Intervention of standardized ethanol leaf extract of Annickia polycarpa, (DC.) Setten and Maas ex I.M. Turner. (Annonaceae), in Plasmodium berghei infested mice produced anti-malaria action and normalized gross hematological indices.

ETHNOPHARMACOLOGICAL RELEVANCE: Malaria is a global public health burden due to large number of annual infections and casualties caused by its hematological complications. The bark of Annickia polycarpa is an effective anti-malaria agent in African traditional medicine. However, there is no standardization parameters for A. polycarpa. The anti-malaria properties of its leaf are also not known. AIM OF THE STUDY: To standardize the ethanol leaf extract of A. polycarpa (APLE) and investigate its anti-malaria properties and the effect of its treatment on hematological indices in Plasmodium berghei infected mice in the Rane's test. MATERIALS AND METHODS: Malaria was induced by inoculating female ICR mice with 1.0 × 107P. berghei-infected RBCs in 0.2 mL (i.p.) of blood. Treatment was commenced 3 days later with APLE 50, 200, 400 mg/kg p.o., Quinine 30 mg/kg i.m. (Standard drug) or sterile water (Negative control) once daily per group for 4 successive days. Anti-malarial activity and gross malaria indices such as hyperparasitemia, mean change in body weight and mean survival time (MST) were determined for each group. Changes in white blood cells (WBCs), red blood cells (RBCs), platelets (PLT) counts, hemoglobin (HGB) concentration, hematocrit (HCT) and mean corpuscular volume (MCV) were also measured in the healthy mice before infection as baseline and on day 3 and 8 after inoculation using complete blood count. Standardization was achieved by UHPLC-MS chemical fingerprint analysis and quantitative phytochemical tests. RESULTS: APLE, standardized to its total alkaloids, phenolics and saponin contents, produced significant (P < 0.05) dose-dependent clearance of mean hyperparasitemia of 22.78 ± 0.93% with the minimum parasitemia level of 2.01 ± 0.25% achieved at 400 mg/kg p.o. on day 8. Quinine 30 mg/kg i.m. achieved a minimum parasitemia level of 6.15 ± 0.92%. Moreover, APLE (50-400 mg/kg p.o.) evoked very significant anti-malaria activity of 89.22-95.50%. Anti-malaria activity of Quinine 30 mg/kg i.m. was 86.22%. APLE also inverse dose-dependently promotes weight gain with the effect being significant (P < 0.05) at 50 mg/kg p.o. Moreover, APLE dose-dependently increased the MST of malaria infested mice with 100% survival at 400 mg/kg p.o. Quinine 30 mg/kg i.m. also produce 100% survival rate but did not promote (P > 0.05) weight gain. Hematological studies revealed the development of leukocytopenia, erythrocytosis, microcytic anemia and thrombocytopenia in the malaria infected mice which were reverted with the treatment of APLE 50-400 mg/kg p.o. or Quinine 30 mg/kg i.m. but persisted in the negative control. The UHPLC-MS fingerprint analysis of APLE led to identification of one oxoaporphine and two aporphine alkaloids (1-3). Alkaloids 1 and 3 are being reported in this plant for the first time. CONCLUSION: These results indicate that APLE possessed significant anti-malaria, immunomodulatory, erythropoietic and hematinic actions against malaria infection. APLE also has the ability to revoke deleterious physiological alteration produced by malaria and hence, promote clinical cure. These properties of APLE are due to its constituents especially, aporphine and oxoaporphine alkaloids.

Protective effects of traditional Tibetan medicine Zuo-Mu-A Decoction () on the blood parameters and myocardium of high altitude polycythemia model rats.

OBJECTIVE: To explore the protective effects of Tibetan medicine Zuo-Mu-A Decoction (, ZMAD) on the blood parameters and myocardium of high altitude polycythemia (HAPC) model rats. METHODS: Forty male Wistar rats were randomly divided into 4 groups by a random number table, including the normal, model, Rhodiola rosea L. (RRL) and ZMAD groups (10 in each group). Every group was raised in Lhasa to create a HAPC model except the normal group. After modeling, rats in the RRL and the ZMAD groups were administered intragastrically with RRL (20 mL/kg) and ZMAD (7.5 mL/kg) once a day for 2 months, respectively; for the normal and the model groups, 5 mL of distilled water was administered intragastrically instead of decoction. Then routine blood and hematologic rheology parameters were taken, levels of erythropoietin and 8-hydroxy-2'-deoxyguanosine (8-OHdG) were tested, and ultrastructural change in the left ventricular myocardium was observed using transmission electron microscopy. RESULTS: Compared with the model group, ZMAD significantly reduced the red blood cell count, hemoglobin levels, whole blood viscosity at low/middle shear rates, plasma viscosity, erythrocyte electrophoretic time, erythropoietin and 8-OHdG levels, and also increased the erythrocyte deformation index (P<0.05). There was no difference in all results between the RRL and the ZMAD groups. The cardiac muscle fibers were well-protected, mitochondrial matrix swelled mildly and ultrastructure changes were less prominent in the ZMAD group compared with the model group. CONCLUSION: ZMAD has significant protective effects on the blood parameters against HAPC, and also has the beneficial effect in protecting against myocardial injury.

Effect of early versus delayed cord clamping on hematological status of preterm infants at 6 wk of age.

OBJECTIVE: To compare the effect of early cord clamping (ECC) vs. delayed cord clamping (DCC) on hematocrit and serum ferritin at 6 wk of life in preterm infants. METHODS: This randomized controlled trial was conducted in the delivery room and neonatal intensive care unit of a tertiary hospital. One hundred preterm infants born between 30 (0)/7 and 36 (6)/7 wk were randomized to either early or delayed cord clamping groups. Parental informed consent was obtained prior to the delivery. In the ECC group, the cord was clamped immediately after the delivery of the baby and in the DCC group; the cord was clamped beyond 2 min after the baby was delivered. Hematocrit and serum ferritin at 6 wk of life were the primary outcomes. Incidence of anemia, polycythemia and significant jaundice were the main secondary outcomes. RESULTS: The mean hematocrit (27.3 ± 3.8 % vs. 31.8 ± 3.5 %, p value 0.00) and the mean serum ferritin (136.9 ± 83.8 ng/mL vs. 178.9 ± 92.8 ng/mL, p value 0.037) at 6 wk of age were significantly higher in the infants randomized to DCC group. The hematocrit on day 1 was also significantly higher in the DCC group (50.8 ± 5.2 % vs. 58.5 ± 5.1 %, p value 0.00). The DCC group required significantly longer duration of phototherapy (55.3 ± 40.0 h vs. 36.7 ± 32.6 h, p value 0.016) and had a trend towards higher risk of polycythemia. CONCLUSIONS: Delaying the cord clamping by 2 min, significantly improves the hematocrit value at birth and this beneficial effect continues till at least 2nd mo of life.

Tiempo de pinzamiento del cordón umbilical y complicaciones neonatales, un estudio prospectivo / Time of cord clamping and neonatal complications, a prospective study

OBJETIVO: Evaluar los efectos del pinzamiento precoz o tardío del cordón umbilical en recién nacidos a término y su correlación con los niveles de hemoglobina, hematocrito, ferritina y ciertas complicaciones neonatales. PACIENTES Y MÉTODOS: Estudio prospectivo en recién nacidos sanos, a término, nacidos por parto eutócico o distócico en nuestro hospital, entre mayo del 2009 y mayo del 2010. Se asignó a los pacientes según el tiempo de pinzamiento: grupo 1 (< 60 s), grupo 2 (1 a <2 min) y grupo 3 (2 a 3 min). Se realizaron análisis al momento del nacimiento y a las 48 h de vida, valorando los niveles de hemoglobina, hematocrito, ferritina y bilirrubina. Se evalúo el riesgo de aparición de policitemia, síndrome distrés respiratorio, fototerapia o ingreso en la Unidad de Cuidados Intensivos neonatal y el tiempo de estancia hospitalaria. RESULTADOS: Se incluyó a 242 pacientes: grupo 1 (g1=80), grupo 2 (g2=31) y grupo 3 (g3=131). Los antecedentes maternos y las características neonatales fueron similares en todas las categorías. El primer análisis demostró diferencias significativas en los niveles de ferritina de aquellos recién nacidos con pinzamiento más tardío (g1: 111 mg/dl, g2: 125 mg/dl, g3: 173 mg/dl; p < 0,01). En el segundo análisis los valores de hemoglobina (g1: 17,3 g/dl, g2: 18,9 g/dl, g3: 19,2 g/dl; p < 0,01), hematocrito (g1: 53,4%, g2: 58%, g3: 59%; p < 0,01) y ferritina (g1: 254 mg/dl, g2: 254,7 mg/dl, g3: 313 mg/dl; p = 0,008), fueron estadísticamente mayores en este mismo grupo. Al evaluar las complicaciones, observamos un aumento significativo en el número de casos de policitemia asintomática en el grupo 3. CONCLUSIONES: El pinzamiento tardío del cordón umbilical se asocia a un aumento en los niveles de hemoglobina, hematocrito y ferritina a las 48 h de vida y en el número de casos de policitemia asintomática

OBJECTIVE: To assess the effects of early or late clamping of the umbilical cord in term newborns, assessing the levels of hemoglobin, hematocrit, and ferritin, and their correlation with some of the complications. PATIENTS AND METHODS: A prospective study of healthy newborns at term or born by dystotic or eutocic delivery in our hospital between May 2009 until May 2010. Patients were assigned according to the time of clamping, group 1 (< 60 seconds), group 2 (1 to < 2 minutes), and group 3 (2 to 3 minutes). Laboratory tests were performed at birth and at 48hours of life, assessing the levels of hemoglobin, hematocrit, ferritin, and bilirubin. The risk of polycythemia, respiratory distress syndrome, neonatal phototherapy or admission to the Intensive Care Unit and the hospital stay, were evaluated. RESULTS: A total of 242 patients were included: group 1 (g1 = 80), group 2 (g2 = 31) y group 3 (g3=131). The background maternal and neonatal characteristics were similar in all sets. The first test showed significant differences in ferritin levels in those infants with delayed clamping (g1: 111 mg/dl, g2: 125 mg/dl, g3: 173 mg/dl; p < 0.01). In the second analysis the values of hemoglobin (g1: 17.3 g/dl, g2: 18.9 g/dl, g3: 19.2 g/dl; p < 0.01), hematocrit (g1: 53.4%, g2: 58%, g3: 59%; p < 0.01) and ferritin (g1: 254 mg/dl, g2: 254.7 mg/dl, g3: 313 mg/dl; p = 0.008) were statistically higher in this group. As regards complications, a significant increase was observed in the number of cases of polycythemia symptoms in group 3. CONCLUSIONS: The late cord clamping is associated with an increase in hematocrit, hemoglobin and ferritin at 48hours of life, as well as an increased risk of polycythemia present with symptoms

Effect of delayed versus early umbilical cord clamping on neonatal outcomes and iron status at 4 months: a randomised controlled trial.

OBJECTIVE: To investigate the effects of delayed umbilical cord clamping, compared with early clamping, on infant iron status at 4 months of age in a European setting. DESIGN: Randomised controlled trial. SETTING: Swedish county hospital. PARTICIPANTS: 400 full term infants born after a low risk pregnancy. INTERVENTION: Infants were randomised to delayed umbilical cord clamping (≥ 180 seconds after delivery) or early clamping (≤ 10 seconds after delivery). MAIN OUTCOME MEASURES: Haemoglobin and iron status at 4 months of age with the power estimate based on serum ferritin levels. Secondary outcomes included neonatal anaemia, early respiratory symptoms, polycythaemia, and need for phototherapy. RESULTS: At 4 months of age, infants showed no significant differences in haemoglobin concentration between the groups, but infants subjected to delayed cord clamping had 45% (95% confidence interval 23% to 71%) higher mean ferritin concentration (117 µg/L v 81 µg/L, P < 0.001) and a lower prevalence of iron deficiency (1 (0.6%) v 10 (5.7%), P = 0.01, relative risk reduction 0.90; number needed to treat = 20 (17 to 67)). As for secondary outcomes, the delayed cord clamping group had lower prevalence of neonatal anaemia at 2 days of age (2 (1.2%) v 10 (6.3%), P = 0.02, relative risk reduction 0.80, number needed to treat 20 (15 to 111)). There were no significant differences between groups in postnatal respiratory symptoms, polycythaemia, or hyperbilirubinaemia requiring phototherapy. CONCLUSIONS: Delayed cord clamping, compared with early clamping, resulted in improved iron status and reduced prevalence of iron deficiency at 4 months of age, and reduced prevalence of neonatal anaemia, without demonstrable adverse effects. As iron deficiency in infants even without anaemia has been associated with impaired development, delayed cord clamping seems to benefit full term infants even in regions with a relatively low prevalence of iron deficiency anaemia. Trial registration Clinical Trials NCT01245296.

Interrelationships between tissue iron status and erythropoiesis during postweaning development following neonatal iron deficiency in rats.

Dietary iron is particularly critical during periods of rapid growth such as in neonatal development. Human and rodent studies have indicated that iron deficiency or excess during this critical stage of development can have significant long- and short-term consequences. Since the requirement for iron changes during development, the availability of adequate iron is critical for the differentiation and maturation of individual organs participating in iron homeostasis. We have examined in rats the effects of dietary iron supplement following neonatal iron deficiency on tissue iron status in relation to erythropoietic ability during 16 wk of postweaning development. This physiological model indicates that postweaning iron-adequate diet following neonatal iron deficiency adversely affects erythroid differentiation in the bone marrow and promotes splenic erythropoiesis leading to splenomegaly and erythrocytosis. This altered physiology of iron homeostasis during postweaning development is also reflected in the inability to maintain liver and spleen iron concentrations and the altered expression of iron regulatory proteins in the liver. These studies provide critical insights into the consequences of neonatal iron deficiency and the dietary iron-induced cellular signals affecting iron homeostasis during early development.

[Clinical implications of high blood viscosity and its reduction bu photo-hemotherapy]. / Klinicheskoe znachenie vysokoí viazkosti krovi i vozmozhnosti ee snizheniia metodami fotogemoterapii.

The paper is concerned with an essential blood characteristic--viscosity which is considered in relation of mechanisms responsible for its normal and abnormal characteristics, its role in the onset of circulatory disorders. High blood viscosity may contribute to development of vascular disturbances. Efficient methods of normalizing high blood viscosity are analyzed with special emphasis on photochemotherapy. Effects of ultraviolet, blue and red laser radiation on blood viscosity are outlined.

[Blood viscosity and cerebral blood flow in aged].

It is well known that there is a close correlation between blood viscosity and blood flow. To clarify any relationship between blood viscosity and regional cerebral blood flow (rCBF) in the elderly, we simultaneously studied both CBF with PET (positron emission tomography) and blood viscosity with viscosimeter before and after phlebotomy in the elderly with various kinds of polycythemia. These subjects consisted of five male cases of secondary polycythemia due to pulmonary fibrosis, one male case of essential erythrocytosis (average age 66.6 +/- 4.6 years old) and one female case of stress polycythemia (47 years old). Before phlebotomy an increase in blood viscosity, decrease in rCBF and regional cerebral metabolic rate of oxygen (rCMRO2) were observed in all cases. After phlebotomy (total amount of 800 to 1,000 ml) blood viscosity rapidly decreased, and both rCBF and rCMRO2 tended to increase. There was a significant negative or positive correlation between CBF and blood viscosity or rCMRO2, respectively. However, no increase in cerebral oxygen transport was observed in any subject after phlebotomy. It was noted that cerebral infarction is not infrequent among elderly visitors to Kusatsu spa, which is characterized by high temperature hot spring water. From the authors' observation of 23 cases of cerebral infarction encountered during the last five years, it is noteworthy that the disease tended to occur more frequently during midnight to morning, specially 3:00 to 6:00. Thus, to clarify the pathogenetic mechanism of the cerebral infarction occurring after bathing in hot spring water, we studied the changes in blood viscosity, blood pressure and coagulation-fibrinolytic system after bathing in hot spring water.(ABSTRACT TRUNCATED AT 250 WORDS)

Identification of some abnormal haemoglobins by fast atom bombardment mass spectrometry and fast atom bombardment tandem mass spectrometry.

The characterization of two abnormal human haemoglobins by fast atom bombardment (FAB) mapping is presented. The first variant, called 'R', exhibits a tryptic FAB map identical to that of normal haemoglobin. However, using Staphylococcus protease V8, a peptide containing the carboxyl end of the beta-chain exhibits a mass shift down to 300 mass units. This clearly indicates the deletion of the two last amino acids of the beta-chain. The second variant, called 'Grenoble', is due to two different modifications of the beta-chain. The location of the Pro----Ser exchange on peptide T5 is achieved by the collisionally activated dissociation mass analyzed ion kinetic energy spectra of the corresponding [MH]+ ion. The m/z value of that peptide indicated a supplementary acid----amide modification, which was located by amino acid sequencing using chemical methods. This work concludes with the necessity of using complementary methods for achieving rapid determinations of abnormal proteins with minute amounts.

[Hemoglobin E: report of the first 2 cases in French subjects]. / Hémoglobinose E: présentation des deux premières observations chez des sujets d'origine française.

Hemoglobin E has been discovered casually in the blood of two French donors: one coming from the Alsace region, the other one coming from the Champagne region. In the two cases, the hemoglobin E is in an heterozygote state and takes the place of 33 per cent of the total haemoglobin in the first and 24 per cent in the second. We investigated in their families and found that other members of these families had hemoglobin E. Each time, it was associated with a microcytosis and polycythaemia without anemy or iron deficiency. The red cells morphology shows many microspherocytes and target-cells. There is no relationship between these two families and the research of an asiatic antecedent proved negative. These two observations give a supplementary proof that the geographic repartition of the hemoglobin E is larger than what we read in the first publications and shows the interest to study the hemoglobin of unexplained polycythaemia with microcytes in the blood of blood donors.