Osmotic Fragility Research Articles

Effects of crude methanol extract of Adansonia digitata fruit pulp on Naja nigricollis venom-induce toxicity in Wistar rats.

This study evaluated the effects of Crude Methanol Extract of Adansonia digitata Fruit Pulp on Naja nigricollis Venom-Induce Toxicity in Wistar rats. A. digitata was extracted using 70% methanol and median lethal dose (LD50) of both the extract and venom were determined using the up-and-down method. Sixty Wistar rats were randomly assigned into 10 groups of 6 rats each and were administered with normal saline, venom only, venom+antivenom, 125mg/kg, 250mg/kg and 500mg/kg crude methanol extract (CME) pre-envenomation, 125mg/kg, 250mg/kg and 500mg/kg CME post-envenomation, and venom+antivenom+250mg/kg CME, respectively. Blood samples were collected 8-h post-envenomation in EDTA and plain sample bottles. Erythrocyte osmotic fragility (EOF) test was carried out on the EDTA blood samples while serum washarvested and used for Malondialdehyde (MDA) and Superoxide Dismutase (SOD) assays. LD50 of the CME and venom was >5,000mg/kg and 0.889mg/kg, respectively. N. nigricollis-induced oxidative stress was evident in group B through increased % haemolysis, MDA and lowered SOD activities. The groups treated with antivenom only, 250mg/kg CME post-envenomation and antivenom +250mg/kg CME significantly (p<0.05) reduced EOF, MDA values and increased SOD. The CME revealed better ameliorative effect than protective via inhibition of EOF, MDA values and increased SOD activity. The CME when administered singly showed more ameliorative properties and the combination of CME with antivenom for protection was not as effective as when compared to single administration.

The Association of Heterozygous β-Thalassemia and β-Spectrin Mutations May Result in a Phenotype of Severe Anemia

Background: The association of mutations in genes related to hereditary anemias (HA) can affect disease phenotype and severity. Understanding HA genetic basis is crucial for avoiding misdiagnosis and for adequate clinical management. β-thalassemias (BT) are characterized by a reduction of β-globulin production caused mainly by point mutations within the HBB gene or its bordering sequences. The imbalance between α and β globin production results in ineffective erythropoiesis and hemolysis. Hereditary spherocytosis (HS) is characterized by abnormalities of erythrocyte membrane proteins, resulting in spheroidal, osmotically fragile erythrocytes with varying degrees of hemolysis and anemia. HS related to spectrin deficiency is clinically heterogeneous, ranging from well-compensated hemolysis to severe anemia. Here we describe two unrelated cases in which patients had a chronic severe hypochromic microcytic hemolytic anemia with elevated HbA2 clinically compatible with BT Intermedia or Major. However, conventional β-globin gene sequencing revealed only BT trait that did not justify the clinical presentation. The extensive research for deletions or other mutations for BT were negative. We then proceeded with additional genetic investigation through exome sequencing. Aims: To investigate additional mutations related to HA that may contribute to a phenotype of severe anemia in patients with a β -Thalassemia trait. Methods: Patients' clinical features and laboratory results were collected by medical charts evaluation. The Institutional Review Board approved this research and patients signed written informed consent. β-globin gene sequencing was initially performed by conventional Sanger sequencing. Exome sequencing was performed utilizing the Illumina HiSeq2500 platform. Results: Patient 1 had a clinical picture of eventual transfusion dependence, moderate hepatic iron overload and splenomegaly. Peripheral blood smears showed hypochromic microcytic red blood cells, together with spherocytes and circulating erythroblasts. Her father was asymptomatic, with a mild hypochromic microcytic hyperproliferative anemia, and her mother had normal hemoglobin and reticulocyte counts. Patient 2 had a clinical presentation of transfusion dependence, extramedullary paravertebral hematopoiesis, moderate hepatic iron overload, mild pulmonary hypertension and leg ulcers. Peripheral blood smears showed hypochromic microcytic red blood cells, together with a moderate number of spherocytes and circulating erythroblasts; clinical and laboratory data from her relatives were not available. Laboratory workup is shown in Table 1. Exome sequencing was performed and, beyond the previously known β-globin mutations, mutations in spectrin gene were found in patient 1 (SPTB p.Leu880Gln) and patient 2 (SPTB p.Asp536Asn). The father of the Patient 1 was diagnosed with a heterozygous mutation in HBB gene, while her mother was diagnosed with a heterozygous SPTB p.Leu880Gln mutation, which did not result in anemia nor in significant hemolysis. Both spectrin mutations were already described in the literature: SPTB p.Leu880Gln was reported in association with a mutation in SPTA1 gene, resulting in a moderate anemia, while SPTB p.Asp536Asn mutation was described in an non-anemic individual. Conclusion: Our results indicate that the association of heterozygous BT and spectrin mutations may result in a phenotype of moderate to severe anemia. These data show that, in unusual severe cases of presumed BT trait, mutations responsible for HS should be investigated. Figure 1View largeDownload PPTFigure 1View largeDownload PPT Close modal

3-Carboxy-4-Methyl-5-Propyl-2-Furanpropionate (CMPF), a Protein-Bound Uremic Solute, Augments RBC Osmotic Fragility

3-Carboxy-4-Methyl-5-Propyl-2-Furanpropionate (CMPF), a Protein-Bound Uremic Solute, Augments RBC Osmotic Fragility

Screening of Extended Family Members of Thalassemia Major Children as a Thalassemia Preventive Strategy.

Thalassemia is considered as the most common single gene disorder worldwide. Preventive measures include identification of thalassemia carriers (traits) through screening, genetic counselling and prenatal diagnosis to reduce the incidence. This study aims at estimating the prevalence of carrier status detection among the extended family members of children having thalassemia major so as to use it as a screening prevention strategy with appropriate counselling. This cross-sectional study was conducted in thalassemia unit of Pediatric Department of a tertiary care teaching hospital over a period of 18 months. Blood samples were collected from 117 extended family members (EFM) of 23 children with thalassemia major to carry out investigations such as Complete Blood Counts (CBC), Naked Eye Single Tube Red Cell Osmotic Fragility Test (NESTROFT), Reticulocyte count, High Performance Liquid Chromatography(HPLC) and serum ferritin. Reports were analysed to find out the prevalence of carriers. Among 117 EFM, 62 (52.9%) were males while 55(47.1%) were females. Mean age distribution in this study was 16.49 years (8.5). Prevalence of thalassemia trait (carrier) was 35%. NESTROFT test was positive in 57(48.7%) participants. The binary logistic regression found only positive NESTROFT test as a predictor (adjusted OR=0.022, P=0.001) of having raised HbA2 (HbA2≥3.5 %). Screening of thalassemia carrier by targeting extended family members of thalassemia major children could yield more carrier cases and targeted counselling could help effectively in decreasing the number of children born with thalassemia major. This strategy could be included in future plan of national prevention programme for thalassemia.

Cadmium chloride generates cytotoxic reactive species that cause oxidative damage and morphological changes in human erythrocytes.

Cadmium chloride (CdCl2) is a widely used industrial compound that exhibits multiple organ toxicity. Cadmium is transported through blood where erythrocytes are exposed to its action. Here the effect of CdCl2 on human erythrocytes was examined under in vitro conditions. Human erythrocytes were treated with 0.01-0.5mM CdCl2 for 24h at 37 °C. Lysates were made from CdCl2 treated and untreated (control) cells and used for further analysis. CdCl2 treatment resulted in marked hemolysis of erythrocytes and oxidation of hemoglobin to methemoglobin. This will result in anemia and also reduce the oxygen carrying ability of erythrocytes. Hemoglobin oxidation was accompanied by degradation of heme and release of free ferrous iron moiety. Further analysis showed elevated lipid hydroperoxides and formation of advanced oxidation protein products along with reduction in total sulfhydryl content, indicating the generation of oxidative stress condition in the cell. Incubation of erythrocytes with CdCl2 enhanced generation of reactive oxygen and nitrogen species, decreased the antioxidant power and inhibited pathways of glucose metabolism. Plasma membrane was damaged as indicated by enhanced osmotic fragility and inhibition of membrane bound enzymes. This was confirmed by electron microscopy which showed formation of echinocytes. These results show that CdCl2 generates reactive species which impair the antioxidant system resulting in oxidative damage to erythrocytes.

Uji Validitas Panel Pemeriksaan Mean Corpuscular Volume, Indeks Mentzer dan Fragilitas Osmotik Tabung Tunggal untuk Skrining Talasemia Minor terhadap Pemeriksaan HbA2 Menggunakan High Performance Liquid Chromatography

Thalassemia is a disease caused by reduced or loss of synthesis of globin chains and then hemoglobin becomes abnormal. Characteristic screening in the population needs to be done to prevent marriage among people with thalassemia minor and reduce the birth of babies with thalassemia mayor. Combining several simple tests such as MCV, Mentzer index and single tube osmotic fragility test can increase the effectiveness of the test for screening of thalassemia minor. The purpose of this study was to determine the sensitivity and specificity of each panel for MCV-Mentzer index, MCV-single tube osmotic fragility, and Mentzer index- single tube osmotic fragility test panel against HbA2 in screening for the thalassemia minor. This study is observational with cross-sectional study design. The study was conducted at the Clinical Pathology Laboratory of Arifin Achmad Hospital Pekanbaru in June-August 2020. There were 27 subjects who had MCV values &lt;80 fl out of 230 screened subjects. Then on 27 research subjects were calculated for Mentzer index, single tube osmotic fragility test and HPLC. The research results show the sensitivity, specificity, positive predictive value and negative predictive value of the MCV-Mentzer index panel for screening of thalassemia minor were 11.8%, 90%, 66.7% and 37.5%, respectively. For the MCV-single tube osmotic fragility panel was, respectively, 70.6%, 20%, 60% and 28.6% and for the Mentzer index- single tube osmotic fragility panel was 11.8%, 90%, 66, 7% and 37.5% respectively. The conclusion is that the MCV-single tube osmotic fragility panel has better sensitivity than the MCV-Mentzer index panel or the Menzer index-single tube osmotic fragility panel that it can be used to screen for thalassemia minor in the premarital population in Pekanbaru.

Novel O-Methylglucoside Derivatives of Flavanone in Interaction with Model Membrane and Transferrin.

Flavonoids were biotransformed using various microorganisms, in order to obtain new compounds with potentially high biological activity. The aim of this work was to determine and compare the biological activity of four novel 6-methylflavanone O-methylglucosides. The tested compounds have the same flavonoid core structure and an attached O-methylglucose and hydroxyl group at different positions of ring A or B. The studies on their biological activity were conducted in relation to phosphatidylcholine membrane, erythrocytes and their membrane, and with human transferrin. These studies determined the compounds’ toxicity and their impact on the physical properties of the membranes. Furthermore, the binding ability of the compounds to holo-transferrin was investigated. The obtained results indicate that used compounds bind to erythrocytes, change their shape and decrease osmotic fragility but do not disrupt the membrane structure. Furthermore, the used compounds ordered the area of the polar heads of lipids and increased membrane fluidity. However, the results indicate the binding of these compounds in the hydrophilic region of the membranes, like other flavonoid glycosides. The used flavanones formed complexes with transferrin without inducing conformational changes in the protein’s structure. The relationship between their molecular structure and biological activity was discussed.

Fresh autologous blood transfusion reverses erythrocytes damage by reducing oxidative stress via promoting mitochondrial metabolism and M2 macrophages polarization

IntroductionSystemic inflammatory and immune suppressive incidents constitute a significant hallmark of red blood cell transfusion's adverse effects. These adverse side effects are seemingly associated with old blood transfusion; therefore, we aim to investigate the underlying mechanism of fresh autologous blood transfusion (fABT) and its subsequent effect in diabetic mice.Material and methodsIn the present study, we utilized 60 Swiss male mice aged 6-8 weeks, categorized into normal, diabetic, and freshly transfused with autologous blood. After treating the mice accordingly, further experimentations took place as we assessed the M1/M2 macrophage polarization concerning CD16/CD32/CD206 cells by flow cytometry, determined the mitochondrial metabolism using Lowry's analysis, measured hepatic oxidative stress using MDA and SOD assays, and examined the erythrocytic oxygen-carrying capacity (Q value) and oxygen consumption rate (OCR) and osmotic fragility.ResultsResults showed that fresh autologous blood transfusion markedly reduced M2 macrophage polarization, enhanced hepatic mitochondrial metabolism, reduced hepatic oxidative stress, and promoted red blood cells' oxygen-carrying capacity while reducing its osmotic fragility and oxygen consumption. Moreover, we found that fABT promoted the IGF2/PI3K signaling pathway, These results indicate the vital necessity of providing fresh autologous blood transfusion during therapy or surgeries.ConclusionsIn conclusion, our results are the first to highlight the underlying mechanism in which how fABT modulates physiological patterns in diabetic animals model, proposing a sound basis for utilizing fABT in clinical applications.

TWO CASE REPORT OF PARVOVIRUS B19 INFECTION IN HIV-INFECTED PATIENTS: DIFFERENT CLINICAL PRESENTATIONS FOLLOWING ACUTE AND PERSISTENT B19V INFECTION

Parvovirus B19 (B19V) infection could cause clinical manifestations depending on the host‘s immunologic and hematologic status. In immunocompetents individuals, clinical manifestations range from asymptomatic, or flu-like symptoms in childhood including a viral exanthem (fifth disease) to a fatal hydrops fetalis in pregnant women. B19V infect red blood cell (RBC) precursors and in immunocompromised hosts, such as HIV-infected patients, it could be manifested as pure red cell aplasia and chronic anemia. B19V may cause transient aplastic crisis (TAC) in patients with increased RBC destruction, including hereditary spherocytosis (HS). To report cases of B19V infection in HIV-positive adults in follow-up at Instituto Nacional de Infectologia Evandro Chagas. Case 1: A 27-year-old black male with HIV since childhood on irregular use of antiretroviral therapy (ART) complained of vertigo, headache, vomiting, fever. In April 2021 he had Hb 1,7 g/dL, Ht 6.5%, reticulocytes 0%, HIV viral load 40,305 cps/mL and CD4 + count 6 cells/μL. Up to October 2021 he required multiple RBC transfusions. Myelogram showed giants proerythroblasts with nuclear inclusions, B19V real time polymerase chain reaction (qPCR) detected 4.3 × 10 10 IU/mL, and B19V anti-IgM was negative. He received intravenous immunoglobulin (IVIG) and stabilized RBC counts. Nine months after treatment, he remained asymptomatic but had 11 cells/ μL CD4+ count and 27,642 HIV cps/mL due to poor adherence to ART. B19V was still detectable in qPCR (1.8 × 10 6 IU/mL), but he did not receive any additional treatment because blood counts were normal. Case 2: A 61-year-old white male with HIV since 2009 on regular use of ART, undetectable HIV viral load, but historically low CD4 + count, presented four weeks of lethargy, and an acute incident of dizziness, diarrhea, and syncope. He exhibited pancytopenia (Hb 5.6 g/dL, Ht 15.2%, WBC 1.48 × 10 9 /L, and platelets 41,000 per mm 3 ), non-immune hemolytic anemic features, jaundice, and palpable spleen. Reticulocyte counts were unavailable. He received transfusions of RBC, the HIV viral load at admission was undetectable, and CD4 + count 127 cells/ μL. The myelogram presented giant proerythroblasts with nuclear inclusions, and bone marrow had 95% cellularity and mild erythroid hyperplasia. B19V anti-IgM was indeterminate, while B19V qPCR detected 8.9 × 10 3 IU/mL, in August 2021. The patient received IVIG and nine months later, showed resolution of pancytopenia, but maintained elevated indirect bilirubin, indicative of chronic hemolysis. B19-DNA was no longer detected by qPCR, hematoscopy showed spherocytes, and the osmotic fragility test suggested RBC membranopathy, revealing an undiagnosed HS. Anemia is a common finding in the HIV setting, is often multifactorial, and represents a significant risk factor for mortality in AIDS. Our study highlights the importance of B19V infection be considered as a part of the differential diagnosis of anemia in this group. Clinicians should search for typical bone marrow and virologic findings. Identification of B19V DNA by qPCR was essential herein because of low sensitivity of serologic tests. Our findings showed that adherence to ART and the status of immunosuppression were crucial to the parvovirus clearance after IVIG. These cases highlight the challenging management of refractory parvovirus disease in immunosuppressed patient.

Effect of myostatin gene mutation on erythrocyte osmotic fragility, hematological parameters and fatty acid composition of serum and erythrocyte membranes in piglets

Fatty acid composition of serum and erythrocyte membrane, erythrocyte osmotic fragility and hematological parameters were estimated with the objective of determining effects of the gene mutation in one-week-old MSTN homozygous mutant (KO, MSTN−/−), heterozygous mutant (MSTN−/+) and wild type (WT, MSTN+/+) piglets (n = 4 each). Erythrocyte osmotic fragility, complete blood count (CBC), and fatty acid composition of serum and erythrocyte membrane were determined by flow cytometric analysis, automated hematology analyzer system, and liquid chromatography, respectively. Mean of median corpuscular fragility (MCF) was lower (P < 0.05, 0.001) in KO than MSTN−/+ and WT piglets. KO piglets had decreased (P < 0.05) white blood cell (WBC) count, lymphocyte (LYM) count, platelet (PLT) count, mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH), red cell distribution width-standard deviation (RDW-SD), red cell distribution width-coefficient volume (RDW-CV), plateletcrit (PCT), mean platelet volume (MPV), platelet distribution width (PDW), platelet-large cell ratio (P-LCR), and an increased red blood cell (RBC) count when compared with MSTN−/+ and WT piglets. The ratios of unsaturated fatty acid (UFA) to saturated fatty acid (SFA) concentrations in serum and erythrocyte membranes of MSTN KO piglets were 2-fold and 4-fold higher compared to WT piglets (P < 0.001), respectively. In conclusion, MSTN KO piglets had a decreased erythrocyte osmotic fragility, and altered hematological profile and fatty acid composition of serum and erythrocyte membranes, as characteristic phenotype.

Storage with ethanol attenuates the red blood cell storage lesion

BackgroundCurrent management of hemorrhagic shock relies on control of surgical bleeding along with resuscitation with packed red blood cells and plasma in a 1-to-1 ratio. Transfusion, however, is not without consequence as red blood cells develop a series of biochemical and physical changes during storage termed “the red blood cell storage lesion.” Previous data has suggested that ethanol may stabilize the red blood cell membrane, resulting in improved deformability. We hypothesized that storage of packed red blood cells with ethanol would alter the red blood cell storage lesion. MethodsMice underwent donation and storage of red blood cells with standard storage conditions in AS-3 alone or ethanol at concentrations of 0.07%, 0.14%, and 0.28%. The red blood cell storage lesion parameters of microvesicles, Band-3, free hemoglobin, annexin V, and erythrocyte osmotic fragility were measured and compared. In additional experiments, the mice underwent hemorrhage and resuscitation with stored packed red blood cells to further evaluate the in vivo inflammatory impact. ResultsRed blood cells stored with ethanol demonstrated decreased microvesicle accumulation and Band-3 levels. There were no differences in phosphatidylserine or cell-free hemoglobin levels. After hemorrhage and resuscitation with packed red blood cells stored with 0.07% ethanol, mice demonstrated decreased serum levels of interleukin-6, macrophage inflammatory protein-1α, keratinocyte chemokine, and tumor necrosis factor α compared to those mice receiving packed red blood cells stored with additive solution-3. ConclusionStorage of murine red blood cells with low-dose ethanol results in decreased red blood cell storage lesion severity. Resuscitation with packed red blood cells stored with 0.07% ethanol also resulted in a decreased systemic inflammatory response in a murine model of hemorrhage.

A case report of hereditary spherocytosis (HS): Approach to diagnosis and management of HS

Hereditary spherocytosis (HS) is a type of congenital hemolytic anemia, in which heterogeneous alterations in one of the six genes that encode for proteins involved in vertical associations which tie the red blood cell (RBC) membrane skeleton to the lipid bilayer causesdysfunction or deficiency of cell membrane protein resulting in spherical-shaped, hyper- dense, and poorly deformable RBCs with a shortened life span. We report a case of HS in a 2-month-old female who presented with severe anemia, jaundice, and hepatosplenomegaly.The peripheral blood smear showed spherocytosis and reticulocytosis. The osmotic fragility was positive and direct antiglobin test was negative. The osmotic fragility test and direct antiglobulin test were positive. She was managed with packed RBCs (PRBCs) transfusionand folic acid supplementation.

Congenital hemolytic anemias due to erythrocyte membrane andenzyme defects

Erythrocyte membrane and enzyme defects are the most common cause of congenital hemolytic anemias in the Central European population. Diagnostics include erythrocyte morphology, special biochemical tests such as osmotic fragility (AGLT) and EMA. For enzymopenic hemolytic anemias, cost-effective biochemical analysis remains the gold standard, supplemented by molecular genetic diagnostics when appropriate. Therapeutically, near complete splenectomy reduces hemolysis significantly for spherocytosis. The residual spleen at least provides a considerable phagocytic function and better response to immunisation and by inference possibly better protection against severe post-splenectomy infection. For pyruvate kinase deficiency, which is not so rare, a new molecular therapy (Mitapivat) is currently being introduced. In G6PD deficiency, there are very few drugs that cause hemolytic crisis. Sudden onset of hemoglobinuria is an early important hallmark of severe hemolytic crisis in G6PD deficiency and these patients should be hospitalized. Aplastic crises in the setting of parvovirus B19 infection occur in all congenital hemolytic anemias. Transfusion is not preventable in most cases. Iron-excreting treatment is required in the rare patients in need of chronic transfusion.

Potential role of blood constituents in pain-relief associated with fibromyalgia treatment with extremely low magnetic field: Spectroscopic investigation

Magnetic field is used as an adjunct therapy for pain control and relief of musculoskeletal pain conditions as Fibromyalgia. It is regarded as more natural and less harmful alternative to analgesic compounds. The exact mechanism underlying these positive effects is still to be determined. Twenty-three patients diagnosed with Fibromyalgia condition were included in this study, and subjected to extremely low magnetic field treatment sessions. The treatment protocol based on exposing patients to gradually increased magnetic field strength that starts with 0.1 μT at the beginning, and increased gradually to 100 μT (3.33 μT/min). UV–Visible, mid-IR and fluorescence characteristics of whole blood, erythrocytes and hemoglobin were investigated; in addition to RBCs-osmotic fragility measurements. The obtained results were analyzed according to control-sham exposed patients. We observed marked changes in the blood (and/or its constituents) absorption spectra of fibromyalgia patients that indicate an enhancement in the energetic pathways and increased hemoglobin-oxygen affinity; in addition, the osmotic fragility measurements show that erythrocytes were characterized by increased elasticity and rehydration. The analyses of infrared spectra show that magnetic field treatment was associated with changes in the erythrocytes skeleton where α-helix component of protein secondary structure is dominant. This study provides scientific evidence that pain relief associated with extremely low magnetic field treatment can be directly related to its systemic effects as well as the enhancement of the cellular activities.

Dietary histidine requirement for fingerlings of Channa punctatus (Bloch)

To estimate the requirement of fingerling Channa punctatus (6.34 ± 0.08 g) for dietary histidine, the present study was conducted by feeding six isoproteic (450 g kg−1 CP) and isoenergetic (14.73 kJ g−1 DE) diets with six histidine levels (2.5, 5, 7.5, 10, 12.5 and 15 g kg−1 diet). The trial diets were fed for 12 weeks in a triplicate group of fish at 08:00, 12:30, and 17:30 h to apparent satiation. Absolute weight gain (AWG; 49.1 g fish−1), daily growth index (DGI, 2.34%), feed conversion ratio (FCR, 1.42), protein retention efficiency (PRE; 28.12), and histidine retention efficiency (HRE; 29.69%) improved significantly with increasing histidine from 2.5 to 12.5 g kg−1 diet. Dietary histidine concentrations considerably affected carcass protein, lipid, and moisture contents. Increment in hematocrit (Hct), hemoglobin (Hb), and red blood cell counts (RBCs) were noted with increasing histidine up to 12.5 g kg−1 after that decreased. In contrast, erythrocyte sedimentation rate (ESR) and erythrocyte osmotic fragility (EOF) showed a reverse pattern. Total Serum protein, superoxide dismutase (SOD), and catalase (CAT) activities responded positively up to 12.5 g kg−1 dietary histidine, whereas malondialdehyde (MDA) aspartate aminotransferase (AST) and alanine aminotransferase (ALT) significantly declined up to 12.5 g kg−1 of dietary histidine. Second-order regression of AWG, FCR, PRE, and HRE against different histidine concentrations showed the requirements at 12.2, 11.7, 12.3, and 11.2 g kg−1, respectively. Dietary inclusion of 12.5 g kg−1 histidine is recommended for fingerling C. punctatus for optimum growth, hematological parameters, and antioxidant activities.

Erythrocyte osmotic fragility is not linked to vitamin C nutriture in adults with well-controlled type 2 diabetes.

Erythrocyte fragility is amplified by oxidative stress and linked to diabetes-specific microvascular disease. Vitamin C supplementation improves glycemic indices in adults with type 2 diabetes (T2D) by improving antioxidant status. This cross-sectional study examined the relationships between vitamin C status and erythrocyte osmotic fragility in adults with or without T2D. Participants provided a fasting blood sample for erythrocyte osmotic fragility testing as a function of hypotonic NaCl concentrations. Additionally, plasma was stabilized with metaphosphoric acid prior to vitamin C analysis using isocratic reverse-phase UV-HPLC separation. Participants were grouped as diagnosed T2D (n = 14; 36% female; 55.5 ± 8.2 y; 31.5 ± 9.0 kg/m2; HbA1c: 7.4 ± 1.9%; plasma vitamin C: 36.0 ± 12.2 μM) or no diabetes (n = 16; 69% female; 38.7 ± 13.5 y; 26.8 ± 6.6 kg/m2; HbA1c: 5.4 ± 0.3%; plasma vitamin C: 34.8 ± 10.9 μM). Participant characteristics differed between groups only for age and hemoglobin A1c (HbA1c; p < 0.05). All hemolysis parameters were in normal ranges for the participants with T2D, and no significant differences in hemolysis parameters were noted between those with or without T2D. However, among participants with T2D, the NaCl concentration eliciting 50% hemolysis was higher for those with low (<7%) vs. high (>7%) HbA1c values (p = 0.037) indicating a slightly higher erythrocyte fragility in the former group. Vitamin C status did not impact any of the hemolysis parameters in adults with or without T2D. Thus, erythrocyte fragility was not elevated in T2D, and vitamin C nutriture was not related to erythrocyte fragility in adults with well-controlled T2D.

Do the diverse environments of Baltic coastal zone affect hematological and biochemical alterations in the blood of mute swans (Cygnus olor)?

Based on studies of the wintering population of mute swans in large urban agglomerations, it is possible to identify and infer the impact of environmental pollution in coastal regions near Baltic Sea on the physiological and biochemical changes in the blood of waterfowls. Hematological and biochemical changes in the blood in relation to chemical elements in their feathers are a useful tool for general ecophysiological conclusions. Hematological changes and blood chemistry in Mute Swan Cygnus olor is particularly environmentally dependent, therefore we examined hematological picture (red blood cells RBC, white blood cells WBC, heterophiles, eosinophils, basophils, monocytes, lymphocytes, hemoglobin, heterophile to lymphocyte ratio H/L, biochemical indicators (glucose, uric acid, total cholesterol, triglycerides, calcium, inorganic phosphorus, magnesium, total protein), stability of erythrocyte membranes, concentration of Ca, P, Mg in the blood, concentration of Al, Zn, Rh, Cu, Ru, Fe, Pb in feathers, in wintering population of 172 mute swans of different age (juvenile, adult) and gender, in three coastal types of areas of Southern Polish zone of Baltic Sea (Słupsk, Gdynia, Sopot). Percentage of changes in the morphological indices in the blood of mute swans with three independent characteristics (environment, sex, age) revealed that hemoglobin content exhibited the highest value (R2 = 53.8%) in the analysis of morphological indices; the effect of RBC, WBC, and basophils was much lower (WBC > RBC > basophils). Male and female erythrocytes from the coastal of Gdynia were more fragile than those of birds from coasts of Sopot and Słupsk. We found that osmotic fragility is altered in juvenile swans from Słupsk area and males from Gdynia area. The consequence was a higher level of hemolyzed erythrocytes in their blood. The effect of type of environment, age-, and sex-related impact on hematological indices and biomarkers of biochemical alterations in the blood of swans and comparison of these data with bioaccumulation of chemical elements in feathers of swans inhabiting 3 types of environment of Baltic coastal zone show significant differences in the hematological and biochemical indices. Albumins and globulins maintain the blood cations balance, however, changes in their concentrations in the blood suggest an impact on physiological mechanisms and body condition of swans.

DEVELOPMENT AND IN VITRO EVALUATION OF VALSARTAN-LOADED RESEALED ERYTHROCYTES

Objective: Resealed erythrocyte technique has attempts for preparation of valsartan loaded reservoir type using red blood cell as a carrier.&#x0D; Methods: The resealed erythrocytes were prepared by using glutaraldehyde as a cross-linking agent by Preswell dilution technique and sodium chloride as the medium has been made to predict mechanism of drug release and absorption based on the order of release. The prepared resealed erythrocytes was characterised as a percentage of cell recovery drug content, osmotic shock, turbulence fragility, osmotic fragility, in vitro drug release studies, and hemoglobin content study.&#x0D; Results: The result of the study showed that the resealed erythrocytes prepared with sodium chloride (9 %) showed biconcave shape, an assay of 6.5±0.4 %, osmotic shock (0.028±0.004 µg/ml), turbulence fragility of 0.228±0.046 µg/ml, and 44.38±5.54 % drug release in 8 h. The drug release kinetics was studied and found that release from spherical matrices, first-order model with non-fickian diffusion with and the dissolution occurs in planes that are parallel to the drug surface pattern.&#x0D; Conclusion: VaL administration could be avoided the number of drawbacks associated with systemic delivery and perhaps maintain a relatively constant plasma level during a lengthy course of treatment.

Effects of Dietary Phenylalanine: Tyrosine Ratio on Growth, DNA/RNA, Serum Biochemistry, Digestive Enzyme Activities and Physiological Responses of Heteropneustes fossilis

Stinging catfish (Heteropneustes fossilis) has high consumer demand due to its high iron and calcium contents. Also, being a lean fish, it is very suitable for people who do not prefer to consume animal fats. In view of its nutritional and therapeutic significance and lack of data on dietary phenylalanine: tyrosine ratio, a dose-response experiment was conducted to determine phe: tyr ratio for H. fossilis fry (4.1±0.3g). Six isonitrogenous (380 g/kg crude protein) and isocaloric (15.3 kJ/g digestible energy) amino acid test diets were prepared by adjusting 26.6 g/kg of total phenylalanine and tyrosine in varying ratios of phenylalanine/tyrosine (30:70, 40:60, 50:50, 60:40, 70:30, 80:20) on molar basis. Quadruplicate groups of fishes were fed with diets indicated for 12-weeks to apparent satiation, thrice daily. Mathematical analyses of Daily Growth Coefficient (DGC%), Specific Growth Rate (SGR%/day), Feed Conversion Ratio (FCR) and Erythrocyte Osmotic Fragility (EOF) as quadratic-broken line regression analyses, exhibited optimum dietary phenylalanine: tyrosine ratio of 64.94: 35.06 corresponding to 15.83 g/kg: 8.55 g/kg on equimolar basis. Data of this study would be of high significance to ensure that optimum phenylalanine and tyrosine ratio of fish is met while using a greater variety of cost-effective dietary protein feedstuffs for sustainable aquafarming of this fish.

Evaluation of selected hematological, biochemical and oxidative stress parameters in stored canine CPDA-1 whole blood

Blood transfusions are mainly given to intensive care patients; therefore, additional complications that could arise from storage lesions in preserved blood should be avoided. It has been shown that human stored red blood cells are subject to changes that are considered to be a number of interdependent processes involving metabolic disarrangement and oxidative stress. The aim of our study was to determine alterations in selected hematological and biochemical parameters and to assess whether and when oxidative stress is a significant phenomenon in stored dog CPDA-1 whole blood. Ten ½ unit bags of whole blood donated from dogs and preserved with CPDA-1 (anticoagulant containing citrate, phosphate, dextrose and adenine) were stored for 5 weeks. Each week, a 9 ml sample was drawn aseptically to measure hematological parameters, selected metabolites, free hemoglobin content, osmotic fragility, antioxidant enzyme activity, total antioxidant capacity, malondialdehyde concentration and protein carbonyl content.The results revealed an MCV decrease in the first week of storage and then a gradual increase; osmotic fragility decreased at that time and remained low throughout the study period. Leukodepletion became significant in the fourth week of storage. The free hemoglobin concentration continuously increased, with the greatest changes observed in the last two weeks of storage. The total antioxidant capacity changed in a reverse manner. Superoxide dismutase and glutathione peroxidase activities decreased from week 0 to week 3, and catalase activity tended to decrease over time. The highest malondialdehyde concentrations in blood supernatant were measured in the first week of storage, and the carbonyl concentration increased after 35 days.Hematological changes and oxidative stress are already present in the first week of storage, resulting in depletion of the antioxidant system and subsequent accumulation of oxidation products as well as erythrocyte hemolysis, which are most pronounced at the end of the storage period.