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1.
Rheumatology (Oxford) ; 58(7): 1239-1244, 2019 07 01.
Artigo em Inglês | MEDLINE | ID: mdl-30726990

RESUMO

OBJECTIVE: Immunoblot (IB) methods are widely used to detect myositis-specific autoantibodies (MSAs); however, false-positive results are common. In this study, we aimed to determine whether associating the anti-nuclear antibody (ANA) IIF pattern may help to improve the specificity of MSA detection by IB in patients with idiopathic inflammatory myositis (IIM). METHODS: Serum samples from 104 patients presenting with muscle weakness/myalgia and positive to at least one MSA by IB (MYOS12 Diver and MIOS7 Diver, D-tek) were tested for ANAs on HEp-2000 cells (Immuno Concepts). The chi-square test was used to analyse the concordance of the MSA result and its corresponding pattern by ANA testing between patients with and without IIM. RESULTS: Eighty-three of the 104 patients had a diagnosis of definite IIM, while in 21 cases, patients were affected by other autoimmune diseases or various non-systemic diseases. Forty nine of 83 (59%) patients in the IIM group and 4/21 (19%) in the non-IIM group showed a concordance between ANA pattern and MSAs by IB (P < 0.001). MSA monopositivity was significantly associated with IIM (91.6%) compared with 61.9% in the non-IIM group (P = 0.0005). CONCLUSIONS: Considering both the MSA result and its corresponding pattern by ANA testing may help to improve the specificity of MSA detection by IB and to confirm the diagnosis of MSA-associated IIM. The monopositivity of MSAs is an important additional tool to validate IB results.


Assuntos
Anticorpos Antinucleares/sangue , Doenças Autoimunes/diagnóstico , Miosite/diagnóstico , Idoso , Algoritmos , Doenças Autoimunes/imunologia , Biomarcadores/sangue , Diagnóstico Diferencial , Feminino , Imunofluorescência/métodos , Humanos , Immunoblotting/métodos , Masculino , Pessoa de Meia-Idade , Miosite/imunologia , Reprodutibilidade dos Testes , Sensibilidade e Especificidade
2.
J Clin Lab Anal ; 27(1): 21-6, 2013 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-23292801

RESUMO

INTRODUCTION: Serum autoantibodies specifically directed toward intracellular cytoskeletal actin filaments (anti-actin antibodies, AAA) were found to be associated with intestinal villous atrophy (IVA) in celiac disease (CD). The aim of this study was to assess IgA-AAA with a commercial test that uses sections of rat intestinal epithelial cells in a well-selected cohort of patients and to evaluate the relationship between the presence of serum IgA-AAA and the severity of intestinal mucosa damage. MATERIALS AND METHODS: Serum samples from 70 CD patients and 150 controls subjects were analyzed retrospectively for the presence of IgA-AAA. RESULTS: The indirect immunofluorescence test that we used has a specificity of 100%; the sensitivity of the test is not high (25.7%). In this study we also show that serum AAA are more frequently positive in CD patients with total IVA (77.8%) and that this association is significant DISCUSSION: IgA-AAA certainly cannot take the place of much more sensitive tests such as a-tTG and EMA in the diagnosis of CD because of their low sensitivity; nonetheless, these antibodies could be determined in a-tTG and/or EMA positive patients who cannot undergo an intestinal biopsy because of a severe contraindication, or in the case of negative consensus regarding endoscopy, or when the histology interpretation is difficult. CONCLUSION: In conclusion, the IFI commercial test with intestinal epithelial cells as substrate offers a useful method for IgA-AAA determination. Serum IgA-AAA positivity is indicative of more severe intestinal histology damage and their assay could be a real help to the clinician, especially in the complicated cases.


Assuntos
Citoesqueleto de Actina/imunologia , Autoanticorpos/sangue , Doença Celíaca/imunologia , Imunoglobulina A/sangue , Adolescente , Adulto , Idoso , Animais , Doença Celíaca/sangue , Distribuição de Qui-Quadrado , Criança , Pré-Escolar , Estudos de Coortes , Feminino , Técnica Indireta de Fluorescência para Anticorpo , Humanos , Masculino , Pessoa de Meia-Idade , Ratos , Sensibilidade e Especificidade
3.
Biomed Pharmacother ; 64(5): 369-72, 2010 May.
Artigo em Inglês | MEDLINE | ID: mdl-20005669

RESUMO

Proteins play a fundamental role in the formation and progression of plaque, but proteomic analysis of plaque as a whole is difficult, due to its heterogeneous cellular composition and an abundance of plasma proteins. Several approaches to this problem are reported in the literature; they include proteomic analysis of vascular tissues, analysis of proteins released by normal and pathological arterial walls, proteomic analysis of vascular cells and proteomic analysis of blood. In a previous study, we proposed a new strategy for studying of proteome of plaque, which permits to select the proteins exclusive to plaque by the constructing of a reference synthetic gel. In the present work, we matched the spots of the reference synthetic gel with the spots of a pool of carotid plaque, in order to select only spots exclusive to plaque from the 2-dimensional electrophoresis of the pool of plaque. We selected some spots between those exclusive and identified them by mass spectrometry. Some proteins identified are involved in transport, others take part in elimination of toxic radicals, others are metabolic enzymes or structural proteins. This study represents an example of application of the new approach which we have proposed: the reference gel of proteome of plaque permits to select, on every sample of interest, only the spots exclusive to plaque; once selected, spots can be identified by mass spectrometry and, being typical of plaque composition, could represent novel markers of lesions and vascular risk.


Assuntos
Aterosclerose/metabolismo , Estenose das Carótidas/metabolismo , Proteoma/análise , Proteômica/métodos , Eletroforese em Gel Bidimensional , Géis , Humanos , Espectrometria de Massas
4.
Nucleosides Nucleotides Nucleic Acids ; 27(6): 624-7, 2008 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-18600516

RESUMO

Free radical excess and oxidative stress are implicated in the formation and progression of atherosclerotic plaque through actions on susceptible vascular cells, such as by activating xanthine oxidase. Purine bases and other antioxidant compounds could play important protective roles in atherogenesis, as could nonenzymatic low molecular weight thiol defenses, not previously evaluated in carotid artery plaque. Therefore, we measured purine catabolites (hypoxanthine, xanthine, uric acid, allantoin) and antioxidant compounds (total sulphydryl groups, homocysteine, cysteine, and glutathione) in advanced carotid artery plaque and found a high ratio of allantoin to uric acid, suggesting a ongoing local oxidative stress.


Assuntos
Antioxidantes/metabolismo , Estenose das Carótidas/metabolismo , Purinas/metabolismo , Idoso , Idoso de 80 Anos ou mais , Cromatografia Líquida de Alta Pressão , Feminino , Humanos , Masculino , Pessoa de Meia-Idade
6.
Int J Immunopathol Pharmacol ; 20(3): 637-42, 2007.
Artigo em Inglês | MEDLINE | ID: mdl-17880777

RESUMO

Atherosclerosis is a complex disease that affects medium and large arteries, leading to the formation and progression of plaque. In this process the proteins play an essential role and as a consequence, proteomic-based strategies examining the protein content of cells or tissues could offer a useful approach for the study of plaque proteins. Due to the heterogeneous cell composition of plaque, proteome analysis of whole lesions is difficult, besides being also complicated by the presence of plasma proteins that cannot be completely eliminated. A good way to study variations in protein expression among series of gels is to construct a synthetic gel. This type of gel is obtained by averaging the positions, shapes and optical densities of spots in a given set of gels. To be included in the synthetic gel, spots must be found in at least three gels. To obtain a profile representative of the proteome of atherosclerotic plaque, cancelling its high variability, we constructed a synthetic gel using an average of ten carotid plaque samples. We then compared it with an equivalent synthetic gel constructed using ten plasma samples from the same carotid surgery patients. For the comparison of two synthetic gels (plasma/plaque) we could discriminate plasma proteins from plaque proteins. Besides analysis of spots common to plasma, the synthetic gel is useful to detect spots exclusive to plaque, thus simplifying a very complex mixture.


Assuntos
Proteínas Sanguíneas/análise , Estenose das Carótidas/metabolismo , Proteômica/métodos , Idoso , Estenose das Carótidas/sangue , Estenose das Carótidas/diagnóstico por imagem , Estenose das Carótidas/cirurgia , Eletroforese em Gel Bidimensional , Endarterectomia das Carótidas , Feminino , Géis/química , Humanos , Concentração de Íons de Hidrogênio , Masculino , Ultrassonografia
7.
Nucleosides Nucleotides Nucleic Acids ; 25(9-11): 1291-4, 2006.
Artigo em Inglês | MEDLINE | ID: mdl-17065109

RESUMO

This study was carried out on carotid artery plaque and plasma of 50 patients. We analyzed uric acid, hypoxanthine, xanthine, and allantoin levels to verify if enzymatic purine degradation occurs in advanced carotid plaque; we also determined free radicals and sulphydryl groups to check if there is a correlation between oxidant status and purine catabolism. Comparing plaque and plasma we found higher levels of free radicals, hypoxanthine, xanthine, and a decrease of some oxidant protectors, such as sulphydryl groups and uric acid, in plaque. We also observed a very important phenomenon in plaque, the presence of allantoin due to chemical oxidation of uric acid, since humans do not have the enzyme uricase. The hypothetical elevated activity of xanthine oxidase in atherosclerosis could be reduced by specific therapies using its inhibitors, such as oxypurinol or allopurinol.


Assuntos
Estenose das Carótidas/sangue , Estenose das Carótidas/metabolismo , Idoso , Idoso de 80 Anos ou mais , Alantoína/sangue , Alopurinol/sangue , Química Clínica/métodos , Feminino , Radicais Livres , Humanos , Hipoxantina/sangue , Masculino , Pessoa de Meia-Idade , Oxidantes/metabolismo , Oxipurinol/sangue , Purinas/metabolismo , Ácido Úrico/sangue , Ácido Úrico/metabolismo , Xantina/sangue
8.
Nucleosides Nucleotides Nucleic Acids ; 23(8-9): 1131-4, 2004 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-15571216

RESUMO

Urate oxidase, or uricase (EC 1.7.3.3), is a peroxisomal enzyme that catalyses the oxidation of uric acid to allantoin. The chemical mechanism of the urate oxidase reaction has not been clearly established, but the involvement of radical intermediates was hypothesised. In this study EPR spectroscopy by spin trapping of radical intermediates has been used in order to demonstrate the eventual presence of radical transient urate species. The oxidation reaction of uric acid by several uricases (Porcine Liver, Bacillus Fastidiosus, Candida Utilitis) was performed in the presence of 5-diethoxyphosphoryl-5-methyl-pyrroline-N-oxide (DEPMPO) as spin trap. DEPMPO was added to reaction mixture and a radical adduct was observed in all cases. Therefore, for the first time, the presence of a radical intermediate in the uricase reaction was experimentally proved.


Assuntos
Espectroscopia de Ressonância de Spin Eletrônica/métodos , Urato Oxidase/química , Animais , Bacillus/metabolismo , Candida/metabolismo , Catálise , Óxidos N-Cíclicos/química , Radicais Livres , Humanos , Peróxido de Hidrogênio/química , Radical Hidroxila , Oxigênio/química , Oxigênio/metabolismo , Marcadores de Spin , Detecção de Spin , Suínos , Ácido Úrico/sangue , Ácido Úrico/química
9.
Horm Metab Res ; 36(9): 614-9, 2004 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-15486812

RESUMO

In previous studies, we found that castration induced interesting morphological and biochemical changes in rat liver. For the present study, we have examined the effects of testosterone on the kinetics of purine nucleotide metabolism with the aim of determining the steps affected by testosterone deficiency. A biomathematical model of purine nucleotide metabolism was used to analyze the many reactions involved. The model simplifies purine nucleotide metabolism to four main steps: 1) de novo synthesis from PRPP to IMP; 2) the inosinic branch point from IMP to GMP or AMP; 3) catabolism of IMP, AMP and GMP to uric acid; 4) RNA and DNA formation from AMP and GMP. We evaluated rate constants from each step from variations in specific radioactivity of metabolites labelled with (14)C-formate, a precursor of de novo synthesis. The model was applied to the liver of normal and castrated rats before and after testosterone treatment. All four steps were slowed after castration, and were not completely restored by androgen administration. The model can give a clear representation of the kinetics of the reactions involved in the liver nucleotide metabolism investigated here, and we propose that a similar approach could be useful whenever a quantitative evaluation of the results obtained in vivo after administration of labelled precursors is required.


Assuntos
Androgênios/farmacologia , Fígado/efeitos dos fármacos , Fígado/metabolismo , Nucleotídeos de Purina/metabolismo , Testosterona/farmacologia , Adenina/metabolismo , Animais , Guanina/metabolismo , Hipoxantina/metabolismo , Masculino , Modelos Biológicos , Orquiectomia , Ratos , Ratos Wistar
10.
Int J Immunopathol Pharmacol ; 17(3 Suppl): 31-3, 2004.
Artigo em Inglês | MEDLINE | ID: mdl-16857104

RESUMO

In this work we determined hypoxanthine (HX), xanthine (X), uric acid (UA), allantoin (ALL) and free radicals in atheromatous plaques to improve the comprehension of oxidative stress, a phenomenon which characterizes the evolution of atherosclerotic lesions. Carotid artery plaque were obtained from subjects undergoing endoarterectomy. Pulverized plaque, extracted by water, was used for analysis of oxidative stress factors (allantoin, uric acid, xanthine, hypoxanthine, free radicals). The peroxidation UA-->ALL was very high in the plaque, as was the level of free radicals. The results show that oxidative degradation of nucleotides, such as LDL oxidation, plays a specific role not only in the progression of atherosclerotic lesions but also in the advanced plaque.


Assuntos
Alantoína/metabolismo , Doenças das Artérias Carótidas/metabolismo , Estenose das Carótidas/metabolismo , Radicais Livres/metabolismo , Estresse Oxidativo , Purinas/metabolismo , Humanos
11.
Clin Chem Lab Med ; 39(6): 501-4, 2001 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-11506461

RESUMO

Different methods have been devised to detect point mutations. Some are very sensitive, detecting mutations even in a background of normal tissue, but none provide information about the percentage of cells with mutant DNA. Here we describe an easy, fast and reliable method, melting temperature analysis, which not only detects point mutations but also provides quantitative information on the percentage of cells with mutant DNA. By this method we detected a G-A transition in codon 12 of the K-ras gene in DNA of subjects with colorectal cancer. The K-ras mutation was found in 9/10 bowel cancers and 8/10 normal adjacent samples. It was also detected in 4/7 stool samples from the same patients. In colorectal cancers, the proportion of K-ras mutant cells was variable: in two the mutant/wild-type DNA ratio was 30/70, in three 50/50, and in four 70/30. Melting temperature analysis was sensitive for the detection of point mutations in bowel cancer and also in apparently normal tissue, providing quantitative information about the percentage of cells with mutant DNA.


Assuntos
Análise Mutacional de DNA/métodos , DNA de Neoplasias/química , DNA de Neoplasias/genética , Desnaturação de Ácido Nucleico , Mutação Puntual , Neoplasias Colorretais/química , Neoplasias Colorretais/genética , Análise Mutacional de DNA/estatística & dados numéricos , Feminino , Genes ras , Humanos , Masculino , Reação em Cadeia da Polimerase/métodos , Sensibilidade e Especificidade , Espectrometria de Fluorescência , Temperatura
13.
Histochem J ; 33(1): 51-7, 2001 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-11352401

RESUMO

The c-erbB2 gene has been found to be amplified in a number of human adenocarcinomas, leading to elevated levels of expression of its encoded product, p185. Mutations in the p53 gene are also common in colorectal carcinomas, brain tumours, leukaemia and lymphomas. In this study, p185 and p53 overexpression was analyzed in colorectal adenomas (22 tubular adenomas and 2 tubulo-villous adenomas) and moderately differentiated adenocarcinomas (n = 22) in order to determine whether there was a relationship between these two proteins. The proteins are encoded by two genes located in the same chromosome. p185 and p53 expression was determined on tissue sections by immunohistochemical staining procedure. Expression of p185 was significantly higher (p < 0.01) in preneoplastic lesions (95.8% of cases) than colorectal cancer (63.6% of cases). p53 showed an inverse pattern to p185, being expressed in 58.3% of benign lesions and 72.7% of adenocarcinomas. These results confirm that p185 overexpression is associated with the early stages of colorectal cancer, whereas p53 is associated with more advanced stages. Although there was no correlation between p185 and p53 expression in premalignant lesions and adenocarcinomas, these two proteins have an important role in the adenoma-carcinoma sequence.


Assuntos
Adenocarcinoma/genética , Adenoma Viloso/genética , Neoplasias Colorretais/genética , Regulação Neoplásica da Expressão Gênica , Genes erbB-2 , Genes p53 , Adenocarcinoma/química , Adenocarcinoma/patologia , Adenoma Viloso/química , Adenoma Viloso/patologia , Idoso , Idoso de 80 Anos ou mais , Neoplasias Colorretais/química , Neoplasias Colorretais/patologia , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Lesões Pré-Cancerosas/química , Lesões Pré-Cancerosas/genética , Lesões Pré-Cancerosas/patologia , Receptor ErbB-2/análise , Proteína Supressora de Tumor p53/análise
14.
Life Sci ; 68(10): 1131-42, 2001 Jan 26.
Artigo em Inglês | MEDLINE | ID: mdl-11228097

RESUMO

In our previous experiments on rat liver we found that 15' after intraperitoneal administration of 14C-formate the specific radioactivity of allantoin was always higher than that of uric acid. The present experiments have been carried out to interpret this unexpected result, which was only observed in liver and we studied: a) the incorporation of 14C-glycine into uric acid and allantoin; b) the effects of two competitive inhibitors of xanthine oxidase and uricase, oxonic acid and allopurinol respectively, on levels of uric acid and allantoin in liver and on their specific radioactivity after administration of labelled precursor. The results suggested: a) that under normal conditions, the formation of allantoin is so fast that it exceedes export from liver to serum, and thus the radioactivity of labelled precursors accumulates in allantoin; b) that when allopurinol or oxonic acid are administered, the rate of export exceeds that of allantoin formation and the incorporation of radioactivity into allantoin is lower; c) that not all the data, however, could be interpreted on this basis, but seems to require the existence of different pools of uric acid, which are transformed separately into allantoin.


Assuntos
Alantoína/metabolismo , Alopurinol/farmacologia , Inibidores Enzimáticos/farmacologia , Fígado/metabolismo , Ácido Oxônico/farmacologia , Nucleotídeos de Purina/metabolismo , Ácido Úrico/metabolismo , Animais , Masculino , Ratos , Ratos Wistar
16.
J Chromatogr B Biomed Sci Appl ; 728(2): 185-92, 1999 May 28.
Artigo em Inglês | MEDLINE | ID: mdl-10406204

RESUMO

Allantoin, uric acid (UA), hypoxanthine (Hx) and xanthine (X) were determined on carotid plaque by capillary zone electrophoresis (CZE) and high-performance liquid chromatography (HPLC). Comparison of the results showed that capillary zone electrophoresis may have similar or even superior analytical performance to HPLC, especially for the determination of allantoin in biological samples.


Assuntos
Arteriosclerose/metabolismo , Artérias Carótidas/metabolismo , Cromatografia Líquida de Alta Pressão/métodos , Eletroforese Capilar/métodos , Purinas/metabolismo , Idoso , Idoso de 80 Anos ou mais , Arteriosclerose/patologia , Artérias Carótidas/patologia , Feminino , Humanos , Masculino , Estresse Oxidativo
17.
Clin Biochem ; 31(7): 523-8, 1998 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-9812171

RESUMO

OBJECTIVES: The HER2 gene has been found amplified in a number of human adenocarcinoma leading to elevated levels of expression of its encoded product, p185 protein. Because little information is available on the tissue and tumor specificity of this gene product, we studied the expression of p185 protein in preneoplastic colon lesions. Adenylosuccinate lyase (ASL, EC 4.3.2.2) is known to increase in malignancies such as colorectal, breast, and prostate cancer. In order to evaluate the potential of ASL as a tumor marker, its activity was determined and compared with the expression of p185. DESIGN AND METHODS: p185 was determined by an immunohistochemical procedure in patients with the preneoplastic lesions. ASL activity was evaluated in intestinal mucosa adjacent to colorectal cancers (patient group A) and in preneoplastic colorectal lesions (group B). The enzyme activity was evaluated in dialyzed supernatants, following the disappearance of substrate (adenylosuccinate AMP-S) and the formation of product (adenosine 5'-monophosphate-AMP), separated by high performance liquid chromatography. RESULTS AND CONCLUSIONS: Increased expression of p185 and elevated ASL activity were observed in tubular and tubulo-villous adenoma and may, therefore, be associated with the early stages of colorectal cancer.


Assuntos
Adenilossuccinato Liase/metabolismo , Colo/patologia , Mucosa Intestinal/metabolismo , Receptor ErbB-2/metabolismo , Adenoma/metabolismo , Adenoma/patologia , Monofosfato de Adenosina/metabolismo , Adenilossuccinato Liase/análise , Adulto , Idoso , Biomarcadores Tumorais , Colo/metabolismo , Neoplasias Colorretais/metabolismo , Neoplasias Colorretais/patologia , Feminino , Humanos , Imuno-Histoquímica , Masculino , Pessoa de Meia-Idade , Receptor ErbB-2/análise
18.
Life Sci ; 62(25): 2257-66, 1998.
Artigo em Inglês | MEDLINE | ID: mdl-9651114

RESUMO

The behaviour of 5'-nucleotidase isoenzymes (ecto-5'-nucleotidase, e-Ns and c-N-II soluble 5'-nucleotidases) was studied in lymphocytes from patients with B-cell chronic lymphocytic leukemia. A strong reduction in ecto- and soluble activities was observed, although the pattern of the three 5'-nucleotidases did not always strictly overlap. A significant decrease (p<0.05) in ecto-5'-nucleotidase, e-Ns and c-N-II was found in B and T populations (B lymphocytes: 1.13, 0.88 and 1.26 nmol/h/10(6) cells versus 95.96, 9.64 and 13.73 nmol/h/10(6) cells in controls; T lymphocytes: 1.31, 0.23 and 0.06 nmol/h/10(6) cells versus 9.25, 1.31 and 2.10 nmol/h/10(6) cells in healthy subjects). The percentage of ecto-5'-nucleotidase-positive cells (CD73+) was reduced in leukemia patients, indicating a lower number of active molecules on the cell surface. The results of RT-PCR analysis showed that the ecto-5'-nucleotidase mRNA of leukemia patients was not defective.


Assuntos
5'-Nucleotidase/sangue , Linfócitos B/enzimologia , Isoenzimas/sangue , Leucemia Linfocítica Crônica de Células B/enzimologia , Linfócitos T/enzimologia , Adenosina/sangue , Trifosfato de Adenosina/sangue , Idoso , Estudos de Casos e Controles , Comunicação Celular/fisiologia , Feminino , Humanos , Hidrólise , Masculino , Pessoa de Meia-Idade , Transdução de Sinais/fisiologia
20.
Rapid Commun Mass Spectrom ; 11(4): 398-404, 1997.
Artigo em Inglês | MEDLINE | ID: mdl-9069642

RESUMO

Fast-atom bombardment (FAB) mass spectrometry, linked with tandem mass spectrometry (MS/MS), was employed for the identification of methylated purine bases in four urinary extracts of healthy subjects and fourteen urinary extracts of patients bearing colorectal tumors. In order to obtain an easy structural identification of the species present in urinary extracts, the MS/MS spectra of MH+ species of twenty nine diagnostically relevant purine bases were studied. Even if definitive quantitative data cannot be obtained by this approach, FAB mass spectra of urine extracts lead to a readily reproducible mapping of endogenous purine bases, allowing a distinction between healthy and sick subjects. Bases such as 9-ethyladenine, N6-2-isopentenyladenine and N6-benzyladenine were detected only in urine samples of colorectal tumor bearing patients. The detection in urine of compounds such as 7-methylguanine and 1-methylguanine, and their increase in the urine of colorectal tumor bearing patients, has been justified either by a more rapid turnover of nucleic acids in tumor tissue or by an increase in the extent of their methylation. The obtained results indicate that the method can be employed for diagnostic purposes.


Assuntos
Mapeamento de Nucleotídeos/métodos , Purinas/urina , Neoplasias Colorretais/urina , Humanos , Metilação , Mapeamento de Nucleotídeos/instrumentação , Espectrometria de Massas de Bombardeamento Rápido de Átomos
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