scholarly journals New advances in thein-vitroculture ofDientamoeba fragilis

Parasitology ◽  
2012 ◽  
Vol 139 (7) ◽  
pp. 864-869 ◽  
Author(s):  
VARUNI S. MUNASINGHE ◽  
D. STARK ◽  
J. T. ELLIS

SUMMARYDientamoeba fragilisis an intestinal protozoan in humans that is commonly associated with diarrhoea and other gastrointestinal complaints. Studies conducted to investigate the biology of this parasite are limited by methods forin vitrocultivation. The objective of this study was to improve a biphasic culture medium, based on the Loeffler's slope, by further supplementation in order to increase the yield of trophozoites in culture. The currentin vitroculture ofD. fragilisis a xenic culture with a mix of bacteria. Three different liquid overlays were evaluated including Earle's balanced salt solution (EBSS), PBS and Dulbecco's modified PBS (DPBS), for their ability to support thein vitrogrowth ofD. fragilistrophozoites. Out of these 3 overlays EBSS gave the highest increase in the trophozoite numbers. The effect of supplementation was analysed by supplementing EBSS with ascorbic acid, ferric ammonium citrate, L-cysteine, cholesterol and alpha-lipoic acid and quantification ofin vitrogrowth by cell counts. A new liquid overlay is here described based upon EBSS supplemented with cholesterol and ferric ammonium citrate that, in conjunction with the Loeffler's slope, supports the growth ofD. fragilistrophozoitesin vitro. This modified overlay supported a 2-fold increase in the numbers of trophozoite in culture from all 4D. fragilisisolates tested, when compared to a PBS overlay. These advances enable the harvest of a larger number of trophozoites needed for further studies on this parasite.

1985 ◽  
Vol 5 (4) ◽  
pp. 595-600
Author(s):  
K K Rao ◽  
D Shapiro ◽  
E Mattia ◽  
K Bridges ◽  
R Klausner

Treatment of K562 cells, a human erythroleukemia cell line, with desferrioxamine raised the levels of the receptor for transferrin (Tf) two- to threefold over that of the control cells. The levels of receptor were reduced by at least 50 and 35% of that of the control in cells treated with diferric Tf and ferric ammonium citrate, respectively. These changes were of total cellular receptors with no alteration in the proportion of receptors found on the cell surface. The half-lives of the receptor were identical in cells treated with desferrioxamine, diferric Tf, or ferric ammonium citrate. Cells metabolically labeled with [35S]methionine showed a 2.5-fold increase in the rate of receptor synthesis when treated with desferrioxamine and a 35 and 65% decrease when treated with ferric ammonium citrate and diferric Tf, respectively. In vitro translations of polyadenylated mRNA isolated from cells incubated with desferrioxamine showed a 2.5-fold increase in translatable mRNA for the receptor, whereas treatment of cells with ferric ammonium citrate and diferric Tf resulted in a 25 and 50% reduction, respectively, in translatable mRNA for this receptor.


1985 ◽  
Vol 5 (4) ◽  
pp. 595-600 ◽  
Author(s):  
K K Rao ◽  
D Shapiro ◽  
E Mattia ◽  
K Bridges ◽  
R Klausner

Treatment of K562 cells, a human erythroleukemia cell line, with desferrioxamine raised the levels of the receptor for transferrin (Tf) two- to threefold over that of the control cells. The levels of receptor were reduced by at least 50 and 35% of that of the control in cells treated with diferric Tf and ferric ammonium citrate, respectively. These changes were of total cellular receptors with no alteration in the proportion of receptors found on the cell surface. The half-lives of the receptor were identical in cells treated with desferrioxamine, diferric Tf, or ferric ammonium citrate. Cells metabolically labeled with [35S]methionine showed a 2.5-fold increase in the rate of receptor synthesis when treated with desferrioxamine and a 35 and 65% decrease when treated with ferric ammonium citrate and diferric Tf, respectively. In vitro translations of polyadenylated mRNA isolated from cells incubated with desferrioxamine showed a 2.5-fold increase in translatable mRNA for the receptor, whereas treatment of cells with ferric ammonium citrate and diferric Tf resulted in a 25 and 50% reduction, respectively, in translatable mRNA for this receptor.


2019 ◽  
Vol 3 (Supplement_1) ◽  
Author(s):  
Cole Guggisberg ◽  
Moon-Suhn Ryu

Abstract Objectives Iron recycled from erythrophagocytosis by macrophages serves as a primary source of systemic iron. NCOA4 mediates ferritin turnover via ferritinophagy. Yet, whether NCOA4 is important in macrophages or erythrophagocytosis-mediated iron recycling remains unclear, and thus was assessed in vitro. Methods J774 cells were employed as an in vitro model of macrophages. Iron studies involved treatments of ferric ammonium citrate (FAC) or an iron chelator, deferoxamine (Dfo). To recapitulate systemic iron recycling and overload, cells were treated with opsonized erythrocytes and minihepcidin, respectively. NCOA4 knock-down was achieved by siRNA transfection. Iron gene responses were measured by qPCR and western analyses, and viable cell counts were colorimetrically determined by CCK8 assays as functional outcomes. Results NCOA4 protein abundance was inversely related to iron availability and ferritin in macrophages. Loss of NCOA4 resulted in impaired ferritin turnover, and led to a reduction in viable cells when combined with iron deficiency. By erythrophagocytosis, a peak in ferritin abundance was observed at 12 h with a subsequent decrease at 24 h. This loss in ferritin was NCOA4-dependent. Minihepcidin caused accumulation of ferritin, along with a repression of NCOA4 in both control and erythrocyte-laden macrophages. Hepcidin activity had no effect on ferritin when NCOA4 was depleted. Conclusions NCOA4 mediates the release of ferritin iron during cellular iron restriction and iron recycling by macrophages. Moreover, our studies suggest that macrophage NCOA4 is integral to systemic iron homeostasis by responding to the iron regulatory hormone, hepcidin. Thus, NCOA4 and ferritinophagy may potentially serve as therapeutic targets for treatments of iron disorders and anemia of chronic disease. Funding Sources Supported by the NIFA, USDA, Hatch project under MIN-18–118 and intramural support to M-S.R.


1992 ◽  
Vol 108 (3) ◽  
pp. 389-396 ◽  
Author(s):  
J. L. Lock ◽  
R. G. Board

SUMMARYA study was made of the persistence of different Salmonella serotypes in hens' egg albumen in vitro at 4, 20 and 30 °C. The majority of serotypes remained viable but did not increase in numbers at 20 and 30 °C for 42 days. At 4 °C many of the serotypes died out.The addition of ferric ammonium citrate on the 42nd day of incubation induced multiplication of organisms incubated at 20 and 30 °C, but not at 4 °C. The pH and glucose concentration of the albumen diminished only when heavy growth occurred.Salmonella enteritidis remained viable on the air cell membrane in vitro for 17 days at 4, 20 and 30 °C. Thirty percent of the organisms also remained motile in albumen for 42 days at 25 °C and up to 5% of the cells remained motile for up to 20 days at 4 °C.


2013 ◽  
Vol 111 (2) ◽  
pp. 261-269 ◽  
Author(s):  
Mary E. Drewnoski ◽  
Perry Doane ◽  
Stephanie L. Hansen

Dissimilatory reduction of sulphate by sulphate-reducing bacteria in the rumen produces sulphide, which can lead to a build-up of the toxic gas hydrogen sulphide (H2S) in the rumen when increased concentrations of sulphate are consumed by ruminants. We hypothesised that adding ferric Fe would competitively inhibit ruminal sulphate reduction. The effects of five concentrations and two sources (ferric citrate or ferric ammonium citrate) of ferric Fe were examinedin vitro(n6 per treatment). Rumen fluid was collected from a steer that was adapted to a high-concentrate, high-sulphate diet (0·51 % S). The addition of either source of ferric Fe decreased (P< 0·01) H2S concentrations without affecting gas production (P= 0·38), fluid pH (P= 0·80) orin vitroDM digestibility (P= 0·38) after a 24 h incubation. Anin vivoexperiment was conducted using eight ruminally fistulated steers (543 (sem12) kg) in a replicated Latin square with four periods and four treatments. The treatments included a high-concentrate, high-sulphate control diet (0·46 % S) or the control diet plus ferric ammonium citrate at concentrations of 200, 300 or 400 mg Fe/kg diet DM. The inclusion of ferric Fe did not affect DM intake (P= 0·21). There was a linear (P< 0·01) decrease in the concentration of ruminal H2S as the addition of ferric Fe concentrations increased. Ferric citrate appears to be an effective way to decrease ruminal H2S concentrations, which could allow producers to safely increase the inclusion of ethanol co-products.


Author(s):  
Moumita Hazra

Background: Anaemia is a global health concern, associated with increased maternal and perinatal mortality, preterm delivery, low birth weight, extreme fatigue and impaired immune system; and controlled by oral haematinics; with a rise in haemoglobin concentration. The objective was to examine the various aspects of pharmacoepidemiology and pharmacohaemovigilance of oral haematinics, among the anaemic women population, in rural India.Methods: This was a multi-centre, retrospective, observational and analytical study of the hospital medical records of 250 anaemic patients, who were allocated into group A of 125 patients within 15-21 years and group B of 125 patients within 22-35 years. The patients were prescribed oral haematinics, containing 60 mg of elemental iron, thrice daily, with meals. The various aspects of pharmacoepidemiology and pharmacohaemovigilance of ferrous ascorbate, ferrous sulphate, ferrous fumarate and ferric ammonium citrate, including patients’ demographic characteristics, anaemic symptoms assessment, prescription patterns, and safety assessment, on 1st, 2nd, 3rd months and follow-up visits, were recorded and thoroughly analysed..Results: In groups A and B, the demographic characteristics of the patients were comparable; ferrous ascorbate was the most commonly prescribed oral haematinic, followed by ferrous sulphate, ferrous fumarate and ferric ammonium citrate, which controlled mild to moderate iron deficiency anaemia, with a gradual significant rise in haemoglobin concentration, in the successive 3 months; and adverse effects were observed to be statistically non-significant in either group.Conclusions: The different aspects of pharmacoepidemiology and pharmacohaemovigilance in the study established that the oral haematinics were reasonably beneficial and safe among the anaemic women population, in rural India.


1959 ◽  
Vol 39 (2) ◽  
pp. 193-201 ◽  
Author(s):  
H. Doornenbal

Haemoglobin levels, haematocrit values and erythrocyte counts were determined at weekly intervals from 3 to 45 days of age for 60 pigs which received iron in the form of: injectable iron-dextran (A); injectable iron-dextran (B); injectable ferric ammonium citrate; oral iron in the form of paste, or sods sprinkled with iron sulphate. The iron-dextran and ferric ammonium citrate compounds were administered at 3 days of age as single injections supplying 100 mgm. of iron and 30 mgm. of ferric ammonium citrate respectively. The paste was administered at 3, 10, 17 and 24 days of age. Sods were fed twice a week during the period of 3 days to 28 days of age.The sod treatment maintained normal blood values while the iron-dextran compounds and the paste resulted in values somewhat below normal, although visible evidence of anaemia was not apparent. Blood values for the group receiving ferric ammonium citrate were extremely low and two pigs on this treatment died at 42 and 60 days of age. Both exhibited severe anaemia.Significant differences were obtained in weaning weights. The heaviest pigs were those receiving sods; the lightest pigs those receiving injectable ferric ammonium citrate. The effect of the different treatments on growth was not apparent until after 21 days of age.


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