Abstract:
We investigated the microbiological and toxicological effects of three Perla black bean... more Abstract: We investigated the microbiological and toxicological effects of three Perla black bean extracts on the growth and culture of selected pathogenic microorganisms, the toxicity over Vero cell lines and an in vivo rat model. Three different solvents were used to obtain Perla black bean extracts. All three Perla black bean extracts were tested for antibacterial and antiparasitic activity and further analysed for intrinsic cytotoxicity (IC50 ). Methanol Perla black bean extract was used for acute toxicity test in rats, with the up-and-down doping method
1. Arch Med Res. 2000 Jul-Aug;31(4 Suppl):S212-3. Entamoeba histolytica is unable to use free cho... more 1. Arch Med Res. 2000 Jul-Aug;31(4 Suppl):S212-3. Entamoeba histolytica is unable to use free cholesterol, phospholipids, and fatty acids under axenic cultivation conditions. Mata-Cárdenas BD, Vargas-Villarreal J, González ...
Entamoeba histolytica phospholipase A and lysophospholipase activities from a vesicular subcellul... more Entamoeba histolytica phospholipase A and lysophospholipase activities from a vesicular subcellular fraction (P30) were analyzed. The products, obtained using specific substrates labeled with14C or3H, indicated the presence of phospholipase A1 and A2 as well as lysophospholipase L1 activities. The enzymes detected could participate in phospholipid metabolism and the alkaline phospholipase A2 may contribute toE. histolytica cytopathogenicity.
The multipurpose method for protein determination (MMPD) is based in the Coomassie brilliant blue... more The multipurpose method for protein determination (MMPD) is based in the Coomassie brilliant blue G-250 binding to immobilized and washed proteins in paper filter disks, and the A600 measurement of the eluted protein-dye complexes. The analysis requires 5-μl samples, put in 7-mm paper filter disks, which can be stored for up to 2 weeks, without sensible changes in their protein content. The A600 is a logarithmic function of the log of bovine serum albumin quantity, between 2.5 and 250 μg with two linear segments used as standard curves; one between 2.5 to 20 μg and the other between 20 and 80 μg. Results with the MMPD were quantitatively comparable with those obtained with the method of Lowry et al., and those reported by other workers, assaying the following material: (i) bovine serum albumin, in doubly distilled water or in presence of several substances that interfere with the current methods; and (ii) total homogenates and their respective trichloroacetic acid-insoluble extracts, which were obtained from several phenol-rich vegetal specimens and complex animal products. The MMPD is proposed as an alternative for protein determination for its versatility and reliability, in both vegetal or animal products, especially when the analysis with traditional methods is made difficult by the presence of natural or added interfering substances and the sample volume is too small to discard them, or when the material must be stored for relatively long periods of time, prior to its processing.
Record Detail. Return to previous page, Save: Title: Effect of agents that modify cAMP levels on ... more Record Detail. Return to previous page, Save: Title: Effect of agents that modify cAMP levels on growth and differentiation of Entamoeba invadens. Author: Valdez Aguirre, A : Villarreal Trevino, L : Vargas Villarreal, J : Mata Cardenas ...
The influence of low-frequency electromagnetic (LF-EM) waves on microorganisms has been a subject... more The influence of low-frequency electromagnetic (LF-EM) waves on microorganisms has been a subject of experimental investigations for more than two decades and the results are promising. In parallel, an interesting procedure known as biophysical-information-therapy or bioresonance therapy (BRT) which in principle is based on LF-EM stimulation, has emerged. BRT was discovered in the late 1980’s but it is still poorly studied. This paper demonstrates that by transferring metronidazole information to water samples by an electronic amplifier (BRT device), the growth of axenically cultured trophozoites of Entamoeba histolytica and Trichomonasvaginalis is significantly inhibited, compared with those cultures treated with non and sham electro-transferred water samples. A positive control of metronidazole, a well-known cytotoxic drug against parasites, was used as a reference.
Sphingomyelinase (SMase) activity was measured in Entamoeba histolytica particulate and soluble s... more Sphingomyelinase (SMase) activity was measured in Entamoeba histolytica particulate and soluble subcellular fractions. The effects on SMase of incubation time, total protein concentration, pH, and several divalent cations were determined. SMase-C and other unidentified esterase activity were detected in soluble and particulate fractions. SMase-C was 94.5–96.0% higher than the unidentified esterase activity. Soluble and insoluble SMase-C specific activities increased with protein dose and incubation time. Soluble and insoluble SMase-C activities were maximum at pH 7.5 and were dependent on Mg2+, Mn2+, or Co2+, and inhibited by Zn2+, Hg2+, Ca2+, and EDTA. SMase-C was active in the pH range of 3–10 and its maximum activity was at pH 7.5. The soluble and insoluble SMases have remarkably similar physicochemical properties, strongly suggesting that E. histolytica has just one isoform of neutral SMase-C that had not been described before and might be essential for E. histolytica metabolism or virulence.
Abstract: We investigated the microbiological and toxicological effects of three Perla black bea... more Abstract: We investigated the microbiological and toxicological effects of three Perla black bean extracts on the growth and culture of selected pathogenic microorganisms, the toxicity over Vero cell lines and an in vivo rat model. Three different solvents were used to obtain Perla black bean extracts. All three Perla black bean extracts were tested for antibacterial and antiparasitic activity and further analysed for intrinsic cytotoxicity (IC50). Methanol Perla black bean extract was used for acute toxicity test in rats, with the up-and-down doping method. All Perla black bean extracts inhibited bacterial growth. Pseudomonas aeruginosa, Proteus vulgaris, Klebsiella oxytoca, Enterococcus faecalis, Staphylococcus aureus, Staphylococcus epidermidis and Listeria monocytogenes showed inhibition, while Escherichia coli and Enterobacter aerogenes did not. Acidified water and acetic acid Perla black bean extract were tested in parasites. The best IC50 was observed for Giardia lamblia, while higher concentrations were active against Entamoeba histolytica and Trichomonas vaginalis. The Vero cells toxicity levels (IC50) for methanol, acidified water and acetic acid Perla black bean extract were [mean ± S.D. (95% CI)]: 275 ± 6.2 (267.9–282.0), 390 ± 4.6 (384.8–395.2) and 209 ± 3.39 (205.6–212.4) µg/ml, respectively. In vivo acute toxicity assays did not show changes in absolute organ weights, gross and histological examinations of selected tissues or functional tests. The acetic acid and methanol Perla black bean extract proved to exhibit strong antibacterial activity and the acidified water Perla black bean extract exerted parasiticidal effects against Giardia lamblia, Entamoeba hystolitica and Trichomonas vaginalis. The three Perla black bean extracts assayed over Vero cells showed very low toxicity and the methanol Perla black bean extract in vivo did not cause toxicity.
Identification of seven chemical factors that favor high-quality Entamoeba histolytica cyst-like ... more Identification of seven chemical factors that favor high-quality Entamoeba histolytica cyst-like structure formation under axenic conditions. ... González-Salazar F, Viader-Salvadó JM, Martínez-Rodríguez HG, Campos-Góngora E, Mata-Cárdenas BD, Said-Fernández S.
The current media for axenic cultivation of Entamoeba histolytica and Entamoeba invadens are supp... more The current media for axenic cultivation of Entamoeba histolytica and Entamoeba invadens are supplemented with bovine or equine serum, which provides several essential nutrients to amoebas. Serum has also been considered an essential component in encystation media for E. invadens. A substitute of serum, PACSR has been described as an alternative for growth of E. histolytica and also maintains growth of E. invadens. When PACSR was used instead of serum for encystation of E. invadens the efficiency was the same as for serum. Our present data show that PACSR can support the growth and induction of encystation of E. invadens strain IP-1.
Several thiosemicarbazone derivatives of 5-nitrothiophene-2-carboxaldehyde were prepared by the s... more Several thiosemicarbazone derivatives of 5-nitrothiophene-2-carboxaldehyde were prepared by the simple process in which N4-thiosemicarbazone moiety was replaced by aliphatic, arylic and cyclic amine. Among these thiosemicarbazones compound 11 showed significant antiamoebic activity whereas compound 3 was more active antitrichomonal than the reference drug.Thiosemicarbazone derivatives were prepared from 5-nitrothiophene-2-carboxaldehyde and tested in vitro against Entamoeba histolytica (strain HK-9), Giardia lamblia (strain IMSS-0989) and Trichomonas vaginalis (strain tv-43). Compound 3 and 11 exhibited better antitrichomonial and antiamoebic activity respectively comparable to that of metronidazole.
Neither phospholipase A1 (PLA A1) nor phospholipase A2 (PLA A2), nor their respective genes, have... more Neither phospholipase A1 (PLA A1) nor phospholipase A2 (PLA A2), nor their respective genes, have been identified in Giardia lamblia, even though they are essential for lipid metabolism in this parasite. A method to identify, isolate, and characterize these enzymes is needed. The activities of PLA A1 and PLA A2 were analyzed in a total extract (TE) and in vesicular (P30) and soluble (S30) subcellular fractions of G. lamblia trophozoites; the effects of several chemical and physicochemical factors on their activities were investigated. The assays were performed using substrate labeled with 14C, and the mass of the 14C-product was quantified. PLA A1 and PLA A2 activity was present in the TE and the P30 and S30 fractions, and it was dependent on pH and the concentrations of protein and Ca2+. In all trophozoite preparations, PLA A1 and PLA A2 activities were inhibited by ethylenediaminetetraacetic acid and Rosenthal's inhibitor. These results suggest that G. lamblia possesses several PLA A1 and PLA A2 isoforms that may be soluble or associated with membranes. In addition to participating in G. lamblia phospholipid metabolism, PLA A1 and PLA A2 could play important roles in the cytopathogenicity of this parasite.
Exposure to extremely low-frequency (ELF) electromagnetic fields appears to result in a number of... more Exposure to extremely low-frequency (ELF) electromagnetic fields appears to result in a number of important biological changes. In the present study, we evaluated the effects of 60 Hz sinusoidal magnetic fields (MF) at magnetic flux densities of 1.0, 1.5 and 2.0 mT on growth and differentiation of the protozoan Entamoeba invadens. We demonstrated an inhibitory growth effect when trophozoite cultures were exposed to 1.5 and 2.0 mT. Furthermore, we found that there was not a synergistic effect in cultures co-exposed to MF and Metronidazole, a cytotoxic drug against amoebic cells. In addition, MF exposure inhibited the encystation process of E. invadens.
The cyst of Entamoeba histolytica is responsible for amebiasis infection. However, no axenic in v... more The cyst of Entamoeba histolytica is responsible for amebiasis infection. However, no axenic in vitro system exists that promotes mass encystation for studying this process of this human-infecting parasite. Cyst-like structures of E. histolytica obtained in this work were induced using TYI-S-33 media in combination with enterobacterias Escherichia coli and Enterococcus faecalis conditioned media, high CO2 tension and histamine. Cyst-like structures showed the same characteristics of a typical E. histolytica cyst: aggregation, resistance to 0.15% sarcosyl for 10 min, high signal of fluorescence under UV light when stained with 10% calcofluor M2r and the surface topology showed a wrinkled wall. In addition these structures are multinucleated with condensed chromatin attached to nuclear membrane, contain big vacuoles and ribonucleoproteic helices in the cytoplasm and also present a thin cell wall. Last all characteristics are all the same as a typical of E. histolytica cyst.
Abstract:
We investigated the microbiological and toxicological effects of three Perla black bean... more Abstract: We investigated the microbiological and toxicological effects of three Perla black bean extracts on the growth and culture of selected pathogenic microorganisms, the toxicity over Vero cell lines and an in vivo rat model. Three different solvents were used to obtain Perla black bean extracts. All three Perla black bean extracts were tested for antibacterial and antiparasitic activity and further analysed for intrinsic cytotoxicity (IC50 ). Methanol Perla black bean extract was used for acute toxicity test in rats, with the up-and-down doping method
1. Arch Med Res. 2000 Jul-Aug;31(4 Suppl):S212-3. Entamoeba histolytica is unable to use free cho... more 1. Arch Med Res. 2000 Jul-Aug;31(4 Suppl):S212-3. Entamoeba histolytica is unable to use free cholesterol, phospholipids, and fatty acids under axenic cultivation conditions. Mata-Cárdenas BD, Vargas-Villarreal J, González ...
Entamoeba histolytica phospholipase A and lysophospholipase activities from a vesicular subcellul... more Entamoeba histolytica phospholipase A and lysophospholipase activities from a vesicular subcellular fraction (P30) were analyzed. The products, obtained using specific substrates labeled with14C or3H, indicated the presence of phospholipase A1 and A2 as well as lysophospholipase L1 activities. The enzymes detected could participate in phospholipid metabolism and the alkaline phospholipase A2 may contribute toE. histolytica cytopathogenicity.
The multipurpose method for protein determination (MMPD) is based in the Coomassie brilliant blue... more The multipurpose method for protein determination (MMPD) is based in the Coomassie brilliant blue G-250 binding to immobilized and washed proteins in paper filter disks, and the A600 measurement of the eluted protein-dye complexes. The analysis requires 5-μl samples, put in 7-mm paper filter disks, which can be stored for up to 2 weeks, without sensible changes in their protein content. The A600 is a logarithmic function of the log of bovine serum albumin quantity, between 2.5 and 250 μg with two linear segments used as standard curves; one between 2.5 to 20 μg and the other between 20 and 80 μg. Results with the MMPD were quantitatively comparable with those obtained with the method of Lowry et al., and those reported by other workers, assaying the following material: (i) bovine serum albumin, in doubly distilled water or in presence of several substances that interfere with the current methods; and (ii) total homogenates and their respective trichloroacetic acid-insoluble extracts, which were obtained from several phenol-rich vegetal specimens and complex animal products. The MMPD is proposed as an alternative for protein determination for its versatility and reliability, in both vegetal or animal products, especially when the analysis with traditional methods is made difficult by the presence of natural or added interfering substances and the sample volume is too small to discard them, or when the material must be stored for relatively long periods of time, prior to its processing.
Record Detail. Return to previous page, Save: Title: Effect of agents that modify cAMP levels on ... more Record Detail. Return to previous page, Save: Title: Effect of agents that modify cAMP levels on growth and differentiation of Entamoeba invadens. Author: Valdez Aguirre, A : Villarreal Trevino, L : Vargas Villarreal, J : Mata Cardenas ...
The influence of low-frequency electromagnetic (LF-EM) waves on microorganisms has been a subject... more The influence of low-frequency electromagnetic (LF-EM) waves on microorganisms has been a subject of experimental investigations for more than two decades and the results are promising. In parallel, an interesting procedure known as biophysical-information-therapy or bioresonance therapy (BRT) which in principle is based on LF-EM stimulation, has emerged. BRT was discovered in the late 1980’s but it is still poorly studied. This paper demonstrates that by transferring metronidazole information to water samples by an electronic amplifier (BRT device), the growth of axenically cultured trophozoites of Entamoeba histolytica and Trichomonasvaginalis is significantly inhibited, compared with those cultures treated with non and sham electro-transferred water samples. A positive control of metronidazole, a well-known cytotoxic drug against parasites, was used as a reference.
Sphingomyelinase (SMase) activity was measured in Entamoeba histolytica particulate and soluble s... more Sphingomyelinase (SMase) activity was measured in Entamoeba histolytica particulate and soluble subcellular fractions. The effects on SMase of incubation time, total protein concentration, pH, and several divalent cations were determined. SMase-C and other unidentified esterase activity were detected in soluble and particulate fractions. SMase-C was 94.5–96.0% higher than the unidentified esterase activity. Soluble and insoluble SMase-C specific activities increased with protein dose and incubation time. Soluble and insoluble SMase-C activities were maximum at pH 7.5 and were dependent on Mg2+, Mn2+, or Co2+, and inhibited by Zn2+, Hg2+, Ca2+, and EDTA. SMase-C was active in the pH range of 3–10 and its maximum activity was at pH 7.5. The soluble and insoluble SMases have remarkably similar physicochemical properties, strongly suggesting that E. histolytica has just one isoform of neutral SMase-C that had not been described before and might be essential for E. histolytica metabolism or virulence.
Abstract: We investigated the microbiological and toxicological effects of three Perla black bea... more Abstract: We investigated the microbiological and toxicological effects of three Perla black bean extracts on the growth and culture of selected pathogenic microorganisms, the toxicity over Vero cell lines and an in vivo rat model. Three different solvents were used to obtain Perla black bean extracts. All three Perla black bean extracts were tested for antibacterial and antiparasitic activity and further analysed for intrinsic cytotoxicity (IC50). Methanol Perla black bean extract was used for acute toxicity test in rats, with the up-and-down doping method. All Perla black bean extracts inhibited bacterial growth. Pseudomonas aeruginosa, Proteus vulgaris, Klebsiella oxytoca, Enterococcus faecalis, Staphylococcus aureus, Staphylococcus epidermidis and Listeria monocytogenes showed inhibition, while Escherichia coli and Enterobacter aerogenes did not. Acidified water and acetic acid Perla black bean extract were tested in parasites. The best IC50 was observed for Giardia lamblia, while higher concentrations were active against Entamoeba histolytica and Trichomonas vaginalis. The Vero cells toxicity levels (IC50) for methanol, acidified water and acetic acid Perla black bean extract were [mean ± S.D. (95% CI)]: 275 ± 6.2 (267.9–282.0), 390 ± 4.6 (384.8–395.2) and 209 ± 3.39 (205.6–212.4) µg/ml, respectively. In vivo acute toxicity assays did not show changes in absolute organ weights, gross and histological examinations of selected tissues or functional tests. The acetic acid and methanol Perla black bean extract proved to exhibit strong antibacterial activity and the acidified water Perla black bean extract exerted parasiticidal effects against Giardia lamblia, Entamoeba hystolitica and Trichomonas vaginalis. The three Perla black bean extracts assayed over Vero cells showed very low toxicity and the methanol Perla black bean extract in vivo did not cause toxicity.
Identification of seven chemical factors that favor high-quality Entamoeba histolytica cyst-like ... more Identification of seven chemical factors that favor high-quality Entamoeba histolytica cyst-like structure formation under axenic conditions. ... González-Salazar F, Viader-Salvadó JM, Martínez-Rodríguez HG, Campos-Góngora E, Mata-Cárdenas BD, Said-Fernández S.
The current media for axenic cultivation of Entamoeba histolytica and Entamoeba invadens are supp... more The current media for axenic cultivation of Entamoeba histolytica and Entamoeba invadens are supplemented with bovine or equine serum, which provides several essential nutrients to amoebas. Serum has also been considered an essential component in encystation media for E. invadens. A substitute of serum, PACSR has been described as an alternative for growth of E. histolytica and also maintains growth of E. invadens. When PACSR was used instead of serum for encystation of E. invadens the efficiency was the same as for serum. Our present data show that PACSR can support the growth and induction of encystation of E. invadens strain IP-1.
Several thiosemicarbazone derivatives of 5-nitrothiophene-2-carboxaldehyde were prepared by the s... more Several thiosemicarbazone derivatives of 5-nitrothiophene-2-carboxaldehyde were prepared by the simple process in which N4-thiosemicarbazone moiety was replaced by aliphatic, arylic and cyclic amine. Among these thiosemicarbazones compound 11 showed significant antiamoebic activity whereas compound 3 was more active antitrichomonal than the reference drug.Thiosemicarbazone derivatives were prepared from 5-nitrothiophene-2-carboxaldehyde and tested in vitro against Entamoeba histolytica (strain HK-9), Giardia lamblia (strain IMSS-0989) and Trichomonas vaginalis (strain tv-43). Compound 3 and 11 exhibited better antitrichomonial and antiamoebic activity respectively comparable to that of metronidazole.
Neither phospholipase A1 (PLA A1) nor phospholipase A2 (PLA A2), nor their respective genes, have... more Neither phospholipase A1 (PLA A1) nor phospholipase A2 (PLA A2), nor their respective genes, have been identified in Giardia lamblia, even though they are essential for lipid metabolism in this parasite. A method to identify, isolate, and characterize these enzymes is needed. The activities of PLA A1 and PLA A2 were analyzed in a total extract (TE) and in vesicular (P30) and soluble (S30) subcellular fractions of G. lamblia trophozoites; the effects of several chemical and physicochemical factors on their activities were investigated. The assays were performed using substrate labeled with 14C, and the mass of the 14C-product was quantified. PLA A1 and PLA A2 activity was present in the TE and the P30 and S30 fractions, and it was dependent on pH and the concentrations of protein and Ca2+. In all trophozoite preparations, PLA A1 and PLA A2 activities were inhibited by ethylenediaminetetraacetic acid and Rosenthal's inhibitor. These results suggest that G. lamblia possesses several PLA A1 and PLA A2 isoforms that may be soluble or associated with membranes. In addition to participating in G. lamblia phospholipid metabolism, PLA A1 and PLA A2 could play important roles in the cytopathogenicity of this parasite.
Exposure to extremely low-frequency (ELF) electromagnetic fields appears to result in a number of... more Exposure to extremely low-frequency (ELF) electromagnetic fields appears to result in a number of important biological changes. In the present study, we evaluated the effects of 60 Hz sinusoidal magnetic fields (MF) at magnetic flux densities of 1.0, 1.5 and 2.0 mT on growth and differentiation of the protozoan Entamoeba invadens. We demonstrated an inhibitory growth effect when trophozoite cultures were exposed to 1.5 and 2.0 mT. Furthermore, we found that there was not a synergistic effect in cultures co-exposed to MF and Metronidazole, a cytotoxic drug against amoebic cells. In addition, MF exposure inhibited the encystation process of E. invadens.
The cyst of Entamoeba histolytica is responsible for amebiasis infection. However, no axenic in v... more The cyst of Entamoeba histolytica is responsible for amebiasis infection. However, no axenic in vitro system exists that promotes mass encystation for studying this process of this human-infecting parasite. Cyst-like structures of E. histolytica obtained in this work were induced using TYI-S-33 media in combination with enterobacterias Escherichia coli and Enterococcus faecalis conditioned media, high CO2 tension and histamine. Cyst-like structures showed the same characteristics of a typical E. histolytica cyst: aggregation, resistance to 0.15% sarcosyl for 10 min, high signal of fluorescence under UV light when stained with 10% calcofluor M2r and the surface topology showed a wrinkled wall. In addition these structures are multinucleated with condensed chromatin attached to nuclear membrane, contain big vacuoles and ribonucleoproteic helices in the cytoplasm and also present a thin cell wall. Last all characteristics are all the same as a typical of E. histolytica cyst.
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Papers by benito mata
We investigated the microbiological and toxicological effects of three Perla black bean extracts on the growth
and culture of selected pathogenic microorganisms, the toxicity over Vero cell lines and an
in vivo
rat model. Three different
solvents were used to obtain Perla black bean extracts. All three Perla black bean extracts were tested for antibacterial and
antiparasitic activity and further analysed for intrinsic cytotoxicity (IC50 ). Methanol Perla black bean extract was used for acute toxicity test in rats, with the up-and-down doping method
We investigated the microbiological and toxicological effects of three Perla black bean extracts on the growth
and culture of selected pathogenic microorganisms, the toxicity over Vero cell lines and an
in vivo
rat model. Three different
solvents were used to obtain Perla black bean extracts. All three Perla black bean extracts were tested for antibacterial and
antiparasitic activity and further analysed for intrinsic cytotoxicity (IC50 ). Methanol Perla black bean extract was used for acute toxicity test in rats, with the up-and-down doping method