Desenvolvimento e aplicação de método GC-MS/MS para análise simultânea de 17 HPAs em material particulado atmosférico

Os hidrocarbonetos policíclicos aromáticos (HPAs) estão associados ao aumento da incidência de diversos tipos de cânceres no homem. Essas moléculas são formadas principalmente na queima incompleta de matéria orgânica, sendo encontradas em todos os compartimentos ambientais. Órgãos regulamentadores das áreas ambiental e de saúde ocupacional consideram 17 HPAs como contaminantes atmosféricos prioritários. Este trabalho apresenta um método para análise simultânea destes HPAs utilizando-se a cromatografia a gás acoplada à espectrometria de massas operando no modo tandem (GC-MS/MS). Os limites de detecção e quantificação do método mostraram-se até 5 vezes inferiores aos obtidos no método GC-MS (SCAN). O método mostrou-se seletivo para análise de HPAs em extratos de amostras de material particulado atmosférico. Uma análise comparativa de dois sistemas de solventes (diclorometano/metanol 4:1 v/v e hexano/acetona 1:1 v/v) para a extração de HPAs, utilizando amostras de material particulado atmosférico, revelou que ambas as misturas de solventes possuem poder de extração semelhante. Os resultados sugerem que é possível realizar extração de HPAs de material particulado atmosférico em ultra-som com a mistura hexano/acetona (1:1), que é menos tóxica em relação à mistura diclorometano/metanol (4:1), bastante utilizada nestas análises, sem perdas significativas na exatidão do método.

Ablação de tumor de canal anal tratado com laser de diodo: seguimento após 17 meses

The Nd:YAG laser is used as the palliative treatment of obstructive and/or hemorrhagic intestinal lesions with an effective but temporary symptomatic relief, with symptoms and signs recurrence after six to eight weeks. This report describes the treatment of a patient bearing a low rectal adenocarcinoma through diode laser ablation and the result after 17 months.

Traumatismo torácico: estudo retrospectivo de 168 casos

OBJETIVO: Avaliar o atendimento ao paciente portador de traumatismo torácico. MÉTODO: Estudo retrospectivo de 168 casos de trauma do tórax, com ênfase na abordagem inicial, conduta operatória e cuidados pós-operatórios. RESULTADOS: Dos 168 pacientes, 120 eram do sexo masculino e a média de idade encontrada foi de 35,5 anos. Dez pacientes foram toracotomizados de urgência, quatro por ferimento cardíaco, quatro devido à lesão de vasos pulmonares, um por lesão do saco pericárdico e outro do pedículo pulmonar. Os demais tiveram suas condições clínicas corrigidas através de simples drenagem torácica do hemitórax atingido. Ocorreram dois óbitos não cirúrgicos e um devido à insuficiência respiratória. CONCLUSÕES: O traumatismo torácico, além de ser muito freqüente, é na maioria das vezes, de fácil resolução. Podem ocorrer, no entanto, lesões torácicas graves envolvendo órgãos vitais que merecem intervenção mais agressiva e perícia técnica, com suporte hospitalar de alto nível.

Hiperplasia adrenal congênita: dosagem da 17-hidroxiprogesterona basal para seleção e casos para realização do teste de estímulo com ACTH sintético

OBJETIVOS: avaliar se os níveis de 17-hidroxiprogesterona podem predizer o resultado do teste de estímulo como diagnóstico de hiperplasia adrenal congênita, forma tardia. MÉTODOS: foram incluídas no estudo e avaliadas retrospectivamente 122 pacientes com suspeita clínica de hiperplasia adrenal congênita forma tardia. Essa suspeita clínica incluía sinais e/ou sintomas de hiperandrogenismo (hirsutismo, acne, pele oleosa, irregularidade menstrual, etc.). Todas as pacientes foram submetidas ao teste de estímulo da adrenal com ACTH sintético 0,25 mg (Synacthen®). Após repouso de 60 minutos as amostras foram colhidas nos tempos basal e 60 minutos após a administração de 0,25 mg de ACTH sintético para dosagem de 17-hidroxiprogesterona, sendo mantido o acesso venoso com catéter heparinizado. Foi utilizado o método de radioimunoensaio para realizar as dosagens séricas da 17-hidroxiprogesterona. A sensibilidade e a especificidade da 17-hidroxiprogesterona basal como teste de rastreamento para hiperplasia adrenal congênita foram medidas, avaliando vários pontos de corte. Curvas ROC foram feitas para analisar a performance do teste, utilizando o software Medcalc®. RESULTADOS: a análise por curva ROC mostrou um ponto de corte de 181 ng/dl acima do qual dever-se-ia realizar o teste de estímulo, bem próximo a 200 ng/dl, mais comumente aceito pela literatura. Níveis séricos da 17-hidroxiprogesterona mais altos que 200 ng/dl têm valores preditivo positivo e negativo de 75% e 100% e acurácia de 98,4% como diagnóstico de hiperplasia adrenal não-clássica. CONCLUSÕES: considerando os dados, sugerimos que pacientes com hiperandrogenismo clínico devam iniciar a investigação com 17-hidroxiprogesterona basal e, caso esta se mostre acima de 181 ng/dl, sigam a investigação com o teste de estímulo com ACTH sintético.

Expressão dos protooncogenes c-fos, c-myc e c-jun em miométrio normal e mioma humanos

OBJETIVO: Comparar a expressão gênica (mRNA) e protéica dos protooncogenes c-fos, c-myc e c-jun em miométrio normal e mioma humanos. MÉTODOS: Foi realizado um estudo do tipo caso-controle. O material foi coletado de 12 pacientes submetidas a histerectomia no Hospital de Clínicas de Porto Alegre. A expressão do mRNA específico para c-myc, c-fos, c-jun e beta-microglobulina foi avaliada pela técnica de RT-PCR, utilizando primers específicos para cada gene. A expressão protéica destes protooncogenes foi avaliada através de Western blot com anticorpos específicos. RESULTADOS: Não houve diferença significativa para expressão gênica desses protooncogenes entre miométrio normal e mioma (c-myc: 0,87 ± 0,08 vs 0,87 ± 0,08, p = 0,952; c-fos: 1,10 ± 0,17 vs 1,01 ± 0,11, p = 0,21; c-jun: 1,03 ± 0,12 vs 0,96 ± 0,09, p = 0,168, respectivamente). Não houve diferença significativa para expressão protéica desses protooncogenes entre miométrio normal e mioma (c-myc: 1,36 ± 0,48 vs 1,53 ± 0,29, p = 0,569; c-fos: 8,85 ± 5,5 vs 6,56 ± 4,22, p = 0,434; e c-jun: 6,47 ± 3,04 vs 5,42 ± 2,03, p = 0,266, respectivamente). CONCLUSÃO: A expressão gênica (transcrição) e a expressão protéica (tradução) dos protooncogenes c-myc, c-fos e c-jun em mioma e miométrio normal são semelhantes.

Aplicação pre-emergente de misturas de alachlor com diuron e cyanazine para o controle de plantas daninhas em algodão "IAC-17"

Para se conhecer a ação de misturas de alachlor com diuron ou com cyanazine para o controle de monotiledôneas e de dicotiledôneas em cultura de algodão 'IAC-17', foi conduzido um experimento de campo, em 1981/82, em Mogi-Mirim, SP, em solo com textura areno-argilosa, onde alachlor a 2,15 kg, 2,58 kg e 3,01 kg/ha foi aplicado em mistura de tanque com diuron, a 1,00 kg/ha ou com cyanazine a 1,00 kg/ha. Os herbicidas que compuseram as misturas foram empregados isoladamente, naquelas mesmas doses. Constaram do experimento ainda, duas testemunhas mantidas sem mato por meio de capinas e uma testemunha sem capina. Os 14 tratamentos foram distribuídos em blocos ao acaso, com quatro repetições. Foram realizadas observações visuais de infestação de mato aos 14, 31, 45, 75, 82, 94 e 100 dias após a aplicação dos herbicidas. Quando, pelo menos, uma parcela do tratamento atingia 25% de reinfestação, todo o tratamento era capinado e mantido no limpo até a colheita. Foram consideradas também a altura das plantas, sua população e a produção do algodão em caroço. Os resultados obtidos mostram que alachlor a 3,01 kg/ha + diuron a 1,00 kg/ha conseguiram bons resultados de controle das plantas daninhas por período de 100 dias enquanto que alachlor a 2,58 kg/ha + diuron a 1,00 kg/ha, e alachlor a 2,58 kg/ha e a 3,01 kg/ha + cyanazine a 1,00 kg/ha persistiram com sua ação de controle por 94 dias. Os tratamentos com alachlor a 3,01 kg/ha, diuron a 1,00 kg/ha, alachlor a 2,15 kg, 2,58 kg e 3,01 kg/ha em mistura com diuron a 1,00 kg/ha, e ainda alachlor a 3,01 kg/ha + cyanazine a 1,00 kg/ha, apresentaram tendências a reduzir a altura das plantas, segundo resultados de suas análises pelo teste de Tukey a 5%, sendo que a testemunha sem capina apresentou altura significativamente inferior aos demais tratamentos. A população e a produção não foram influenciadas pelos tratamentos com herbicidas, sendo semelhantes à testemunha capinada e superiores à testemunha sem capina.

Comparação entre diferentes concentrações e épocas de aplicação de cloreto de mepiquat, cloreto de clorocolina e ethephon em algodoeiro (Gossypium hirsutum L. cv. (AC - 17)

No ano agríco la de 1978/79 instalou-se um ensaio na área experimental da Escola Superior de Agricultura "Luiz de Queiroz", USP, em Piracicaba, SP, tendo por objetivo estudar os efeitos de reguladores vegetais, em diferentes doses e épocas de aplicação, sobre a cultura do algodoeiro (Gossypium hirsutum L. ). Os tratamentos utilizados foram: cloreto de mepiquat (DPC) nas concentrações de 84, 167 e 250ppm, cloreto de clorocolina (CCC) nas dosagens de 250, 380 e 450ppm e ethephon (CEPA) nas concentrações 1670, 3340 e 6880ppm, sendo cada substância aplicada 51, 65 e 143 dias após a germinação, respectivamente. CCC e DPC tornaram mais compacta a arquitetura das plantas de algodoeiro, assim como promoveram reduções nos parâmetros da análise de crescimento. O número de ramos produtivos e de maçãs não foi afetado por esses reguladores. Foram notadas algumas variações nos parâmetros de produtividade, sendo que CCC, DPC e CEPA tenderam a aumentar o peso médio de algodão por planta . Esses produtos não depreciaram os caracteres agronômicos do algodão, nem as características tecnológicas.

Floristic diversity of two crystalline rocky outcrops in the Brazilian northeast semi-arid region

Floristic composition and structure of vegetation were studied in two rocky outcrop areas in the semi-arid region of northeastern Brazil. From April 2007 to September 2008, 18 monthly field trips were carried out. Vascular plants were randomly collected throughout the outcrop areas. For structural analysis, 30 plots of 1 × 1 m were set in the vegetation islands. The checklist presented combines 211 species (69 families and 168 genera), although only 56 species were collected in the plots. Fabaceae (18 spp.; 8.5%), Asteraceae (17 spp.; 8%), Orchidaceae (13 spp.; 6.1%), Euphorbiaceae (13 spp.; 6.1%), Bromeliaceae (10 spp.; 4.7%), and Poaceae (eight spp.; 3.8%) are the richest families. Overall, 1,792 shrub and herbaceous specimens were counted in the plots. The Shannon-Wiener (H) diversity index values were 2.572 and 2.547 nats individual-1. The species that presented the highest absolute abundance values (number of plants) had low frequencies in the plots and vice-versa. The biological spectrum had a high proportion of phanerophytes and therophytes, followed by cryptophytes, chamaephytes, and hemicryptophytes. The studied flora shares floristic components similar to other rocky outcrop areas of the semi-arid region in northeastern Brazil, including in relation to dominant groups in the vegetation structure.

TP53 mutations and loss of heterozygosity of chromosome 17 in colorectal tumors

The incidence of TP53 point mutations and loss of heterozygosity (LOH) of chromosome 17 in colorectal tumors was determined in a group of Brazilian patients. We screened DNA samples from tumors and distal normal mucosa of 39 patients with colorectal cancer, for TP53 mutations by PCR-SSCP (single-strand conformation polymorphism) analysis. Chromosome 17 LOH was investigated using six PCR-based polymorphic markers and one VNTR probe. TP53 mutations were demonstrated in 15/39 of the cases. Mutations were distributed among all exons examined (five to eight), the majority of them being G/C to A/T transitions. LOH of chromosome 17p and 17q was detected in 70 and 46% of the tumors, respectively. There was a significant association between TP53 mutations and LOH in chromosome 17p (P = 0.0035) and 17q (P = 0.03). Although no correlation was observed between TP53 genetic alterations and clinical/ pathological characteristics, the association of TP53 mutations with loss of both chromosome 17 arms may indicate that TP53 inactivation provokes an unstable phenotype in tumor cells in colorectal tumors.

Percutaneous 17ß-estradiol replacement therapy in hypertensive postmenopausal women

The present study evaluated the short-term effects of percutaneous 17ß-estradiol on blood pressure, metabolic profile and hormonal levels in postmenopausal women with systemic arterial hypertension. After a wash-out period of 15 days, 10 hypertensive patients were treated with guanabenz acetate to control blood pressure, followed by 17ß-estradiol in the form of hydroalcoholic gel administered for 21 of 28 days of each cycle, for 3 cycles. Patients were evaluated before, during and 2 months after estrogen administration. Systolic and diastolic blood pressure or heart rate did not present any significant change in any patient when compared to those periods with the antihypertensive drug only (pretreatment period and 60 days after estrogen therapy was discontinued). Plasma biological markers of hepatic estrogenic action (plasma renin activity, antithrombin III, triglycerides, total cholesterol and lipoproteins) also remained unchanged during the study. Hormone treatment was effective, as indicated by the relief of menopausal symptoms, a decrease in FSH levels (73.48 ± 27.21 to 35.09 ± 20.44 IU/l, P<0.05), and an increase in estradiol levels (15.06 ± 8.76 to 78.7 ± 44.6 pg/ml, P<0.05). There was no effect on LH (18.0 ± 9.5 to 14.05 ± 8.28 IU/l). Hormone levels returned to previous values after estrogen treatment was discontinued. The data indicate that short-term percutaneous 17ß-estradiol replacement therapy, at the dose used, seems to be a safe hormone therapy for hypertensive menopausal women. Nevertheless, a controlled, prospective, randomized clinical assay with a larger number of subjects is needed to definitely establish both the beneficial and harmful effects of hormone replacement therapy in hypertensive women

NADPH-diaphorase activity in area 17 of the squirrel monkey visual cortex: neuropil pattern, cell morphology and laminar distribution

We studied the distribution of NADPH-diaphorase activity in the visual cortex of normal adult New World monkeys (Saimiri sciureus) using the malic enzyme "indirect" method. NADPH-diaphorase neuropil activity had a heterogeneous distribution. In coronal sections, it had a clear laminar pattern that was coincident with Nissl-stained layers. In tangential sections, we observed blobs in supragranular layers of V1 and stripes throughout the entire V2. We quantified and compared the tangential distribution of NADPH-diaphorase and cytochrome oxidase blobs in adjacent sections of the supragranular layers of V1. Although their spatial distributions were rather similar, the two enzymes did not always overlap. The histochemical reaction also revealed two different types of stained cells: a slightly stained subpopulation and a subgroup of deeply stained neurons resembling a Golgi impregnation. These neurons were sparsely spined non-pyramidal cells. Their dendritic arbors were very well stained but their axons were not always evident. In the gray matter, heavily stained neurons showed different dendritic arbor morphologies. However, most of the strongly reactive cells lay in the subjacent white matter, where they presented a more homogenous morphology. Our results demonstrate that the pattern of NADPH-diaphorase activity is similar to that previously described in Old World monkeys

Dose-dependent effects of 17-ß-estradiol on pituitary thyrotropin content and secretion in vitro

We studied the basal and thyrotropin-releasing hormone (TRH) (50 nM) induced thyrotropin (TSH) release in isolated hemipituitaries of ovariectomized rats treated with near-physiological or high doses of 17-ß-estradiol benzoate (EB; sc, daily for 10 days) or with vehicle (untreated control rats, OVX). One group was sham-operated (normal control). The anterior pituitary glands were incubated in Krebs-Ringer bicarbonate medium, pH 7.4, at 37oC in an atmosphere of 95% O2/5% CO2. Medium and pituitary TSH was measured by specific RIA (NIDDK-RP-3). Ovariectomy induced a decrease (P<0.05) in basal TSH release (normal control = 44.1 ± 7.2; OVX = 14.7 ± 3.0 ng/ml) and tended to reduce TRH-stimulated TSH release (normal control = 33.0 ± 8.1; OVX = 16.6 ± 2.4 ng/ml). The lowest dose of EB (0.7 µg/100 g body weight) did not reverse this alteration, but markedly increased the pituitary TSH content (0.6 ± 0.06 µg/hemipituitary; P<0.05) above that of OVX (0.4 ± 0.03 µg/hemipituitary) and normal rats (0.46 ± 0.03 µg/hemipituitary). The intermediate EB dose (1.4 µg/100 g body weight) induced a nonsignificant tendency to a higher TSH response to TRH compared to OVX and a lower response compared to normal rats. Conversely, in the rats treated with the highest dose (14 µg/100 g body weight), serum 17-ß-estradiol was 17 times higher than normal, and the basal and TRH-stimulated TSH release, as well as the pituitary TSH content, was significantly (P<0.05) reduced compared to normal rats and tended to be even lower than the values observed for the vehicle-treated OVX group, suggesting an inhibitory effect of hyperestrogenism. In conclusion, while reinforcing the concept of a positive physiological regulatory role of estradiol on the TSH response to TRH and on the pituitary stores of the hormone, the present results suggest an inhibitory effect of high levels of estrogen on these responses

Horizontal projections of area 17 in Cebus monkeys: metric features, and modular and laminar distribution

Metric features and modular and laminar distributions of intrinsic projections of area 17 were studied in Cebus apella. Anterogradely and retrogradely labeled cell appendages were obtained using both saturated pellets and iontophoretic injections of biocytin into the operculum. Laminar and modular distributions of the labeled processes were analyzed using Nissl counterstaining, and/or cytochrome oxidase and/or NADPH-diaphorase histochemistry. We distinguished three labeled cell types: pyramidal, star pyramidal and stellate cells located in supragranular cortical layers (principally in layers IIIa, IIIb a, IIIb ß and IIIc). Three distinct axon terminal morphologies were found, i.e., Ia, Ib and II located in granular and supragranular layers. Both complete and partial segregation of group I axon terminals relative to the limits of the blobs of V1 were found. The results are compatible with recent evidence of incomplete segregation of visual information flow in V1 of Old and New World primates

Acute promyelocytic leukemia: the study of t(15;17) translocation by fluorescent in situ hybridization, reverse transcriptase-polymerase chain reaction and cytogenetic techniques

Acute promyelocytic leukemia (AML M3) is a well-defined subtype of leukemia with specific and peculiar characteristics. Immediate identification of t(15;17) or the PML/RARA gene rearrangement is fundamental for treatment. The objective of the present study was to compare fluorescent in situ hybridization (FISH), reverse transcriptase-polymerase chain reaction (RT-PCR) and karyotyping in 18 samples (12 at diagnosis and 6 after treatment) from 13 AML M3 patients. Bone marrow samples were submitted to karyotype G-banding, FISH and RT-PCR. At diagnosis, cytogenetics was successful in 10 of 12 samples, 8 with t(15;17) and 2 without. FISH was positive in 11/12 cases (one had no cells for analysis) and positivity varied from 25 to 93% (mean: 56%). RT-PCR was done in 6/12 cases and all were positive. Four of 8 patients with t(15;17) presented positive RT-PCR as well as 2 without metaphases. The lack of RT-PCR results in the other samples was due to poor quality RNA. When the three tests were compared at diagnosis, karyotyping presented the translocation in 80% of the tested samples while FISH and RT-PCR showed the PML/RARA rearrangement in 100% of them. Of 6 samples evaluated after treatment, 3 showed a normal karyotype, 1 persistence of an abnormal clone and 2 no metaphases. FISH was negative in 4 samples studied and 2 had no material for analysis. RT-PCR was positive in 4 (2 of which showed negative FISH, indicating residual disease) and negative in 2. When the three tests were compared after treatment, they showed concordance in 2 of 6 samples or, when there were not enough cells for all tests, concordance between karyotype and RT-PCR in one. At remission, RT-PCR was the most sensitive test in detecting residual disease, as expected (positive in 4/6 samples). An incidence of about 40% of 5' breaks and 60% of 3' breaks, i.e., bcr3 and bcr1/bcr2, respectively, was observed.