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Biomedical subjects

M A Ferreira

Publications and source records attributed to M A Ferreira.

At least 19 recordsLinked to original sources

Analysis of heterocyclic aromatic amines in foods by gas chromatography-mass spectrometry as their tert.-butyldimethylsilyl derivatives.

A derivatization method for the analysis of 12 heterocyclic aromatic amines (HAs) in food, by gas chromatography-electron impact mass spectrometry, was developed. The amines are derivatized in a one-step reaction with N-methyl-N-(tert.-butyldimethylsilyl)trifluoroacetamide. The derivatives are characterized by easy-to-interpret mass spectra due to the prominent ion [M-57]+ by loss of a tert.-butyldimethylsilyl group, allowing quantification in the selected-ion monitoring mode at the picogram level. The effect of temperature, time, and reagents on the formation of the derivatives was monitored in detail. Quality parameters were evaluated in the optimum working conditions. This derivatization method is not applicable to the pyridoimidazoles Glu-P-1 and Glu-P-2 and to the beta-carboline harman due to incompletely derivatization. The instability of the imidazolquinoline and imidazoquinoxaline derivatives, requiring their injection on the same working day, is a further drawback. This simple, rapid and accurate derivatization procedure is suitable for routine analysis, as illustrated by the analysis of some common foods.

Amines↗

Quantification of short-chain free fatty acids in "Terrincho" ewe cheese: intravarietal comparison.

Headspace solid-phase microextraction coupled with gas chromatography-mass spectrometry was employed for the quantification of volatile free fatty acids (FFA) in "Terrincho" ewe cheese. Solid-phase microextraction quantitative analysis was feasible under equilibrium situations as long as the conditions of agitation and the adsorption time were held constant. An excellent linear relationship between the amount of the adsorbed analyte and its initial concentration in the sample matrix was obtained when an adequate amount of sample was chosen. Thus, quantification was possible if biases due to competition or linear range excesses were controlled. Solid-phase microextraction sampling was carried out at 65 degrees C, and a fiber coated with an 85-micro/m polyacrylate film was chosen. After equilibration at 65 degrees C for 40 min, the fiber was exposed to the headspace above the sample for 20 min and then inserted into the gas chromatograph. The evolution of the volatile FFA during Terrincho ewe cheese ripening was analyzed for a 60-d period. An overall increase in FFA contents was verified up to 30 d of ripening. Between 30 and 45 d most FFA did not suffer significant changes. All FFA increased significantly by the 60-d ripening period. The excessive lipolysis observed at 60 d of ripening may result in the presence of off-flavors. Principal component analysis performed for intravarietal comparison of volatile FFA composition of 19 Terrincho cheeses, analyzed at 30 ripening days, enabled discrimination between cheeses produced at five different dairy plants.

Animals↗

Gas chromatographic-mass spectrometric quantification of 4-(5-)methylimidazole in roasted coffee after ion-pair extraction.

A GC-MS method is described for quantification of 4-(5-)methylimidazole (4MI) in coffee. Although tested, GC-flame ionization detection proved inadequate for this purpose due to the complexity of the coffee matrix. The developed method was based on ion-pair extraction with bis-2-ethylhexylphosphate and derivatization with isobutylchloroformate. Quantification was carried out by the standard addition method using 2-ethylimidazole as internal standard. Reproducibility data from the complete procedure are presented. Mean recoveries were higher than 98%. The method was applied to green and roasted coffee samples from the two most important varieties, arabica and robusta, and to commercial "torrefacto" coffee blends. 4MI was not detected in the green coffee samples analysed and ranged from 0.307 to 1.241 mg/kg in roasted samples.

Coffee↗

Solid-phase microextraction in combination with GC/MS for quantification of the major volatile free fatty acids in ewe cheese.

This work describes a method for quantification of the major free fatty acids of ewe cheese that contribute to its distinct and strongly marked flavor. A headspace SPME method in combination with GC/MS was used for the extraction, identification, and quantification of butanoic, hexanoic, octanoic and decanoic acids in ewe cheeses. The method used for sample preparation was simple. A fiber coated with 85-microm polyacrylate film was chosen to extract the free fatty acids. To perform a reliable quantification, several factors were taken into consideration for reliable quantification, namely, (i) the influence of addition of water, of an electrolyte or of a hygroscopic salt, on the release of free fatty acids from the matrix; (ii) the linear relationship between the amount of analyte adsorbed by the SPME polymer film and the initial concentration of the analyte in the cheese sample; and (iii) the competition for adsorption by fiber. Water removal with sodium sulfate promoted a more efficient extraction of volatile free fatty acids; biases due to competition or linear range excesses were controlled by choosing the appropriate amount of sample for each ewe cheese. The method of standard additions was used with success for the quantification of free fatty acids. Calibration curves that were constructed for the major short-chain free fatty acids (butanoic, hexanoic, octanoic, and decanoic acids) spiked into cheese followed linear relationships with highly significant (p < 0.001) correlation coefficients (r > 0.999). Coefficients of variation of <7.9% indicated that the technique was reproducible. A marked increase in concentration of short-chain free fatty acids was observed during cheese ripening, ranging from 0.35 to 9.33 mg/100 g for butanoic acid, 0.363 to 4.34 mg/100 g for hexanoic acid, 0.343 to 2.0 mg/100 g for octanoic acid, and 1.291 to 3.85 mg/100 g for decanoic acid. The limits of quantification were registered at levels of parts per million. The absolute quantification of butanoic acid was also carried out by using isotope dilution assays (IDA). The levels of acid obtained with this method were similar to those obtained by the standard additions method.

Animals↗

Morphological and ultrastructural changes occurring during degeneration of goat preantral follicles preserved in vitro.

The present work has investigated the morphological and ultrastructural changes occurring during degeneration of goat preantral follicles preserved in vitro and showed quantitative data about the distribution of follicular degeneration types in the control and after preservation in coconut water solution or Braun-Collins solution at different temperatures (4, 20 or 39 degrees C) and incubation times (4, 12 or 24h). At the slaughterhouse, the pair of ovaries of each animal was divided into 19 fragments. One ovarian fragment was immediately fixed (control: Time 0). The other 18 fragments were randomly distributed in tubes containing 2ml of coconut water or Braun-Collins solution at 4, 20 or 39 degrees C and stored for 4, 12 or 24h. Normal preantral follicles exhibited a healthy oocyte surrounded by one or more well-organized layers of granulosa cells. The ooplasm contained numerous rounded or elongated mitochondria with continuous mitochondrial membranes. Golgi complexes were rare. Both smooth and rough endoplasmic reticulum were observed, either as isolated aggregations or complex associations with mitochondria and vesicles. Degenerated preantral follicles in the control tissue exhibited pycnotic nuclei of the oocyte, vacuolated ooplasm and normal granulosa cells. This kind of degeneration also predominated significantly (P<0.05) after preservation at 4 degrees C. In contrast, after preservation at 20 or 39 degrees C a significant predominance (P<0.05) of preantral follicles showing a retracted oocyte and swollen granulosa cells was observed. These follicles showed large irregularity of the oocyte and nuclear outlines. The ooplasm exhibited moderate proliferation of the endoplasmic reticulum and mitochondria showed disappearance of most of the cristae and damage to the mitochondrial membrane. Some follicles had numerous vacuoles in the ooplasm. Granulosa cells were spread and a low density of organelles was observed. The alterations in follicular structure progressed with an increase of temperature from 20 to 39 degrees C as well as with an increase of the incubation time from 4 to 12, or 24h. In conclusion, the present study shows for the first time that initial proliferation of the endoplasmic reticulum and damage to mitochondria are the first signs of degeneration in goat preantral follicles during storage in vitro.

Animals↗

Effects of storage time and temperature on atresia of goat ovarian preantral follicles held in M199 with or without indole-3-acetic acid supplementation.

Maintenance of follicular quality after removal and during transport of ovaries is necessary for studies on development of preantral follicles in vitro. The present work investigated the effectiveness of M199 and M199IAA for preservation of goat preantral follicles in ovarian tissue. At the slaughterhouse, the ovarian pair of each animal was divided into 19 fragments. One ovarian fragment was immediately fixed (control--Time 0). The other 18 fragments were randomly distributed in M199 or M1991AA at 4, 20 or 39 degrees C and stored for 4, 12 or 24 h. Histological analysis showed that storage of ovarian fragments in either solution at 20 or 39 degrees C significantly reduced the percentage of normal preantral follicles when compared with the control, in all cases except after preservation in M199IAA at 20 degrees C for 4 h. In contrast, preservation at 4 degrees C, in either solution, kept the percentage of normal preantral follicles at control values. Reduced cellular metabolism may explain why the best preservation of preantral follicles was at 4 degrees C. The addition of IAA to the TCM 199 was effective for goat preantral follicle preservation at 20 degrees C for 4 h.

Animals↗

Analysis of phenolic compounds in Spanish Albrariño and Portuguese Alvarinho and Loureiro wines by capillary zone electrophoresis and high-performance liquid chromatography.

The concentration of different phenolic compounds was measured in Spanish Albariño and Portuguese Alvarinho and Loureiro white wines by capillary zone electrophoresis (CZE), in order to characterize them. Although all samples presented the same qualitative pattern (characterized by tyrosol; (-)-epicatechin; syringic acid; ferulic acid; p-coumaric acid; caffeic acid, gallic acid; 3,4-dihydroxybenzoic acid; cis-coumaroyl tartaric acid (COUTA); trans-COUTA; trans-caffeoyl tartaric acid (CAFTA), and hydroxycinnamic esters), some quantitative differences were observed. When samples were analyzed by high-performance liquid chromatography (HPLC), in order to compare the results obtained by both techniques, no significant qualitative or quantitative differences were obtained. Nevertheless, CZE proved to be a more convenient technique for the routinary analyses of these wines, due to better separation of the different compounds, better peak shapes, and higher speed than HPLC.

Chromatography, High Pressure Liquid↗

Structural characterisation of flavonoids and flavonoid-O-glycosides extracted from Genista tenera by fast-atom bombardment tandem mass spectrometry.

Genista tenera is a plant native to the Madeira Island (Portugal). From the ethanol extract of its powdered aerial parts, two flavones, three isoflavones and one 7-O-glucosyl isoflavone were isolated. A mass spectrometric study of these compounds was performed using liquid secondary ion mass spectrometry (LSIMS) in combination with high-energy collision-induced dissociation (CID) and tandem mass spectrometry (MS/MS). Characteristic fragmentation patterns were observed in all the investigated compounds; the loss of small neutral species from the protonated molecules was useful for identifying the presence of specific functional groups in the A- and B-rings. In order to help to establish the proposed structures, NMR and UV studies were also performed.

Flavonoids↗

Effects of phenolic propyl esters on the oxidative stability of refined sunflower oil.

The oxidative stability of refined sunflower oil in the presence and in the absence of propyl caffeate (PC), propyl hydrocaffeate (PHC), propyl ferulate (PF), and propyl isoferulate (PI) has been evaluated according to the Rancimat method. The antioxidant activity of the phenolic derivatives was compared with that obtained with native [alpha-tocopherol (alpha-TOH)] and synthetic [propyl gallate (PG)] antioxidants. The results allow the establishment of a decreasing order of antioxidant power: PG > PHC > PC >> alpha-TOH > PI > PF. The oxidative stability was improved neither by the addition of PF nor by a supplement of alpha-TOH. Moreover, a positive antioxidant effect was obtained for PC that was placed between those of alpha-TOH and PG. The antioxidant activity of PHC was higher than that of its analogue (PC). A dose-dependent effect was observed for PG, PHC, and PC. A chain-breaking mechanism was proposed for the antioxidant activity of propyl phenolic esters because the same ranking order of efficacy was obtained for their antiradical activities evaluated by using the 2,2-diphenyl-1-picrylhydrazyl radical method.

Antioxidants↗

Differential effects of thalidomide on angiogenesis and tumor growth in mice.

Thalidomide, clinically used as an antiinflammatory and antitumoral drug, inhibited sponge-induced angiogenesis when administered systemically (100 mg/kg(-1)) in mice. However, it failed to inhibit solid Ehrlich tumor in the same mouse strain. We have used functional, biochemical and histological parameters to assess neovascularization and fibrovascular tissue infiltration of the mice sponge granuloma. The neovascularization growth as detected by development of blood flow and hemoglobin content extracted from the implants showed that thalidomide inhibited fibrovascular tissue formation by 40%. The functional and biochemical parameters correlated well with the histological study. Thalidomide had no inhibitory effect in the development of Ehrlich tumor. The detection of this selective action using the same animal strain bearing two different processes, supports the hypothesis that rather than species specificity, thalidomide is tissue specific. This approach may be used to identify the specificity of other therapeutic agents against distinct angiogenesis-dependent diseases.

Angiogenesis Inhibitors↗

Renal cell carcinoma and malignant phase hypertension.

Many cross-sectional and follow-up studies of large numbers of patients with hypertension have demonstrated an increased prevalence and mortality from renal cancer. We report the details of three patients with renal cell carcinoma from a series of 254 consecutive patients with malignant phase hypertension, an excess over the expected number reported from several large published series with non-malignant hypertension. In view of this excess we investigated the prevalence of hypertension in a series of 192 consecutive patients who presented with a diagnosis of renal cell carcinoma, in comparison with a local unselected population screening survey. Hypertension was found in 43% of the renal carcinoma patients and 20% of the local population, also a clear excess. The mechanism of the association between renal cancer and malignant and non-malignant hypertension is unclear.

Adult↗

Prognostic value of simple measurement of chronic damage in renal biopsy specimens.

BACKGROUND: A simple method of measurement of chronic damage in renal biopsy specimens would be useful in clinical management, prognosis, comparisons between different centres and trials. METHODS: An interactive image analysis system was used to outline and measure areas of chronic damage in 247 renal biopsy specimens to give an index of chronic damage, expressed as a percentage of cortical cross-sectional area. Prognostic value was analysed by the Kaplan-Meier method to study time between biopsy and onset of permanent dialysis. RESULTS: There was no significant bias between measurements by the same observer or different observers. The index of chronic damage ranged from 0 to 90%. Increasing severity of chronic damage was associated with shortened renal survival. Each increase of 10% in the index increased the hazard ratio of risk of permanent dialysis by 1.5 times (95% confidence interval 1.4-1.7, P<0.001). CONCLUSIONS: A simple measure of chronic damage was a powerful indicator of prognosis. This is likely to be clinically useful in routine practice and trials.

Biopsy↗

Evidence of type II pneumocyte apoptosis in the pathogenesis of idiopathic pulmonary fibrosis (IFP)/usual interstitial pneumonia (UIP).

BACKGROUND/AIMS: The pathogenesis of idiopathic pulmonary fibrosis (IPF)/usual interstitial pneumonia (UIP), a chronic and incurable human respiratory disease, is not well established. This study was designed to investigate whether the apoptosis of type II pneumocytes could be the precipitating factor in the pathogenesis of IPF. METHODS: Nineteen specimens obtained by retrospective review of the medical and pathological records of 55 patients with IPF, four normal subjects, and 10 disease control lungs were analysed. The selected specimens had normal alveoli with intervening patchy scarring of the lung parenchyma, fulfilling the pathological criteria for UIP. To identify individual cells undergoing apoptosis in the normal alveoli, electron microscopy and in situ end labelling of fragmented DNA were performed on paraffin was embedded sections using digoxigenin-11-dUTP and the enzyme terminal deoxynucleotidyl transferase. RESULTS: Apoptosis was detected in the normal alveoli of 17 of the 19 patients with IPF/UIP and was absent in the controls. Electron microscopy demonstrated apoptotic changes in type II pneumocytes. These results indicate that apoptotic type II pneumocyte death occurs in normal alveoli of IPF/UIP and could be the principal cause of several events that account for the histological, clinical, and functional alterations seen in IPF/UIP. CONCLUSIONS: In conclusion, numerous type II pneumocytes from the normal alveoli of most patients with IPF/UIP actively undergo programmed cell death. This finding may shed new light on the pathogenesis of this disease, with implications mainly for the treatment of affected patients.

Aged↗

Bilateral endogenous endophthalmitis associated with infective endocarditis: case report.

Bilateral endogenous endophthalmitis is a rare condition initiated by infection by microbes in the bloodstream, such as those arising from a foci of infective endocarditis. We report a case and discuss the diagnostic aspects and the clinical outcome of a patient with characteristic findings of the disease. The patient was a 49 year old white male who had a metallic aortic valve implanted 7 months previously, and who presented to the hospital with 10 days of fever, cough and dyspnea, then diarrhea and mental confusion. On the second day of hospitalization, he experienced sudden loss of vision in both eyes. A Gram-positive coccobacillus was isolated from the bloodstream, he was treated with fluoroquinolone with disappearance of fever, decreased ocular inflammation, and improvement in his vision to light perception. He later underwent valve replacement surgery but died during the procedure. We review the occurrence of ocular signs and symptoms and their importance in patients with endocarditis.

Blood↗

HPLC/UV analysis of proteins in dairy products using a hydrophobic interaction chromatographic column.

High-performance liquid chromatography using a Chrompack P-300-RP column containing a polystyrene-divinylbenzene copolymer-based packing was examined to analyze bovine milk protein components. The separation of major raw-milk proteins could be performed rapidly and reliably with this HPLC/UV method. The determinations were performed in the linear ranges of 0.01-2.0 mg/ml for alpha-lactalbumin, 0.04-2.5 mg/ml for caseins and 0.02-2.0 mg/ml for beta-lactoglobulin. The validity of the method was verified. Since the chromatographic column enabled the quantification of only "native" milk proteins, the extent of denaturation and loss of milk proteins could be examined. Thus, evaluation of heat-induced proteins denaturation was carried out in raw milk heated for 5 min at pre-determined temperatures.

Animals↗

[The most common conditions in a neurology specialty clinic].

OBJECTIVE: To present the most frequent diagnosis among patients referred for neurological evaluation to estimate their labour capacities at the unit of National Institute of Social Security (INSS), Florianópolis-SC, southern Brazil. METHOD: Review of all medical records of 108 patients evaluated between October 97 and May 98. The sample was submitted to judicious medico-legal assessment to define their final diagnosis. RESULTS: Neurological evaluation disclosed as the commonest disorders, in decreasing order of frequency: epilepsy, rheumatic diseases, psychiatric illnesses, neurological disorders related to chronic alcoholism, head trauma syndrome and cerebrovascular diseases. CONCLUSION: Neurological disorders may be responsible for important disability among workers in our society. However, the potential for social rehabilitation, often underestimated, must be considered. Moreover, diverse non-neurological conditions used to be sent for neurological evaluation.

Ambulatory Care↗

Combined ion-pair extraction and gas chromatography-mass spectrometry for the simultaneous determination of diamines, polyamines and aromatic amines in Port wine and grape juice.

An accurate and very sensitive method which allows for the simultaneous determination of the diamines (1,3-diaminopropane, putrescine and cadaverine), of the polyamines (spermidine and spermine), and of the aromatic amines (beta-phenylethylamine and tyramine) found in Port wines and corresponding grape juices is presented. Sample clean-up consisted of the extraction of the amines with the ion-pairing reagent bis-2-ethylhexylphosphate dissolved in chloroform followed by a back-extraction with 0.1 M HCl. The hydrochloric extract obtained was dried and the amines were further derivatized with heptafluorobutyric anhydride and analyzed by GC-MS in the selected ion-monitoring mode, with a total run time of 18 min. Under the adopted conditions, the extraction of all the studied compounds was almost complete and the obtained extracts were free of potential interferents present in the samples, namely sugars, and most of the amino acids and polyphenols. Via the use of a set of five selected internal standards (amphetamine, [2H8]putrescine, 1,7-diaminoheptane, norspermidine and norspermine), the data obtained from the linearity, repeatability and recovery experiments were very good for all the compounds assayed. The corresponding limits of detection were invariably below 10 microg l(-1). The method was successfully applied to measure the content of biogenic amines in twelve young and five aged Port wine samples, eleven grape juice samples as well as in ten Portuguese red and white table wines. Results are presented and briefly discussed.

Amines↗