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1.
Int J Mol Sci ; 25(8)2024 Apr 15.
Artículo en Inglés | MEDLINE | ID: mdl-38673944

RESUMEN

It is important to search for cytostatic compounds in order to fight cancer. One of them could be 2'-methylthiamine, which is a thiamine antimetabolite with an additional methyl group at the C-2 carbon of thiazole. So far, the cytostatic potential of 2'-methylthiamine has not been studied. We have come forward with a simplified method of synthesis using commercially available substrates and presented a comparison of its effects, as boosted by oxythiamine, on normal skin fibroblasts and HeLa cancer cells, having adopted in vitro culture techniques. Oxythiamine has been found to inhibit the growth and metabolism of cancer cells significantly better than 2'-methylthiamine (GI50 36 and 107 µM, respectively), while 2'-methylthiamine is more selective for cancer cells than oxythiamine (SI = 180 and 153, respectively). Docking analyses have revealed that 2'-methylthiamine (ΔG -8.2 kcal/mol) demonstrates a better affinity with thiamine pyrophosphokinase than thiamine (ΔG -7.5 kcal/mol ) and oxythiamine (ΔG -7.0 kcal/mol), which includes 2'-methylthiamine as a potential cytostatic. Our results suggest that the limited effect of 2'-methylthiamine on HeLa arises from the related arduous transport as compared to oxythiamine. Given that 2'-methylthiamine may possibly inhibit thiamine pyrophosphokinase, it could once again be considered a potential cytostatic. Thus, research should be carried out in order to find the best way to improve the transport of 2'-methylthiamine into cells, which may trigger its cytostatic properties.


Asunto(s)
Simulación del Acoplamiento Molecular , Oxitiamina , Humanos , Células HeLa , Oxitiamina/farmacología , Oxitiamina/química , Oxitiamina/metabolismo , Tiamina/farmacología , Tiamina/análogos & derivados , Tiamina/química , Antineoplásicos/farmacología , Antineoplásicos/química , Proliferación Celular/efectos de los fármacos , Fibroblastos/efectos de los fármacos , Fibroblastos/metabolismo , Simulación por Computador
2.
Molecules ; 28(22)2023 Nov 10.
Artículo en Inglés | MEDLINE | ID: mdl-38005248

RESUMEN

Cytostatic and pro-apoptotic effects of selenium steroid derivatives against HeLa cells were determined. The highest cytostatic activity was shown by derivative 4 (GI50 25.0 µM, almost complete growth inhibition after three days of culture, and over 97% of apoptotic and dead cells at 200 µM). The results of our study (cell number measurements, apoptosis profile, relative expression of apoptosis-related APAF1, BID, and mevalonate pathway-involved HMGCR, SQLE, CYP51A1, and PDHB genes, and computational chemistry data) support the hypothesis that tested selenosteroids induce the extrinsic pathway of apoptosis by affecting the cell membrane as cholesterol antimetabolites. An additional mechanism of action is possible through a direct action of derivative 4 to inhibit PDHB expression in a way similar to steroid hormones.


Asunto(s)
Citostáticos , Humanos , Células HeLa , Citostáticos/farmacología , Apoptosis , Colesterol/metabolismo
3.
Int J Mol Sci ; 24(18)2023 Sep 16.
Artículo en Inglés | MEDLINE | ID: mdl-37762487

RESUMEN

Magnetite and gallium substituted cuboferrites with a composition of GaxFe3-xO4 (0 ≤ x ≤ 1.4) were fabricated by thermal decomposition from acetylacetonate salts. The effect of Ga3+ cation substitution on the structural and thermomagnetic behavior of 4-12 nm sized core-shell particles was explored by X-ray and neutron diffraction, small angle neutron scattering, transmission electron microscopy, Mössbauer spectroscopy, and calorimetric measurements. Superparamagnetic (SPM) behavior and thermal capacity against increasing gallium concentration in nanoferrites were revealed. The highest heat capacity typical for Fe3O4@Ga0.6Fe2.4O4 and Ga0.6Fe2.4O4@Fe3O4 is accompanied by a slight stimulation of fibroblast culture growth and inhibition of HeLa cell growth. The observed effect is concentration dependent in the range of 0.01-0.1 mg/mL and particles of Ga0.6Fe2.4O4@Fe3O4 design have a greater effect on cells. Observed magnetic heat properties, as well as interactions with tumor and healthy cells, provide a basis for further biomedical research to use the proposed nanoparticle systems in cancer thermotherapy (magnetic hyperthermia).

4.
J Enzyme Inhib Med Chem ; 37(1): 876-894, 2022 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-35296203

RESUMEN

With increasing number of immunocompromised patients as well as drug resistance in fungi, the risk of fatal fungal infections in humans increases as well. The action of echinocandins is based on the inhibition of ß-(1,3)-d-glucan synthesis that builds the fungal cell wall. Caspofungin, micafungin, anidulafungin and rezafungin are semi-synthetic cyclic lipopeptides. Their specific chemical structure possess a potential to obtain novel derivatives with better pharmacological properties resulting in more effective treatment, especially in infections caused by Candida and Aspergillus species. In this review we summarise information about echinocandins with closer look on their chemical structure, mechanism of action, drug resistance and usage in clinical practice. We also introduce actual trends in modification of this antifungals as well as new methods of their administration, and additional use in viral and bacterial infections.


Asunto(s)
Antifúngicos/farmacología , Aspergillus/efectos de los fármacos , Candida/efectos de los fármacos , Diseño de Fármacos , Equinocandinas/farmacología , Antifúngicos/química , Aspergillus/metabolismo , Candida/metabolismo , Pared Celular/efectos de los fármacos , Pared Celular/metabolismo , Equinocandinas/química , Glucanos/antagonistas & inhibidores , Glucanos/metabolismo , Pruebas de Sensibilidad Microbiana , Estructura Molecular
5.
J Enzyme Inhib Med Chem ; 36(1): 122-129, 2021 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-33187452

RESUMEN

Oxythiamine (OT) and 3-deazathiamine (DAT) are the antimetabolites of thiamine. The aim of study was to compare the effects of OT and DAT pyrophosphates (-PP) on the kinetics of mammalian pyruvate dehydrogenase complex (PDHC) and the in vitro culture of HeLa cells. The kinetic study showed that 3-deazathiamine pyrophosphate (DATPP) was a much stronger competitive inhibitor (Ki = 0.0026 µM) of PDHC than OTPP (Ki = 0.025 µM). Both Ki values were much lower versus K m for thiamine pyrophosphate (0.06 µM). However, DATPP added to the culture medium for the HeLa cells culture did not hamper the rate of cell growth and showed not significant impact on the viability of the cells, whereas OTPP and OT showed a significant cytostatic effect. The differences between the thiamine antivitamins in their effect on cell growth in vitro may be due to differences in physicochemical properties and difficulty in DAT transport across the cell membrane.


Asunto(s)
Antineoplásicos/farmacología , Inhibidores Enzimáticos/farmacología , Complejo Piruvato Deshidrogenasa/antagonistas & inhibidores , Tiamina/farmacología , Antineoplásicos/síntesis química , Antineoplásicos/química , Apoptosis/efectos de los fármacos , Proliferación Celular/efectos de los fármacos , Relación Dosis-Respuesta a Droga , Ensayos de Selección de Medicamentos Antitumorales , Inhibidores Enzimáticos/síntesis química , Inhibidores Enzimáticos/química , Células HeLa , Humanos , Estructura Molecular , Complejo Piruvato Deshidrogenasa/metabolismo , Relación Estructura-Actividad , Tiamina/análogos & derivados , Tiamina/química , Células Tumorales Cultivadas
6.
Mycologia ; 111(4): 624-631, 2019.
Artículo en Inglés | MEDLINE | ID: mdl-31322986

RESUMEN

Malassezia pachydermatis causes infections of the skin and mucous membranes, especially in individuals with metabolic, hormonal, and immunological disorders. The search for M. pachydermatis properties that differentiate isolates from healthy and infected animals may result in the identification of typically commensal and potentially pathogenic strains within the entire species. We aimed to determine and compare protein profiles of M. pachydermatis strains isolated from 30 dogs with clinical symptoms of otitis externa and 34 dogs without symptoms of any disease. Two-dimensional gel electrophoresis was applied, and proteins distinguishing the two groups of strains were identified by liquid chromatography coupled with tandem mass spectrometry. Significant differences were found between potentially pathogenic and commensal isolates. The most significant finding was the presence of nicotinamide adenine dinucleotide phosphate (NADP)-dependent mannitol dehydrogenase and ketol-acid reductoisomerase among M. pachydermatis strains obtained from dogs with otitis externa. Nevertheless, it is not clear whether they are associated directly with the pathogenicity or they play the role of fungal allergen. On the basis of these findings, we can conclude that there may be two distinct groups of M. pachydermatis strains-one typically commensal and the other with properties that enhance the infection process. These results may be used for more precise diagnosis and identification of potentially pathogenic strains in the future.


Asunto(s)
Dermatomicosis/veterinaria , Enfermedades de los Perros , Otitis Externa/microbiología , Animales , Dermatomicosis/diagnóstico , Dermatomicosis/microbiología , Dermatomicosis/terapia , Enfermedades de los Perros/diagnóstico , Enfermedades de los Perros/microbiología , Enfermedades de los Perros/terapia , Perros , Electroforesis en Gel Bidimensional , Proteínas Fúngicas , Malassezia/clasificación , Malassezia/metabolismo , Malassezia/patogenicidad , Otitis Externa/diagnóstico , Otitis Externa/terapia
7.
Vet Dermatol ; 29(6): 476-e160, 2018 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-30251451

RESUMEN

BACKGROUND: Malassezia pachydermatis is an opportunistic yeast involved in skin and ear canal infections of dogs and cats. Reports suggest that strains of M. pachydermatis resistant to commonly used antifungal agents may be emerging. Therefore, new therapeutic strategies should be explored. OBJECTIVES: The synergistic effect of oxythiamine (OT) and ketoconazole (KTC) was analysed using a reference strain and field isolates (n = 66) of M. pachydermatis. Hydrogel formulations containing these components also were evaluated. METHODS AND MATERIALS: The minimum inhibitory concentrations (MICs) and minimum fungicidal concentrations (MFCs) of OT, KTC and their mixtures were determined by a broth macrodilution method. The antifungal effects of hydrogel formulations were determined by a plate diffusion method. RESULTS: The MIC and MFC values of OT were in the range 0.08 × 103 to 10 × 103  mg/L. All M. pachydermatis strains showed higher susceptibility to KTC (MICs and MFCs in the range 0.04-0.32 mg/L). Formulations that combined OT and KTC showed a synergistic effect for all tested isolates (n = 66). Hydrogels that contained OT at a concentration of 10 × 103 or 20 × 103  mg/L and KTC at the concentration of 0.1 × 103  mg/L showed a stronger effect than a commercially available product with KTC alone (20 × 103  mg/L). CONCLUSIONS AND CLINICAL IMPORTANCE: Synergy of these drugs may allow for successful topical treatment which utilizes lower doses of KTC without changing its therapeutic effectiveness. Hydrogel formulations proved to be attractive drug carriers for potential topical use.


Asunto(s)
Antifúngicos/uso terapéutico , Dermatomicosis/veterinaria , Enfermedades de los Perros/microbiología , Cetoconazol/uso terapéutico , Malassezia , Otitis Externa/veterinaria , Oxitiamina/uso terapéutico , Animales , Antifúngicos/administración & dosificación , Dermatomicosis/tratamiento farmacológico , Enfermedades de los Perros/tratamiento farmacológico , Perros , Sinergismo Farmacológico , Quimioterapia Combinada , Hidrogel de Polietilenoglicol-Dimetacrilato/administración & dosificación , Cetoconazol/administración & dosificación , Malassezia/efectos de los fármacos , Pruebas de Sensibilidad Microbiana/veterinaria , Otitis Externa/tratamiento farmacológico , Otitis Externa/microbiología , Oxitiamina/administración & dosificación
8.
Mycologia ; 110(4): 666-676, 2018.
Artículo en Inglés | MEDLINE | ID: mdl-30130476

RESUMEN

Malassezia pachydermatis causes infections of the skin and mucous membranes, especially in animals. It is commonly accepted that symptom manifestation depends on the physiological status of the host (different metabolic, hormonal, and immunological disorders). However, it should be considered whether distinct strains of M. pachydermatis could have different pathogenic potential and maintain opposite relations with the host, such as commensalism or parasitism. The scope of this study was to explore the population structure, genetic diversity, and phylogenetic relationships of M. pachydermatis strains isolated from dogs with clinical symptoms of otitis externa and from healthy dogs in order to investigate their relationships and evolutionary history. For all tests, a group of 30 strains derived from dogs with otitis externa and 34 strains from healthy dogs were used. The level of genetic diversity was initially assessed by polymerase chain reaction (PCR)-based random amplification of polymorphic DNA (RAPD-PCR), whereas evolutionary history was assessed by comparison of the nucleotide sequences of the internal transcribed spacer ITS1 region of nuclear rDNA. RAPD-PCR fingerprinting revealed a high level of genetic polymorphism in both tested groups (85% of unique profiles), but clinical isolates usually grouped together with other strains from otitis externa cases. Sequencing analysis identified 17 distinct genotypes with 59 polymorphic sites within both populations; however, putatively virulent strains were more closely related, indicating a probable correlation between the genotype and the virulence potential. Therefore, the hypothesis that M. pachydermatis virulence depends solely on the host's properties should be reconsidered including evolutionary and epidemiological data.


Asunto(s)
Enfermedades de los Perros/microbiología , Variación Genética , Malassezia/genética , Malassezia/patogenicidad , Otitis Externa/veterinaria , Animales , ADN Intergénico , ADN Ribosómico , Enfermedades de los Perros/epidemiología , Perros/microbiología , Genotipo , Malassezia/aislamiento & purificación , Malassezia/fisiología , Otitis Externa/epidemiología , Otitis Externa/microbiología , Filogenia , Reacción en Cadena de la Polimerasa , Polimorfismo Genético , Técnica del ADN Polimorfo Amplificado Aleatorio , Simbiosis , Virulencia
9.
Biosci Rep ; 38(1)2018 02 28.
Artículo en Inglés | MEDLINE | ID: mdl-29208764

RESUMEN

Thiamine plays a very important coenzymatic and non-coenzymatic role in the regulation of basic metabolism. Thiamine diphosphate is a coenzyme of many enzymes, most of which occur in prokaryotes. Pyruvate dehydrogenase and 2-oxoglutarate dehydrogenase complexes as well as transketolase are the examples of thiamine-dependent enzymes present in eukaryotes, including human. Therefore, thiamine is considered as drug or diet supplement which can support the treatment of many pathologies including neurodegenerative and vascular system diseases. On the other hand, thiamine antivitamins, which can interact with thiamine-dependent enzymes impeding their native functions, thiamine transport into the cells or a thiamine diphosphate synthesis, are good propose to drug design. The development of organic chemistry in the last century allowed the synthesis of various thiamine antimetabolites such as amprolium, pyrithiamine, oxythiamine, or 3-deazathiamine. Results of biochemical and theoretical chemistry research show that affinity to thiamine diphosphate-dependent enzymes of these synthetic molecules exceeds the affinity of native coenzyme. Therefore, some of them have already been used in the treatment of coccidiosis (amprolium), other are extensively studied as cytostatics in the treatment of cancer or fungal infections (oxythiamine and pyrithiamine). This review summarizes the current knowledge concerning the synthesis and mechanisms of action of selected thiamine antivitamins and indicates the potential of their practical use.


Asunto(s)
Diseño de Fármacos , Tiamina Pirofosfato/metabolismo , Tiamina/metabolismo , Amprolio/química , Amprolio/metabolismo , Antimetabolitos/uso terapéutico , Transporte Biológico , Humanos , Oxitiamina/antagonistas & inhibidores , Oxitiamina/metabolismo , Piritiamina/antagonistas & inhibidores , Piritiamina/metabolismo , Tiamina/antagonistas & inhibidores , Tiamina/síntesis química , Tiamina Pirofosfato/química
10.
Mycoses ; 59(1): 20-7, 2016 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-26559663

RESUMEN

Malassezia pachydermatis can cause infections of the skin and mucous membranes, especially in animals. It becomes a problem also in medicine. It is considered that metabolic disorders as well as hormonal and immunological status of the host promote diseases caused by M. pachydermatis. Here we consider whether specific features of fungi could also favour infections. We checked whether there are differences in lipid profiles between strains obtained from dogs with otitis externa and strains obtained from healthy dogs. Lipid profiles were determined using thin layer chromatography and gas chromatography-mass spectrometry. All analyses were carried out on 32 strains derived from dogs with otitis externa and 31 strains isolated from dogs without symptoms of disease. The results show that strains isolated from dogs without symptoms of otitis externa are characterised by a higher content of fatty acids. They contain significantly more behenic and lignoceric acids on medium without addition of lipids, and more oleic acid and total monounsaturated fatty acids on medium with lipids supplementation. These strains have also a higher content of esters of ergosterol and triglycerides. Data obtained show differences which may be specific features of M. pachydermatis-specific strains related to the ability of infection, which could be not directly related of the host condition.


Asunto(s)
Dermatomicosis/veterinaria , Enfermedades de los Perros/microbiología , Lípidos/análisis , Malassezia/química , Malassezia/aislamiento & purificación , Otitis Externa/veterinaria , Animales , Dermatomicosis/microbiología , Perros , Ácidos Grasos/análisis , Otitis Externa/microbiología
11.
Mycoses ; 59(2): 108-16, 2016 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-26691773

RESUMEN

Severe skin diseases and systemic fungaemia are caused by Malassezia pachydermatis and Candida albicans respectively. Antifungal therapies are less effective because of chronic character of infections and high percentage of relapses. Therefore, there is a great need to develop new strategies of antifungal therapies. We previously found that oxythiamine decreases proliferation of yeast (Saccharomyces cerevisiae), therefore we suggest that thiamine antivitamins can be considered as antifungal agents. The aim of this study was the comparison of thiamine antivitamins (oxythiamine, amprolium, thiochrome, tetrahydrothiamine and tetrahydrooxythiamine) inhibitory effect on the growth rate and energetic metabolism efficiency in non-pathogenic S. cerevisiae and two potentially pathogenic species M. pachydermatis and C. albicans. Investigated species were cultured on a Sabouraud medium supplemented with trace elements in the presence (40 mg l(-1)) or absence of each tested antivitamins to estimate their influence on growth rate, enzyme activity and kinetic parameters of pyruvate decarboxylase and malate dehydrogenase of each tested species. Oxythiamine was the only antivitamin with antifungal potential. M. pachydermatis and S. cerevisiae were the most sensitive, whereas C. albicans was the least sensitive to oxythiamine action. Oxythiamine can be considered as supportive agent in superficial mycoses treatment, especially those caused by species from the genus Malassezia.


Asunto(s)
Antifúngicos/farmacología , Antimetabolitos/farmacología , Candida albicans/efectos de los fármacos , Candidiasis Cutánea/tratamiento farmacológico , Dermatomicosis/tratamiento farmacológico , Malassezia/efectos de los fármacos , Tiamina/antagonistas & inhibidores , Antifúngicos/uso terapéutico , Antimetabolitos/uso terapéutico , Candida albicans/crecimiento & desarrollo , Candidiasis Cutánea/microbiología , Dermatomicosis/microbiología , Fungemia/tratamiento farmacológico , Fungemia/microbiología , Humanos , Malassezia/crecimiento & desarrollo , Pruebas de Sensibilidad Microbiana , Oxitiamina/farmacología , Oxitiamina/uso terapéutico , Saccharomyces cerevisiae/efectos de los fármacos , Saccharomyces cerevisiae/crecimiento & desarrollo
12.
FEBS J ; 280(24): 6412-42, 2013 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-24004353

RESUMEN

Bringing a knowledge of enzymology into research in vivo and in situ is of great importance in understanding systems biology and metabolic regulation. The central metabolic significance of thiamin (vitamin B1 ) and its diphosphorylated derivative (thiamin diphosphate; ThDP), and the fundamental differences in the ThDP-dependent enzymes of metabolic networks in mammals versus plants, fungi and bacteria, or in health versus disease, suggest that these enzymes are promising targets for biotechnological and medical applications. Here, the in vivo action of known regulators of ThDP-dependent enzymes, such as synthetic structural analogs of the enzyme substrates and thiamin, is analyzed in light of the enzymological data accumulated during half a century of research. Mimicking the enzyme-specific catalytic intermediates, the phosphonate analogs of 2-oxo acids selectively inhibit particular ThDP-dependent enzymes. Because of their selectivity, use of these compounds in cellular and animal models of ThDP-dependent enzyme malfunctions improves the validity of the model and its predictive power when compared with the nonselective and enzymatically less characterized oxythiamin and pyrithiamin. In vitro studies of the interaction of thiamin analogs and their biological derivatives with potential in vivo targets are necessary to identify and attenuate the analog selectivity. For both the substrate and thiamin synthetic analogs, in vitro reactivities with potential targets are highly relevant in vivo. However, effective concentrations in vivo are often higher than in vitro studies would suggest. The significance of specific inihibition of the ThDP-dependent enzymes for the development of herbicides, antibiotics, anticancer and neuroprotective strategies is discussed.


Asunto(s)
Modelos Animales de Enfermedad , Diseño de Fármacos , Enzimas/metabolismo , Metabolismo/fisiología , Tiamina Pirofosfato/metabolismo , Animales , Enzimas/química , Humanos , Tiamina Pirofosfato/química
13.
J Physiol Biochem ; 68(3): 345-51, 2012 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-22274913

RESUMEN

Thyroid diseases are one of the most common metabolic disorders in the human population. In this work, we present data concerning changes in the activity and kinetic parameters of several enzymes associated with both anabolic (glucose-6-phosphate dehydrogenase-G6PDH, EC 1.1.1.49; 6-phosphogluconate dehydrogenase-6PGDH, EC 1.1.1.44; malic enzyme-ME, EC 1.1.1.40; and isocitrate dehydrogenase-IDH, EC 1.1.1.42) and catabolic (NAD-dependent malate dehydrogenase-NAD-MDH, EC 1.1.1.37; and lactate dehydrogenase-LDH, EC 1.1.1.27) processes under conditions of hypothyroidism and T(3) treatment. Hypothyroidism was induced in rats by the surgical removal of the thyroid gland. T(3)-treated rats were injected by T(3) (0.5 mg T(3)/kg body weight daily during 10 days). We have found that T(3) treatment caused an increase of NAD-MDH activity as well as heart hypertrophy whereas the activity of LDH increased in the direction of pyruvate reduction. Moreover, we observed increased activity of both enzymes in the liver. These results confirm earlier observation concerning the relevance of oxidative metabolism in the heart under T(3) treatment. Hypothyroidism resulted in changes in the LDH activity in the heart whereas NAD-MDH activity did not change. Moreover, our data show that T(3) treatment caused an increase of G6PDH, 6PGDH, and ME activities in the liver. We also observed a decrease of IDH activity in both organs, whereas hypothyroidism caused the opposite effect. This data indicate that either deficiency or excess of thyroid hormones can prove to be particularly dangerous for the physiology of the heart muscle by disturbing bioenergetic and anabolic processes.


Asunto(s)
Hipotiroidismo/metabolismo , Hígado/enzimología , Miocardio/enzimología , Triyodotironina/metabolismo , Animales , Glucosafosfato Deshidrogenasa/metabolismo , Corazón , Hipotiroidismo/enzimología , Cinética , L-Lactato Deshidrogenasa/metabolismo , Hígado/metabolismo , Masculino , Miocardio/metabolismo , Fosfogluconato Deshidrogenasa/metabolismo , Ratas , Ratas Wistar
14.
Mycoses ; 55(3): e106-13, 2012 May.
Artículo en Inglés | MEDLINE | ID: mdl-22066764

RESUMEN

Malassezia pachydermatis and Candida albicans are fungi involved in the skin diseases and systemic infections. The therapy of such infections is difficult due to relapses and problems with pathogen identification. In our study, we compare the fatty acids profile of M. pachydermatis, C. albicans and S. cerevisiae to identify diagnostic markers and to investigate the effect of oxythiamine (OT) on the lipid composition of these species. Total fatty acid content is threefold higher in C. albicans and M. pachydermatis compared with S. cerevisiae. These two species have also increased level of polyunsaturated fatty acids (PUFA) and decreased content of monounsaturated fatty acids (MUFA). We noted differences in the content of longer chain (>18) fatty acids between studied species (for example a lack of 20 : 1 in S. cerevisiae and 22 : 0 in M. pachydermatis and C. albicans). OT reduces total fatty acids content in M. pachydermatis by 50%. In S. cerevisiae, OT increased PUFA whereas it decreased MUFA content. In C. albicans, OT decreased PUFA and increased MUFA and SFA content. The results show that the MUFA to PUFA ratio and the fatty acid profile could be useful diagnostic tests to distinguish C. albicans, M. pachydermatis and S. cerevisiae, and OT affected the lipid metabolism of the investigated species, especially M. pachydermatis.


Asunto(s)
Candida albicans/metabolismo , Dermatomicosis/microbiología , Ácidos Grasos/metabolismo , Malassezia/metabolismo , Oxitiamina/farmacología , Saccharomyces cerevisiae/metabolismo , Candida albicans/química , Candida albicans/efectos de los fármacos , Ácidos Grasos/análisis , Humanos , Malassezia/química , Malassezia/efectos de los fármacos , Saccharomyces cerevisiae/química , Saccharomyces cerevisiae/efectos de los fármacos
15.
Postepy Hig Med Dosw (Online) ; 65: 447-69, 2011 Jul 06.
Artículo en Polaco | MEDLINE | ID: mdl-21734329

RESUMEN

For over 70 years thiamine (vitamin B1) has aroused the interest of biologists, biochemists and medical doctors because of its multilateral participation in key biochemical and physiological processes. The thiamine molecule is composed of pyrimidine and thiazole rings which are linked by a methylene bridge. It is synthesized by microorganisms, fungi and plants, whereas animals and humans have to obtain it from food. There are several known forms of vitamin B1 inside cells: free thiamine, three phosphate esters (mono-, di-, and triphosphate), and the recently found adenosine thiamine triphosphate. Thiamine has a dual, coenzymatic and non-coenzymatic role. First of all, it is a precursor of thiamin diphosphate, which is a coenzyme for over 20 characterized enzymes which are involved in cell bioenergetic processes leading to the synthesis of ATP. Moreover, these enzymes take part in the biosynthesis of pentose (required for the synthesis of nucleotides), amino acids and other organic compounds of cell metabolism. On the other hand, recent discoveries show the non-coenzymatic role of thiamine derivatives in the process of regulation of gene expression (riboswitches in microorganisms and plants), the stress response, and perhaps so far unknown signal transduction pathways associated with adverse environmental conditions, or transduction of nerve signals with participation of thiamine triphosphate and adenosine thiamine triphosphate. From the clinical point of view thiamine deficiency is related to beri-beri, Parkinson disease, Alzheimer disease, Wernicke-Korsakoff syndrome and other pathologies of the nervous system, and it is successfully applied in medical practice. On the other hand, identifying new synthetic analogues of thiamine which could be used as cytostatics, herbicides or agents preventing deficiency of vitamin B1 is currently the major goal of the research. In this paper we present the current state of knowledge of thiamine and its derivatives, indicating the participation of these compounds in the regulation of cell metabolism at both the coenzymatic and non-coenzymatic level.


Asunto(s)
Células/efectos de los fármacos , Células/metabolismo , Tiamina/metabolismo , Tiamina/farmacología , Humanos
16.
Postepy Biochem ; 57(3): 304-13, 2011.
Artículo en Polaco | MEDLINE | ID: mdl-22235656

RESUMEN

Enzymes control the course of biochemical reactions. The enzymes involved in bioenergetic processes play most important role in cell metabolism. One of them is 2-oxoglutarate dehydrogenase complex (OGDHC), the key regulatory enzyme of Krebs cycle. Krebs cycle integrates basic metabolic pathways of carbohydrates, fatty acids and amino acids during catabolic as well as anabolic reactions. Due to the key position of OGDHC in mitochondrial metabolism, its activity is controlled by many factors. Allosteric regulation by positive effectors (ADP, Pi, Ca2+, Mn2+) of the complex is very important. These effectors strongly enhances affinity of the first component of OGDHC to 2-oxoglutarate. Moreover there are negative effectors (ATP, NADH, succinyl-CoA) which affect all three enzymes of the complex. Regulation of biosynthesis of individual components of the complex by activation or inactivation of genes expression is very important for proper OGDHC activity too. Activity of OGDHC also depends on posttranslational modifications of its components. All of this control processes maintain OGDHC activity on adequate level and prevent the complex against its excessive action.


Asunto(s)
Ciclo del Ácido Cítrico/fisiología , Complejo Cetoglutarato Deshidrogenasa/metabolismo , Mitocondrias/metabolismo , Animales , Regulación de la Expresión Génica , Humanos
17.
Can J Microbiol ; 54(9): 734-41, 2008 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-18772936

RESUMEN

Candida albicans and Malassezia pachydermatis cause human and animal infections of the skin and internal organs. We compare the properties of two enzymes, pyruvate decarboxylase (PDC) and malate dehydrogenase (MDH), from these species and from Saccharomyces cerevisiae cultivated under aerobic and anaerobic conditions to find differences between the enzymes that adapt pathogens for virulence and help us in searching for new antifungal agents. Malassezia pachydermatis did not show any growth under anaerobic conditions, as opposed to C. albicans and S. cerevisiae. Under aerobic conditions, C. albicans showed the highest growth rate. Malassezia pachydermatis, contrary to the others, did not show any PDC activity, simultaneously showing the highest MDH activity under aerobic conditions and a Km value for oxaloacetate lower than S. cerevisiae. Candida albicans and S. cerevisiae showed a strong decrease in MDH activity under anaerobic conditions. Candida albicans shows four different isoforms of MDH, while M. pachydermatis and S. cerevisiae are characterized by two and three isoforms. Candida albicans shows about a twofold lower activity of PDC but, simultaneously, almost a threefold lower Km value for pyruvate in comparison with S. cerevisiae. The PDC apoform share under aerobic conditions in C. albicans was 47%, while in S. cerevisiae was only 26%; under anaerobic conditions, the PDC apoform decreased to 12% and 8%, respectively. The properties of enzymes from C. albicans show its high metabolic flexibility (contrary to M. pachydermatis) and cause easy switching between fermentative and oxidative metabolism. This feature allows C. albicans to cause both surface and deep infections. We take into consideration the use of thiamin antimetabolites as antifungal factors that can affect both oxidative and fermentative metabolism.


Asunto(s)
Proteínas Fúngicas/química , Proteínas Fúngicas/metabolismo , Hongos/enzimología , Malato Deshidrogenasa/química , Malato Deshidrogenasa/metabolismo , Piruvato Descarboxilasa/química , Piruvato Descarboxilasa/metabolismo , Aerobiosis , Anaerobiosis , Candida albicans/química , Candida albicans/enzimología , Candida albicans/genética , Proteínas Fúngicas/genética , Hongos/química , Hongos/genética , Humanos , Cinética , Malassezia/química , Malassezia/enzimología , Malassezia/genética , Malato Deshidrogenasa/genética , Micosis/microbiología , Piruvato Descarboxilasa/genética , Saccharomyces cerevisiae/química , Saccharomyces cerevisiae/enzimología , Saccharomyces cerevisiae/genética
18.
Can J Microbiol ; 51(10): 833-9, 2005 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-16333342

RESUMEN

Oxythiamine is an antivitamin derivative of thiamine that after phosphorylation to oxythiamine pyro phosphate can bind to the active centres of thiamine-dependent enzymes. In the present study, the effect of oxythiamine on the viability of Saccharomyces cerevisiae and the activity of thiamine pyrophosphate dependent enzymes in yeast cells has been investigated. We observed a decrease in pyruvate decarboxylase specific activity on both a control and an oxythiamine medium after the first 6 h of culture. The cytosolic enzymes transketolase and pyruvate decarboxylase decreased their specific activity in the presence of oxythiamine but only during the beginning of the cultivation. However, after 12 h of cultivation, oxythiamine-treated cells showed higher specific activity of cytosolic enzymes. More over, it was established by SDS-PAGE that the high specific activity of pyruvate decarboxylase was followed by an increase in the amount of the enzyme protein. In contrast, the mitochondrial enzymes, pyruvate dehydrogenase and 2-oxoglutarate dehydrogenase complexes, were inhibited by oxythiamine during the entire experiment. Our results suggest that the observed strong decrease in growth rate and viability of yeast on medium with oxythiamine may be due to stronger inhibition of mitochondrial pyruvate dehydrogenase than of cytosolic enzymes.


Asunto(s)
Antimetabolitos/farmacología , Oxitiamina/farmacología , Saccharomyces cerevisiae/enzimología , Saccharomyces cerevisiae/crecimiento & desarrollo , Tiamina Pirofosfato/metabolismo , Recuento de Colonia Microbiana , Medios de Cultivo , Citosol/enzimología , Complejo Cetoglutarato Deshidrogenasa/efectos de los fármacos , Complejo Cetoglutarato Deshidrogenasa/metabolismo , Mitocondrias/enzimología , Piruvato Descarboxilasa/efectos de los fármacos , Piruvato Descarboxilasa/metabolismo , Complejo Piruvato Deshidrogenasa/efectos de los fármacos , Complejo Piruvato Deshidrogenasa/metabolismo , Saccharomyces cerevisiae/efectos de los fármacos , Transcetolasa/efectos de los fármacos , Transcetolasa/metabolismo
19.
Ital J Biochem ; 53(4): 131-4, 2004 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-15997904

RESUMEN

Earlier it was noted that purified pyruvate dehydrogenase complex (PDC) produced by "Sigma" usually contains almost saturating amounts of thiamine pyrophosphate (ThPP). In this communication we present the observation that the endogenous ThPP coupled to PDC is dephosphorylated while staying at -10 degrees C, because in the enzyme preparation thiamine monophosphate and un-phosphorylated thiamine appear (HPLC determination). Under the same conditions exogenous ThPP is not dephosphorylated despite contact with the PDC preparation. This may suggest that interactions of some active groups of the enzyme with molecules of endogenous ThPP leads to break-up of the phosphoesters bonds, and destruction of the coenzyme. Decrease of PDC activity during storage is not in proportion with the degree of ThPP dephosphorylation. However the observed instability of PDC activity may be a consequence of the spontaneous process of its coenzyme autodestruction.


Asunto(s)
Complejo Piruvato Deshidrogenasa/metabolismo , Complejo Piruvato Deshidrogenasa/normas , Tiamina Pirofosfato/química , Tiamina Pirofosfato/metabolismo , Animales , Almacenaje de Medicamentos , Congelación , Miocardio/enzimología , Porcinos , Tiamina Monofosfato/análisis
20.
J Basic Microbiol ; 43(6): 522-9, 2003.
Artículo en Inglés | MEDLINE | ID: mdl-14625902

RESUMEN

Oxythiamin is one of the antivitamin derivatives of thiamin which, after phosphorylation, can be bound to the catalytic centre of thiamin-dependent enzymes and inhibit these enzymes. In this work the influence of oxythiamin on the growth rate, survival and the activity of pyruvate decarboxylase of Saccharomyces cerevisiae (s288c) was investigated. Oxythiamin decreased both the growth rate and survival ability of yeast cells. Moreover, in three-day-old cultures on a medium with oxythiamin, an increase of pyruvate decarboxylase activity was observed. This unusual effect may be in response to the earlier inhibition of pyruvate decarboxylase. A high concentration of pyruvate in the cell extracts taken from the medium with oxythiamin was found. This accumulation of pyruvate could provide for enhanced biosynthesis of the pyruvate decarboxylase apoform and an increase of enzyme activity.


Asunto(s)
Oxitiamina/farmacología , Piruvato Descarboxilasa/metabolismo , Saccharomyces cerevisiae/efectos de los fármacos , Antimetabolitos/farmacología , Piruvato Descarboxilasa/biosíntesis , Ácido Pirúvico/metabolismo , Saccharomyces cerevisiae/química , Saccharomyces cerevisiae/enzimología , Saccharomyces cerevisiae/crecimiento & desarrollo
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