Trace-element analysis by charged-particle-induced reactions as a tool for physicians and chemists

The object of this article is to apprise physicians and chemists of nuclear analytical techniques and, in particular, of ion beam analysis (PIXE and PIGE) for the purpose of application to the clinical diagnostic method. The feasibility of the technique, sampling, and sample preparation for trace element analysis in biological and biomedical samples has been described previously (1–3). Analysis data from normal human blood samples and biomedical samples by ion beam reactions have been compared at the end. Emphasis will be placed on the use of the analytical technique on determination of the range of trace and toxic elements in human blood samples.     

Determination of total arsenic concentrations in nails by inductively coupled plasma mass spectrometry

The analysis of trace elements in biological samples will extend our understanding of the impact that environmental exposure to these elements has on human health. Measuring arsenic content in nails has proven useful in studies evaluating the chronic body burden of arsenic. In this study, we developed methodology with inductively coupled plasma-mass spectrometry (ICP-MS) for the determination of total arsenic in nails. We assessed the utility of the washing procedures for removing surface contamination. Four types of preanalysis treatments (water bath, sonication, water bath plus sonication, and control) after sample decomposition by nitric acid were compared to evaluate the digestion efficiencies. In addition, we studied the stability of the solution over 1 wk and the effect of acidity on the arsenic signal. Arsenic content in the digested solution was analyzed by using Ar-N₂ plasma with Te as the internal standard. The results suggest that washing once with 1% Triton Χ-100 for 20 min for cleaning nail samples prior to ICP-MS analysis is satisfactory. Repeated measurement analysis of variance revealed that there was no significant difference among the various sample preparation techniques. Moreover, the measurements were reproducible within 1 wk, and acidity seemed to have no substantial influence on the arsenic signal. A limit of detection (on the basis of three times the standard deviation of the blank measurement) of 7 ng As/g toenail was achieved with this system, and arsenic recoveries from reference materials (human hair and nails) were in good agreement (95–106% recovery) with the certified/reference values of the standard reference materials. ICP-MS offers high accuracy and precision, as well as highthroughput capacity in the analysis of total arsenic in nail samples.     

Trace element content in fingernails and hair of a nonindustrialized US control population

The concentrations of 17 elements in the nail and hair of 117 subjects from a nonindustrialized environment were determined by instrumental neutron activation analysis (INAA). A new method of statistical treatment that allows for more meaningful use of detection limit values was used to process the concentration data. Geometric means and standard errors are presented for each element, along with a summary of the effects of age, sex, and treatment on the concentration of each element. For nails, these data represent the first comprehensive study for several important elements. Correlations for each element between hair and nail were determined. With few exceptions, concentrations of nonessential trace elements were positively correlated in hair and nail, whereas concentrations of essential elements showed no correlations. The factors affecting concentrations and control levels must be considered in studying alterations in disease states.     

PIXE analysis of some medicinal plants used in cleaning teeth in southwestern Nigeria

Sixteen medicinal plants used for cleaning teeth in southwestern Nigeria were analyzed using the particle-induced X-ray emission (PIXE) technique. These medicinal plants have other therapeutic uses, but the emphasis and main focus were on their beneficial effects on human teeth. PIXE measurements were carried out using 1.8 MeV collimated proton beam from the 2.5 MV AN 2000 Van de Graaff accelerator at the Istituto Nazionale di Fisica Nucleare, Laboratori Nazionali di Legnaro, Padova Italy. The results showed the presence of 18 different elements at different concentrations, 2 plants had traces of gold, and none of the plants contained any toxic heavy metals. The metabolic roles of the detected elements were discussed especially as they affected the formation, growth, development, and protection of human teeth. The results of this study are presented and discussed.     

Application of PIXE analysis to the study of the regional distribution of trace elements in normal human brain

A particle-induced X-ray emission (PIXE) analysis method is presented, which allows measurement of eight elements (i.e., K, Ca, Mn, Fe, Cu, Zn, Se, and Rb) in human brain samples of only a few mg dry weight. The precision and accuracy of the method were investigated by analyzing animal brain matter with both PIXE and instrumental neutron activation analysis (INAA). The method was applied to measure the 8 elements in 46 different regions of 3 human brains. The sections analyzed originated from either the left or the right cerebral hemisphere, brain stem, and cerebellum. For one of the brains, sections were also analyzed from 26 corresponding regions of both hemispheres. For all elements, similar concentrations were found in the corresponding areas of the left and right sides of the brain. The concentrations (in μg/g dry weight) of the elements K, Fe, Cu, Zn, Se, and Rb were consistently higher in cortical structures than in white matter. Deep nuclei and brain stem, which have a mixed composition, showed intermediate values for K, Zn, Se, and Rb. A hierarchical cluster analysis indicated that the various brain regions clustered into two large groups, one comprising gray and mixed matter regions and the other, white and mixed matter brain areas.     

Multielement proton-induced X-ray emission analysis of betel leaves,betel nuts,and lime

Proton-induced X-ray emission (PIXE) method was employed to study the concentration of heavy elements in betel leaves, betel nuts, and mineral lime consumed in Bangladesh. The samples were collected from different parts of Bangladesh and analyzed by the thicktarget external beam technique of the PIXE method. The samples were exposed to the proton beam as 1-mm thick pellets and irradiated with 2.0-MeV protons having 20-nA beam intensity. The concentration of some 15 elements (K, Ca, Ti, V, Cr, Mn, Fe, Ni, Cu, Zn, Se, Br, Rb, Sr, and Pb) was measured by comparison with a calibration curve constructed from the NBS orchard leaf standard SRM 1571. The validity of the procedure has been established by comparative measurements of Cu and Zn with atomic absorption spectrophotometry. The significance of the results is discussed in view of their implications in health and disease.     

Applications of ion beam analysis to calcified tissue research

Various nondestructive ion beam analysis techniques have been developed and applied to study the concentrations of fluorine and other trace elements in calcified tissues. Fluorine has been determined by prompt gamma activation analysis through the F(?,αγ)O reaction. This method is quick, convenient, and sensitive, and can also be applied to measure fluorine depth distribution nondestructively in teeth and bone samples. By the application of this technique, fluorine concentrations have been determined in a number of teeth with known histories and bones of experimental animals. Moreover, F-depth-distributions to 10-μm depths have also been obtained in a number of selected human and animal teeth. Carbon was measured by activation analysis with He-3 ions, and the technique of Proton-Induced X-ray Emission (PIXE) has been developed to simultaneously determine Ca, P, and trace elements in well-documented teeth. Dental hard tissues, i.e., enamel, dentine, cementum, and their junctions, as well as different parts of the same tissue, have been examined separately. A number of elements, Na, Mg, Al, P, Cl, Ca, Cr, Mn, Fe, Ni, Cu, Zn, Sr, Br, Rb and Pb, have been determined in these dental tissues. The concentrations of some of these elements vary considerably in different teeth and in various parts of the same tooth. The special advantages of these nuclear techniques for studying metabolic bone diseases, renal diseases, and the effect of Pb on childrens' intelligence are discussed.     

甲真菌病患者甲微生物群分析

目的研究甲真菌病患者甲微生物群的构成,为进一步阐明甲真菌病的发病机制提供线索。方法本研究共纳入47例甲真菌病患者及7例健康志愿者。取患者病甲的甲屑进行真菌镜检及培养鉴定;提取患者病甲、患者对侧健甲及健康人甲的DNA,对真菌rDNA ITS区及细菌16S rDNA V3-V4区PCR扩增,分析微生物群构成,并进行α多样性、β多样性、Simper分析及Spearman相关性分析。结果甲真菌病患者病甲的微生物群构成与对侧健甲及健康人甲存在差异。患者病甲的真菌菌群的丰富度高于对侧健甲。患者对侧健甲细菌菌群的多样性高于病甲及健康人甲,且患者对侧健甲细菌菌群的构成与患甲和健康人甲均有部分重叠。结论患者对侧健甲的细菌菌群构成具有由健康人甲向患者病甲转变的趋势。且在患者病甲的微生物群中,某些真菌和细菌菌属可能具有相关性。     

Hydration of human nails investigated by NIR-FT-Raman spectroscopy.

The human nail, although it is usually stable against outer influences, becomes soft and flexible after soaking in water. Frequent washing increases brittleness of nails. Hydration of nails is thought to be the most important factor influencing the physical properties of nails and possibly acts through changes in keratin structure. Here NIR-FT-Raman has been used to examine molecular structural changes of intact moisten nails. Raman spectra were obtained both in vitro from nail samples and in vivo before and after soaking in water. The water uptake of normal nail samples during the first 15 min was reflected in the increasing intensity ratio of the nu(OH)/nu(CH(2)) bands. A saturating effect appeared soon after 10 min which is explained by a defined water holding capacity. R(nu) representation of the low frequency range of the Raman spectra showed that mainly bound water is found both in dry and in wet nails. This implies water-protein interaction. Protein backbone vibration at 932 cm(-1) indicating alpha-helical proteins increased in intensity in the wet nails. The nu(S-S) which is sensitive to changes in conformation of proteins showed a 4% higher intensity. Additional protein-water interactions could lead to a slight change of the dihedral angle of the C-S-S-C bonds and to geometric changes in coiling behavior of the alpha-helical protein. Suggesting a separation between matrix proteins and fiber proteins giving them a greater freedom of flexibility. The in vivo spectra detected from the distal part of the nail resembled spectra in vitro. Raman spectra of the proximal part of the nail showed that it was fully saturated with water. The proximal part of the nail did not show changes in water content and protein structure during nail moisturizing in the Raman spectra. Our results suggest that the softening of the nail following hydration may be due to changed matrix protein molecular structure induced by water.     

Feasibility of Breast Cancer Screening by PIXE Analysis of Hair

To reveal the role of key elements present in the hair of breast cancer patients on cancer development, the levels of a number of elements in scalp hair samples of 82 people including healthy individuals, people suffering from benign breast disease, and breast cancer patients were measured by PIXE analysis. Pellets of hair samples were prepared and bombarded by 2.2 MeV proton beam of a 3-MV Van de Graaff accelerator. The number of incident ions hitting the sample was indirectly measured using the RBS spectrum of a thin Ag film placed in the beam path. The concentrations of S, Cl, K, Ca, Fe, and Cu in the hair of healthy individuals were in agreement with those observed in the hair of hyperplasia and cancer patients within standard deviations. However, a lower average level of zinc was found in samples from hyperplasia and breast cancer patients. Strong positive correlations were found between iron and potassium as well as between calcium and potassium in the cancer patients. These results could be of significance in the screening for breast cancer.     

Level of Trace Elements (Copper,Zinc, Magnesium and Selenium) and Toxic Elements (Lead and Mercury) in the Hair and Nail of Children with Autism

Autism is a multi-factorial pathology observed in children with altered levels of essential and elevated levels of toxic elements. There are also studies reporting a decrease in nutritional trace elements in the hair and nail of autistic children with healthy controls; moreover, bioelements have been shown to play an important role in the central nervous system. Therefore, the purpose of the present study was to assess the levels of trace elements like copper (Cu), zinc (Zn), magnesium (Mg), and selenium (Se) and toxic elements like mercury (Hg), and lead (Pb) in the hair and nail samples of autistic children and to evaluate whether the level of these elements could be correlated with the severity of autism. The subjects of the study were 45 autistic children with different grades of severity (low (LFA), medium (MFA), and high (HFA) functioning autism) according to Childhood Autism Rating Scale, n = 15 children in each group and 50 healthy children (age and sex matched). The boys and girls ratio involved in this study was 4:1, and they were 4-12 years of age. The study observed a valid indication of Cu body burden in the autistic children. The children with different grades of autism showed high significance (p < 0.001) in the level of copper in their hair and nail samples when compared to healthy controls. The level of Cu in the autistic children could be correlated with their degree of severity (more the Cu burden severe is autism). The study showed a significant elevation (p < 0.001) in the levels of toxic metals Pb and Hg in both hair and nail samples of autistic children when compared to healthy control group. The elevation was much pronounced in LFA group subjects when compared among autistic groups MFA and HFA. The levels of trace elements Mg and Se were significantly decreased (p < 0.001) in autistic children when compared to control. The trace element Zn showed significant variation in both hair and nails of LFA group children when compared to control group and other study groups. The significant elevation in the concentration of Cu, Pb, and Hg and significant decrease in the concentration of Mg and Se observed in the hair and nail samples of autistic subjects could be well correlated with their degrees of severity.     

Biological monitoring of lead and cadmium in human hair and nail and their correlations with biopsy materials, age and exposure

Hair and fingernails of exposed and unexposed subjects were analyzed for their lead (Pb) and cadmium (Cd) contents by atomic absorption spectrophotometer with graphite furnace and air-acetylene flame. Hair and nail Pb concentrations in occupationally exposed subjects ranged between 1.020-409.726 and 8.130-765.306 microg/g and in environmentally unexposed subjects 0.123-25.160 and 1.076-65.613 microg/g, respectively. Similarly, hair and nail Cd concentrations in occupationally exposed subjects ranged between 0.014-22.086 and 0.214-35.714 microg/g and in environmentally unexposed subjects 0.113-1.627 and 0.028-8.108 microg/g, respectively. A significant correlation was observed between Pb hair and nail concentrations in exposed subjects at P < 0.05, as compared to unexposed subjects and Cd hair and nail in exposed, as well as unexposed subjects. With respect to exposure, levels of Pb in hair and nails were found to be significant in exposed subjects, compared to unexposed ones and levels of Cd were significant only in nails of exposed ones. With respect to age, no significant correlation was found between hair and nail Pb and Cd concentrations in both exposed and unexposed subjects.     

Trace Elements Concentration in Nails and Association with Airway Inflammation in Adolescents

Asthma is a multifactorial disease and its severity varies with the inflammatory status of the patient. It has been hypothesized that deficiency of trace elements has an important role in the etiology of asthma. The aim of the study was to evaluate the concentration of trace elements in nails of adolescents with different degrees of airway inflammation. We assessed the prevalence of active and severe asthma in adolescents (n?=?106) through the ISAAC questionnaire. Subjects were allocated into two different groups according to the value of fraction exhaled nitric oxide (Fe_NO) measured. Also, nail specimens were collected in order to determine the concentration of accumulated trace elements (Cu, Zn, Mn, Se) by ICP-MS. It was performed Student’s t test to evaluate statistical differences between inflammatory groups considering normally distributed data. On the other hand, when data presented non-normal distribution, Mann-Whitney U test was utilized. Spearman coefficients were calculated to investigate correlations between Fe_NO and mineral concentrations. Exhaled NO was increased in male subjects. We found association between active asthma and elevated exhaled NO. There was no significant difference in Cu (4.40 vs. 4.52), Zn (84.66 vs. 79.48), Mn (0.59 vs. 0.76), and Se (0.18 vs. 0.19) concentrations (μg/g) among distinct Fe_NO groups as well as was not observed correlation between exhaled NO levels and any element. Deficiency of trace elements in nails were not associated with increased Fe_NO.     

Nail Antioxidant Trace Elements Are Inversely Associated with Inflammatory Markers in Healthy Young Adults

Antioxidant intake may be linked to a reduction of the chronic low-grade inflammatory state related to obesity and several accompanying disorders such as insulin resistance, cardiovascular diseases, and metabolic syndrome. So, the aim of this study was to evaluate the potential associations between nail trace elements and several indicators in healthy young adults, emphasizing on the putative effect of antioxidant trace element intake on inflammation-related marker concentrations. This study enrolled 149 healthy young adults, whose anthropometrical and blood pressure values as well as lifestyle features were analyzed. Fasting blood samples were collected for the biochemical and inflammation-related measurements (C-reactive protein, tumor necrosis factor-alpha (TNF-α), interleukin (IL)-6, IL-18, and homocysteine). Nail samples were collected for the analysis of selenium, zinc, and copper concentrations. Our results showed that nail selenium was negatively associated with IL-18; nail zinc concentrations were inversely related to circulating IL-6, IL-18, and TNF-α, whereas nail copper (Cu) and Cu/selenium values were negatively correlated with homocysteine levels and the Cu/zinc ratio was unaffected. In conclusion, nail content on some trace elements related to antioxidant defense mechanisms seems to be associated with several inflammation-related markers linked to chronic diseases in apparently healthy young adults, which is of interest to understand the role of antioxidant intake.     

Are fingernail lead levels a reliable biomarker of lead internal dose?

BackgroundPotentially toxic elements, such as lead, can bioaccumulate and alter human physiology. Human biomonitoring is an essential tool to evaluate chemical exposures in different biological matrices (blood, urine, saliva, nails, and hair). Of these biological matrices, nails are favorable for their ease of sampling, transport and storage. The aim of this study was to investigate possible correlations between blood lead levels (BLL) and washed and non-washed fingernail lead levels (FLL) in 55 adults living in a lead-contaminated area.MethodVenous blood and fingernail (thumbs and forefingers) samples were collected. Nails from the left hand were washed with Triton X-100 (0.5 % m/v) and HNO₃ solution, while nails from the right hand were not submitted to the pre-analytical procedures. Samples were analyzed by graphite furnace atomic absorption spectrometry, and pairwise correlations were used to correlate lead concentrations between BLL and FLL; nails from fingers of the same hand and between washed and unwashed fingernails. Principal component analysis was performed and scatter diagrams were plotted to investigate correlations.ResultsA non-significant positive correlation was found between BLL and washed forefinger nails lead (r = 0.219, p = 0.112) and between BLL and thumbnail lead levels (r = 0.182, p = 0.191). Comparison of fingernails from the same hand (thumb and forefinger), showed that lead concentrations of non-washed nails varied widely, even on analyses of transversal fragments from the same nail. Lead levels in non-washed forefinger nails were not correlated with non-washed thumbnails (r = 0.169, p = 0.219). Conversely, washed thumb and forefinger nails were found to be correlated (r = 0.39, p = 0.003). Washed and non-washed nails were also found to be correlated (p < 0.0001).ConclusionIn conclusion, the results showed that non-washed nails are not a reliable biomarker for lead exposure. Although washing nails before analysis may reduce external contamination, the correlation of lead concentrations between fingers is poor for fingernail lead levels to serve as an internal dose biomarker to lead exposure. In addition, levels in washed nails were not significantly correlated with blood lead levels. Fingernail lead levels seem to serve as an indicator of lead exposure sources in contact with the individual, but not as a reliable biomarker of internal dose.     

Low Concentrations of Selenium and Zinc in Nails are Associated with Childhood Asthma

The purpose of this study was to investigate possible associations between Zn, Se, Cu, Mn, and Co concentrations in nails and asthma in a young population from a Southern Brazil city. Additionally, correlations between these chemical elements among asthmatic and non-asthmatic children were evaluated. Before nail collection (n?=?165), children were asked to complete the International Study of Asthma and Allergies in Childhood questionnaire. The concentrations of trace elements were determined by inductively coupled plasma mass spectrometry. The chi-square test was used to evaluate the association between element concentrations in nails and the respiratory outcome. To evaluate correlations between the elements, we used the Spearman correlation test. For all tests, the significance level was set at 95% (P?≤?0.05). Children included in the highest quartile of nail Se and Zn concentration presented a fivefold decrease in the prevalence ratio of asthma while children in the lowest Se range presented an almost 2.5-fold increase in the asthma prevalence ratio. There were weak to strong correlations between Cu vs. Zn, Cu vs. Co, Cu vs. Se, Zn vs. Se, Zn vs. Mn, and Mn vs. Co in both asthmatic and non-asthmatic children. Interestingly, non-asthmatics also presented correlations between Co vs. Se and Zn. Taken together, our results clearly demonstrated an association between concentrations of selenium and zinc and childhood asthma and the usefulness of nail as a noninvasive matrix to detect minerals imbalance in asthma patients.     

致病真菌对甲侵袭性的体外初步研究

目的观察不同致病真菌在体外对甲板的侵袭能力。方法将分离自甲真菌病患者的致病真菌包括白念珠菌、红色毛癣菌和短帚霉,接种到沙堡培养基中,同时将灭菌甲板埋植入接种处。第28d时,取出甲板,进行病理切片,观察真菌对甲板的侵袭能力。结果病理显示接种于红色毛癣菌及短帚霉的甲板内可见菌丝生长,而接种于白念珠菌的甲板内无菌丝生长。结论皮肤癣菌和霉菌可以在甲板内侵袭生长,而白念珠菌对甲板无侵袭能力。     

Trace elemental analysis of cancer-afflicted intestine by PIXE technique

Trace elemental analysis was carried out in the biological samples of cancer-afflicted intestine using the particle-induced X-ray emission technique (PIXE). A 2-MeV proton beam was employed to excite the samples. From the present results, it can be seen that the concentration of the elements Cr, Fe, and Ni are higher in the cancerous tissue of the intestine than those observed in the normal tissue, whereas the concentration levels of the element Zn is slightly lower in the cancer tissue of intestine than that observed in the normal tissue. The concentrations of S, Cl, K, Ca, Ti, Mn, Co, and Cu in the cancer tissue of the intestine are in agreement with those observed in the normal tissues within standard deviations. The present results support the previous observations that Ni and Cr are carcinogenic agents. The observed slightly low levels of zinc in the cancer tissue of the intestine suggest that zinc could possibly inhibit the tumor growth and development of neoplastic transformation. For correctly assessing the role played by the trace elements in initiating, promoting, or inhibiting cancer in various organs, there is a need for the acquisition of more data by trace elemental analysis from several investigations of this type undertaken in different regions.     

Matrix effects in PIXE analysis of aerosols and ashes

A comparison of instrumental neutron activation analysis (INAA) and proton-induced X-ray emission (PIXE) results for sizefractionated atmospheric aerosols (“coarse” and “fine” fractions with an equivalent aerodynamic diameter of 2–10 Μm and < 2 Μm, respectively, or the PM10 fraction) showed that PIXE yielded significantly lower results for the PM10 and coarse fractions, especially for elements with a low Z resulting from a particle size effect. Somewhat lower PIXE results were also obtained for the fine fraction of atmospheric aerosols. A correction is also needed for irregularly shaped deposits of combustion aerosols collected by a cascade impactor in 11 size fractions ranging from 0.016 to 14.3 Μm, as well as for thick samples of fly and bottom ashes. An equivalent layer thickness (ELT) model is proposed to correct the matrix effects in PIXE. The approaches for the calculation of ELT using a comparison of PIXE and INAA results or by comparing PIXE results obtained using two different incident proton beam energies (1.31 and 2.35 MeV) are described. The correction for the ash pellets and irregular deposits are also discussed.