Oxidative and osmotolerant effects in Salvator merianae (Squamata: Teiidae) red blood cells during hibernation / Efeitos de oxidação e osmotolerância dos eritrócitos de Salvator merianae (Squamata: Teiidae) durante a hibernação

Abstract Hibernation is a natural condition of animals that lives in the temperate zone, although some tropical lizards also experience hibernation annually, such as the lizard native from South America, Salvator merianae, or "tegu" lizard. Even though physiological and metabolic characteristic associated with hibernation have been extensively studied, possible alterations in the red blood cells (RBC) integrity during this period remains unclear. Dehydration and fasting are natural consequences of hibernating for several months and it could be related to some cellular modifications. In this study, we investigated if the osmotic tolerance of RBCs of tegu lizard under hibernation is different from the cells obtained from animals while normal activity. Additionally, we indirectly investigated if the RBCs membrane of hibernating tegus could be associated with oxidation by quantifying oxidized biomolecules and the activity of antioxidant enzymes. Our findings suggest that RBCs are more fragile during the hibernation period, although we did not find evidence of an oxidative stress scenario associated with the accentuated fragility. Even though we did not exclude the possibility of oxidative damage during hibernation, we suggested that an increased RBCs volume as a consequence of hypoosmotic blood during hibernation could also affect RBCs integrity as noted.

Resumo A hibernação é uma condição natural dos animais que vivem na zona temperada, embora alguns lagartos tropicais também experenciem hibernação anualmente, como é o caso do lagarto nativo da América do Sul, Salvator merianae ou "teiú". Embora as características fisiológicas e metabólicas associadas à hibernação tenham sido amplamente estudadas, possíveis alterações na integridade das hemácias durante esse período ainda permanecem obscuras. A desidratação e o jejum são consequências naturais da hibernação por vários meses e podem estar relacionadas a algumas modificações celulares. Neste estudo, investigamos se a tolerância osmótica de hemácias do lagarto teiú sob hibernação são diferentes das células obtidas de animais em atividade normal. Além disso, investigamos indiretamente por meio da quantificação de biomoléculas oxidadas e da atividade de enzimas antioxidantes se a membrana das hemácias dos teiús em hibernação poderia estar associada à oxidação. Nossos resultados sugerem que as hemácias possuem maior fragilidade durante o período de hibernação, embora não tenhamos encontrado evidências de um cenário de estresse oxidativo associado à essa fragilidade acentuada. Embora não tenhamos excluído a possibilidade de dano oxidativo durante a hibernação, sugerimos que um aumento no volume das hemácias como consequência de sangue hipoosmótico durante a hibernação também poderia afetar a integridade de hemácias, tal como foi observado.

Oxidative and osmotolerant effects in Salvator merianae (Squamata: Teiidae) red blood cells during hibernation

Abstract Hibernation is a natural condition of animals that lives in the temperate zone, although some tropical lizards also experience hibernation annually, such as the lizard native from South America, Salvator merianae, or tegu lizard. Even though physiological and metabolic characteristic associated with hibernation have been extensively studied, possible alterations in the red blood cells (RBC) integrity during this period remains unclear. Dehydration and fasting are natural consequences of hibernating for several months and it could be related to some cellular modifications. In this study, we investigated if the osmotic tolerance of RBCs of tegu lizard under hibernation is different from the cells obtained from animals while normal activity. Additionally, we indirectly investigated if the RBCs membrane of hibernating tegus could be associated with oxidation by quantifying oxidized biomolecules and the activity of antioxidant enzymes. Our findings suggest that RBCs are more fragile during the hibernation period, although we did not find evidence of an oxidative stress scenario associated with the accentuated fragility. Even though we did not exclude the possibility of oxidative damage during hibernation, we suggested that an increased RBCs volume as a consequence of hypoosmotic blood during hibernation could also affect RBCs integrity as noted.

Resumo A hibernação é uma condição natural dos animais que vivem na zona temperada, embora alguns lagartos tropicais também experenciem hibernação anualmente, como é o caso do lagarto nativo da América do Sul, Salvator merianae ou teiú. Embora as características fisiológicas e metabólicas associadas à hibernação tenham sido amplamente estudadas, possíveis alterações na integridade das hemácias durante esse período ainda permanecem obscuras. A desidratação e o jejum são consequências naturais da hibernação por vários meses e podem estar relacionadas a algumas modificações celulares. Neste estudo, investigamos se a tolerância osmótica de hemácias do lagarto teiú sob hibernação são diferentes das células obtidas de animais em atividade normal. Além disso, investigamos indiretamente por meio da quantificação de biomoléculas oxidadas e da atividade de enzimas antioxidantes se a membrana das hemácias dos teiús em hibernação poderia estar associada à oxidação. Nossos resultados sugerem que as hemácias possuem maior fragilidade durante o período de hibernação, embora não tenhamos encontrado evidências de um cenário de estresse oxidativo associado à essa fragilidade acentuada. Embora não tenhamos excluído a possibilidade de dano oxidativo durante a hibernação, sugerimos que um aumento no volume das hemácias como consequência de sangue hipoosmótico durante a hibernação também poderia afetar a integridade de hemácias, tal como foi observado.

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 causes dysfunction 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) transfusion and folic acid supplementation

Prevention and amelioration of erythrocyte instability observed under deficiency of vitamin B12 alone or combined with micronutrient limitation through dietary supplementation with Chlorella and Spirulina

Micronutrient rich microalgae, Chlorella and Spirulina, could be natural food supplements to overcome the micronutrient deficiency, increasingly recognised as a global health issue. In two independent experiments, the Spirulina and Chlorella were evaluated as prophylactic and ameliorative dietary supplements of vitamin B12. Erythrocyte stability (relative osmotic fragility and haemolysis percentage), haematological parameters, micronutrient deficiency (serum levels of iron, zinc), plasma vitamin B12 and vitamin B12 biomarker (methylmalonic acid) were analysed. The deficient groups receiving Spirulina and Chlorella as prophylactic dietary supplements showed a 1.34 to 1.41 folds increase in serum iron and a 2.13 to 2.19 folds increase in plasma vitamin B12, compared to B12 deficient group. Supplementation of Spirulina to ameliorate vitamin B12 deficiency combined with micronutrient limitation showed an increase of 1.14 folds and 1.2 folds in serum iron and zinc respectively and 1.51 folds in plasma vitamin B12 compared to the deficient group. The relative osmotic fragility of erythrocytes in deficient experimental animals was 17 to 45% higher compared to the control. The osmotic fragility and deformation in the morphology of erythrocytes observed under vitamin B12 deficiency, alone or in combination with micronutrient limitation, were prevented and ameliorated on dietary supplementation with the microalgal biomass.

Stability of refrigerated whole blood samples for osmotic fragility test

ABSTRACT Background: The osmotic fragility test (OFT), conventionally used for assisting the diagnosis of many erythrocyte disorders, is a manual and time-consuming analysis not daily performed in many medical laboratories. This study was aimed at defining the stability of whole blood samples used for assessing erythrocyte osmotic resistance. Methods: Twenty-one consecutive routine whole blood samples collected into 5.4 mg K2EDTA were tested immediately after collection (day 0) and at different time intervals afterward (day 1, 2, 3, 4, 7, 10 and 14) after storage at 4 °C. The OFT was performed with the Osmored Monotest (1.3% glycerol; Eurospital, Trieste, Italy). Results at the different time points were compared with those obtained at day 0 and with the reference change value (i.e., 33%). Results: The median value of both hyperosmolar and hyposmolar resistance increased from baseline, reaching statistical significance at day 7 for hyperosmolar resistance and at day 1 for hyposmolar resistance, respectively. The median relative increase of hemolysis percentage values become greater than the reference change value at day 3 for hyposmolar resistance, while this limit was never overcome for hyperosmolar resistance. A significant inverse association was found between the mean increase in hyperosmolar resistance and the baseline value of hyperosmolar resistance (r = −0.92), mean corpuscular volume (MCV; r = −0.46) or mean corpuscular hemoglobin (MCH; r = −0.44), as well as between the mean increase in hyposmolar resistance and the baseline value of hyposmolar resistance (r = −0.86), or patient age (r = −0.56). Conclusions: The sample stability seems critical for the OFT. Whole blood specimens should not be stored refrigerated at 4 °C for >2 days before testing.

Experience with NESTROFT for screening for thalassemia trait/ minor: evaluation against CBC and HPLC in a high prevalence region in Saurashtra, Gujarat, India

Background: Hemoglobinopathies pose a significant health burden in India. Prevention programmes can significantly reduce this burden. Although sophisticated methods of screening for β thalassemia trait are available, a cheap and simple method is beneficial for population screening. Although the Naked Eye Single Tube Red Cell Osmotic Fragility Test (NESTROFT) has been evaluated in many studies, sample sizes were small in some and many earlier studies have not done complete blood count (CBC) and High-Performance Liquid Chromatography (HPLC) in all the cases. We evaluate the suitability of NESTROFT for detection of β-thalassemia trait in a high prevalence region in Saurashtra, Gujarat.Methods: Here, 1000 unrelated individuals were studied. NESTROFT, CBC and estimation of HbA2 and HbF or other hemoglobin variants were done by HPLC.Results: Prevalence of β thalassemia trait was 7.8% in this population. NESTROFT showed an overall sensitivity and specificity of 94.87 and 85.38 respectively for the detection of β thalassaemia trait. Using red cell indices (MCH <27 pg and MCV <80 fl), One β thalassemia trait with normal indices would have been missed. Among twelve individuals with other hemoglobinopathies (HbS, HbD, HbE, δβ thalassemia trait or HPFH), seven had a positive NESTROFT while three had normal MCV & MCH values.Conclusions: NESTROFT is a cost-effective sensitive test which does not require any equipment and can be done in remote areas. It remains a useful first line screening test when large populations have to be screened.

Cryohemolysis, erythrocyte osmotic fragility, and supplementary hematimetric indices in the diagnosis of hereditary spherocytosis

BACKGROUND: Hereditary spherocytosis (HS) is a chronic hemolytic anemia characterized by microspherocytes in the peripheral blood and increased erythrocyte osmotic fragility (EOF). This study evaluated the cryohemolysis test (CHT); initial hemolysis (IH); immediate and incubated hemolysis percentage in 5.5 g/L NaCl (H5.5); mean corpuscular hemoglobin concentration (MCHC); red blood cell distribution width (RDW); and Hb/MCHC, Hb/RDW, and MCHC/RDW ratios for the diagnosis of HS. METHODS: Data from 13 patients with HS were evaluated at the Instituto de Bioquímica Aplicada and compared with data from 14 unaffected individuals and 11 patients with anemia due to another etiology. Total blood and reticulocyte counts, CHT, and immediate and incubated EOF were performed in all subjects; sensitivity, specificity, efficiency, and Youden index (YI) were calculated. RESULTS: Eight patients with HS had MCHC ≥345 g/L, 10 had RDW ≥14.5%, 12 had IH >5.0 g/L, 11 had immediate H5.5 ≥5%, and 13 had incubated H5.5 ≥50% (the cut-off value to consider HS). The efficiency and YI were: immediate H5.5 (0.94–0.85), incubated H5.5 (0.89–0.82), IH (0.89–0.78), MCHC (0.87–0.62), CHT (0.84–0.54), and Hb/MCHC (0.71–0.56), respectively. The calculated ratios could distinguish subjects with HS from unaffected individuals (P 0.05). CONCLUSION: Although the CHT and supplementary hematimetric indexes were useful to differentiate individuals with SH from healthy controls, they cannot distinguish from anemias of other etiology. CHT and MCHC, in addition to EOF, are recommended for diagnosing HS patients because of their low cost and efficiency.

Effects of hypoxia exposure for different time on structure and function of red blood cells in rats / 中国病理生理杂志

AIM:To investigate the effects of hypoxia exposure on the structure and function of erythrocytes in rats at different time.METHODS:Male SD rats(n=40)were randomly divided into 5 groups,normal control group,1-week hypoxia group ,2-week hypoxia group ,3-week hypoxia group and 4-week hypoxia group ,with 8 rats per group.The rats in hypoxia groups were placed in the simulated 5800 m of high altitude in a hypobaric chamber for different time.The values of detected blood ,erythrocyte deformation index ,erythrocyte osmotic fragility ,erythrocyte oxygen dissociation ,e-rythrocyte apoptosis and bone marrow biopsy were determined.RESULTS:Compared with normal control group ,the red blood cell count ,hemoglobin content ,mean corpuscular volume and mean corpuscular hemoglobin significantly increased(P<0.01).Eversion rate of phosphatidylserine of erythrocytes increased.Oxygen half-saturation of hemoglobin increased(P<0.05).Bone marrow erythroid proliferation increased.The erythrocyte deformation index and erythrocyte osmotic fra-gility decreased significantly(P<0.01).In addition,oxygen dissociation curves shifted to the right.CONCLUSION:In the early stage of hypoxia ,compared with normal control group ,the changes of erythrocyte structure and function increase the oxygen supply to the tissue and are conducive to adapting to the plateau.However ,with the extension of hypoxia ,ex-cessive erythrocytosis results in thrombosis ,microcirculation disturbance and aggravating tissue hypoxia.

Esferocitosis hereditaria: experiencia clínica y diagnóstica en Argentina / Hereditary spherocytosis: clinical and diagnostic experience in Argentina / Esferocitose hereditária: experiência clínica e diagnóstica na Argentina

Los objetivos del presente estudio fueron: a) Analizar las características demográficas y clínicas de nuestra población al diagnóstico; b) Evaluar si las pruebas más recientes presentan ventajas sobre las tradicionales; c) Confirmar la frecuencia de las distintas deficiencias de proteínas de membrana; d) Establecer la relación entre severidad y resultado de las pruebas o tipo de deficiencia. Se analizaron 359 individuos estudiados desde 2007, cuando se incorporaron criohemólisis hipertónica (CH), citometría de flujo con eosina-5'- maleimida (5'EMA-CF), FOE por citometría de flujo (FOE-CF) y electroforesis de proteínas de membrana (SDS-PAGE) al estudio de laboratorio clásico, fragilidad osmótica eritrocitaria (FOE) y autohemólisis (AH). Criterios diagnósticos para Esferocitosis Hereditaria (ESH): esferocitos en frotis y dos pruebas positivas. Se identificaron 174 pacientes con ESH y 22 portadores sanos. El 74,9% eran menores de 12 años. La transmisión fue dominante en el 83,1% de los casos. Tuvieron manifestaciones neonatales 89,1%. Las pruebas con mayor sensibilidad fueron CH (92,0%), FOE diferida (91,1%) y 5'EMA-CF (88,5%). En los 125 pacientes en quienes se realizaron CH, 5'EMA-CF y FOE-CF se observó que todos tenían al menos una prueba positiva; 122 (97,6%) tuvieron dos o tres positivas. Las deficiencias más frecuentes fueron ankirina y espectrina. No hubo diferencia en el resultado de las pruebas entre los subgrupos de severidad. Se concluye que las deficiencias más frecuentes en Argentina son ankirina y espectrina, coincidiendo con otras poblaciones latinoamericanas. El uso simultáneo de CH, 5'EMA-CF y FOE-CF permite diagnosticar más del 97% de los casos. La incidencia de manifestaciones neonatales es elevada.

The aims of this study were (a) to assess demographic and clinical aspects of our population at diagnosis; (b) to evaluate diagnostic accuracy of hypertonic cryohemolysis (HC), eosin-5'-maleimide flow cytometry (EMA-FC) and flow cytometric osmotic fragility (OF-FC) in relation to standard screening tests osmotic fragility (OF) and autohemolysis (AH); (c) to confirm the previously reported prevalence of membrane proteins defects; and (d) to assess the relationship between severity of anemia and results of confirmatory tests. Since 2007, the following tests were available in our laboratory: OF, AH, HC, EMA-FC, OF-FC and SDS-PAGE of membrane proteins. Diagnostic criteria for hereditary spherocytosis were spherocytes in blood smear plus ≥2 positive tests. Data from 359 individuals were analyzed: 174 HS patients and 22 silent carriers were detected; 74.9% of patients were less than 12 years old; 83.1% of them showed a dominant inheritance pattern; antecedent of neonatal jaundice/anemia was registered in 89.1%. Tests with higher sensitivity were: HC (92.0%), incubated OF (91.1%), and EMA-FC (88.5%). HC, EMA-FC and OF-FC were simultaneously performed on 125 patients: each of them had at least 1 positive test; 122 (97.6%) had 2 or 3 positive tests. Ankyrin and spectrin were the most frequently found protein deficiencies. Comparison of test results in relation to severity of anemia showed no difference between groups. It can be concluded that compared toother Latin American countries, ankyrin and spectrin were the most frequent protein deficiencies. Simultaneous performing of HC, EMA-FC and OF-FC enabled diagnosing HS in more than 97% of patients. A high incidence of neonatal jaundice/anemia was observed.

Os objetivos do presente estudo foram: a) analisar as características demográficas e clínicas de nossa população ao diagnóstico; b) Avaliar se as provas mais recentes apresentam vantagens sobre as tradicionais; c) Confirmar a frequência das diversas deficiências de proteínas de membrana; d) Establecer a relação entre severidade e resultado das provas ou tipo de deficiência. Foram analisados 359 indivíduos estudados desde 2007, quando se incorporaram crio-hemólise hipertônica (CH), citometria de fluxo com eosina-5'-maleimida (5'EMA-CF), FOE por citometria de fluxo (FOE-CF) e eletroforese de proteínas de membrana (SDS-PAGE) ao estudo de laboratório clássico - fragilidade osmótica eritrocitária (FOE) e auto-hemólise (AH). Critérios diagnósticos para ESH: esferócitos em esfregaço e duas provas positivas. Foram identificados 174 pacientes com ESH e 22 portadores sadios. 74,9% eram menores de 12 anos. A transmissão foi dominante em 83,1%. Tiveram manifestações neonatais 89,1%. As provas com maior sensibilidade foram CH (92,0%), FOE diferida (91,1%) e 5'EMA-CF (88,5%). Nos 125 pacientes aos quais lhes realizaram CH, 5'EMA-CF e FOE-CF se observou que todos tinham no mínimo uma prova positiva; 122 (97,6%) tiveram duas ou três positivas. As deficiências mais frequentes foram anquirina e espectrina. Não houve diferença no resultado das provas entre os subgrupos de severidade. Conclui-se que as deficiências mais frequentes na Argentina são anquirina e espectrina, as quais coincidem com outras populações latinoamericanas. O uso simultâneo de CH, 5'EMA-CF e FOE-CF permite diagnosticar mais de 97% dos casos. A incidência de manifestações neonatais é elevada.

Grape extract protects against ionizing radiation-induced membrane alterations in Red Blood Cells.

Ionizing radiation causes damage to biomolecules in living cells through oxidative stress by excessive generation of reactive oxygen species (ROS) from radiolysis of body water. Blood and its components including the cells are exposed to a significant dose of radiation during irradiation. Grapes (Vitis vinifera L.) contain several bioactive phytochemicals and are rich source of antioxidants. Therefore, we hypothesized that the grape extracts would offer protection against the ionizing radiation-induced damage of the red blood cells (RBCs). To test our hypothesis, in the current study we investigated the radio-protective actions of extract of four different grape (Vitis vinifera) cultivars, namely Flame seedless (Black grapes), Kishmish chorni (Black with reddish brown), Red globe (Red) and Thompson seedless mutant (Sonaka, Green) against the g-irradiation-induced oxidative stress leading to the structural alteration in the RBC membrane in vitro. Freshly drawn blood samples from healthy volunteers itself or mixed with grape extracts from seed, skin or pulp of each cultivar were irradiated at 4 Gy after one hour of treatment. -irradiation for one hour did not change the hematological parameters. The average osmotic fragility (H50) and the maximum rate of hemolysis (dH/dC)max increased after the -irradiation. The confocal microscopic and atomic force microscopic (AFM) studies showed that irradiation induced transformation of RBC from biconcave cells to echinocytes, altered their surface roughness and the vertical distance. The grape extracts did not alter the viability of human erythrocytes. Our results suggested that the grape extract pretreatment ameliorated the ionizing radiation-induced alterations at a dose of 4 Gy in human erythrocytes in vitro. Moreover, protection offered by the seed extract was significantly better than that that of skin or pulp of the same cultivar. Furthermore, the protective action of grape extract depends on its source (seed, skin or pulp) as well on cultivars.

Esferocitosis hereditaria: Revisión. Parte I. Historia, demografía, etiopatogenia y diagnóstico / Hereditary spherocytosis. Review. Part I. History, demographics, pathogenesis, and diagnosis

La esferocitosis hereditaria es la anemia hereditaria más frecuente en nuestro país luego de la talasemia menor. En este artículo, se revisan aspectos históricos, demográficos, genéticos y etiopatogénicos de la enfermedad, y se describen las pruebas de laboratorio para su diagnóstico. Se remarca el comportamiento de la enfermedad en nuestra población y se detallan las deficiencias proteicas predominantes en nuestro país. Se enfatiza sobre las nuevas técnicas de laboratorio actualmente disponibles, con alta sensibilidad y especificidad, que permiten realizar un diagnóstico más temprano con volúmenes de muestra mucho menores que los necesarios para las pruebas convencionales.

Hereditary spherocytosis is the most frequent hereditary anemia excluding beta thalassemia in Argentina. Historical, demographic, genetic and pathogenic aspects of the disease are reviewed, and confirmatory laboratory tests are described. Special characteristics on the outcome of the disease in our population and prevalent protein deficiencies in our country are described. Emphasis is given on new available laboratory tests, which allow an earlier diagnosis using volume of blood samples significantly smaller than required for conventional tests.

Andrographolide effect on both Plasmodium falciparum infected and non infected RBCs membranes

Objective: To explore whether its antiplasmodium effect of andrographolide is attributed to its plausible effect on the plasma membrane of both Plasmodium falciparum infected and non-infected RBCs. Methods: Anti-plasmodium effect of andrographolide against Plasmodium falciparum strains was screened using the conventional malaria drug sensitivity assay. The drug was incubated with uninfected RBCs to monitor its effect on their morphology, integrity and osmotic fragility. It was incubated with the plasmodium infected RBCs to monitor its effect on the parasite induced permeation pathways. Its effect on the potential of merozoites to invade new RBCs was tested using merozoite invasion assay. Results: It showed that at andrographolide was innocuous to RBCs at concentrations approach its therapeutic level against plasmodia. Nevertheless, this inertness was dwindled at higher concentrations. Conclusions: In spite of its success to inhibit plasmodium induced permeation pathway and the potential of merozoites to invade new RBCs, its anti-plasmodium effect can't be attributed to these functions as they were attained at concentrations higher than what is required to eradicate the parasite. Consequently, other mechanisms may be associated with its claimed actions.

Discussion of Improved Examination Method of Osmotic Fragility Test / 现代检验医学杂志

Objective To verify result consistency of the improved examination method and the standard operation method with the erythrocyte osmotic fragility test (OFT).Methods The samples and reagents were reduced half volume and used different testing equipment to evaluate methods for OFT.Selected 100 samples that the brittleness were increased and de-creased (50 samples positive and negative respectively),used a kind of improved examination method and traditional test method to evaluate the consistency.Results The detection result of improved examination method and the detection results of the manufacturer standard method were consistent (t=1.660 8,P >0.05).So,there was no significant difference of con-trast between two groups of data.Conclusion Improved mothed OFT alternatives to traditional OFTscreening method could be faster,more objectively the results of clinical service,and could effectively reduce reagent and manpower cost,improve the efficiency of work.Therefore,this method could be used to groups detection and lack of equipment for the primary care of hospital screening thalassemia desease method is preferred.

Antisickling Activity and Membrane Stabilizing Effect of Anthocyanins Extracts from Adansonia digitata L. Barks on Sickle Blood Cells.

Background: Parts of baobab tree (Adansonia digitata) including especially the barks are commonly used for their medicinal properties. Aims: The aim of this work is to evaluate the antisickling activity of baobab tree barks, which are used in Congolese traditional medicine to manage Sickle Cell Disease. Study Design: Baobab tree barks was extracted with water by maceration. Phytochemical tests were conducted with standard procedures. Antisickling activity and the minimum concentration of extract required to normalize sickled cells was determined by Emmel test. Place and Duration of Study: This work was done at department of chemistry, Science Faculty, University of Kinshasa (DR Congo), between November 2012 and February 2013. Methodology: The barks collected from Adansonia digitata were dried and powdered. A chemical screening was perform and extraction of anthocyanins done. Antisickling activity was evaluated by Emmel test, membrane stability by osmotic fragility test and Fe3+ evolution by following solution absorbance at 630nm. The rate of sickle cell shape normalization was determined at different plant aqueous extract concentrations in order to determine the minimal concentration of extract required to normalize sickle cells. The shape modification was quantitatively evaluated from the values of parameters such as surface, radius and perimeters of sickle blood cells before or after treatment with plant extract using Motic software. Results: The aqueous extract of Adansonia digitata showed an antisickling activity with a maximal normalization rate of 65.7% and a minimal concentration required to normalize sickled cells of 5.0mg/mL. The cell surface, perimeter and radius were significantly different before and after treatment with plant extract. The chemical screening showed the presence of polyphenols among which anthocyanins. The biological activity of this plant would be due to these pigments. The anthocyanins extract have also shown a stabilization effect on sickle blood red cells membranes and a reduction of methemoglobin to hemoglobin effect Conclusion: The results obtained show significant antisickling activity of Adansonia digitata barks thus justifying the use of this plant by traditional healers in Congolese traditional medicine in the management of Sickle Cell Disease.

Alleviation of PEGylated Puerarin on Erythrocyte Hemolysis Induced by Puerarin in Glucose-6-phosphate Dehydrogenase-deficient Rats / 中草药·英文版

Objective: To explore and analyze the reducing hemolytic effects of PEGylated puerarin (PEG-PUE) on erythrocytes induced by PUE in glucose-6-phosphate dehydrogenase (G6PD)-deficient rats. Methods: The rat model with G6PD-deficiency was established via sc injecting 1% acetylphenyl-hydrazine. Then the G6PD-deficient erythrocyte suspension obtained from this rat model was used to evaluate the hemolytic effects of PUE and the reducing hemolytic effects of PEG-PUE via hemolytic activity and erythrocyte osmotic fragility assay. Results: It was found that PUE could cause a serious hemolysis to the erythrocyte suspension with the increase of drug concentration and the prolongation of drug incubation time, the hemolytic rate of PUE was up to 40%, while the addition of PEG-PUE to the erythrocyte suspension revealed no significant hemolysis. Additionally, the result of erythrocyte osmotic fragility indicated that PEG-PUE exerted a slight effect on the erythrocyte membranes, and the NaCl concentration that induced 50% hemolysis (32 mmol/L) was about one-third PUE. Conclusion: These results demonstrate that PEG-PUE could play a significant role in reducing the side effect of hemolysis induced by PUE. The low hemolytic activity of PEG-PUE makes it a favorable candidate for in vivo tests and PEG-PUE could also provide the useful insight for the further formulation development as an innovative drug. © 2013 Tianjin Press of Chinese Herbal Medicines.

Heparina e K3EDTA como anticoagulantes para tambaqui (Colossoma macropomum Cuvier, 1816) / Heparin and K3EDTA as anticoagulants for tambaqui (Colossoma macropomum Cuvier, 1816)

Este estudo avaliou a eficácia da heparina sódica e do EDTA tripotássico como anticoagulantes e seus efeitos sobre os parâmetros hematológicos de tambaqui (Colossoma macropomum). Foram utilizados dez indivíduos de tambaqui com médias de 384,9 ± 85,71 g de peso e 27,90 ± 2,10 cm de comprimento total para avaliação da heparina 5.000 UI e 100 UI, bem como do K3EDTA 10 por cento. Foram analisados a inibição da coagulação por 10 h, eritrograma e teste de fragilidade osmótica dos eritrócitos. Os resultados foram submetidos à análise de variância e as médias comparadas pelo teste de Tukey (P < 0,05). Heparina 5000 UI, heparina 100 UI e K3EDTA 10 por cento foram eficazes na prevenção da coagulação por mais de 10 h, no entanto o EDTA tripotássico causou hemólise desde os primeiros momentos. No eritrograma não foi observada diferença (P > 0,05) na contagem de eritrócitos, hematócrito, taxa de hemoglobina e CHCM, no entanto, houve aumento do VCM (P < 0,05) nas amostras acondicionados com K3EDTA 10 por cento. Este anticoagulante causou incremento (P < 0,01) na fragilidade osmótica dos eritrócitos quando comparado com a heparina pura, heparina diluída, e grupo controle. A utilização da heparina como anticoagulante é mais apropriada para tambaqui (Colossoma macropomum), visto que foi eficiente na prevenção da coagulação por mais de 10 h, sem ocasionar hemólise, ou alterações nos parâmetros hematológicos e na fragilidade osmótica dos eritrócitos.

The efficacy of sodium heparin and tripotassium EDTA as anticoagulant and their effect on the hematological parameters of tambaqui (Colossoma macropomum) were evaluated in this study. Ten fish weighing 384.9 ± 85.71 g and measuring 27.90 ± 2.10 cm were used for heparin 5.000 IU, heparin 100 IU and K3EDTA 10 percent evaluation. Clotting inhibition after 10 h, erythrogram and osmotic fragility of erythrocytes were observed. The results were submitted to variance analysis and means compared by Tukey test (P < 0.05). Heparin 5.000 IU, heparin 100 IU and K3EDTA 10 percent were effective in preventing coagulation for more than 10 h. However, tripotassium EDTA caused hemolysis since first moments. In erythrogram there was no difference (P > 0.05) in erythrocyte count, hematocrit, hemoglobin and MCHC. On the other hand, an increase in MCV (P < 0.05) in samples kept with K3EDTA10 percent was observed. This anticoagulant provoked a significant increase (P < 0.01) in the osmotic fragility of erythrocytes when compared to pure heparin, diluted heparin and the control group. Heparin as an anticoagulant is more appropriate for tambaqui since it was effective in preventing coagulation for more than 10 h, without causing hemolysis, changes on hematological parameters or osmotic fragility of erythrocytes.

Avaliação da influência de variáveis de processo sobre a cinética de desidratação osmótica da polpa de abacate (Persea americana L.) / Evaluation of the influence of process variables on osmotic dehydration kinetics of avocado pulp (Persea americana L.)

O abacate é uma das frutas tropicais cultivadas em diversas regiões do mundo e possui alto valor nutricional, porém é instável a deteriorações. A desidratação osmótica é uma técnica útil na conservação de frutas e vegetais pode ser uma alternativa para estabilização da polpa do abacate. Porém, é necessário estudar o comportamento físico da polpa da fruta frente a esta técnica. Desta forma, o objetivo deste trabalho foi avaliar a influência das variáveis de processo na desidratação osmótica da polpa do abacate. A desidratação osmótica foi realizada com auxílio de um planejamento experimental fatorial com as seguintes variáveis: concentrações das soluções de sacarose (30 e 60% p/p), espessura do corte (0,5 e 1 cm) e temperatura de imersão (25 e 45 °C) tendo por variáveis de resposta a perda de água (PA), ganho de sólidos (GS) e relação ganho de sólidos/perda de água (GS/PA) em 200 minutos de processo. Os resultados obtidos mostraram que o nível máximo de perda de água (PA), de 55,53%, ocorreu na menor concentração de açúcar (30%), maior temperatura (45 °C) e menor espessura (0,5cm). O nível mínimo de PA (14,5%) foi observado tanto na combinação de maior concentração de açúcar (60%), espessura (1 cm) e temperatura (45 °C) como nas condições de menor temperatura (25 °C), menor concentração de sacarose (30%) e maior espessura (1cm). A espessura foi o parâmetro que mais influenciou o processo em relação às respostas (GS, PA e GS/PA) estudadas. A temperatura também obteve significância para a resposta GS/PA, e a concentração de sacarose não apresentou influência significativa, nas condições em que a pesquisa foi realizada.

Avocado is one kind of tropical fruit grown in different regions in the world, and it has high nutritional value, but it is unstable to damage. Osmotic dehydration is a useful technique for the conservation of fruit and vegetables, and it can be an alternative for the stabilization of avocado pulp. However, it is necessary to study the physical behavior of the fruit pulp in the face of this technique. Thus, the aim of this study was to evaluate the influence of process variables on avocado pulp osmotic dehydration. Osmotic dehydration was carried out with help of a factorial-experimental planning with the following variables: concentrations of sucrose solutions (30 and 60% w/w), section thickness (0.5 and 1cm) and immersion temperature (25 and 45 °C), in which the response variables are water loss (WL), gain of solids (GS) and gain of solids/water loss relation (GS/WL) in 200 minutes of process. The results showed that the highest level of water loss (WL), 55.53%, occurred at the lowest sucrose concentration (30%), highest temperature (45 °C) and smallest thickness (0.5cm). The minimum level of WL (14.5%) was observed both in the combination of higher sugar concentration (60%), thickness (1cm) and temperature (45 °C) and lower temperature (25 °C), lower concentration of sucrose (30%) and greater thickness (1cm). The thickness was the parameter that most influenced the process concerning the responses analyzed (GS, WL, and GS/WL). The temperature was also significant for the GS/WL response, and the sucrose concentration had no significant influence in the conditions in which the survey was conducted.

Biomedical studies on lipid peroxidation and erythrocyte fragility during the process of aging

Objective: To investigate oxidative stress, hemoglobin percentage and erythrocyte osmotic fragility in various aging groups. Methods: A total of 200 healthy volunteers of both genders between age group 20-65 years were selected by random method. Determination of hemoglobin percentage was done employing modified cyanide method of Dacie and Lewis. The erythrocyte lysis was observed in hypotonic solution of buffered saline at varying concentrations and optical density was measured at 540 nm. The extent of lipid peroxidation in form of malondialdehyde was measured by thiobarbituric acid method. Results: The study found a significant decrease in hemoglobin percentage, increase in erythrocyte osmotic fragility and increased lipid peroxidation in form of malondialdehyde with increasing age. Conclusions: Supplementation of antioxidants may prevent the oxidative injury in elderly group of subjects.

Biomedical studies on lipid peroxidation and erythrocyte fragility during the process of aging

<p><b>OBJECTIVE</b>To investigate oxidative stress, hemoglobin percentage and erythrocyte osmotic fragility in various aging groups.</p><p><b>METHODS</b>A total of 200 healthy volunteers of both genders between age group 20-65 years were selected by random method. Determination of hemoglobin percentage was done employing modified cyanide method of Dacie and Lewis. The erythrocyte lysis was observed in hypotonic solution of buffered saline at varying concentrations and optical density was measured at 540 nm. The extent of lipid peroxidation in form of malondialdehyde was measured by thiobarbituric acid method.</p><p><b>RESULTS</b>The study found a significant decrease in hemoglobin percentage, increase in erythrocyte osmotic fragility and increased lipid peroxidation in form of malondialdehyde with increasing age.</p><p><b>CONCLUSIONS</b>Supplementation of antioxidants may prevent the oxidative injury in elderly group of subjects.</p>

Effect of the extract of Ricinus communis L. on the osmotic fragility, labeling of red blood cells with Technetium-99m and morphology of the cells

The aim of this study was to evaluate the influence of the proteic extract of R. communis on the cell physiology by the osmotic fragility, labeling of the blood elements with the 99mTc and cell morphology. To evaluate the osmotic fragility, the blood samples of the Wistar rats were incubated with the concentrations of R. communis and with the solutions of NaCl (0.4; 0.7; 0.9 percent). In the labeling of the blood elements procedure, the rat blood was treated with a solution of Tc-99m and TCA at 5 percent, determining the rate of radioactivity ( percentATI) in the plasma (P) and in the red blood cells (RBC). The soluble and insoluble fractions of the plasma were also evaluated. The cells morphology submitted to the extract was evaluated by the optical microscopy (x40). The results indicated that the rate of the hemolysis increased in the presence of 0.125 mg/mL of the extract. There was a decay of 49.69 percent in the rate of ATI in the insoluble fraction of the cells, with the morphological alterations in the red blood cells. These results suggested that the extract changed the capability of binding of the red blood cells due to the stannous ion oxidation, modifying the cells structure.

Produtos naturais são usados freqüentemente por muitas pessoas no tratamento do câncer. O Ricinus communis L é uma Euforbiaceae que apresenta propriedades laxativas, purgativas e antitumorais. O objetivo deste trabalho é estudar a influência da fração protéica do extrato hidroalcoólico de R. communis L. na fisiologia celular através da fragilidade osmótica, da marcação de elementos sanguíneo com 99mTc e da morfologia celular. Para avaliar a fragilidade osmótica, amostras de sangue de ratos Wistar foram incubadas com concentrações de R. communis e com soluções de NaCl (0,4; 0,7; 0,9 por cento). No procedimento de marcação de elementos sanguíneos, as amostras de sangue foram tratadas com solução de Tc-99m e TCA à 5 por cento, determinando o percentual de radioatividade ( por centoATI) no plasma (P) e nas células vermelhas (RBC); as frações solúvel e insolúvel do plasma também foram avaliadas. A morfologia das células submetidas ao extrato foi avaliada por microscopia óptica (x40). Os resultados indicam que o extrato na concentração de 0,125 mg/mL provoca hemólise de 49,69 por cento , no por cento ATI na fração insolúvel das células, ocorrendo alterações morfológicas das células sangüíneas. Esses resultados sugerem que o extrato radiomodifica a ligação do 99mTc às células vermelhas. Isto pode ser devido à oxidação do íon estanoso, a um processo de competição com os sítios de ligação do 99mTc ou por modificação das estruturas da membrana.