolysaccharide attenuated oxidative stress and arthritis in experimental animals

Introduction: Methyl jasmonate is a plant stress hormone indicated in the treatment of various cancers. The purpose of the current study is to investigate the anti-arthritic and antioxidant activity in experimental animals. Materials and Methods: Methyl jasmonate was screened in two doses (20 mg/kg and 40 mg/kg) against lipopolysaccharide (LPS) induced arthritis in rats. The assessment of arthritis such as paw thickness, paw volume, and body weight was assessed on 0th, 7th, 14th, 21st, and 28th days. The percent change in paw thickness, body weight, and arthritis index was calculated on 7th, 14th, 21st, and 28th days. The estimation of serum marker enzymes such as alkaline phosphatase, alanine transaminase, and aspartate aminotransferase was carried out on 14th and 28th days. The antioxidant enzyme such as superoxide dismutase, catalase, GST, glutathione peroxidase, and lipid peroxidation activity in articular cartilage tissues was carried out on the 28th day. White blood cell (WBC), red blood cell (RBC), and erythrocyte sedimentation rate were determined on the 28th day. Free radical scavenging activity was carried out using 2,2-diphenyl-1-picrylhydrazyl (DPPH) method and alpha-tocopherol used as a standard for comparison. After completion of the treatment period, tarsal-metatarsal joints were subjected to histopathology. Results: MJ significantly reduces arthritis which was evident from the assessment of arthritis such as paw volume, paw thickness, arthritis index, and body weight. Significant reduction in lipid peroxidation and serum marker enzymes was observed in MJ-treated rats as compared to the disease control group. Significant antioxidant activity was exhibited by MJ-treated group. Dose-dependent DPPH radical scavenging was observed which assess free radical scavenging potential of MJ. Erythrocyte sedimentation rate and WBC count are significantly reduced whereas RBC count was compensated similarly to control compared to the diseased group. Conclusion: Methyl jasmonate exhibits significant anti-arthritic and antioxidant activity through a reduction in paw volume, thickness, serum marker enzyme, ESR, WBC count, increase in RBC count, and gain in body weight. The reduction in inflammation and cell damage may be due to antioxidant, free radical scavenging, and antilipid peroxidative effect which is supported by histopathological studies.

Hematology reference intervals for adult cows in France using the Sysmex XT-2000iV analyzer

In order to develop bovine hematology reference intervals (RIs) in accordance with new international recommendations, we analyzed 156 blood specimens of healthy adult dairy and beef cows from 32 farms with a Sysmex XT-2000iV analyzer, and by manual scoring of platelet clumps and white blood cell (WBC) differential. We established RIs by the nonparametric method, and examined effects of age, production type (beef vs. dairy), and stage of lactation. RIs could not be determined for platelet count and indices because clumps were observed in 80% of specimens. Optical and impedance red blood cell (RBC) counts were similar, although statistically different. RIs for analyzer and manual WBC differentials were not different except for lymphocyte concentration, the subpopulations of which were counted manually. Hematocrit was higher in beef than dairy cattle, and hemoglobin was lower in early lactation. Increases in RBC count, mean corpuscular volume, mean corpuscular hemoglobin, and RBC distribution width were noted with increasing age, along with decreases in WBC count, neutrophils, and lymphocytes. Most RIs in our study, with the exception of neutrophils, were similar to those previously reported using a flow cytometry analyzer.

Role Of Transferrin In Recovery From Phlebotomy In Iron Replete Or Iron Deficient Mice

The most common reason for deferral of blood donation is low hemoglobin concentration and anemia. Furthermore, repeated donations, especially in young females, may lead to iron deficiency, drop of hemoglobin, and consequent iron deficiency anemia. Recent data suggests that iron deficiency, in the absence of anemia, may be associated with symptoms in otherwise healthy individuals. Our previous findings suggested that additional transferrin ameliorates anemia in mouse models of β-thalassemia; this effect appears driven by an increase in the number of circulating red blood cell (RBC) by extending RBC survival (Li Nat Med 2010). We hypothesized that exogenous transferrin increases RBC count and enables a more rapid recovery after phlebotomy. To test this hypothesis, C57 mice were fed an iron replete (35ppm iron) and iron deficient (2.5ppm iron) diet for 3 months to simulate the human condition. Mice were phlebotomized an average of 220ul of blood weekly for four weeks. Additionally, all mice received intraperitoneal (IP) injection of either of 10mg of human apo-transferrin (apo-Tf) (200 ul volume) or 200ul PBS daily Monday – Friday for 4 weeks (in total 20 days of injection). Mice (n=4/group) were sacrificed and samples analyzed at the end of the 4 weeks. RBC survival was evaluated using an in vivo biotin avidin assay previously described (Li Nat Med 2010). We analyzed all RBC indices using ADVIA 120 (Bayer). MCV decreased in phlebotomized mice on 2.5ppm compared to 35ppm diet (p=0.012) and after apo-Tf injection (p<0.05 in 2.5ppm diet, and p<0.001 in 35ppm diet). Reticulocyte counts increased by 50% in apo-Tf treated compared to PBS treated phlebotomized mice on 2.5ppm diet (p=0.006). Apo-Tf injections also resulted in an increase in RBC counts in phlebotomized mice on 35ppm diet (p=0.056). Furthermore, using antibodies to CD44 and TER119 in flow cytometry (Chen PNAS 2010), we assessed the proportion of erythroid precursors at all stages of terminal erythroid differentiation. Apo-Tf injections result in more erythroid precursors in spleen and bone marrow (p<0.05) in phlebotomized mice on 2.5ppm or 35ppm diets without changes in the degree of apoptosis or quantity of reactive oxygen species. Mice on 2.5ppm diet had smaller spleens relative to 35ppm diet (0.002 vs. 0.004 spleen / body weight, p=0.05) that increased as a consequence of apo-Tf injection (0.003 vs. 0.002 spleen / body weight, p=0.02). No differences in spleen size or spleen / body weight is observed in apo-Tf treated mice on 35ppm diet. Immunohistochemistry of spleen samples using anti-TER119 antibodies revealed more extramedullary erythropoiesis in all apo-Tf treated mice. Lastly, no difference in RBC survival was observed in mice as a consequence of diet or injection. Taken together, our data suggest that exogenous apo-Tf in recurrently phlebotomized mice on iron deficient diet stimulates erythropoiesis without a direct effect on RBC survival. This data provides initial evidence that exogenous apo-Tf accelerates recovery from phlebotomy especially in iron replete conditions, without evidence of harm in iron deficiency. The molecular mechanisms underlying the effect of iron availability and iron delivery on different stages of erythroid differentiation requires further analysis. Disclosures:No relevant conflicts of interest to declare.

Effect of Melatonin on Some Hematological Parameters and Immune Status of Broiler Chicks

Melatonin, the pineal gland hormone, has been shown to play a principle role in maintenance of health and well being of man and animals in all stages of life. The aim of the present study was to investigate the effect of melatonin on some hematological parameters and immune status of broiler chicks. For this purpose sixty-four, three weeks-old chicks of Hubbard strain weighing 500-633g were used. Birds were kept on lighting regimen of 12 hours natural light to 12 hours darkness. Chicks were randomly allocated into four equal groups of 16 birds each. The first group was kept as a control group and fed starter ration sprayed with ethanol saline without melatonin from the 4th to 8th weeks of age. The second, third and fourth groups were fed on the starter ration sprayed with ethanol saline containing 20, 30 and 40 mg melatonin / kg ration, respectively from the 4th to 8th weeks of age. Blood samples were obtained from the tibial vein of all birds at the end of the 1st week of treatment and then every week up to the end of the experiment. The obtained blood were used for determination of red blood cells (RBCs) count, hemoglobin (Hb) concentration, packed cell volume (PCV) as well as total and differential leukocytic count. Tissue samples from liver, spleen, thymus and bursa of Fabricious were taken for histopathological examination. The results revealed that melatonin in the three doses produced a significant increase in RBCs count, PCV, Hb concentration, total leukocytic count and lymphocyte percentage which clarifies the effect of melatonin on improving the health and immune status of the chicks under investigation. The histopathological study in the present investigation revealed that melatonin treatment induced a lymphoid hyperplasia in the liver, spleen, and bursa of Fabricious of all treated groups. This lymphoid hyperplasia might be another explanations for the immunostimulant effect of melatonin. In conclusion melatonin was found to improve the health and immune status of broiler chicks.

Changes in Red Blood Cells Membrane Protein Composition during Hemodialysis Procedure

Our aim was to evaluate the influence of the hemodialysis (HD) procedure in red blood cells (RBC) membrane protein composition. We evaluated hematological data (RBC count, hemoglobin concentration, and hematimetric indices) and RBC membrane protein composition (linear and exponential gradient polyacrylamide gel electrophoresis in the presence of sodium dodecylsulfate [SDS-PAGE] followed by densitometry analysis of RBC membrane proteins) before and immediately after the HD procedure in 20 patients (10 responders and 10 non-responders to recombinant human erythropoietin therapy [rhEPO]) and 26 healthy controls. Before HD, patients presented anaemia and significant changes in membrane protein composition, namely, a statistically significant reduction in spectrin associated with a significant increase in bands 6, as well as an altered membrane protein interaction (protein 4.1/spectrin, protein 4.1/band 3, protein 4.2/band 3 and spectrin/band 3). After HD, we found that patients showed a statistically significant increase in RBC count and hemoglobin, a further and statistically significant decrease in spectrin, an increase in band 3, and an altered spectrin/band 3 ratio. When comparing responders and non-responders patients after HD, we found that the non-responders presented a trend to a higher reduction in spectrin. Our data suggest that HD procedure seems to contribute to a reduction in spectrin, which is normally associated with a reduction in RBC deformability, being that reduction in spectrin is higher in non-responder patients.

Erythropoietic and anti‐obesity effects of Garcinia cambogia (bitter kola) in Wistar rats

The anti-obesity and erythropoietic effects of crude ethanolic extracts of Garcinia cambogia (bitter kola) seeds on Wistar rats (Rattus norvegicus) were investigated. The rats were divided into three dosage groups: A (0 mg/kg of body weight), B (200 mg/kg) and C (400 mg/kg). Weight changes, plasma lipoprotein levels and the lipid profile of the liver, gastrointestinal system and adipose tissue were monitored as indices for anti-obesity, while the RBC (red blood cell) count (assessed by using a haemocytometer) was monitored as a measure of erythropoiesis. The extract was administered by gavage for 5 weeks. The results for each test group was compared statistically with those for the control (P<0.05). Analysis of the results showed a significant increase in RBC counts in both test groups and a decrease in weights of experimental animals. There was a dose-dependent decrease in the plasma level of very-low-density lipoprotein and a dose-dependent increase in the level of chylomicrons. There was a slight, but significant, decrease in the level of high-density lipoprotein and a significant increase in the level of LDL (low-density lipoprotein). There was significant dose-dependent decrease in the TAG (triacylglycerol) pool of adipose tissue and the liver of the test groups, but a significant increase in the TAG pool of the gastrointestinal system. The increase in the TAG pool of the gastrointestinal system is possibly compensatory. The results therefore confirm that ethanolic extracts of G. cambogia seeds have both haematologically enhancing and anti-obesity effects. The decrease in the high-density-lipoprotein level and an increase in the LDL level may play an important role in cardiovascular disease.

Effect of a high-altitude expedition to a Himalayan peak (Pumori, 7,161 m) on plasma and erythrocyte antioxidant profile.

The effects of a high-altitude exposure were studied in six mountaineers who spent 3 weeks at an altitude range between 5,250 and 7,161 m after 1 week in an acclimatization trek (2,800-5,250 m). Blood drawn from the antecubital vein was collected at sea level 1 day before and 1 day after the expedition to analyse some haematological variables [haemoglobin (Hb), haematocrit (Htc) and red blood cell (RBC) count], erythrocyte antioxidant enzyme activity [superoxide dismutase (SOD), glutathione peroxidase (GPx) and glutathione reductase (Gr)] and membrane fatty acid profile [mono-unsaturated fatty acids (MUFA), polyunsaturated fatty acids (PUFA), saturated fatty acids (SFA), trans fatty acids (TRANS)]. Moreover, total antioxidant status (TAS), thiobarbituric acid reactive substances (TBARS), thiol protein groups (SH), SOD, GPx and Gr were measured in plasma. High-altitude exposure induced polycythaemia, with significant increases in RBC count (5.26%), Hb concentration (4.83%) and Htc (6.26%). Furthermore, a significant increase in plasma TBARS, SOD and Gr was observed after the expedition, whereas SH, TAS and GPx decreased. Erythrocyte glutathione-cycle-related antioxidant enzyme activity was upregulated, whereas SOD activity was maintained after the expedition. In addition, despite the unchanged (MUFA+PUFA)/SFA ratio, the membrane erythrocyte fatty acid content showed a significant increase in PUFAs and a decrease in TRANS, suggesting enhanced membrane fluidity. In conclusion, it seems that high-altitude exposure, besides quantitative variations in RBC expression, induced plasma oxidative stress and damage, and significant changes in erythrocyte components, namely in antioxidant enzyme activity and membrane fatty acid profile that might modify RBC functionality.

Hematological parameters of five species of marine fishes

1. 1. Blood oxygen capacity (BOC). hemoglobin (Hb) content, hematocrit (Hct). red blood cell (RBC) count, and pH were measured in five species of marine fishes which varied in activity from sedentary to fast swimming. 2. 2. BOC and Hb content were correlated with the level of activity for a species. RBC and Hct varied between species and were more weakly correlated with levels of activity. Blood pH did not vary between the species studied. 3. 3. These data suggest that increased BOC is correlated with high levels of activity in fish and that this increase is due to elevated Hb content of the blood. The increased Hb is generally accompanied by an increase in RBC count, so that the average corpuscular hemoglobin (ACH) remains the same.