Dynamics of Morphological Indicators of Blood of Piglets under the Influence of Iron Clathrochelate Complex and Cyanocobalamin

Iron deficiency anemia is one of the most common non-contagious diseases of piglets. Veterinary antianemic drugs have several drawbacks, so finding new medicines is an important current task for scientists. Therefore, the present study investigated the antianemic effect of iron (IV) clathrochelate in the organism of piglets. The subsequent studies included the exploration of its antianemic actions, particularly in combination with cyanocobalamin when this combination was administered to sows for prophylaxis in piglets. The experiment was carried out on 30 suckling piglets during the period of their detention with sows. According to the method of analogue groups, two groups of control (I) and experimental (II, each containing 15 animals) were formed and they were studied for 30 days. The piglets from five sows (three from each) were selected for the experimental group. During the pregnancy of these sows, 10 ml of 10% solution of iron (IV) IV clathrochelate and solution of cyanocobalamin were injected intramuscularly twice 7 and 14 days before their expected farrowing. For prevention of iron deficiency anemia, the traditional solution of iron dextran was administered once intramuscularly to piglets of the control group. The investigative material included the blood samples of piglets considering the dynamics of probable changes in the number of erythrocytes, hemoglobin content and hematocrit, and other morphological indicators and blood indices of piglets. The dynamics of changes in erythrocyte count, hemoglobin content, hematocrit, leukocytes and platelets, indices of blood almost did not differ from the dynamics of these values when using only 10% solution of iron (IV) clathrochelate for pregnant sows. The proposed scheme for the prophylaxis of iron deficiency anemia in piglets, involving simultaneous intramuscular injections of IV clathrochelate and cyanocobalamin to pregnant sows, is somewhat inferior to the previous preventive measures, which included only the intramuscular injections of iron (IV) IV clathrochelate, but it can be recommended as highly effective.

Optical control of the interface between gold surface and blood cell samples

The optical properties of blood (spectra of the extinction coefficient, k, refractive index, n, etc.) carry important diagnostic information and are usually monitored using bulk samples. In this work, attention is drawn to the interface between the blood volume and the surface of glass or thin gold films on it, where the refractive index may differ from the bulk one. We draw attention to the relationship between two effects – SPR and TIR. It is shown that if the named effects are measured for two different external media 0 and 1 with different refractive indices, then the values of the angles SPR and TIR will be linearly related by the empirical formula SPR1=SPR0+TIR1- TIR0)*K, where the coefficient K depends on the thickness of the transition layer di between the surface and the volume of the liquid medium (suspension). Numerical calculation of K (di) for gold films shows that K = 1.6 at di = 0 and monotonically decreases to 0.01 with an increase in di to 300 nm (and further to 0). Measurement of the angular dependences of reflection, R(), on (1) 100% hematocrit blood samples, (2) hemolyzed samples and (3) washed erythrocytes with dilutions with a buffer solution. It was shown that all samples exhibit a minimum SPR, but the TIR angle can be measured only for blood samples with destroyed membranes (hemolyzed), buffer solution and plasma. The n-value for hemolyzed blood is 1.3505, which is indicative of a low hemoglobin content in the sample. At the same time, di for a sample of 100% hematocrit was 60-105 nm, which indicates a strong deformation of erythrocytes in the form of polyhedrocytes and their dense packing after centrifugation. Washing the cells with a buffer increases di to 280 nm and more and practically eliminates blood cells from the SPR sensitivity region. The reason for this may be that in the blood of 100% hematocrit, erythrocytes are in the form of polyhedrocytes tightly adhering to the gold surface, while as a result of washing and diluting with a buffer solution, the cells relax back into discocytes. As a result, the containing hemoglobin erythrocyte cytoplasm moves away from the surface at a distance di> 300 nm into the suspension volume and leaves the area of the enhanced plasmon-polariton field.

Arsenite exposure inhibits the erythroid differentiation of human hematopoietic progenitor CD34+ cells and causes decreased levels of hemoglobin

Arsenic exposure poses numerous threats to human health. Our previous work in mice has shown that arsenic causes anemia by inhibiting erythropoiesis. However, the impacts of arsenic exposure on human erythropoiesis remain largely unclear. We report here that low-dose arsenic exposure inhibits the erythroid differentiation of human hematopoietic progenitor cells (HPCs). The impacts of arsenic (in the form of arsenite; As3+) on red blood cell (RBC) development was evaluated using a long-term culture of normal human bone marrow CD34+-HPCs stimulated in vitro to undergo erythropoiesis. Over the time course studied, we analyzed the expression of the cell surface antigens CD34, CD71 and CD235a, which are markers commonly used to monitor the progression of HPCs through the stages of erythropoiesis. Simultaneously, we measured hemoglobin content, which is an important criterion used clinically for diagnosing anemia. As compared to control, low-dose As3+ exposure (100 nM and 500 nM) inhibited the expansion of CD34+-HPCs over the time course investigated; decreased the number of committed erythroid progenitors (BFU-E and CFU-E) and erythroblast differentiation in the subsequent stages; and caused a reduction of hemoglobin content. These findings demonstrate that low-dose arsenic exposure impairs human erythropoiesis, likely by combined effects on various stages of RBC formation.

A Monoclonal Antibody Targeting ALK2 As a Potential Therapeutic Agent for Anemia of Inflammation

Background: Iron homeostasis is primarily regulated by hepcidin, a hormone predominantly expressed in the liver. Hepcidin activates the degradation of the transmembrane iron exporter ferroportin, thereby downregulating the release of iron from cells. Hepcidin expression is, at least partly, regulated in response to signaling of the type I TGF-β receptor ALK2, via SMAD2/3 phosphorylation. IL-6, which is commonly elevated in chronic kidney disease (CKD) and other inflammatory conditions, upregulates hepcidin expression and reduces serum iron bioavailability. As a result, chronic inflammatory conditions are often accompanied by secondary anemia of inflammation (AI). We have previously demonstrated that ALK2 inhibition suppressed hepcidin expression in rodents, monkeys and healthy humans. We further described that administration of a selective small molecule ALK2 kinase inhibitor (KTI-2338) reversed changes in hepcidin and iron in a mouse model of CKD, supporting the potential benefit of ALK2 inhibition in AI. Another approach to targeting ALK2 signaling is use of a neutralizing antibody. KTI-018 is a neutralizing ALK2 antibody with high affinity and selectivity for ALK2. This biologic has been demonstrated to reduce serum hepcidin and increase serum iron in healthy non-human primates. Aims: To further elucidate the specific contribution of ALK2 signaling as a driver in AI, and to determine the therapeutic potential of the antibody in this type of anemia, we assessed the effect of KTI-018 in the CKD mouse model. Methods: The study was conducted with 6-week-old male C57Bl/6 mice. Mice in the CKD cohort (CKD) were treated with once daily oral administration of adenine, a compound that metabolizes to 2,8-dihydroxyadenine, forming crystals in the proximal tubular epithelia and causing inflammation and fibrosis in the kidneys. Mice in the control cohort (healthy) received once daily oral administration of vehicle. Upon confirmation of disease, the CKD cohort was subdivided into two groups. The treatment group received twice weekly intraperitoneal treatment with KTI-018 (CKD-KTI-018), and the control group received tris-buffered saline (CKD-TBS). Healthy mice received TBS only. All mice were maintained on their assigned daily adenine or vehicle regimen. At day 53, the study was terminated and hematologic parameters, serum hepcidin, iron, and IL-6 levels were assessed. Results: After 42 days of adenine or vehicle administration, serum hepcidin, serum iron, and hematologic parameters were assessed in representative cohorts of CKD-TBS and healthy mice. The CKD-TBS cohort experienced changes associated with anemia of inflammation as compared to the healthy mice, including increased hepcidin, decreased serum iron, and decreased hematologic parameters. The differences between the healthy and CKD-TBS groups were maintained through the duration of the study. At study termination, CKD-TBS mice had increased serum IL-6 levels (218%), elevated serum hepcidin (149%), and reduced serum iron (-30%) as compared to the healthy mice. Laboratory findings characteristic of anemia were present in the CKD-TBS group, including decreased red blood cells (-6.1%), hemoglobin (-13.2%), and reticulocyte hemoglobin content (-9.3%) as compared to healthy mice. In contrast, CKD-KTI-018 mice had decreased serum hepcidin (-25%) and increased serum iron (59%) as compared to CKD-TBS mice. This restoration of serum iron corresponded to improvements in red blood cells, hemoglobin, and reticulocyte hemoglobin content, which were increased by 7.6%, 9.6%, and 6.7%, respectively, in the CKD-KTI-018 mice as compared to the CKD-TBS mice. These results demonstrate that, by decreasing serum hepcidin, KTI-018 increased the bioavailability of iron, which led to the restoration of hematologic parameters and appeared to reverse AI in mice. Discussion: In this study, a neutralizing ALK2 antibody decreased serum hepcidin, increased serum iron and consequently reversed AI in a mouse model of CKD. These results support the role of ALK2 signaling in AI and suggest that inhibition of ALK2 may be a potential treatment approach for anemia resulting from CKD and other chronic inflammatory diseases. Future studies will explore if ALK2 inhibition may prevent or treat progression of CKD itself, and the role that ALK2 inhibition may play in other chronic inflammatory conditions associated with elevated hepcidin. Disclosures Medeiros: Keros Therapeutics: Current Employment, Current equity holder in publicly-traded company. Backus: Keros Therapeutics: Current Employment, Current equity holder in publicly-traded company. Materna: Keros Therapeutics: Current Employment, Current equity holder in publicly-traded company. Fisher: Keros Therapeutics: Current Employment, Current equity holder in publicly-traded company. Lachey: Keros Therapeutics: Current Employment, Current equity holder in publicly-traded company, Membership on an entity's Board of Directors or advisory committees. Seehra: Keros Therapeutics: Current Employment, Current equity holder in publicly-traded company.

Investigation of the correlation between immature reticulocyte fraction and reticulocyte hemoglobin content with common tests for iron deficiency anemia

Reticulocyte hemoglobin content (Ret-He, the hemoglobin within reticulocytes or immature red blood cells) and immature reticulocyte fraction (IRF, the immature fraction of the absolute-reticulocyte-count) are tests that provide insight into erythropoiesis and iron status earlier than conventional iron studies offering the added benefit of not being acute-phase-reactants. Studies have shown that Ret-He is a diagnostic marker for iron-deficiency-anemia (IDA), but fewer studies have investigated IRF. Our laboratory is currently planning to report these parameters when reticulocyte is ordered. Since these are new parameters, we wanted to investigate their overall correlation with complete blood count (CBC) and other iron studies to gain a better appreciation of their utility in our patient population. The aim of this study was to compare the overall correlation of Ret-He and IRF with seven tests used in the evaluation of IDA. To our knowledge these parameters have not all been directly correlated within a single study. CBC and reticulocytes were quantified using XN 9000 hematology analyzers (Sysmex Corporation), ferritin (DXI 800, Beckman Coulter Inc.), and % iron-saturation (measured using total iron-binding-capacity (TIBC)=transferrin*1.18 on Cobas 6000, Roche Diagnostics). Two de-identified cohorts of patients undergoing physician-ordered reticulocyte testing were used for this analysis. Dataset 1 (DS1): (N=2026 from Mayo Clinic Florida) had Ret-He and IRF compared to absolute-reticulocyte-count (Ret), ferritin and % iron saturation. Dataset 2 (DS2): (N=3990 from Mayo Clinic Rochester) had Ret-He and IRF compared to the red-cell-indices of the CBC including hemoglobin (Hgb), mean-corpuscular-volume (MCV), mean-corpuscular-hemoglobin (MCH), and mean-corpuscular-hemoglobin-concentration (MCHC). Correlation coefficients were calculated using Spearman rank-order (ρ) wherein values below +/-0.39 are weak, between +/-0.40-0.59 are considered moderate, and values above +/-0.60 are considered strong. For DS1, Ret-He demonstrated the following correlations: Ret (ρ=0.01), ferritin (ρ=0.33), % iron saturation (ρ=0.63). IRF demonstrated: Ret (ρ=0.46), ferritin (ρ=-0.05), % iron saturation (ρ=-0.22). For DS2, Ret-He demonstrated the following correlations: Hgb (ρ=0.17), MCV (ρ=0.64), MCH (ρ=0.74), MCHC (ρ=0.56). IRF demonstrated Hgb (ρ=-0.41), MCV (ρ=0.10), MCH (ρ=0.04), MCHC (ρ=-0.11). Ret-He and IRF demonstrated different correlative profiles suggesting they may have differing uses. Ret-He was strongly positively-correlated with % iron saturation, MCV, MCH and moderately positively-correlated with MCHC. These positive-correlations are consistent with relationships established in the literature. Interestingly, Ret-He was only weakly correlated with ferritin, possibly owing to ferritin being an acute-phase-reactant. IRF had a moderate positive correlation with Ret and moderate inverse correlation with Hgb. Both of these IRF relationships are consistent with other reports, but both relationships have not been shown in the same study before, preventing direct comparison until now. The literature suggests IRF may have more potential in monitoring treatment than in diagnosis. One limitation of these datasets is their lack of clinical correlation such as established iron-deficiency, anemia status, or treatment information.

Reticulocyte Hemoglobin Content as a Best Indicator of Iron Deficiency in Female Patients with Diffuse Non-Scarring Hair Loss

Background and objective: Iron deficiency is a well-documented cause of diffuse non-scarring hair loss. We aimed to find the best representative laboratory parameter for iron deficiency. Methods:This was a cross-sectional observational study conducted on 51 female patients with diffuse non-scarring hair loss and iron deficiency state. Iron deficiency was diagnosed as serum ferritin below 30 ng/ml, TSAT below 20% or CHr below 29 pg. Results: Among 51 female patients with diffuse non-scarring hair loss with laboratory proven iron deficiency; low CHrwas reported in 50 (98%) patients, low TSAT was reported in 43 (84.3%) patients, low serum ferritin was reported in 28 (55%). Conclusion:The reticulocyte hemoglobin content (CHr) shows the highest frequency of iron deficiency in patients with diffuse hair loss and iron deficiency state.

EFFECT OF ENROFLOXACIN ON RED BLOOD CELL INDICES OF DUCKLINGS

In this study we present the results of our research into the effect of Enrofloxacin (a fluoroquinolone drug) on red blood cell indices of ducklings, breed Bashkir duck. The ducklings of the experiment group received Enrofloxacin at a concentration of 200 g/L via drinking water for 10 consecutive days. Blood samples from all the ducklings were collected by cardiac puncture on Day 1, Day 3, Day 5, Day 7, Day 9 and Day 11 after the withdrawal of the drug. After evaluating red blood cell indices, including RBC count, hematocrit, color index of blood, erythrocyte sedimentation rate, hemoglobin content and erythrocyte indices we observed short-term reliable changes in blood of ducklings in the experiment group. We registered the most dynamic changes in RBC count of Group II on Day 1, Day 9 and Day 11, when it increased by 16%, 14% and 16% respectively whereas on Day 3 we registered a single decrease by 8% as compared to the control group. We marked ambiguous variations in color index of blood, MCV and MHC. Throughout the experiment we noted the most pronounced changes in practically every value only on Day 5, when we registered increases in RBC count, hemoglobin content, color index of blood, hematocrit, mean corpuscular volume and mean hemoglobin content of duckling blood. In spite of the fact that these singular changes may reflect certain intensification of erythropoiesis, on the whole they do not reflect any negative effect of Enrofloxacin on physiological status of ducklings.

Establishing of the mono- and polyinvasion impact on some morpho-functional indices in wild boars

In the paper is described the mono- and poly-invasions impact on some morpho-functional indices in wild boars. So, in result of the investigation of hematological indices in uninfested mono- and poly-parasitized wild boars, it was established that both the indices of hemoglobin content, of hematocrit, erythrocyte’s number, thrombosis time and ESR (erythrocyte sedimentation rate) vary and are more increased in the I group with uninfested wild boars compared to mono - and poly-parasitized ones. It has been established that at infested boars with S. papillosus from the I group, and in those infested with D. lanceolatum from the II group there is a decrease of hemostatic indices, but their maximum decrease is highlighted in the IV group with wild boars infested with Dicrocoelium lanceolatum, Strongyloides papillosus, Metastrongylus elongatus and Eimeria debliecki. This decrease is due to eliminated exotoxins by parasites, which contain anticoagulants and hemolyzers and which neutralize the fibrinogen, thrombin, Ca+ ions and vitamin K properties from the body.