ABSTRACT
BACKGROUND: Oral manifestations are common in neurofibromatosis 1 (NF1), and include jaws and teeth alterations. Our aim was to investigate the craniomaxillofacial morphology of Brazilian children, adolescents and adults with NF1 using cone beam computed tomography. MATERIAL AND METHODS: This study was conducted with 36 Brazilian individuals with NF1 with ages ranging from 4 to 75. The participants were submitted to anamnesis, extra and intraoral exam and cephalometric analysis using cone beam computed tomography. Height of the NF1 individuals was compared to the length of jaws and skull base. The results of the cephalometric measurements of the NF1 group were compared with a control group paired by age, gender and skin color. RESULTS: Individuals with NF1 had lower maxillary length (p<0.0001), lower mandibular length (p<0.0001), lower skull base length (p<0.0001). In children and adolescents, the mandible was more posteriorly positioned (p=0.01), when compared with the control group. There was no association between jaws and skull base length with the height of the individuals with NF1. CONCLUSIONS: Brazilian children, adolescents and adults with NF1 have short mandible, maxilla and skull base. Moreover, children and adolescents present mandibular retrusion.
Subject(s)
Cone-Beam Computed Tomography , Jaw/diagnostic imaging , Jaw/pathology , Neurofibromatosis 1/complications , Skull Base/diagnostic imaging , Skull Base/pathology , Adolescent , Adult , Aged , Brazil , Case-Control Studies , Cephalometry , Child , Child, Preschool , Female , Humans , Male , Middle Aged , Prospective Studies , Young AdultABSTRACT
Neurofibromatosis type 1 (NF1) is a common autosomal genetic disorder with a prevalence of 1 in 3,000 births. NF1 is a complex syndrome characterized by many abnormalities and may affect all organ systems. Oral manifestations of NF1 occur frequently, but reports including NF1 children with facial plexiform neurofibromas and oral alterations are scant. Facial plexiform neurofibroma may cause asymmetry, disfigurement and usually arises from the trigeminal nerve. The aim of this paper is to to report three pediatric NF1 cases with facial plexiform neurofibroma presenting with oral manifestations, which were evaluated clinically and radiographically, and also to briefly review the literature. Patients presented with changes in the oral soft tissues, jaws, and teeth ipsilateral to the tumor.
Subject(s)
Dentofacial Deformities/diagnosis , Facial Neoplasms/diagnosis , Neurofibroma, Plexiform/diagnosis , Neurofibromatosis 1/diagnosis , Child , Female , Gingival Neoplasms/diagnosis , Gingival Overgrowth/diagnosis , Humans , Macroglossia/diagnosis , Male , Malocclusion/diagnosis , Mandible/abnormalities , Mandibular Condyle/abnormalities , Tongue Neoplasms/diagnosisABSTRACT
We estimated the genetic gains of the 12th cycle of reciprocal recurrent selection for maize traits of agronomic interest. We used 23 ISSR molecular markers in an attempt to maximize genetic variability among and within populations based on selection of S(1) progenies. To this end, 138 full-sib families were evaluated in a randomized block design in two environments (the municipalities of Campos dos Goytacazes and Itaocara, in the State of Rio de Janeiro, Brazil), with replications within sets. Direct selection for grain yield was used for the selection of the families. To assess genetic diversity among and within populations, we examined plants produced from part of the S(1s) seeds from the parents that originated the 42 full-sib families that were selected from the agronomic traits. Direct selection for grain yield provided good gains for the traits evaluated, with estimated improvement of -0.87 days for days to flowering, 0.35 plants, 1.79 ears per plot, 0.58 g per 100-grain weight, 308.21 g ear weight per plot, and 261.83 kg/ha grain yield. Application of molecular markers at the stage of superior progeny selection led to increased genetic distance among populations, which is a very important factor for utilization of heterosis and providing greater longevity to the reciprocal recurrent selection program.
Subject(s)
Crosses, Genetic , Selection, Genetic , Zea mays/genetics , Flowers/genetics , Flowers/physiology , Genetic Markers , Genetics, Population , Genotype , Microsatellite Repeats/genetics , Phylogeny , Polymorphism, Genetic , Principal Component Analysis , Quantitative Trait, HeritableABSTRACT
We examined the effect of incorporation of molecular markers on variability between and within populations in order to maximize heterotic effects and longevity of a maize reciprocal recurrent selection program. Molecular variability was quantified by inter-simple sequence repeat (ISSR) markers between and within the maize populations Cimmyt and Piranão in the 10th cycle of a reciprocal recurrent selection program. Forty-two S(1) progenies of each population were analyzed, these being families of full-sibs selected according to their agronomic traits. Thirteen primers were selected, which produced 140 bands; 114 of them were polymorphic and 26 monomorphic. Based on UPGMA grouping analysis and by genetic distances, it was possible to identify "contaminant" progenies. These progenies belong to the Piranão or Cimmyt groups, but cluster in the opposite heterotic group. Identification of "contaminant" progenies is relevant for selection, because, besides identifying genotypes that should be eliminated at the recombination stage, it allows increased heterosis expression in crosses between more genetically distinct individuals. After the elimination of the "contaminant" progenies and those that were allocated between the heterotic groups, a new statistical analysis was carried out, which demonstrated increased genetic distances between the populations. It was concluded that the application of molecular markers in reciprocal recurrent selection programs allows the optimization of the monitoring of genetic variability within and between populations, favoring recombination between more distant progenies, besides ensuring increased longevity of the reciprocal recurrent selection program.
Subject(s)
Breeding , DNA, Plant/genetics , Genetic Variation , Hybrid Vigor , Sequence Analysis, DNA , Zea mays/genetics , DNA Primers/genetics , Genetic Markers , Selection, Genetic , Zea mays/growth & developmentABSTRACT
This in vivo study investigated the genotoxicity of two dental bonding agents: Adper Single Bond Plus and Prime&Bond 2.1. The somatic mutation and recombination test (SMART) in Drosophila melanogaster was applied to analyse their genotoxicity expressed as homologous mitotic recombination, as well as point and chromosomal mutation. SMART detects the loss of heterozygosity of marker genes expressed phenotypically on the fly's wings. This fruit fly has extensive genetic homology to mammals, which makes it a suitable model organism for genotoxic investigations. Adper Single Bond Plus induced statistically significant increases in the frequency of total spots at the highest concentration tested, while Prime&Bond 2.1 was positive at all concentrations tested. The mechanistic basis underlying the genotoxicity of Adper Single Bond Plus relies on mitotic recombination alone, and was different from that of Prime&Bond 2.1, which showed evidence of the contribution of both recombination and mutational events. These findings indicate that both adhesives are inducers of toxic-genetic events, with the mitotic recombination being the main mechanism of action. The clinical significance of these observations has to be interpreted with data obtained in other bioassays.
Subject(s)
Acetone/toxicity , Bisphenol A-Glycidyl Methacrylate/toxicity , Dental Bonding , Drosophila melanogaster/drug effects , Mutagens/toxicity , Polymethacrylic Acids/toxicity , Animals , Female , Heterozygote , Larva/drug effects , MaleABSTRACT
Abacavir (ABC), zidovudine (AZT), and lamivudine (3TC) are nucleoside analog reverse transcriptase inhibitors (NRTIs) widely used as combination-based antiretroviral therapy against human immunodeficiency virus. Despite effective viral suppression using NRTI combinations, genotoxic potential of NRTIs can be increased when administered in combination. This study investigated the toxic and genotoxic potential of ABC when administered alone or in combination with AZT and/or 3TC using the somatic mutation and recombination test in Drosophila melanogaster. This test simultaneously evaluated two events related to carcinogenic potential: mutation and somatic recombination. The results indicated that ABC was responsible for toxicity when administered alone or in combination with AZT and/or 3TC. In addition, all treatment combinations increased frequencies of mutation and somatic recombination. The combination of AZT/3TC showed the lowest genotoxic activity compared to all combinations with ABC. Therefore, our results indicated that ABC was responsible for a significant portion of genotoxic activity of these combinations. Somatic recombination was the main genetic event observed, ranging from 83.7% to 97.7%.
Subject(s)
Anti-HIV Agents/toxicity , Dideoxynucleosides/toxicity , Drosophila melanogaster/drug effects , Lamivudine/toxicity , Zidovudine/toxicity , Animals , DNA Damage , Drosophila melanogaster/genetics , Drug Synergism , Mutation , Recombination, GeneticABSTRACT
Production of IFN-gamma guarantees helpful T cell-mediated immunity against Mycobacterium tuberculosis infection. We have evaluated the in vitro immune responses to M. tuberculosis antigens using IFN-gamma production among 43 Brazilian tuberculosis (TB) patients prior to and after specific treatment, and 18 community controls. Peripheral blood mononuclear cells (PBMC) were cultivated in the presence either of purified protein derivative, ferritin, 10 kDa, 38 kDa, MPT59, Ag85A or Ag85B. Also, the two M. tuberculosis and M. bovis heat-shock proteins (hsp) 65 and 70 kDa were compared, and 5 day supernatants were harvested for cytokine detection by ELISA. The results showed that the overall profile of primary PBMC in response to most M. tuberculosis antigens was well correlated, since high IFN-gamma levels were induced by Ag85A, Ag85B, 38 kDa, ferritin and 10 kDa, as well as M. tuberculosis hsp65 in TB patients. In addition, analysis was carried out of the in vitro expression of activation molecules on lymphocytes, as CD25 and CD69 expression assessed in 17 TB patients showed induction on CD4+ T cells by Ag85B. Overall, significantly low responses were found in untreated, in comparison with the treated TB patients. Furthermore, internal community but not healthy control individuals have higher immune responses than do TB patients.
Subject(s)
Antigens, Bacterial/immunology , Interferon-gamma/biosynthesis , Mycobacterium tuberculosis/immunology , T-Lymphocytes/immunology , Tuberculosis/immunology , Adolescent , Adult , Aged , Brazil , CD4-Positive T-Lymphocytes/immunology , CD8-Positive T-Lymphocytes/immunology , Female , Humans , In Vitro Techniques , Male , Middle AgedABSTRACT
BACKGROUND: The hallmark of neurofibromatosis type 1 (NF1) is the development of multiple neurofibromas. Solitary neurofibroma may occur in an individual who does not have NF1, but multiple neurofibromas tend to develop only in those with NF1. It has been suggested that hormones may influence the neurofibromas of patients with NF1. The evidence that hormones may influence the growth of neurofibromas comes mainly from the observation that localised neurofibromas of patients with NF1 commonly grow during puberty and pregnancy. Because growth hormone (GH) concentrations increase during puberty, it is possible that GH influences the growth of these tumours. AIMS: To investigate the presence of GH receptors (GHRs) in neurofibromas. METHODS: By means of immunohistochemistry, the presence of GHRs was investigated in two groups of patients: 16 patients without NF1 with solitary neurofibromas (group A) and 10 patients with NF1 with localised neurofibromas (group B). RESULTS: Six of the 16 patients in group A had neurofibromas that were immunopositive for GHR, whereas nine of the 10 patients in group B were immunopositive. CONCLUSIONS: Most patients with NF1 have localised neurofibromas that express GHR. This suggests that GH may play some role in the development of localised neurofibromas in patients with NF1.
Subject(s)
Biomarkers, Tumor/analysis , Neurofibromatosis 1/metabolism , Receptors, Somatotropin/analysis , Adolescent , Adult , Child , Contraindications , Female , Growth Disorders/complications , Growth Disorders/drug therapy , Growth Hormone , Humans , Immunohistochemistry/methods , Male , Neurofibroma/chemistry , Neurofibromatosis 1/complicationsABSTRACT
To investigate the effects of tannic acid (TA) on ring-X chromosome loss, Drosophila melanogaster females exposed to different TA concentrations were crossed with untreated, methyl methanesulfonate (MMS)- or mitomycin C (MMC)-treated males which carried a ring-X chromosome. Progeny were analyzed for loss of the ring-X. The results of this in vivo study showed that TA had no suppressing effect on chromosome loss occurring spontaneously or after induction by MMS in mature spermatozoa. In contrast, TA caused a significant increase in the frequency of MMC-induced ring-X loss. The increase caused by this co-mutagenic effect reached values of 34, 33 and 40% at TA concentrations of 10, 25 and 50 mM, respectively. These increments may reflect the action of TA on a uvrABC-type enzyme which, by increasing the double-strand breaks (DSBs), somehow interferes with the post-replicational repair responsible for the final DSB correction.
Subject(s)
Chromosome Deletion , Drosophila melanogaster/drug effects , Hydrolyzable Tannins/toxicity , Mutagens/toxicity , X Chromosome/drug effects , Animals , Antimutagenic Agents/pharmacology , Chi-Square Distribution , DNA Repair/drug effects , Drosophila melanogaster/genetics , Drug Synergism , Female , Male , Methyl Methanesulfonate/toxicity , Mitomycin/toxicity , Mutagenicity Tests , Ring Chromosomes , Spermatozoa/drug effectsABSTRACT
Using a short-term bulk culture protocol designed for an intracellular-staining method based on a flow cytometry approach to the frequencies of cytokine-producing cells from tuberculosis and leprosy patients, we found distinct patterns of T cell subset expression. The method also reveals the profile of peak cytokine production and can provide simultaneous information about the phenotype of cytokine-producing cells, providing a reliable assay for monitoring the immunity of these patients. The immune response of Mycobacterium leprae and purified protein derivative (PPD) in vitro to a panel of mycobacteria-infected patients from an endemic area was assessed in primary mononuclear cell cultures. The kinetics and source of the cytokine pattern were measured at the single-cell level. IFN-gamma-, TNF-alpha-, IL-4- and IL-10-secreting T cells were intracytoplasmic evaluated in an attempt to identify M. leprae- and PPD-specific cells directly from the peripheral blood. The analysis by this approach indicated that TNF-alpha was the first (8 h) to be produced, followed by IFN-gamma (16 h), IL-10 (20 h) and IL-4 (24 h), and double-staining experiments confirmed that CD4+ were a greater source of TNF-alpha than of CD8+ T cells (P < 0.05). Both T cell subsets secreted similar amounts of IFN-gamma. We conclude that the protocol permits rapid evaluation of cytokine production by different T cell populations. The method can also be used to define immune status in non-infected and contact individuals.
Subject(s)
CD4-Positive T-Lymphocytes/immunology , CD8-Positive T-Lymphocytes/immunology , Cytokines/biosynthesis , Leprosy/immunology , Mycobacterium leprae/immunology , Tuberculosis, Pulmonary/immunology , Cytoplasm/immunology , Flow Cytometry , Humans , Interferon-gamma/biosynthesis , Interleukin-10/biosynthesis , Interleukin-4/biosynthesis , Tuberculin/immunology , Tumor Necrosis Factor-alpha/biosynthesisABSTRACT
The term neurofibromatosis (NF) is used for a group of genetic disorders that primarily affect the cell growth of neural tissues. Neurofibromatosis type 1 (NF1), also known as von Recklinghausen's disease, is the most common type of NF and accounts for about 90% of all cases. It is one of the most frequent human genetic diseases, with a prevalence of one case in 3,000 births. The expressivity of NF1 is extremely variable, with manifestations ranging from mild lesions to several complications and functional impairment. Oral manifestations can be found in almost 72% of NF1 patients. A case of a NF1 patient with a gingival neurofibroma in the attached gingiva of the lingual aspect of the lower central incisors is presented. The lesion was nodular, with sessile base, non-ulcerated, non-painful, with normal colour and measured 1 cm in diameter. An excisional biopsy of the oral lesion was performed. Histopathological and immunohistochemical analysis confirmed the clinical hypothesis of neurofibroma. Because NF1 is one of the most common genetic diseases and oral manifestations are very common, dentists should be aware of the characteristics of this disease.
Subject(s)
Gingival Neoplasms/pathology , Neurofibromatosis 1/pathology , Biopsy , Female , Humans , Immunohistochemistry , Incisor/pathology , Mandible , Middle Aged , Neurofibroma/pathologyABSTRACT
Nucleoside reverse-transcriptase inhibitor (NRTI) drugs are a major component of highly-active antiretroviral therapy (HAART). NRTI combinations have been demonstrated as producing a sustained reduction in plasma viremia with an increased CD4 count, thereby showing clear clinical benefits. Therefore, the secondary effects caused by the combination of two NRTIs, mainly those related to amplification of genotoxic effects, due to increased risk of DNA damage caused by these drugs, should be carefully examined. We employed the standard version of the wing SMART in Drosophila melanogaster to obtain more detailed knowledge about the genotoxic profile of NRTI combinations of AZT+ddI, AZT+3TC and AZT+d4T. Our results showed that all combinations increased the frequencies of induction of mutant spots. The combinations AZT+ddI and AZT+3TC were shown to induce recombination rates ranging from 86.38% to 98.36% while AZT+d4T showed a large discrepancy between recombination and mutation percentages. The combination index demonstrated that 3TC and d4T produced antagonism while ddI showed synergistic effects in combination with AZT.
Subject(s)
Antiviral Agents/adverse effects , DNA Damage/drug effects , Drosophila melanogaster/drug effects , Animals , Anti-HIV Agents/adverse effects , CD4 Lymphocyte Count , Didanosine/adverse effects , Dose-Response Relationship, Drug , Drosophila melanogaster/metabolism , Drug Combinations , Female , Lamivudine/adverse effects , Male , Mitosis , Mutation , Recombinant Proteins , Wings, Animal/drug effects , Zidovudine/adverse effectsABSTRACT
Multidrug-resistant tuberculosis (MDR-TB) is known as having a poor prognosis with a weak response to therapy and very high death rates. The aim of this work was to assess the immune response to the RD1-encoded antigen ESAT-6 of Mycobacterium tuberculosis in MDR-TB patients and compare to non-resistant (NR) TB patients and healthy controls (HC). Evaluation of interferon (IFN)-gamma production showed that, although 55% of the MDR patients were responsive to ESAT-6, they produced lower IFN-gamma levels (553 +/- 11 pg/ml) when compared to NR-TB (1179 +/- 163 pg/ml; P < 0.05) but not to controls (412 +/- 65.7 pg/ml). Differences in the response to ESAT-6 and to its overlapping peptides mixture were also significant between MDR versus treated pulmonary NR-TB. Furthermore, a very low rate of response to PPD (23.5%) and to Ag85B (33.3%) was noted in MDR-TB patients as compared to the other groups. To determine the inflammatory response in patients' groups, detection of tumour necrosis factor (TNF)-alpha was assessed in their sera before and during chemotherapy. Mean TNF-alpha levels in MDR-TB (43.8 +/- 9 pg/ml) paralleled those found in treated pulmonary, and it was significantly different (P < 0.05) from the values found in untreated NR and HC. Interestingly, secretion of IFN-gamma and TNF-alpha were predominant in MDR patients who presented with bilateral pulmonary lesions and lung cavitation. The present data indicate that the overall immune response to mycobacterial antigens is decreased in resistant TB and the major role inflammatory cytokines may play in perpetuating pulmonary tissue damage.
Subject(s)
Interferon-gamma/analysis , Tuberculosis, Multidrug-Resistant/immunology , Tumor Necrosis Factor-alpha/analysis , Adult , Antigens, Bacterial/immunology , Antitubercular Agents/therapeutic use , Bacterial Proteins , Case-Control Studies , Cells, Cultured , Female , Humans , Leukocytes, Mononuclear/immunology , Male , Middle Aged , Statistics, Nonparametric , Tuberculosis, Multidrug-Resistant/blood , Tuberculosis, Multidrug-Resistant/drug therapyABSTRACT
In this study, the taxanes, paclitaxel and docetaxel were investigated for genotoxicity in the wing spot test of Drosophila melanogaster. These relatively new drugs are used in cancer therapy and show great promise in the treatment of a variety of cancers. Their major cellular target is the alpha,beta-tubulin dimer but, unlike other spindle poisons, they stabilize microtubules by a shift towards assembly, producing nonfunctional microtubule bundles. The Drosophila wing Somatic Mutation and Recombination Test (SMART) provides a rapid means to evaluate agents able to induce gene mutations and chromosome aberrations, as well as rearrangements related to mitotic recombination. We applied the standard version of SMART (with normal bioactivation) and a variant version with increased cytochrome P450-dependent biotransformation capacity. In the standard assay, docetaxel was found to be aneuploidogenic; this was effectively abolished by a high cytochrome P450-dependent detoxification capacity. This suggests, as previously reported, the involvement of this family of enzymes in the detoxification of docetaxel rather than in its activation. In contrast, paclitaxel was clearly non-genotoxic at the same (millimolar) concentrations as used for docetaxel in both crosses. The weak responsiveness of SMART assays to aneugenic compounds, the weaker ligand and assembly action of paclitaxel and the more rapid reversibility of the microtubules formed with this compound, may have caused the negative response observed in the present study.
Subject(s)
Bridged-Ring Compounds/toxicity , Drosophila melanogaster/drug effects , Drosophila melanogaster/genetics , Mutagenicity Tests/methods , Paclitaxel/analogs & derivatives , Paclitaxel/toxicity , Taxoids , Angiogenesis Inhibitors/toxicity , Animals , Antineoplastic Agents, Phytogenic/toxicity , Biotransformation/drug effects , Biotransformation/genetics , Docetaxel , Drosophila melanogaster/cytology , Female , Genetic Markers/drug effects , Larva/drug effects , Larva/genetics , Male , Radiation-Sensitizing Agents/toxicityABSTRACT
Tannic acid (TA) was tested for genotoxic activity in three different assays (1-3) in Drosophila melanogaster by feeding of larvae or adult flies. TA did not induce sex-linked recessive lethals (1) nor sex-chromosome loss, mosaicism or non-disjunction (2) in male germ cells. In the wing somatic mutation and recombination test (SMART) (3) TA was found to be toxic for larvae of the high bioactivation cross and produced a weak positive response. These results suggest that this compound, when administered orally to larvae or adults of D. melanogaster, is not mutagenic and clastogenic in male germ cells, but weakly genotoxic in somatic cells of the wing imaginal disk.
Subject(s)
Drosophila melanogaster/genetics , Hydrolyzable Tannins/toxicity , Mutagens/toxicity , Animals , Female , Genes, Insect/drug effects , Genes, Lethal , Genes, Recessive , Larva , Male , Mosaicism , Mutagenicity Tests , Nondisjunction, Genetic , Recombination, Genetic , Sex Chromosome AberrationsABSTRACT
Apesar de o cloreto ser um exame amplamente disponível e solicitado em instituições de saúde, a maior pane dos médicos crê ser difícil interpretar seus resultados, estabelecer correlações com outros parâmetros laboratoriais associados e tomar decisões terapêuticas baseadas no mesmo. Devido à complexidade de sua homeostase e à intrincada correlação com o status hidroeletrolítico e ácido-básico, poucos médicos se sentem efetivamente aptos a aproveitar a valiosa informação clínica que ele pode revelar. Isto é agravado pela rarefeita bibliografia objetiva sobre o tema. Os autores desta revisão não foram capazes de encontrar nenhum capítulo especificamente dedicado ao cloreto nos principais livros-texto de Fisiologia, Clínica Médica e Nefrologia, disponíveis, à exceção de três revisões bibliográficas no Medline. Não obstante, tentamos organizar a informação tão claramente quanto possível, com o objetivo de tornar o cloreto uma ferramenta útil aos nossos colegas profissionais de saúde.
Even though chloride is a widely available and requested test in health institutions, most part of physicians find it difficult to interpret its results, establish correlations with other laboratory linked parameters and take therapeutic decisions based on it. Due to the complexity of its homeostatic balance and intrincated correlation to hydroelectrolytic and acid base status, few doctors feel actually able to fully profit from the valuable clinical information it can unfold. This is aggravated by the scarce objective bibliography on the issue. The authors of this review were not able to find any chapters specifically dedicated to chloride on major Physiology, Internal Medicine and Nephrology textbooks, but only three reviews on Medline. Nevertheless, we managed to organize the information as clearly as possible with the aim of making chloride test an useful tool to our fellow health professionals.
Subject(s)
Clinical Laboratory Techniques , Chlorides/physiology , Chlorides/blood , Acid-Base Imbalance/complications , Acid-Base Imbalance/diagnosis , Acid-Base Imbalance/etiology , Acid-Base Equilibrium/physiology , Acidosis/diagnosis , Acidosis/etiology , Alkalosis/diagnosis , Alkalosis/etiology , Bromides/adverse effects , Metabolism/physiologyABSTRACT
Using a short-term bulk culture protocol designed for an intracellular-staining method based on a flow cytometry approach to the frequencies of cytokine-producing cells from tuberculosis and leprosy patients, we found distinct patterns of T cell subset expression. The method also reveals the profile of peak cytokine production and can provide simultaneous information about the phenotype of cytokine-producing cells, providing a reliable assay for monitoring the immunity of these patients. The immune response of Mycobacterium leprae and purified protein derivative (PPD) in vitro to a panel of mycobacteria-infected patients from an endemic area was assessed in primary mononuclear cell cultures. The kinetics and source of the cytokine pattern were measured at the single-cell level. IFN-gamma-, TNF-alpha-, IL-4- and IL-10-secreting T cells were intracytoplasmic evaluated in an attempt to identify M. leprae- and PPD-specific cells directly from the peripheral blood. The analysis by this approach indicated that TNF-alpha was the first (8 h) to be produced, followed by IFN-gamma (16 h), IL-10 (20 h) and IL-4 (24 h), and double-staining experiments confirmed that CD4+ were a greater source of TNF-alpha than of CD8+ T cells (P < 0.05). Both T cell subsets secreted similar amounts of IFN-gamma. We conclude that the protocol permits rapid evaluation of cytokine production by different T cell populations. The method can also be used to define immune status in non-infected and contact individuals.