RESUMO
On the basis of recent observations it is supposed that seminal fluids may contain--mainly in hydroxymethyl groups--formaldehyde (HCHO) and quaternary ammonium compounds as potential HCHO generators, therefore, preliminary investigations were carried out for the identification of these compounds in pig seminal fluids using OPLC, HPLC and MALDI MS techniques. The fresh pig seminal fluid was frozen in liquid nitrogen, powdered and aliquots (0.25 g) were treated with 0.7 ml ethanolic dimedone solution. The suspension was centrifuged and the clear supernatant was used for analysis by OPLC or after dilution with HPLC or MALDI MS technique. After OPLC separation of formaldemethone the fully N-methylated compounds which are stayed on the start point were separated by OPLC using an other eluent system. It has been established that the HCHO is really a normal component of the pig seminal fluid, as well. It can be isolated and identified in dimedone adduct form. The measurable amount of HCHO depended on the concentration applied of dimedone. According to OPLC and MALDI MS investigations L-carnitine is the main quaternary ammonium compound in pig seminal fluid which can generate a protection of the sperm cells against environmental and other influences. Considerable differences have been found among individuals concerning concentrations of quaternary ammonium compounds in the seminal fluid of pigs.
Assuntos
Alcaloides/análise , Carnitina/análise , Formaldeído/análise , Sêmen/química , Suínos , Animais , Cromatografia Líquida de Alta Pressão , Masculino , Metilação , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por MatrizRESUMO
The effect of heat shock (40 degrees C) on the formaldehyde cycle has been studied in European Turkey oak (Quercus cerris L.) acorns germinated to a 10% increase in mass. Hydroxy-methyl groups bonded to sulfur, oxygen and nitrogen atoms were made to react with dimedone and the derivative obtained (formaldemethone), which represented the endogenous formaldehyde level, was determined by high performance liquid chromatography. Qualitative alterations of methyl donors and acceptors in the response of acorns to the heat shock have been mapped by MALDI (matrix assisted laser desorption ionization) mass analysis. In the first experiment the acorns were prevented from withering by wrapping them in aluminium foil and in the second they were not. The relatively high temperature of the acorns wrapped in aluminium foil was the dominant stress effect and the role of withering was subsidiary. Alteration of the endogenous formaldehyde level in the seed-leaves reflected the phases of the stress syndrome. If the withering were not hindered, two local minima in the alteration of endogenous formaldehyde level were found. First, the increase in temperature decreased the endogenous formaldehyde level and after a local maximum a repeated local minimum was observed as a delayed response. It is presumed that the second minimum was induced by the decreasing water amount becoming more and more significant in the seed-leaves.
Assuntos
Formaldeído/metabolismo , Resposta ao Choque Térmico , Árvores/embriologia , Cromatografia Líquida de Alta Pressão , Sementes , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz , Espectrofotometria UltravioletaRESUMO
Acorns of Quercus cerris L., after saturation with water and storage at -20 degrees C, were studied for changes in their contents of endogenous formaldehyde and its potential precursor and generator compounds. For the measurement of formaldehyde, after conversion to formaldemethone and some methyl acceptor and donor substances, high performance liquid chromatography (HPLC) were used. First, the amount of formaldehyde was drastically decreased. Having reached a minimum value within three to five days of the beginning of low temperature storage, a higher steady-state than the control acorns was recorded. Trigonelline and gamma-amino-butyric acid in seedleaf extracts were identified by matrix assisted laser desorption-ionization mass spectrometry (MALDI-MS).
Assuntos
Temperatura Baixa , Formaldeído/metabolismo , Germinação , Árvores/metabolismo , Cromatografia Líquida de Alta Pressão , Sementes , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz , Árvores/fisiologiaRESUMO
The effect of mercury accumulation on the formaldehyde cycle of different pig tissues resulted by a single dose of mercury (0.4 mg mercury in Hg(II)-chloride form, 500 kBq Hg-203/animal) has been studied. Daily mercury excretion was tracked, and having reached the steady-state mercury level of the body (10th day), samples were taken from the liver, kidney and muscle (musculus longissimus dorsi). After reaction with dimedone the endogenous formaldehyde levels in the samples were measured by high performance liquid chromatography. Our results show that the endogenous formaldehyde level decreased by more than 50% in the liver and the kidney, where the average mercury accumulation was the highest (1017 Bq/100 g and 625 Bq/100 g, respectively). In contrast, the muscle tissues, with a low mercury level (139 Bq/100 g), responded to the stress effect by about a 30% increase in their endogenous formaldehyde level.
Assuntos
Formaldeído/metabolismo , Cloreto de Mercúrio/metabolismo , Animais , Rim/metabolismo , Fígado/metabolismo , Masculino , Cloreto de Mercúrio/farmacocinética , Músculos/metabolismo , Suínos , Distribuição TecidualRESUMO
S-adenosyl-L-methionine serves as a methyl donor in virtually all of the vast number of enzymatic transmethylation reactions including DNA methylation. On the basis of our former experiences we questioned the formation of a methyl cation or methyl radical in the enzymatic transmethylation reactions. The formation of formaldehyde from the methyl moiety of S-adenosyl-L-methionine has been demonstrated. It became increasingly evident that there is a formaldehyde cycle in biological systems in which the formation of the methyl group of L-methionine takes place through formaldehyde and the formation of formaldehyde from S-adenosyl-L-methionine is linked to different enzymatic transmethylation reactions. It is also known that during demethylation processes both formaldehyde and demethylated compound can be formed. The abnormalities of the originally controlled formaldehyde cycle and the uncontrolled enzymatic production of formaldehyde from endogenous and/or exogenous substrates may be potential risk factors in pathogenesis of different disorders. The formaldehyde generator and capturer molecules may potentially normalise these abnormal processes. Trans-resveratrol (trans-3,5,4'-trihydroxystilbene), which is as phytoalexin, occurs naturally in grapes and a variety of medicinal plants. According to our present observations it is a natural concentration-dependent formaldehyde capture molecule. It would seem that elimination of the uncontrolled formaldehyde with resveratrol may exert a double effect in biological systems. The elimination of formaldehyde with resveratrol (first step) may cause a cardioprotective effect and the reaction products between resveratrol and formaldehyde (second step) may act as a chemopreventive factor against cancer.
Assuntos
Formaldeído/química , Formaldeído/metabolismo , Metionina/biossíntese , Metilação , Modelos QuímicosRESUMO
The phytoalexin resveratrol (3,5,4'-trihydroxy-trans-stilbene) and formaldehyde (as its dimedone adduct, formaldemethone) have been identified and measured in the extracts of parts of white and blue grapes as well as in white and red wines by overpressured layer chromatography (OPLC), high performance liquid chromatography (HPLC) and from matrix assisted laser desorption/ionization mass spectrometric data. It has been established that the level of resveratrol was very high in skin and in some cases in the stem. Blue grape varieties and red wines always contained a considerably higher amount of resveratrol than white grapes and wines. The measurable level of formaldehyde as well as the resveratrol content was always parallelly high in the same parts of the berries, however, the formaldehyde level was higher in white grapes than in blue ones. The simultaneous occurrence of resveratrol and formaldehyde gives a possibility for interaction between these two special molecules, consequently, hydroxymethyl derivatives of resveratrol can be formed. These resveratrol derivatives may be responsible for special biological activities of resveratrol in grapes and dietetically (cardioprotective effect and chemopreventive effect against cancer) in the human organism.
Assuntos
Formaldeído/análise , Rosales/química , Estilbenos/análise , Cromatografia Líquida de Alta Pressão , Resveratrol , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz , Espectrofotometria UltravioletaRESUMO
Taking into consideration the unquestionable intracellular occurrence of formaldehyde (HCHO) and its generators in cells of plant, animal and human organisms as well as in body fluids it was resolved to determine their levels in hard tissues of physiologically and pathologically changed teeth. The aim of the work was to determine the relationship between the level of HCHO and the levels of its generators in pathologically changed teeth, mainly carietic teeth as tooth caries is still a serious and commonly occurring problem. The occurrence of HCHO (captured as its dimedone adduct) and some of its potential generators was demonstrated in the hard tissues of healthy and pathological human teeth by means of OPLC, HPLC and MS analyses. It was established that the measurable level of HCHO was increased in the carietic teeth in comparison with healthy ones. In the case of paradontic tooth sample, a dramatic increase of HCHO was observed and at the same time the level of betaines was decreased considerably. The obtained results give a new insight into the pathology of hard tissues of teeth in strong correlation with the phases of stress syndrome.