In Vitro Exchange of β-Sitosterol between Erythrocytes and Plasma of Rats

The erythrocytes and plasma of rats were not labeled equally with sterols even after feeding plant sterols for 2 months.

When erythrocytes and plasma were labeled in vivo with radioactive sterols, the in vitro exchange of cholesterol between cells and plasma was considerably greater than that of β-sitosterol. The dependence of the transfer on plasma lecithin : cholesterol acyltransferase was much less with β-sitosterol.

More labeled β-sitosterol existed in high density lipoprotein and less in very-low density and low density lipoproteins than cholesterol, when plasma was labeled in vivo. A similar distribution pattern was observed when plasma was incubated with labeled erythrocytes. These results suggest that an extra ethyl group in the side chain of the molecule substantially influences the metabolic behavior of the sterols.     

Agglutinins from marine macroalgae of the southeastern United States

Protein extracts from 22 species of marine macroalgae from Florida and North Carolina were compared for their abilities to agglutinate sheep and rabbit erythrocytes. Protein extracts from 21 algal species agglutinated rabbit erythrocytes compared to 19 for sheep erythrocytes. However, agglutination by brown algal extracts was variable. The agglutination produced by protein extracts from Dictyota dichotoma could be blocked by addition of polyvinylpyrrolidone. Protein extracts from North Carolina macroalgae were also tested against five bacterial species. Three of these agglutinated bacterial cells. Ulva curvata and Bryopsis plumosa agglutinated all five species. Protein extracts from five species of Florida algae were tested for their effects on mitogenesis in mouse splenocytes and human lymphocytes. Gracilaria tikvahiae HBOI Strain G-5, Ulva rigida and Gracilaria verrucosa HBOI Strain G-16S stimulated mitogenesis in mouse splenocytes, while Gracilaria tikvahiae HBOI Strain G-16stimulated mitogenesis in human lymphocytes.     

Effects of β‐carotene on antioxidant status in rats with chronic alcohol consumption

This study examined the effects of β‐carotene on antioxidant status in rats with chronic alcohol consumption. At the beginning of experiment (week 0), according to both the plasma aspartate aminotransferase (AST) and alanine aminotransferase (ALT) activities, rats (n = 24) were divided into 3 groups and fed with a standard diet (group C), a diet containing ethanol (group E), or a diet containing ethanol and β‐carotene (group E+B). After 10 weeks, plasma AST and ALT, fat accumulation in the liver, antioxidant enzyme activities in erythrocytes and the liver, malondialdehyde (MDA), and α‐tocopherol and retinol in plasma and hepatic samples were analyzed. The chronic alcohol diet significantly increased AST and ALT levels in plasma, and these changes were prevented by supplementing the diet with β‐carotene. Glutathione (GSH) in erythrocytes and in the liver was significantly elevated in rats fed with a diet containing β‐carotene. The results indicate that β‐carotene supplementation can prevent ethanol‐induced liver damage and increase GSH concentrations in erythrocytes and the liver. Copyright © 2009 John Wiley & Sons, Ltd.     

Analysis of the agglutinating activity from unicellular algae

Agglutinating activity often varies both between and within the algal species assayed. However, it is difficulty to interpret such variation without further analysis. We report a statistical analysis of agglutinating activities against human, cow, sheep, and pig erythrocytes, using cell extracts from 43 taxa (strains) of freshwater microalgae. Most of the extracts agglutinated erythrocytes from at least one of the sources, but pig erythrocytes appeared to be most suitable for the detection of agglutination reactions. Chlorella cell extracts preferentially agglutinated human erythrocytes, whereas extracts of other taxa were less active against mammalian erythrocytes. Cluster analysis generated four distinct subclusters of taxa, characterized by different specificities for antigens or carbohydrate receptors on the erythrocytes. Principal component analysis further separated the agglutination characteristics of Chlamydomonas from Chlorella on the first two components. Specificity for pig erythrocytes accounted for most of the clustering or grouping of algal taxa in multivariate analysis. However, clustering or grouping patterns of Chlorella species on haemagglutinating activity resembled that based on DNA sequences, revealing a possible genetic connection of agglutinins and their biochemical characteristics in algal cells. Variability of agglutination reactions among the algae investigated is simplified and interpreted most easily using multivariate analysis.     

Agglutination of human and animal erythrocytes in marine unicellular algae

Ethanol extracts of 12 marine unicellular algae were assayed for agglutinating activity against native and enzyme-treated human and animal erythrocytes. All of the algae assayed agglutinated at least one group of normal erythrocytes from humans. Notably, all algal extracts agglutinated erythrocytes of hemophilia patients arising from coagulation disorders. Meanwhile, all algae had a strong reaction with monkey erythrocytes, but to a lesser extent or not at all with sheep erythrocytes. Both trypsin and pronase improved the detection of most algal agglutinins and caused a drastic increase in hemagglutinating activity after treatment for 2 h or more. However, hemagglutinating activity decreased or disappeared completely when two extracts of different algal species were combined. Journal of Industrial Microbiology & Biotechnology (2000) 24, 262–266. Received 24 September 1999/ Accepted in revised form 06 January 2000     

Membrane cholesterol is required for activity of <Emphasis Type="Italic">Vibrio vulnificus </Emphasis>cytolysin

Vibrio vulnificus cytolysin (VVC) forms a pore in the plasma membrane and induces cytolysis of various cells including erythrocytes, neutrophil and endothelial cells. The cytolytic activity of VVC is inhibited by exogenously added cholesterol, suggesting that membrane cholesterol might be required for VVC cytolytic activity. However, there is no direct evidence that membrane cholesterol is involved in VVC-induced cytolysis. Herein we demonstrate that membrane cholesterol is required for binding of VVC to the plasma membrane. Membrane cholesterol depletion with methyl-β-cyclodextrin inhibited VVC-induced K⁺ release, 2-deoxy glucose release and Ca²⁺ influx, which are indicators of VVC pore formation. The cholesterol depletion-induced blockage of VVC cytolysis was due to the inhibition of VVC binding to membrane. These findings suggest that interaction with cholesterol is required for activity of VVC. Hong-Nu Yu, and Young-Rae Lee have equally contributed to this study.     

Functional Characteristics of <Emphasis Type="Italic">Lactobacillus</Emphasis><Emphasis Type="Italic">fermentum</Emphasis> F1

In this study, Lactobacillus fermentum (L. fermentum) F1 reduced cholesterol 48.87%. The strain was screened from cattle feces using an API 50 CHL system and the 16S rRNA sequence contrasting method. L. fermentum F1 showed acid and bile tolerance, and antimicrobial activity against pathogenic Escherichia coli and Staphylococcus aureus. L. fermentum F1 deconjugated 0.186 mM of free cholalic acid after it was incubated at 37°C in 0.20% sodium taurocholate (TCA) broth for 24 h. Heat-killed and resting cells of L. fermentum F1 showed small amounts of cholesterol removal (6.85 and 25.19 mg/g, respectively, of dry weight) compared with growing cells (33.21 mg/g of dry weight). The supernatant fluid of the broth contained 50.85% of the total cholesterol, while the washing buffer and cell extracts had 13.53 and 35.39%, respectively. These findings suggest that L. fermentum F1 may remove cholesterol by co-precipitating with deconjugated bile salt, assimilating with cells and by incorporation into cellular membranes. Cholesterol assimilated by cells held 72.0% of the reduced cholesterol, while 21.65% of the reduced cholesterol was coprecipitated with deconjugated bile salt and 5.89% was adsorbed into the surface of the cells.     

Screening and preliminary characterization of hemagglutinins in Vietnamese marine algae

Extracts from 44 species of Vietnamese marine algae, including 15 Chlorophyta, 18 Rhodophyta and 11 Phaeophyta species, were examined for hemagglutination activity with a variety of different animal and human erythrocytes that were untreated or treated with enzymes. Almost all extracts showed activity toward at least one type of erythrocytes, although those from three Chlorophyta and two Rhodophyta species showed no hemagglutination with any type of erythrocytes examined. Strong activity was detected in extracts from two Chlorophyta (Anadyomene plicata and Avrainvillea erecta) and four Rhodophyta species (Gracilaria eucheumatoides, Gracilaria salicornia, Kappaphycus alvarezii, and Kappaphycus striatum) with enzyme-treated rabbit and sheep erythrocytes. The hemagglutinins of seven Chlorophyta and eight Rhodophyta species were examined for sugar-binding specificity, pH- and temperature-stability, and divalent cation-independency of hemagglutination using ammonium sulfate-precipitates prepared from their extracts. In a hemagglutination-inhibition test with various monosaccharides and glycoproteins, none of the hemagglutinins had affinity for monosaccharides, except the Codium arabicum and Gracilaria euchematoides hemagglutinins, whose activities were inhibited by both N-acetyl-d-galactosamine and N-acetyl-d-glucosamine. On the other hand, all of the hemagglutinins activities were inhibited by some glycoproteins. The inhibition profiles with glycoproteins were different depending on hemagglutinin species, and suggest the presence of lectins specific for high mannose N-glycans, complex N-glycans, or O-glycans. The activities of these algal hemagglutinins were stable over a wide range of pH and temperature, and independent of the presence of divalent cations. These results indicate that Vietnamese marine algae are a good source of novel and useful lectins.     

A new screening for hemagglutinins from Vietnamese marine macroalgae

Aqueous extracts from 42 species of Vietnamese marine macroalgae, including 17 Chlorophyta, 22 Rhodophyta, and three Phaeophyta species, were examined for hemagglutination activity using native and enzyme-treated different animal and human erythrocytes. All extracts agglutinated at least one type of erythrocytes tested. Strong activity was detected in extracts from four Chlorophyta (Caulerpa serulata var. boryana, Caulerpa sertularioides f. longipes, Halimeda velasquezii, and Halimeda discoidea) and two Rhodophyta species (Gelidiella acerosa and Titanophora pulchra) with enzyme-treated rabbit and horse erythrocytes. The hemagglutinins of some active species were examined for sugar-binding specificity, pH, temperature stability, and divalent cation independency using ammonium sulfate precipitates prepared from their extracts. In a hemagglutination–inhibition test with various monosaccharides and glycoproteins, none of the hemagglutinins had affinity for monosaccharides. The activity of the hemagglutinins was inhibited by some glycoproteins tested. The inhibition profiles with glycoproteins were different depending on hemagglutinin species, suggesting the presence of lectins specific for complex N-glycans, high mannose N-glycans or O-glycans. On the other hand, the activities of almost all algal hemagglutinins were stable over a wide range of pH and temperature, and independent of the presence of divalent cations, except Gelidiopsis scoparia hemagglutinin, its activity was dependent on the presence of divalent cations. These results suggest that Vietnamese marine macroalgae may be good sources of useful lectins for many biological applications.     

Effect of <Emphasis Type="Italic">Gymnema montanum</Emphasis> leaves on red blood cell resistance to oxidative stress in experimental diabetes

The aim of the present study was to evaluate the protective effect of Gymnema montanum on red blood cell (RBC) membrane in diabetic rats during lipid peroxidation. Ethanol extract of G. montanum leaves (GLEt) was administered orally to alloxan-induced diabetic rats for 3 weeks, and the effects on blood glucose, insulin, lipid peroxidation markers, thiobarbituric acid reactive substances, hydroperoxides in plasma and antioxidant enzymes including superoxide dismutase, catalase and glutathione peroxidase activities in erythrocytes were studied. Administration of GLEt to diabetic animals at doses of 50, 100, and 200 mg/kg body weight lowered elevated blood glucose levels by 24, 35, and 66%, respectively, relative to untreated diabetic rats. In comparison, treatment with the known antidiabetic drug, glibenclamide (600 μg/kg body weight) decreased blood glucose concentrations by 51%. Plasma insulin concentrations were increased in the diabetic rat by 73% with GLEt (200 mg/kg body weight) and 45% with glibenclamide (600 μg/kg body weight). Although a significant decrease in the lipid peroxidation markers was observed in plasma on treatment with GLEt and glibenclamide, the RBC antioxidant levels were increased significantly in diabetic rats. Furthermore, erythrocytes from the GLEt-treated animals were found to be more resistant to H₂O₂-induced peroxidation than that of untreated diabetic animals. The chemical characterization of the polyphenolics of the extract showed the presence of gallic acid (5.29% w/w), resveratrol (2.2% w/w), and quercetin (16.6% w/w). The results of this study suggest that G. montanum may be useful for the control, management, and prevention of oxidative stress associated with diabetes.     

Hemagglutinins (lectins) in fruit bodies of Japanese higher fungi

Extracts from fruit bodies of 110 species of Japanese fungi were examined with trypsinized human and rabbit erythrocytes. More than 80% of the extracts showed the hemagglutination activities, a higher proportion than reported previously. Over half of species that had been reported to be inactive exhibited hemagglutination. Among them, some extracts showed human blood group specific-hemagglutination; A-specific,Panellus serotinus, Psathyrella piluliformis, Cantharellus cibarius andStropharia rugosoannulata; B-, O-specific,Gyroporus castaneus andPanellus stypticus; and Ospecific,Linderia bicolumnata andPhallus impudicus. Twenty-one species were reactive toward only rabbit erythrocytes. Several species exhibited very high hemagglutination activity. The results suggested that some of these Japanese fungi would be promising sources of lectins.     

Hemagglutination and Erythrophagocytosis Associated with the Anemia of Plasmodium berghei Infections of Rats*

SYNOPSIS. The presence of anemia that often seems excessive for the amount of parasitemia in Plasmodium berghei infections had led to the suggestion that autoimmunity might be in part responsible for the anemia. In another erythrocytic infection, Anaplasma marginale of cattle, the association of erythrophagocytosis, autohemagglutination and anemia with infection has led to the suggestion that autoimmunization may occur in anaplasmosis. The possibility that similar findings might be present in P. berghei infections of rats has been investigated. Groups of rats infected with P. berghei were examined at 2–3 day intervals during the course of infection. Red blood cell counts, hematocrit values and percentages of parasitized erythrocytes were determined. The rats were bled at intervals and the sera tested for agglutinins for trypsinized rat erythrocytes. Other infected rats were killed, and their spleens and bone marrow were examined for evidence of erythrophagocytosis. Parasitemia reached a peak on the 9th day of infection and became subpatent by the 14th. The greatest depression in erythrocyte numbers occurred on the 11th day, and the counts remained below normal until the 23rd day. Phagocytized erythrocytes, predominantly uninfected, were found in the phagocytes of the spleen and bone marrow from the 5th through the 21st day. Agglutinins for trypsinized normal rat erythrocytes were present in the sera of the rats in titers as high as 1:64 from the 5th through 14th day of infection. Lower agglutinin titers (1:8) were found from time to time in sera of rats made anemic by repeated bleedings. It is not clear whether these agglutinins are responsible for erythrophagocytosis; however, the fact that predominantly uninfected erythrocytes were phagocytized suggests that the erythrocytes might have been opsonized by an autoantibody associated with the P. berghei infections.     

Pantothenic Acid Metabolism During Avian Malaria Infection: Pantothenate Kinase Activity in Duck Erythrocytes and in Plasmodium lophurae*

SYNOPSIS. The initial enzymatic reaction in the pathway of coenzyme A genesis from pantothenic acid, i.e., the conversion of pantothenic acid to phosphopantothenic acid catalyzed by pantothenic acid kinase, was found in extracts of avian erythrocytes but not in extracts of Plasmodium lophurae or intact parasites. The activity of the kinase was significantly higher in extracts prepared from normal duck erythrocytes than those infected with P. lophurae. The apparent absence of pantothenic acid kinase in P. lophurae is corroborative support for nutritional studies(7,9) which suggest that the avian malarial parasite is dependent for its supply of coenzyme A on the host cell.     

Fragility of Erythrocytes from Chicks and Rats Fed Dilauryl Succinate and Related Compounds

Susceptibility to hydrogen peroxide of the erythrocytes from chicks and rats fed dilauryl succinate and related compounds with and without supplementation of dl-α-tocopheryl acetate was determined.

Dilauryl succinate, lauryl alcohol, n-decyI alcohol, myristyl alcohol, and lauraldehyde were confirmed to make the erythrocytes from the chicks fragile. Supplemented dl-α-tocopheryl acetate of 200 mg per kg of diet completely prevented the hemolysis induced by these compounds. Dilauryl succinate also makes the rat’s erythrocytes fragile and supplemented dl-α-tocopheryl acetate prevented the hemolysis of the rats, but ethoxyquin was not. The symptoms of encephalomalacia in the chick is preceded by increased hemolysis value of the erythrocytes, and this hemolysis value dropped after the appearance of encephalomalacia.     

In vitro and in vivo activity of extracts from the soft coral <Emphasis Type="Italic">Lobophytum pauciflorum</Emphasis> on trophozoites of <Emphasis Type="Italic">Entamoeba histolytica</Emphasis>

Methanolic extracts from the marine coral Lobophytum pauciflorum exhibited an in vitro amebicidal activity against trophozoites of Entamoeba histolytica and were found to cure caecal amoebiasis in 85% of infected rats. This activity was restricted to the n-butanol fraction, from which three pure compounds were isolated.Original Russian Text Copyright © 2004 by Biologiya Morya, Ghoshal, Lakshmi.     

(+)‐Z‐Bisdehydrodoisynolic Acid Ameliorates Obesity and the Metabolic Syndrome in Female ZDF Rats

Objective: The putative selective estrogen receptor modulator (+)‐Z‐bisdehydrodoisynolic acid (Z‐BDDA) has been found to improve cardiovascular risk in rodents. The objective of this study was to investigate the effectiveness of (+)‐Z‐BDDA compared with the antidiabetic drug, rosiglitazone, in treating obesity and risk factors associated with the metabolic syndrome. Research Methods and Procedures: Female Zucker Diabetic Fatty rats were randomly assigned to three treatment groups for 29 weeks: control (C), 1.8 mg (+)‐Z‐BDDA/kg diet [control diet + (+)‐Z‐BDDA (CB)], or 100 mg rosiglitazone/kg diet [control diet + rosiglitazone (CR)]. At sacrifice, physiological, biochemical, and molecular parameters were examined. Results: CB animals gained less weight and exhibited a decrease in total body lipids (p < 0.05) as compared with C or CR rats. Body weight and total body lipids were the highest in CR rats (p < 0.05). Liver weights in CB and CR rats were lower (p < 0.05) than in C rats, whereas kidney weights were lower in CB (p < 0.05) than in C and CR animals. Fasting plasma glucose was lower (p < 0.05) in the CB and CR animals when compared with C animals. C rats exhibited the highest concentration of total plasma cholesterol, and CR‐treated rats exhibited the lowest concentration. Plasma triglycerides followed the same pattern as plasma cholesterol. Histomorphometry of heart vasculature revealed that CB and CR treatments produced a significant shift from small to large venules and arterioles compared with C (p < 0.05). Liver expression profiles of peroxisome proliferator‐activated receptor (PPAR) α, PPARγ, and PPAR‐regulated genes revealed encouraging CB‐induced effects. Discussion: These results suggest that (+)‐Z‐BDDA may have applications in treating obesity and complications associated with the metabolic syndrome.     

LIBS-Based Detection of Antioxidant Elements in Seeds of <Emphasis Type="Italic">Emblica officinalis</Emphasis>

The aim of the study was to determine the effect of the elements of the extract of seed from Emblica officinalis on antioxidant enzymes and osmotic fragility of erythrocytes membrane in normal as well as streptozotocin-induced severely diabetic albino Wister rats. The results revealed that the untreated diabetic rats exhibited increase in oxidative stress as indicated by significantly diminished activities of free radical scavenging enzymes such as catalase (CAT) and superoxide dismutase (SOD) by 37.5% (p < 0.001) and 18.6% (p < 0.01), respectively. However, the E. officinalis seed extract treatment showed marked improvements in CAT and SOD activities by 47.09% (p < 0.001) and 21.61% (p < 0.001), respectively. The enhanced lipid peroxidation by 30.87% (p < 0.001) in erythrocytes of untreated diabetic rats was significantly accentuated in the extract treated animals by 23.72% (p < 0.001). The erythrocytes showed increased osmotic fragility due to diabetes in terms of hemolysis. It attained the normal level in diabetic treated group. The findings thus suggest that E. officinalis seed extract has the potential to be exploited as an agent to boost the antioxidant system in the diabetic animal model. Laser-induced breakdown spectroscopy has been used as an analytical tool to detect major and minor elements like Mg, Fe, Na, K, Zn, Ca, H, O, C, and N present in the extract. The higher concentration of Ca (II), Mg (II) and Fe (II) as reflected by their intensities are responsible for the antioxidant potential of E. officinalis.     

<Emphasis Type="Italic">In vivo</Emphasis> and <Emphasis Type="Italic">in vitro</Emphasis> antioxidant effects of three Veronica species

The aim of the present study was to examine the antioxidant activity of three Veronica species (Plantaginaceae). The antioxidant potential of various extracts obtained from aerial flowering parts was evaluated by DPPH-free (1,1-diphenyl-2-picrylhydrazyl-free) radical scavenging activity and ferric-reducing antioxidant power assays. Considerable antioxidant activity was observed in the plant samples (FRAP values ranged from 0.97 to 4.85 mmol Fe²⁺/g, and DPPH IC₅₀ values from 12.58 to 66.34 μg/ml); however, these levels were lower than the activity of the control compound butylated hydroxytoluene (BHT) (FRAP: 10.58 mmol Fe²⁺/g; DPPH IC₅₀: 9.57 μg/ml). Also, the in vivo antioxidant effects were evaluated in several hepatic antioxidant systems in rats (activities of glutathione peroxidase, glutathione reductase, peroxidase, catalase, xanthine oxidase, glutathione content and level of thiobarbituric acid reactive substances) after treatment with different Veronica extracts, or in combination with carbon tetrachloride (CCl₄). Pretreatment with 100 mg/kg b.w. of Veronica extracts inhibited CCl₄-induced liver injury by decreasing TBA-RS level, increasing GSH content, and bringing the activities of CAT and Px to control levels. The present study suggests that the extracts analyzed could protect the liver cells from CCl₄-induced liver damage by their antioxidative effect on hepatocytes.     

Effect of Vaccinium ashei reade Leaf Extracts on Lipid Metabolism in Obese OLETF Rats

The effects of a hot water extract and fractional extracts from rabbiteye blueberry (Vaccinium ashei reade) leaves (BBL) on lipid metabolism were studied in obese Otsuka Long-Evans Tokushima Fatty (OLETF) rats. Feeding the hot water extract and fractional extracts from BBL alleviated hepatic triglyceride accumulation in the rats. Additionally, feeding with the flavonol glycoside (FG) and proanthocyanidin (PA) fractions lowered serum cholesterol levels in the obese rats. The results from measurements of the hepatic enzyme activity indicate that the hypolipidemic effects of the hot water extract and the PA fraction might be attributable to enhanced lipolysis in the liver. The reduced serum levels of C-reactive protein, an inflammatory cytokine, by the chlorogenic acid + rutin fraction and FG fraction might be associated with alleviating the metabolic abnormalities in obese rats. These results indicate that the BBL extracts, and especially FG and PA, exerted hypolipidemic effects on obese OLETF rats and suggest that an infusion of BBL can be useful as a dietary hypolipidemic component.     

Hemolytic activity of Rhizopus nigricans and Rhizopus arrhizus

Rhizopus nigricans andR. arrhizus were cultured in modified Sabouraud broth and synthetic dextrose-glutamate medium for periods of two to eight weeks. The culture filtrates and extracts of fungous mats were examined for the presence ofin vitro hemolytic activity. The culture filtrates exhibited little hemolytic activity but all fungous mat extracts produced hemolysis of sheep, rabbit, guinea pig and human erythrocytes. Guinea pig erythrocytes were in general most readily hemolyzed and sheep erythrocytes least readily hemolyzed.

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Zusammenfassung Rhizopus nigricans undR. arrhizus sind in modifizierter Sabouraud's Brühe und in synthetischem Dextroseglucamate-Nährboden für zwei bis acht Wochen gezüchtet worden. Das Kulturfiltrat und der Extrakt des Pilzmyzeliums sind für das Vorhandensein der in-vitro hämolytischen Aktivität untersucht. Das Kulturfiltrat zeigte eine geringe hämolytische Aktivität, jedoch verursachten alle Pilzmyzeliumextrakte eine Hämolyse in Erythrozyten von Schaf, Kaninchen, Meerschweinchen und Menschen. Meerschweinchenerythrozyten sind im allgemeinen am leichtesten, Schaferythrozyten am wenigsten hämolysiert.