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Guimarães, D., M. L. Carvalho, V. Geraldes, I. Rocha, and J. P. Santos. "Study of lead accumulation in bones of Wistar rats by X-ray fluorescence analysis: aging effect." Metallomics 4 (2012): 66. AbstractWebsite

The accumulation of lead in several bones of Wistar rats with time was determined and compared Q3 for the different types of bones. Two groups were studied: a control group (n = 20), not exposed to lead and a contaminated group (n = 30), exposed to lead from birth, first indirectly through
mother’s milk, and then directly through a diet containing lead acetate in drinking water (0.2%). Rats age ranged from 1 to 11 months, with approximately 1 month intervals and each of the collections had 3 contaminated rats and 2 control rats. Iliac, femur, tibia–fibula and skull have been analysed by energy dispersive X-ray fluorescence technique (EDXRF). Samples of formaldehyde used to preserve the bone tissues were also analysed by Electrothermal Atomic Absorption (ETAAS), showing that there was no significant loss of lead from the tissue to the preservative. The bones mean lead concentration of exposed rats range from 100 to 300 mg g 1 while control rats never exceeded 10 mg g 1. Mean bone lead concentrations were compared and
the concentrations were higher in iliac, femur and tibia–fibula and after that skull. However, of all the concentrations in the different collections, only those in the skull were statistically Q4 significantly different (p o 0.05) from the other types of bones. Analysis of a radar chart also allowed us to say that these differences tend to diminish with age. The Spearman correlation test applied to mean lead concentrations showed strong and very strong positive correlations between
all different types of bones. This test also showed that mean lead concentrations in bones are negatively correlated with the age of the animals. This correlation is strong in iliac and femur and very strong in tibia–fibula and skull. It was also shown that the decrease of lead accumulation with age is made by three plateaus of accumulation,

Guimarães, Diana, Maria Luisa Carvalho, Vera Geraldes, Isabel Rocha, Luís Cerqueira Alves, and Jose Paulo Santos. "Lead in liver and kidney of exposed rats: Aging accumulation study." Journal of Trace Elements in Medicine and Biology 26 (2012): 285. AbstractWebsite

The concentration of lead in liver and kidneys of Wistar rats, fed with lead since fetal period in relation to their age and to a control group, was determined. A group of rats was exposed to lead acetate (n=30) in drinking water and the other group was exposed to normal water (n=20). Samples were collected from rats aging between 1 and 11 months and were analyzed by Energy Dispersive X-ray Fluorescence (EDXRF) without any chemical preparation. The EDXRF results were assessed by the PIXE (Proton Induced X-ray Emission) technique. The formaldehyde used to preserve the samples was also analyzed by ETAAS (Electro-Thermal Atomic Absorption Spectrometry) in order to verify if there was any loss of lead from the samples to the formaldehyde. We found that the loss was not significant (<2%). Concerning the mean values of the lead concentration measured in the contaminated soft tissues, in liver they range from 6 to 22μgg(-1), and in kidneys from 44 to 79μgg(-1). The control rats show, in general, values below the EDXRF detection limit (2μgg(-1)). The ratio kidney/liver ranges from 2 to 10 and is strongly positively correlated with the age of the animals. A Spearman correlation matrix to investigate the correlation between elemental concentrations and the dependence of these concentrations with age showed that there is a strong positive correlation with age for lead in the liver but not in the kidney. The correlation matrix showed also that the concentration of lead in these two soft tissues is not correlated. The lead accumulation in liver is made by different plateaus that strongly decrease with age. It was verified the existence of two levels of accumulation in kidney, not very highlighted, which might be indicative of a maximum accumulation level for lead in kidney.

Guimarães, D., J. P. Santos, M. L. Carvalho, G. Vale, H. M. Santos, V. Geraldes, I. Rocha, and J. L. Capelo. "Ultrasonic energy as a tool to overcome some drawbacks in the determination of lead in brain tissue and urine of rats." Talanta 86 (2011): 442-446. AbstractWebsite

An ultrasonic assisted solid–liquid extraction method was developed to determine the level of lead in the brain and urine of rats. Lead was determined by electrothermal atomic absorption spectrometry with longitudinal-Zeeman background correction. Several analytical drawbacks were addressed and overcome, namely small brain sample mass and the formation of precipitate in the urine samples. Utrasonication provided by an ultrasonic probe succeeded in extracting lead from brain samples. Furthermore, it was demonstrated that the formation of a precipitate lowered the lead content in the liquid phase of the urine. Lead was back extracted from the precipitate to the liquid phase with the aid of ultrasonic energy and acidifying the urine with 10% v/v nitric acid. A microwave-assisted acid digestion protocol was used to check the completeness of the lead extraction. The within bath and between bath precision was 5% (n&#xa0;=&#xa0;9) and 7% (n&#xa0;=&#xa0;3) respectively. The limit of quantification was 1.05&#xa0;μg&#xa0;g−1 for brain samples and 2.1&#xa0;μg&#xa0;L−1 for urine samples. A total of 6 samples of urine and 12 samples of brain from control rats and another 6 samples of urine and 12 samples of brain from rats fed with tap water rich in lead acetate were used in this research. Lead levels in brain and urine from exposed rats ranged from1.9&#xa0;±&#xa0;0.2&#xa0;μg&#xa0;g−1 to 3.5&#xa0;±&#xa0;0.2&#xa0;μg&#xa0;g−1 and from 752&#xa0;±&#xa0;56&#xa0;μg&#xa0;L−1 to 60.9&#xa0;±&#xa0;1.2&#xa0;mg&#xa0;L−1 respectively. Statistically significant differences of levels of lead in brain and urine were found between exposed and non exposed rats.

Guimarães, D., M. L. Carvalho, M. Becker, A. von Bohlen, V. Geraldes, I. Rocha, and J. P. Santos. "Lead concentration in feces and urine of exposed rats by X-ray Fluorescence and Electrothermal Atomic Absorption Spectrometry." X-Ray Spectrometry In press (2011). Abstract
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Guimarães, D., M. L. Carvalho, M. Becker, A. von Bohlen, V. Geraldes, I. Rocha, and J. P. Santos. "Lead concentration in feces and urine of exposed rats by x-ray fluorescence and electrothermal atomic absorption spectrometry." X-Ray Spectrometry 41 (2012): 80. AbstractWebsite

Measurements made in feces and urine of Wistar rats exposed to lead acetate (n = 20) in drinking water since the fetal period were compared with those obtained from a control group (n = 20) in order to assess the age influence on Pb excretion. The measurements were made in different collections of rats aging between 1 and 11 months. To determine the Pb content of the samples, total reflection X-ray fluorescence (TXRF) and electrothermal atomic absorption spectrometry (ETAAS) were used for the urine samples and energy dispersive X-ray fluorescence (EDXRF) was used for the feces.The results show high concentrations of Pb being eliminated from the organism by urine and feces in contaminated rats. Values vary from (600`140)mgl1 to (5 460`115)mgl1 in urine and from (4 500`300)mgg1 to (11 400`3 300)mgg1 in dry feces. The control rats show, in general, low lead concentrations or below detection limits. The fecal/urinary ratio was studied. It was shown to be about three to four orders of magnitude and positively correlated with time. It was verified in feces and urine that excretion decreases with the animal age and that this decrease is made by different levels of excretion. The excretions of Pb in urine and in feces are positively correlated.A good agreement was found between the results obtained with TXRF and ETAAS for urine samples. This work also stresses the suitability of these techniques in the study of Pb intoxication.

Guimarães, D., M. L. Carvalho, M. Becker, A. von Bohlen, V. Geraldes, I. Rocha, and J. P. Santos. "Lead concentration in feces and urine of exposed rats by x-ray fluorescence and electrothermal atomic absorption spectrometry." X-Ray Spectrom. 41 (2012): 80. Abstract

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Guimarães, Diana, Maria Luisa Carvalho, Vera Geraldes, Isabel Rocha, Luís Cerqueira Alves, and Jose Paulo Santos. "Lead in liver and kidney of exposed rats: Aging accumulation study." J. Trace Elem. Med Biol. 26 (2012): 285. AbstractWebsite

The concentration of lead in liver and kidneys of Wistar rats, fed with lead since fetal period in relation to their age and to a control group, was determined. A group of rats was exposed to lead acetate (n=30) in drinking water and the other group was exposed to normal water (n=20). Samples were collected from rats aging between 1 and 11 months and were analyzed by Energy Dispersive X-ray Fluorescence (EDXRF) without any chemical preparation. The EDXRF results were assessed by the PIXE (Proton Induced X-ray Emission) technique. The formaldehyde used to preserve the samples was also analyzed by ETAAS (Electro-Thermal Atomic Absorption Spectrometry) in order to verify if there was any loss of lead from the samples to the formaldehyde. We found that the loss was not significant (<2%). Concerning the mean values of the lead concentration measured in the contaminated soft tissues, in liver they range from 6 to 22μgg(-1), and in kidneys from 44 to 79μgg(-1). The control rats show, in general, values below the EDXRF detection limit (2μgg(-1)). The ratio kidney/liver ranges from 2 to 10 and is strongly positively correlated with the age of the animals. A Spearman correlation matrix to investigate the correlation between elemental concentrations and the dependence of these concentrations with age showed that there is a strong positive correlation with age for lead in the liver but not in the kidney. The correlation matrix showed also that the concentration of lead in these two soft tissues is not correlated. The lead accumulation in liver is made by different plateaus that strongly decrease with age. It was verified the existence of two levels of accumulation in kidney, not very highlighted, which might be indicative of a maximum accumulation level for lead in kidney.

Guimarães, D., M. L. Carvalho, V. Geraldes, I. Rocha, and J. P. Santos. "Study of lead accumulation in bones of Wistar rats by X-ray fluorescence analysis: aging effect." Metallomics In Press (2011): DOI: 10.1039/c1mt00149c. AbstractWebsite

The accumulation of lead in several bones of Wistar rats with time was determined and comparedQ3 for the different types of bones. Two groups were studied: a control group (n = 20), not exposedto lead and a contaminated group (n = 30), exposed to lead from birth, first indirectly throughmother’s milk, and then directly through a diet containing lead acetate in drinking water (0.2%).Rats age ranged from 1 to 11 months, with approximately 1 month intervals and each of thecollections had 3 contaminated rats and 2 control rats. Iliac, femur, tibia–fibula and skull havebeen analysed by energy dispersive X-ray fluorescence technique (EDXRF). Samples offormaldehyde used to preserve the bone tissues were also analysed by Electrothermal AtomicAbsorption (ETAAS), showing that there was no significant loss of lead from the tissue to thepreservative. The bones mean lead concentration of exposed rats range from 100 to 300 mg g 1while control rats never exceeded 10 mg g 1. Mean bone lead concentrations were compared andthe concentrations were higher in iliac, femur and tibia–fibula and after that skull. However, ofall the concentrations in the different collections, only those in the skull were statisticallyQ4 significantly different (p o 0.05) from the other types of bones. Analysis of a radar chart alsoallowed us to say that these differences tend to diminish with age. The Spearman correlation testapplied to mean lead concentrations showed strong and very strong positive correlations betweenall different types of bones. This test also showed that mean lead concentrations in bones arenegatively correlated with the age of the animals. This correlation is strong in iliac and femur andvery strong in tibia–fibula and skull. It was also shown that the decrease of lead accumulationwith age is made by three plateaus of accumulation,