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Hiroshi Tachibana - One of the best experts on this subject based on the ideXlab platform.

  • Comparison of hemolytic activity of the intermediate subunit of Entamoeba histolytica and Entamoeba dispar lectins.
    PloS one, 2017
    Co-Authors: Kentaro Kato, Xunjia Cheng, Takashi Makiuchi, Hiroshi Tachibana
    Abstract:

    Galactose and N-acetyl-D-galactosamine-inhibitable lectin of Entamoeba histolytica has roles in pathogenicity and induction of protective immunity in rodent models of amoebiasis. Recently, the intermediate subunit of the lectin, Igl1, of E. histolytica has been shown to have hemolytic activity. However, the corresponding lectin is also expressed in a non-virulent species, Entamoeba dispar, and another subunit, Igl2, is expressed in the protozoa. Therefore, in this study, we compared the activities of Igl1 and Igl2 subunits from E. histolytica and E. dispar using various regions of recombinant Igl proteins expressed in Escherichia coli. The recombinant E. dispar Igl proteins had comparable hemolytic activities with those of E. histolytica Igl proteins. Furthermore, Igl1 gene-silenced E. histolytica trophozoites showed less hemolytic activity compared with vector-transfected trophozoites, indicating that the expression level of Igl1 protein influences the activity. These results suggest that the lower hemolytic activity in E. dispar compared with E. histolytica reflects the lower expression level of Igl1 in the E. dispar parasite.

  • Primary structure, expression and localization of two intermediate subunit lectins of Entamoeba dispar that contain multiple CXXC motifs.
    Parasitology, 2007
    Co-Authors: Hiroshi Tachibana, Xunjia Cheng, S. Kobayashi, Y. Okada, Johbu Itoh, T. Takeuchi
    Abstract:

    We have recently identified 2 surface proteins in Entamoeba histolytica as intermediate subunits of galactose- and N-acetyl-D-galactosamine-inhibitable lectin (EhIgl1 and EhIgl2) ; these proteins both contain multiple CXXC motifs. Here, we report the molecular characterization of the corresponding proteins in Entamoeba dispar, which is neither pathogenic nor invasive. Two Igl genes encoding 1110 and 1106 amino acids (EdIgl1 and EdIgl2) were cloned from 2 strains of E. dispar. The amino acid sequence identities were 79% between EdIgl1 and EdIgl2, 75-76% between EdIgl1 and EhIgl1, and 73-74% between EdIgl2 and EhIgl2. However, all the CXXC motifs were conserved in the EdIgl proteins, suggesting that the fold conferred by this motif is important for function. Comparison of the expression level of the Igl genes by real-time RT-PCR showed 3-5 times higher expression of EdIgl1 compared to EdIgl2. Most EdIgl1 and EdIgl2 proteins were co-localized on the surface and in the cytoplasm of trophozoites, based on confocal microscopy. However, a different localization of Edigll and EdIgl2 in intracellular vacuoles and a different level of phenotypic expression of the two Igls were also observed. These results demonstrate that Igls are important proteins even in non-pathogenic amoeba and that Igll and Igl2 may possess different functions.

  • Entamoeba histolytica and Entamoeba dispar infections in cynomolgus monkeys imported into Japan for research.
    Parasitology research, 2005
    Co-Authors: Jun-ichiro Takano, Hiroshi Tachibana, Toyoko Narita, Toshiyuki Shimizu, Hirofumi Komatsubara, Keiji Terao, Koji Fujimoto
    Abstract:

    Three hundred and three stool samples of cynomolgus monkeys (Macaca fascicularis) imported from China and the Philippines were examined for Entamoeba histolytica/Entamoeba dispar infections. Microscopy detected E. histolytica/E. dispar cysts in 41 samples. Positive rates were higher in the monkeys from China (37.5%) than in the monkeys from the Philippines (3.7%). PCR analysis of 25 samples successfully cultured from the cysts demonstrated that 24 were E. dispar, one of the samples from China was E. histolytica. The one sample was also identified as E. histolytica by an antigen detection kit, although the monkey was asymptomatic and serology was negative. To our knowledge, this is the first report of E. histolytica isolation from cynomolgus monkeys based on the discrimination between E. histolytica and E. dispar.

  • axenic cultivation of Entamoeba dispar in newly designed yeast extract iron gluconic acid dihyrdoxyacetone serum medium
    Journal of Parasitology, 2005
    Co-Authors: Seiki Kobayashi, Ali Haghighi, Hiroshi Tachibana, Eiko Imai, Shaden A. Khalifa, Tsutomu Takeuchi
    Abstract:

    Yeast extract–iron–gluconic acid–dihyrdoxyacetone-serum medium that allows axenic cultivation of Entamoeba dispar was designed based on casein-free yeast extract-iron-serum (YI-S) medium, and the usefulness of the medium was assessed. The main differences from YI-S medium are replacement of glucose by gluconic acid, addition of dihydroxyacetone and d-galacturonic acid monohydrate, and sterilization by filtration. This medium promoted the axenic growth of 5 strains of E. dispar (2 strains of nonhuman primate isolates and 3 strains of human isolates). In addition, to clarify the biological basis for the growth of E. dispar in this medium, analyses of relevant enzymes on the glycolytic pathway of the amoebae as well as of the protozoans that are the best culture supplement for amoebae are being performed.

  • AXENIC CULTIVATION OF Entamoeba dispar IN NEWLY DESIGNED YEAST EXTRACT–IRON–GLUCONIC ACID–DIHYRDOXYACETONE–SERUM MEDIUM
    The Journal of parasitology, 2005
    Co-Authors: Seiki Kobayashi, Ali Haghighi, Hiroshi Tachibana, Eiko Imai, Shaden A. Khalifa, Tsutomu Takeuchi
    Abstract:

    Yeast extract–iron–gluconic acid–dihyrdoxyacetone-serum medium that allows axenic cultivation of Entamoeba dispar was designed based on casein-free yeast extract-iron-serum (YI-S) medium, and the usefulness of the medium was assessed. The main differences from YI-S medium are replacement of glucose by gluconic acid, addition of dihydroxyacetone and d-galacturonic acid monohydrate, and sterilization by filtration. This medium promoted the axenic growth of 5 strains of E. dispar (2 strains of nonhuman primate isolates and 3 strains of human isolates). In addition, to clarify the biological basis for the growth of E. dispar in this medium, analyses of relevant enzymes on the glycolytic pathway of the amoebae as well as of the protozoans that are the best culture supplement for amoebae are being performed.

Tsutomu Takeuchi - One of the best experts on this subject based on the ideXlab platform.

  • axenic cultivation of Entamoeba dispar in newly designed yeast extract iron gluconic acid dihyrdoxyacetone serum medium
    Journal of Parasitology, 2005
    Co-Authors: Seiki Kobayashi, Ali Haghighi, Hiroshi Tachibana, Eiko Imai, Shaden A. Khalifa, Tsutomu Takeuchi
    Abstract:

    Yeast extract–iron–gluconic acid–dihyrdoxyacetone-serum medium that allows axenic cultivation of Entamoeba dispar was designed based on casein-free yeast extract-iron-serum (YI-S) medium, and the usefulness of the medium was assessed. The main differences from YI-S medium are replacement of glucose by gluconic acid, addition of dihydroxyacetone and d-galacturonic acid monohydrate, and sterilization by filtration. This medium promoted the axenic growth of 5 strains of E. dispar (2 strains of nonhuman primate isolates and 3 strains of human isolates). In addition, to clarify the biological basis for the growth of E. dispar in this medium, analyses of relevant enzymes on the glycolytic pathway of the amoebae as well as of the protozoans that are the best culture supplement for amoebae are being performed.

  • AXENIC CULTIVATION OF Entamoeba dispar IN NEWLY DESIGNED YEAST EXTRACT–IRON–GLUCONIC ACID–DIHYRDOXYACETONE–SERUM MEDIUM
    The Journal of parasitology, 2005
    Co-Authors: Seiki Kobayashi, Ali Haghighi, Hiroshi Tachibana, Eiko Imai, Shaden A. Khalifa, Tsutomu Takeuchi
    Abstract:

    Yeast extract–iron–gluconic acid–dihyrdoxyacetone-serum medium that allows axenic cultivation of Entamoeba dispar was designed based on casein-free yeast extract-iron-serum (YI-S) medium, and the usefulness of the medium was assessed. The main differences from YI-S medium are replacement of glucose by gluconic acid, addition of dihydroxyacetone and d-galacturonic acid monohydrate, and sterilization by filtration. This medium promoted the axenic growth of 5 strains of E. dispar (2 strains of nonhuman primate isolates and 3 strains of human isolates). In addition, to clarify the biological basis for the growth of E. dispar in this medium, analyses of relevant enzymes on the glycolytic pathway of the amoebae as well as of the protozoans that are the best culture supplement for amoebae are being performed.

  • Asymptomatic cyst passers of Entamoeba histolytica but not Entamoeba dispar in institutions for the mentally retarded in Japan.
    Parasitology International, 2000
    Co-Authors: Hiroshi Tachibana, Seiki Kobayashi, Kouichi Nagakura, Yoshimasa Kaneda, Tsutomu Takeuchi
    Abstract:

    Entamoeba histolytica/Entamoeba dispar was isolated from 50 asymptomatic amebic cyst passers in three institutions for the mentally retarded in Kanagawa Prefecture, Japan. To distinguish between E. histolytica and E. dispar, the isolates were analyzed by PCR, reactivity to monoclonal antibodies, and zymodemes. All isolates were identified as E. histolytica. The results lead us to conceive that, in Japan, E. histolytica is predominant even in asymptomatic cyst passers.

  • Cultivation of Entamoeba dispar: growth-promoting effect of ferredoxin.
    Archives of Medical Research, 2000
    Co-Authors: Seiki Kobayashi, Ali Haghighi, Hiroshi Tachibana, Eiko Imai, Tsutomu Takeuchi
    Abstract:

    We designed a medium (YIGADHA-S) (1) for the trial of axenic cultivation of Entamoeba dispar based on the caseinfree YI-S medium (YI-S) (2,3). YIGADHA-S is different from YI-S in that glucose is replaced by gluconic acid and dihydroxyacetone, and its sterilization is carried out by filtration. Consequently, one strain of E. dispar (CYNO16:TPC) was adapted to this YIGADHA-S and grew stably. However, in this YIGADHA-S culture system, other strains of E. dispar still required a culture associate, such as 10% formalin fixed or autoclaved Pseudomonas aeruginosa or Crithidia fasciculata. Moreover, we recently found that over 18 different cells of animal or plant origin, which were autoclaved (121 8 C, 15 min), could also support the growth of E. dispar. These studies led us to search for a property common to every cell examined (e.g., protozoan, mammalian, and plant cells) for supporting the growth of E. dispar in YIGADHA-S. These cells had either mitochondria, hydrogenosome, or mitochondria and chloroplast. On the other hand, some protozoan cells [ E. histolytica , E. dispar , E. moshkovskii (Laredo), E. invadens (IP-1), Giardia lamblia (Portland I)], and human erythrocytes, which lacked mitochondria or mitochondria-like organelles, did not support the growth of E. dispar in this medium. Based on these findings, we speculated that some substance existing commonly in mitochondria, hydrogenosomes, chloroplasts, and bacteria might support the growth of E. dispar. The present study was attempted to test the growth-promoting effect of a plant nonheme iron–sulfur protein (ferredoxin) on E. dispar in culture. Materials and Methods

  • Entamoeba dispar: cultivation with sterilized Crithidia fasciculata.
    The Journal of eukaryotic microbiology, 1998
    Co-Authors: Seiki Kobayashi, Hiroshi Tachibana, Eiko Imai, Tatsushi Fujiwara, Tsutomu Takeuchi
    Abstract:

    Four isolates of Entamoeba dispar identified by their hexokinase and phosphoglucomutase isoenzyme profile and by their failure to react with Entamoeba histolytica-specific monoclonal antibody (4G6) could be grown in either Diamond's BI-S-33 medium, newly developed BCSI-S (Biosate cysteine starch iron-serum) medium, or casein-free YI-S medium in the presence of Crithidia fasciculata (ReF-1:PRR) sterilized by heating 56 degrees C for 30 min and subsequent incubation with 1% hydrogen peroxide for 24 hours at 4 degrees C. After the cultures were maintained for over 50 passages, the amebae were identified as E. dispar by isoenzyme analysis, polymerase chain reaction with E. histolytica- and E. dispar-specific primers, i.e. p11 plus p12 and p13 plus p14, respectively, and by negative reactivity with monoclonal antibody 4G6. The flagellates added to the culture were judged to be metabolically inactive based on the results of nuclear magnetic resonance spectroscopy, electron microscopy, and polarographic analysis. All of these findings suggest that E. dispar can grow in vitro with metabolically inactive C. fasciculata as a culture associate.

Seiki Kobayashi - One of the best experts on this subject based on the ideXlab platform.

  • axenic cultivation of Entamoeba dispar in newly designed yeast extract iron gluconic acid dihyrdoxyacetone serum medium
    Journal of Parasitology, 2005
    Co-Authors: Seiki Kobayashi, Ali Haghighi, Hiroshi Tachibana, Eiko Imai, Shaden A. Khalifa, Tsutomu Takeuchi
    Abstract:

    Yeast extract–iron–gluconic acid–dihyrdoxyacetone-serum medium that allows axenic cultivation of Entamoeba dispar was designed based on casein-free yeast extract-iron-serum (YI-S) medium, and the usefulness of the medium was assessed. The main differences from YI-S medium are replacement of glucose by gluconic acid, addition of dihydroxyacetone and d-galacturonic acid monohydrate, and sterilization by filtration. This medium promoted the axenic growth of 5 strains of E. dispar (2 strains of nonhuman primate isolates and 3 strains of human isolates). In addition, to clarify the biological basis for the growth of E. dispar in this medium, analyses of relevant enzymes on the glycolytic pathway of the amoebae as well as of the protozoans that are the best culture supplement for amoebae are being performed.

  • AXENIC CULTIVATION OF Entamoeba dispar IN NEWLY DESIGNED YEAST EXTRACT–IRON–GLUCONIC ACID–DIHYRDOXYACETONE–SERUM MEDIUM
    The Journal of parasitology, 2005
    Co-Authors: Seiki Kobayashi, Ali Haghighi, Hiroshi Tachibana, Eiko Imai, Shaden A. Khalifa, Tsutomu Takeuchi
    Abstract:

    Yeast extract–iron–gluconic acid–dihyrdoxyacetone-serum medium that allows axenic cultivation of Entamoeba dispar was designed based on casein-free yeast extract-iron-serum (YI-S) medium, and the usefulness of the medium was assessed. The main differences from YI-S medium are replacement of glucose by gluconic acid, addition of dihydroxyacetone and d-galacturonic acid monohydrate, and sterilization by filtration. This medium promoted the axenic growth of 5 strains of E. dispar (2 strains of nonhuman primate isolates and 3 strains of human isolates). In addition, to clarify the biological basis for the growth of E. dispar in this medium, analyses of relevant enzymes on the glycolytic pathway of the amoebae as well as of the protozoans that are the best culture supplement for amoebae are being performed.

  • Entamoeba dispar: Molecular characterization of the galactose/N-acetyl-D-galactosamine lectin
    Experimental parasitology, 2001
    Co-Authors: Dylan R. Pillai, Seiki Kobayashi, Kevin C. Kain
    Abstract:

    Amebiasis contributes to approximately 50 million cases of life-threatening dysentery worldwide. Comparison of the lectins from Entamoeba histolytica (pathogenic) and Entamoeba dispar (nonpathogenic) was undertaken to elucidate the differential roles of this molecule in invasion versus colonization. Surface lectin was less abundant on axenic E. dispar than on axenic E. histolytica, commensurate with differences in lectin (heavy and light subunits) RNA when assessed by semiquantitative RT-PCR. The 1G7 epitope, which falls within the immunodominant and immunoprotective cysteine-rich region (480-900), was absent on axenic E. dispar. Indirect immunofluorescence, transient transection of COS7, and immunoprecipitation demonstrated that the 1G7 epitope was conserved in the nonpathogenic lectin homologue but not exposed on live E. dispar trophozoites. Hgl2 (E. histolytica) and Dhgl2 (E. dispar) lectin homologues demonstrated comparable high-affinity binding to multivalent GalNAc(19) BSA. These data provide evidence for relative gene and conformational regulation of the E.dispar lectin.

  • Asymptomatic cyst passers of Entamoeba histolytica but not Entamoeba dispar in institutions for the mentally retarded in Japan.
    Parasitology International, 2000
    Co-Authors: Hiroshi Tachibana, Seiki Kobayashi, Kouichi Nagakura, Yoshimasa Kaneda, Tsutomu Takeuchi
    Abstract:

    Entamoeba histolytica/Entamoeba dispar was isolated from 50 asymptomatic amebic cyst passers in three institutions for the mentally retarded in Kanagawa Prefecture, Japan. To distinguish between E. histolytica and E. dispar, the isolates were analyzed by PCR, reactivity to monoclonal antibodies, and zymodemes. All isolates were identified as E. histolytica. The results lead us to conceive that, in Japan, E. histolytica is predominant even in asymptomatic cyst passers.

  • Cultivation of Entamoeba dispar: growth-promoting effect of ferredoxin.
    Archives of Medical Research, 2000
    Co-Authors: Seiki Kobayashi, Ali Haghighi, Hiroshi Tachibana, Eiko Imai, Tsutomu Takeuchi
    Abstract:

    We designed a medium (YIGADHA-S) (1) for the trial of axenic cultivation of Entamoeba dispar based on the caseinfree YI-S medium (YI-S) (2,3). YIGADHA-S is different from YI-S in that glucose is replaced by gluconic acid and dihydroxyacetone, and its sterilization is carried out by filtration. Consequently, one strain of E. dispar (CYNO16:TPC) was adapted to this YIGADHA-S and grew stably. However, in this YIGADHA-S culture system, other strains of E. dispar still required a culture associate, such as 10% formalin fixed or autoclaved Pseudomonas aeruginosa or Crithidia fasciculata. Moreover, we recently found that over 18 different cells of animal or plant origin, which were autoclaved (121 8 C, 15 min), could also support the growth of E. dispar. These studies led us to search for a property common to every cell examined (e.g., protozoan, mammalian, and plant cells) for supporting the growth of E. dispar in YIGADHA-S. These cells had either mitochondria, hydrogenosome, or mitochondria and chloroplast. On the other hand, some protozoan cells [ E. histolytica , E. dispar , E. moshkovskii (Laredo), E. invadens (IP-1), Giardia lamblia (Portland I)], and human erythrocytes, which lacked mitochondria or mitochondria-like organelles, did not support the growth of E. dispar in this medium. Based on these findings, we speculated that some substance existing commonly in mitochondria, hydrogenosomes, chloroplasts, and bacteria might support the growth of E. dispar. The present study was attempted to test the growth-promoting effect of a plant nonheme iron–sulfur protein (ferredoxin) on E. dispar in culture. Materials and Methods

Anton M. Polderman - One of the best experts on this subject based on the ideXlab platform.

  • short communication prevalence of Entamoeba histolytica and Entamoeba dispar in northern ghana
    Tropical Medicine & International Health, 2003
    Co-Authors: Jaco J Verweij, Eric A. T. Brienen, Fieke Oostvogel, Alexis Nangbeifubah, Juventus B Ziem, Anton M. Polderman
    Abstract:

    Summary Since the redescription of the potentially invasive Entamoeba histolytica, separating it from the morphologically identical non-invasive Entamoeba dispar, there is a need for the reassessment of epidemiological data on amoebiasis. In this context we conducted a descriptive survey on the presence of E. histolytica and E. dispar in a rural area in northern Ghana. We found a high prevalence (39.8%) of the E. histolytica/E. dispar complex with microscopy, but E. histolytica and E. dispar-specific DNA amplification using real-time polymerase chain reaction identified only one E. histolytica case and revealed a considerably higher prevalence of E. dispar (82.8%).

  • Differentiation of Entamoeba histolytica and Entamoeba dispar Cysts Using Polymerase Chain Reaction on DNA Isolated from Faeces with Spin Columns
    European Journal of Clinical Microbiology & Infectious Diseases, 2000
    Co-Authors: Jaco J Verweij, Eric A. T. Brienen, J. Blotkamp, A. Aguirre, Anton M. Polderman
    Abstract:

    Since Entamoeba histolytica and Entamoeba dispar were formally recognized as two different species at the World Health Organization (WHO)/Pan American Health Organization (PAHO)/United Nations Educational, Scientific and Cultural Organization (UNESCO) meeting in Mexico City in 1997, the specific differentiation of the two morphologically identical species would seem relevant in clinical diagnosis. Several polymerase chain reaction (PCR)-based methods have been described and used successfully, but methods for DNA isolation from cysts in stool samples are time-consuming and problematic due to inhibitory factors in faeces. The use of the slightly modified QIAamp tissue method (Qiagen, Germany) for DNA isolation was evaluated in 657 unpreserved faecal samples from cases of suspected Entamoeba histolytica/Entamoeba dispar infection. In only 1.7% of the cases was PCR hampered by inhibitors present in the faeces. The DNA isolation procedure was found to be rapid, simple and one that could easily be implemented in a routine diagnostic setting. In 98.8% of Entamoeba histolytica/Entamoeba dispar cyst-positive faecal samples, the true identity of the cysts could be determined using PCR specific for Entamoeba histolytica and Entamoeba dispar, respectively.

Eiko Imai - One of the best experts on this subject based on the ideXlab platform.

  • axenic cultivation of Entamoeba dispar in newly designed yeast extract iron gluconic acid dihyrdoxyacetone serum medium
    Journal of Parasitology, 2005
    Co-Authors: Seiki Kobayashi, Ali Haghighi, Hiroshi Tachibana, Eiko Imai, Shaden A. Khalifa, Tsutomu Takeuchi
    Abstract:

    Yeast extract–iron–gluconic acid–dihyrdoxyacetone-serum medium that allows axenic cultivation of Entamoeba dispar was designed based on casein-free yeast extract-iron-serum (YI-S) medium, and the usefulness of the medium was assessed. The main differences from YI-S medium are replacement of glucose by gluconic acid, addition of dihydroxyacetone and d-galacturonic acid monohydrate, and sterilization by filtration. This medium promoted the axenic growth of 5 strains of E. dispar (2 strains of nonhuman primate isolates and 3 strains of human isolates). In addition, to clarify the biological basis for the growth of E. dispar in this medium, analyses of relevant enzymes on the glycolytic pathway of the amoebae as well as of the protozoans that are the best culture supplement for amoebae are being performed.

  • AXENIC CULTIVATION OF Entamoeba dispar IN NEWLY DESIGNED YEAST EXTRACT–IRON–GLUCONIC ACID–DIHYRDOXYACETONE–SERUM MEDIUM
    The Journal of parasitology, 2005
    Co-Authors: Seiki Kobayashi, Ali Haghighi, Hiroshi Tachibana, Eiko Imai, Shaden A. Khalifa, Tsutomu Takeuchi
    Abstract:

    Yeast extract–iron–gluconic acid–dihyrdoxyacetone-serum medium that allows axenic cultivation of Entamoeba dispar was designed based on casein-free yeast extract-iron-serum (YI-S) medium, and the usefulness of the medium was assessed. The main differences from YI-S medium are replacement of glucose by gluconic acid, addition of dihydroxyacetone and d-galacturonic acid monohydrate, and sterilization by filtration. This medium promoted the axenic growth of 5 strains of E. dispar (2 strains of nonhuman primate isolates and 3 strains of human isolates). In addition, to clarify the biological basis for the growth of E. dispar in this medium, analyses of relevant enzymes on the glycolytic pathway of the amoebae as well as of the protozoans that are the best culture supplement for amoebae are being performed.

  • Cultivation of Entamoeba dispar: growth-promoting effect of ferredoxin.
    Archives of Medical Research, 2000
    Co-Authors: Seiki Kobayashi, Ali Haghighi, Hiroshi Tachibana, Eiko Imai, Tsutomu Takeuchi
    Abstract:

    We designed a medium (YIGADHA-S) (1) for the trial of axenic cultivation of Entamoeba dispar based on the caseinfree YI-S medium (YI-S) (2,3). YIGADHA-S is different from YI-S in that glucose is replaced by gluconic acid and dihydroxyacetone, and its sterilization is carried out by filtration. Consequently, one strain of E. dispar (CYNO16:TPC) was adapted to this YIGADHA-S and grew stably. However, in this YIGADHA-S culture system, other strains of E. dispar still required a culture associate, such as 10% formalin fixed or autoclaved Pseudomonas aeruginosa or Crithidia fasciculata. Moreover, we recently found that over 18 different cells of animal or plant origin, which were autoclaved (121 8 C, 15 min), could also support the growth of E. dispar. These studies led us to search for a property common to every cell examined (e.g., protozoan, mammalian, and plant cells) for supporting the growth of E. dispar in YIGADHA-S. These cells had either mitochondria, hydrogenosome, or mitochondria and chloroplast. On the other hand, some protozoan cells [ E. histolytica , E. dispar , E. moshkovskii (Laredo), E. invadens (IP-1), Giardia lamblia (Portland I)], and human erythrocytes, which lacked mitochondria or mitochondria-like organelles, did not support the growth of E. dispar in this medium. Based on these findings, we speculated that some substance existing commonly in mitochondria, hydrogenosomes, chloroplasts, and bacteria might support the growth of E. dispar. The present study was attempted to test the growth-promoting effect of a plant nonheme iron–sulfur protein (ferredoxin) on E. dispar in culture. Materials and Methods

  • Entamoeba dispar: cultivation with sterilized Crithidia fasciculata.
    The Journal of eukaryotic microbiology, 1998
    Co-Authors: Seiki Kobayashi, Hiroshi Tachibana, Eiko Imai, Tatsushi Fujiwara, Tsutomu Takeuchi
    Abstract:

    Four isolates of Entamoeba dispar identified by their hexokinase and phosphoglucomutase isoenzyme profile and by their failure to react with Entamoeba histolytica-specific monoclonal antibody (4G6) could be grown in either Diamond's BI-S-33 medium, newly developed BCSI-S (Biosate cysteine starch iron-serum) medium, or casein-free YI-S medium in the presence of Crithidia fasciculata (ReF-1:PRR) sterilized by heating 56 degrees C for 30 min and subsequent incubation with 1% hydrogen peroxide for 24 hours at 4 degrees C. After the cultures were maintained for over 50 passages, the amebae were identified as E. dispar by isoenzyme analysis, polymerase chain reaction with E. histolytica- and E. dispar-specific primers, i.e. p11 plus p12 and p13 plus p14, respectively, and by negative reactivity with monoclonal antibody 4G6. The flagellates added to the culture were judged to be metabolically inactive based on the results of nuclear magnetic resonance spectroscopy, electron microscopy, and polarographic analysis. All of these findings suggest that E. dispar can grow in vitro with metabolically inactive C. fasciculata as a culture associate.

  • Entamoeba dispar : Cultivation with sterilized Crithidia fasciculata : Host-parasite interactions in amebiasis
    Journal of Eukaryotic Microbiology, 1998
    Co-Authors: Seiki Kobayashi, Hiroshi Tachibana, Eiko Imai, Tatsushi Fujiwara, Tsutomu Takeuchi
    Abstract:

    Four isolates of Entamoeba dispar identified by their hexokinase and phosphoglucomutase isoenzyme profile and by their failure to react with Entamoeba histolytica-specific monoclonal antibody (4G6) could be grown in either Diamond's BI-S-33 medium, newly developed BCSI-S (Biosate cysteine starch iron-serum) medium, or casein-free YI-S medium in the presence of Crithidia fasciculata (ReF-1:PRR) sterilized by heating 56° C for 30 min and subsequent incubation with 1% hydrogen peroxide for 24 hours at 4° C. After the cultures were maintained for over 50 passages, the amebae were identified as E. dispar by isoenzyme analysis, polymerase chain reaction with E. histolytica- and E. dispar-specific primers, i.e. p11 plus p12 and p13 plus p14, respectively, and by negative reactivity with monoclonal antibody 4G6. The flagellates added to the culture were judged to be metabolically inactive based on the results of nuclear magnetic resonance spectroscopy, electron microscopy, and polarographic analysis. All of these findings suggest that E. dispar can grow in vitro with metabolically inactive C. fasciculata as a culture associate.