scholarly journals THE DEVELOPMENT OF LEISHMANIA DONOVANI IN VITRO AT 37°C

1953 ◽  
Vol 97 (2) ◽  
pp. 177-188 ◽  
Author(s):  
William Trager
Keyword(s):  
Human Serum ◽  
Human Erythrocyte ◽  
Small Proportion ◽  
Culture Medium ◽  
Leishmania Donovani ◽  
Cell Extract ◽  
Rabbit Serum ◽  
Red Cell

Suspensions of leishmanias from the spleen of hamsters infected with Leishmania donovani were placed in culture flasks and incubated at 37°C. In a medium of human erythrocyte extract and human serum there appeared within a day or two aflagellate forms resembling leishmanias but larger, as well as other aflagellate forms more nearly resembling rounded leptomonads. These intermediate forms multiplied during the first 4 days of culture. They then slowly died off, despite frequent renewal of the culture medium. Sometimes a small proportion of motile, typical leptomonads also appeared in such cultures. Leptomonads from cultures maintained at 28°C., when placed in the human red cell extract-human serum medium and incubated at 37°C., survived at least 4 days. For both types of effect, human serum could be replaced by normal hamster serum but not by rabbit serum. Nicotinamide, added to the human red cell extract-human serum medium at a concentration of 400 mg. per 100 ml., completely prevented the development of intermediate forms from leishmanias and brought about the rapid death of leptomonads at 37°C.

scholarly journals Esterases in Serum-Containing Growth Media Counteract Chloramphenicol Acetyltransferase Activity In Vitro

1999 ◽  
Vol 43 (3) ◽  
pp. 655-660 ◽  
Author(s):  
Charles D. Sohaskey ◽  
Alan G. Barbour
Keyword(s):  
Human Serum ◽  
Horse Serum ◽  
Chloramphenicol Acetyltransferase ◽  
Rabbit Serum ◽  
Growth Media ◽  
Chloramphenicol Resistance ◽  
E Coli ◽  
Cell Extracts

ABSTRACT The spirochete Borrelia burgdorferi was unexpectedly found to be as susceptible to diacetyl chloramphenicol, the product of the enzyme chloramphenicol acetyltransferase, as it was to chloramphenicol itself. The susceptibilities of Escherichia coli and Bacillus subtilis, as well as that ofB. burgdorferi, to diacetyl chloramphenicol were then assayed in different media. All three species were susceptible to diacetyl chloramphenicol when growth media were supplemented with rabbit serum or, to a lesser extent, human serum. Susceptibility ofE. coli and B. subtilis to diacetyl chloramphenicol was not observed in the absence of serum, when horse serum was used, or when the rabbit or human serum was heated first. In the presence of 10% rabbit serum, a strain of E. colibearing the chloramphenicol acetyltransferase (cat) gene had a fourfold-lower resistance to chloramphenicol than in the absence of serum. A plate bioassay for chloramphenicol activity showed the conversion by rabbit, mouse, and human sera but not bacterial cell extracts or heated serum of diacetyl chloramphenicol to an inhibitory compound. Deacetylation of acetyl chloramphenicol by serum components was demonstrated by using fluorescent substrates and thin-layer chromatography. These studies indicate that esterases of serum can convert diacetyl chloramphenicol back to an active antibiotic, and thus, in vitro findings may not accurately reflect the level of chloramphenicol resistance by cat-bearing bacteria in vivo.

Comparison of results obtained with human serum and a protein solution as a supplement for in vitro fertilization culture medium

1992 ◽  
Vol 58 (3) ◽  
pp. 637-639 ◽  
Author(s):  
Gerritdina J. Huisman ◽  
Nadia M. Lo-A-Njoe ◽  
Albert Th. Alberda ◽  
Robert A. Leerentveld ◽  
Arie Verhoeff ◽  
...  
Keyword(s):  
In Vitro Fertilization ◽  
Human Serum ◽  
Culture Medium ◽  
Protein Solution ◽  
Vitro Fertilization ◽  
Comparison Of Results

scholarly journals Soluble Asialoglycoprotein Receptors Reflect the Apoptosis of Hepatocytes

2002 ◽  
Vol 11 (5) ◽  
pp. 407-415 ◽  
Author(s):  
Tetsuji Kakegawa ◽  
Hirohiko Ise ◽  
Nobuhiro Sugihara ◽  
Toshio Nikaido ◽  
Naoki Negishi ◽  
...  
Keyword(s):  
Cell Death ◽  
Liver Tissue ◽  
Liver Diseases ◽  
Culture Medium ◽  
Apoptotic Cell ◽  
Rabbit Serum ◽  
Mouse Serum ◽  
Apoptosis And Necrosis

Cell death is thought to take place through at least two distinct processes: apoptosis and necrosis. There is increasing evidence that dysregulation of the apoptotic program is involved in liver diseases. However, there is no method to simply evaluate apoptosis in the liver tissue at present. It has been reported that the expression of asialoglycoprotein receptors (AGPRs) increases with apoptosis, but there is no report until now that investigates the influence of soluble AGPRs on apoptosis of hepatocytes. Soluble AGPRs have been reported to be present in human serum under physiological conditions. In the present study, in order to investigate the correlation between apoptosis of hepatocytes and soluble AGPR, mouse soluble AGPRs were detected using SDS-PAGE and Western blot analysis was conducted using anti-extracellular mouse hepatic lectin-1 (Ex-MHL-1) antiserum (polyclonal rabbit serum). The mouse soluble AGPRs were present in culture medium and mouse serum when hepatocytes were damaged. The soluble AGPRs increased proportionately, as the number of dead hepatocytes increased. In addition, soluble AGPRs existed more when apoptotic cell death was observed in in vitro and in vivo than when necrotic cell death was observed. The extracellular moiety of MHL-1 exists in the culture medium and mouse serum as a soluble AGPR, but the detailed mechanism of releasing soluble AGPR from hepatocytes has not been revealed yet. We described the first evidence for the relation between quantity of soluble AGPRs with two kinds of cell death: necrosis and apoptosis. Based on the results of our study, soluble AGPRs might become a new marker of apoptosis in the liver tissue and be useful for clinical diagnosis and treatment for liver diseases.

Partial purification of antigens collected during in vitro cultivation of the larval stages of Taenia pisiformis

Parasitology ◽  
1976 ◽  
Vol 72 (3) ◽  
pp. 269-279 ◽  
Author(s):  
M. D. Rickard ◽  
J. C. Katiyar
Keyword(s):  
Protective Immunity ◽  
Culture Medium ◽  
Serum Proteins ◽  
Skin Tests ◽  
Rabbit Serum ◽  
Partial Purification ◽  
In Vitro Cultivation ◽  
Normal Rabbit Serum ◽  
Skin Reactions

SummaryLarvae of Taenia pisiformis were cultured in vitro in medium containing 2·5, 5 or 20% (v/v) of normal rabbit serum (NRS). Greatest development occurred in 20% NRS, and the potency of antigens collected in medium from each culture tested by intradermal (i/d) skin tests in infected rabbits paralleled the in vitro growth rate of larvae. ‘Culture’ antigens from 5% NRS stimulated good immunity in rabbits to a challenge infection with T. pisiformis eggs, although they were poorly reactive in skin tests.T. pisiformis larvae were also cultured in 10% (v/v) of nitrates of serum reduced to one-half of its volume by passage through 300 000 MW cut oif (XM300F) or 100000 MW cut off (XM100F) ultrafiltration membranes. Larvae cultured using XM300F had growth rates comparable with those cultured in 20% NRS, and the antigens released into the culture medium had equal potency in i/d tests and in stimulating protective immunity in rabbits. Larvae did not develop in XM100F orproduce skin-reactive or protective antigens.Crude ‘culture’ antigen from cultures in 20% NRS was separated into 4 fractions by nitration on Sephadex G200. All of these fractions gave i/d skin reactions in infected rabbits. Fraction 3 (F3) was the most active, but was shown by acrylamide gel electrophoresis and immunoelectrophoresis to be highly contaminated with rabbit serum proteins. F3 was separated into fractions on DEAE-Sephadex A50, and the third fraction from this was as active as the original culture medium in i/d skin tests, but had only 5% of the original protein concentration. Electrophoresis demonstrated few serum contaminants, and 2 indistinct protein bands that were not present in a similar fraction of NRS.Neither Sephadex G200 F3 nor DEAE-Sephadex F3 stimulated protective immunity in rabbits, suggesting that antigens stimulating immunity against the establishment of T. pisiformis in rabbits and those provoking cell-mediated immune reactions may be different.

scholarly journals TECHNIQUE OF CULTIVATING HUMAN TISSUES IN VITRO

1916 ◽  
Vol 24 (4) ◽  
pp. 367-372 ◽  
Author(s):  
Robert A. Lambert
Keyword(s):  
Human Serum ◽  
Human Plasma ◽  
Culture Medium ◽  
Mechanical Injury ◽  
Human Tissues ◽  
Tissue Cultures ◽  
Active Growth ◽  
Homologous Serum

1. Unmodified human plasma is not a satisfactory culture medium for human tissues owing to the susceptibility of human fibrin to digestion by tissue ferments. The necessary framework is thus destroyed before the cells begin to migrate. The difficulty can be overcome by adding to human plasma or serum a small quantity of fowl or pigeon plasma, the fibrin of which is highly resistant to digestion. Human tissues have been propagated in this medium for several months through subcultures, and growth in vitro can probably be maintained indefinitely. 2. Human tissues show no greater sensitiveness to changes in temperature and mechanical injury associated with preparation of cultures than those of lower animals. They may be preserved in an ordinary ice box at 10–15°C. as long as 6 or 8 days. Tissues obtained at operation give best results, but pieces of organs removed at autopsy 1 to 4 hours after death sometimes show active growth. 3. The presence of normally existing iso-antibodies (agglutinins and hemolysins) in human serum is without influence on the growth of human tissues in vitro. In other words, autogenous serum has no advantage in tissue cultures over homologous serum.

scholarly journals STUDIES ON CONDITIONS AFFECTING THE SURVIVAL IN VITRO OF A MALARIAL PARASITE (PLASMODIUM LOPHURAE)

1941 ◽  
Vol 74 (5) ◽  
pp. 441-462 ◽  
Author(s):  
William Trager
Keyword(s):  
Liver Extract ◽  
Cell Extract ◽  
Malarial Parasite ◽  
Red Cell ◽  
High Potassium ◽  
Plasmodium Lophurae ◽  
Parasite Plasmodium ◽  
Chick Embryo Extract ◽  
Very High

The survival of Plasmodium lophurae in vitro, at temperatures of 39.5–42°C., is favored by a balanced salt solution having a high potassium content, by aeration but not by a very high oxygen tension, by an optimal density of parasites per cubic millimeter, by frequent renewal of the suspending medium, by concentrated red cell extract, by optimal concentrations of plasma or serum, of chick embryo extract, of glucose or glycogen, and of glutathione, and probably by yeast extract and a very low concentration of liver extract. In the best preparations, as judged by infectivity, more than 40 per cent of the parasites were alive on the 3rd day, more than 20 per cent on the 4th day, perhaps 1 per cent on the 5th day, and about 0.05 per cent on the 6th day. Evidence was obtained that the parasites had multiplied during the 1st day of incubation.

scholarly journals Prospective isolation of human erythroid lineage-committed progenitors

2015 ◽  
Vol 112 (31) ◽  
pp. 9638-9643 ◽  
Author(s):  
Yasuo Mori ◽  
James Y. Chen ◽  
John V. Pluvinage ◽  
Jun Seita ◽  
Irving L. Weissman
Keyword(s):  
Bone Marrow ◽  
Myelodysplastic Syndrome ◽  
Human Erythrocyte ◽  
Molecular Mechanisms ◽  
Expression Patterns ◽  
Red Cell ◽  
Developmental Pathway ◽  
Erythroid Development ◽  
Adult Bone

Determining the developmental pathway leading to erythrocytes and being able to isolate their progenitors are crucial to understanding and treating disorders of red cell imbalance such as anemia, myelodysplastic syndrome, and polycythemia vera. Here we show that the human erythrocyte progenitor (hEP) can be prospectively isolated from adult bone marrow. We found three subfractions that possessed different expression patterns of CD105 and CD71 within the previously defined human megakaryocyte/erythrocyte progenitor (hMEP; Lineage− CD34+ CD38+ IL-3Rα− CD45RA−) population. Both CD71− CD105− and CD71+ CD105− MEPs, at least in vitro, still retained bipotency for the megakaryocyte (MegK) and erythrocyte (E) lineages, although the latter subpopulation is skewed in differentiation toward the erythroid lineage. Notably, the proliferative and differentiation output of the CD71intermediate(int)/+ CD105+ subset of cells within the MEP population was completely restricted to the erythroid lineage with the loss of MegK potential. CD71+ CD105− MEPs are erythrocyte-biased MEPs (E-MEPs) and CD71int/+ CD105+ cells are EPs. These previously unclassified populations may facilitate further understanding of the molecular mechanisms governing human erythroid development and serve as potential therapeutic targets in disorders of the erythroid lineage.

scholarly journals THE ESSENTIAL PARTICIPATION OF AN ENZYME IN THE INHIBITION OF GROWTH OF TUBERCLE BACILLI BY SPERMINE

1953 ◽  
Vol 97 (3) ◽  
pp. 327-343 ◽  
Author(s):  
James G. Hirsch
Keyword(s):  
Guinea Pig ◽  
Culture Medium ◽  
Culture Media ◽  
Enzymatic Reaction ◽  
Rabbit Serum ◽  
Inhibition Of Growth ◽  
Plasma Fraction ◽  
Bovine Plasma ◽  
Chemical Contaminant

Spermine inhibits the multiplication of tubercle bacilli in vitro only if another tissue substance is present in the culture medium. This activating substance occurs as a chemical contaminant of the albumin in commercial bovine plasma fraction V ordinarily added to culture media; it is also found in whole bovine and sheep serum and in aqueous extracts of the guinea pig kidney. No detectable spermine activator is contained in whole human, guinea pig, or rabbit serum. The serum constituent which renders spermine inhibitory for the growth of acid-fast bacteria has the properties of a protein of the alpha globulin classification. This protein is an enzyme which acts on spermine. Presumably, some product of the enzymatic reaction exerts a toxic effect on mammalian tubercle bacilli.

scholarly journals Lymphocyte-mediated lysis of antibody coated human red cells in the presence of human serum

Blood ◽  
1979 ◽  
Vol 53 (6) ◽  
pp. 1197-1202
Author(s):  
RJ Kurlander ◽  
WF Rosse
Keyword(s):  
Human Serum ◽  
Culture Medium ◽  
Human Albumin ◽  
Peripheral Blood Lymphocytes ◽  
Red Cells ◽  
The Other ◽  
High Concentration ◽  
The Third ◽  
Human Red Cells

When peripheral blood lymphocytes and human red cells coated with IgG were incubated in vitro in culture medium, antibody-dependent lymphocyte-mediated lysis was observed. This lysis was markedly inhibited by the addition of purified monoclonal IgG1 (1000 microgram/ml) to the culture medium. In contrast, lysis by lymphocytes of sensitized red cells in the presence of undiluted human serum was equal to or greater than lysis in medium alone, even in the presence of IgG1 at 1000 microgram/ml, despite the high concentration of IgG in human serum (6000--19,000 microgram/ml). Serum heated to 56 degrees C for 30 min also restored lysis in the presence of IgG1. When serum was separated into three fractions by passage through a Sephadex G-200 column, the third fraction, which contained proteins with a molecular weight of less than 100,000 d (but neither of the other two fractions nor purified human albumin), restored lymphocyte-mediated lysis in the presence of IgG1.

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