Mitose como fator prognóstico para biópsia de linfonodo sentinela em melanoma fino

Paciente, sexo feminino, 23 anos, com melanoma extensivo superficial em dorso, Breslow 0,35 mm, Clark II, sem ulcerações e com 2 mitoses / mm². Foi submetida à ampliação de margem e biópsia de dois linfonodos sentinela (axila esquerda). O exame anatomopatológico mostrou micrometástases, no seio subcapsular de ambos. Seguindo a recomendação do American Joint Commitee on Cancer 2009, a paciente foi submetida à linfadenectomia axilar total, sem outros linfonodos metastáticos. A aplicação da dermatoscopia vem permitindo maior precisão diagnóstica de melanoma cutâneo, contribuindo para maior proporção de melanoma fino ao diagnóstico. A taxa mitótica foi incluída como um importante fator prognóstico para melanomas finos pelo American Joint Commitee on Cancer 2009, sugerindo biópsia para esses pacientes

Solubilization of proteins from human lymph node tissue and two-dimensional gel storage.

In the present study, we compared six different solubilization buffers and optimized two-dimensional electrophoresis (2-DE) conditions for human lymph node proteins. In addition, we developed a simple protocol for 2-D gel storage. Efficient solubilization was obtained with lysis buffers containing (a) 8 M urea, 4% CHAPS (3-[(3-cholamidopropyl) dimethylammonio]-1-propanesulfonate), 40 mM Tris base, 65 mM DTT (dithiothreitol) and 0.2% carrier ampholytes; (b) 5 M urea, 2 M thiourea, 2% CHAPS, 2% SB 3-10 (N-decyl-N,N-dimethyl-3-ammonio-1-propanesulfonate), 40 mM Tris base, 65 mM DTT and 0.2% carrier ampholytes or (c) 7 M urea, 2 M thiourea, 4% CHAPS, 65 mM DTT and 0.2% carrier ampholytes. The optimal protocol for isoelectric focusing (IEF) was accumulated voltage of 16,500 Vh and 0.6% DTT in the rehydration solution. In the experiments conducted for the sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), best results were obtained with a doubled concentration (50 mM Tris, 384 mM glycine, 0.2% SDS) of the SDS electrophoresis buffer in the cathodic reservoir as compared to the concentration in the anodic reservoir (25 mM Tris, 192 mM glycine, 0.1% SDS). Among the five protocols tested for gel storing, success was attained when the gels were stored in plastic bags with 50% glycerol. This is the first report describing the successful solubilization and 2D-electrophoresis of proteins from human lymph node tissue and a 2-D gel storage protocol for easy gel handling before mass spectrometry (MS) analysis.

Influence of apoptosis on the cutaneous and peripheral lymph node inflammatory response in dogs with visceral leishmaniasis

In canine visceral leishmaniasis (CVL), the abnormalities most commonly observed in clinical examination on the animals are lymphadenomegaly and skin lesions. Dogs are the main domestic reservoir for the protozoon Leishmania (L.) chagasi and the skin is the main site of contamination by the vector insect. Some protozoa use apoptosis as an immunological escape mechanism. The aim of this study was to correlate the presence of apoptosis with the parasite load and with the inflammatory response in the skin and lymph nodes of dogs naturally infected with Leishmania (L.) chagasi. Thirty-three dogs from the municipality of Araçatuba (São Paulo, Brazil) were used, an endemic area for CVL. Muzzle, ear and abdominal skin and the popliteal, subscapular, iliac and mesenteric lymph nodes of symptomatic (S), oligosymptomatic (O) and asymptomatic (A) dogs were analyzed histologically. The parasite load and percentage apoptosis were evaluated using an immunohistochemical technique. Microscopically, the lymph nodes presented chronic lymphadenitis and the skin presented plasmacytic infiltrate and granulomatous foci in the superficial dermis, especially in the ear and muzzle regions. The inflammation was most severe in group S. The parasite load and apoptotic cell density were also greatest in this group. The cause of the lymphoid atrophy in these dogs was correlated with T lymphocyte apoptosis, thus leaving the dogs more susceptible to CVL. The peripheral lymph nodes presented the greatest inflammatory response. Independent of the clinical picture, the predominant inflammatory response was granulomatous and plasmacytic, both in the skin and in the peripheral lymph nodes. The ear skin presented the greatest intensity of inflammation and parasite load, followed by the muzzle skin, in group S. The ear skin area presented a non-significant difference in cell profile, with predominance of macrophages, and a significant difference from group A to groups O and S. It was seen that in these areas, there were high densities of parasites and cells undergoing apoptosis, in group S. The association between apoptosis and parasite load was not significant in the lymph nodes, but in the muzzle regions and at the ear tips, a positive correlation was seen between the parasite load and the density of cells undergoing apoptosis. The dogs in group S had the highest parasite load and the greatest number of apoptotic cells, thus suggesting that the parasite had an immune evasion mechanism, which could be proven statistically in the skin. © 2012 Elsevier B.V.

Papel da ultrassonografia axilar e da punção aspirativa por agulha fina na seleção de pacientes para a biópsia do linfonodo sentinela

Pós-graduação em Ginecologia, Obstetrícia e Mastologia - FMB

Estudo anátomo-clínico do linfonodo sentinela no câncer de mama

Pós-graduação em Ginecologia, Obstetrícia e Mastologia - FMB

Punção aspirativa de linfonodo axilar guiada pela ultrassonografia é eficaz como método de predição de acometimento linfonodal em pacientes com câncer de mama?

OBJETIVO: Avaliar a viabilidade e acurácia diagnóstica da ultrassonografia pré-operatória combinada com biopsia por agulha fina (US-PAAF) e do exame clínico da axila em pacientes com câncer de mama.MÉTODOS: Neste estudo prospectivo 171 axilas de pacientes com câncer de mama foram avaliadas pelo exame clínico e ultrassonografia (US) com e sem biopsia por agulha fina (PAAF). Os linfonodos com espessura cortical maior que 2,3 mm na ultrassonografia foram considerados suspeitos e submetidos a US-PAAF.RESULTADOS: A análise de regressão logística não mostrou correlação estatisticamente significativa entre exame clínico e axilas positivas no exame patológico. Em relação à avaliação axilar com US, o risco de achados anatomopatológicos positivos aumentou 12,6 vezes, valor Kappa de Cohen foi de 0,12 para exame clínico, 0,48 para US e 0,80 para US-PAAF. A acurácia foi de 61,4% para o exame clínico, 73,1% para os US e 90,1% para US-PAAF. Análise Receiver Operating Chracteristics (ROC) mostrou que uma espessura de 2,75 mm cortical correspondeu à mais elevada sensibilidade e especificidade na predição metástase axilar (82,7 e 82,2%, respectivamente).CONCLUSÕES: A US combinada com aspiração por agulha fina é mais precisa que o exame clínico na avaliação do status axilar no pré-operatório em mulheres com câncer de mama. Aquelas que são US-PAAF positivo podem ser direcionadas para esvaziamento linfonodal axilar imediatamente, e somente aqueles que são US-PAAF negativos devem ser considerados para biópsia de linfonodo sentinela.

Knowledge and practice of women regarding cervical cancer in a primary health care unit

Objectives: evaluating the level of information about the examination of uterine cervical cancer and its association with sociodemographic variables in women of a health care unit in the city of Bauru, São Paulo, Brazil. Methods: we conducted a cross-sectional descriptive study with 370 women aged 25 to 59, through structured interviews in their own homes; we used descriptive statistics and the χ2 test. Results: 40.5% of the women had not undergone the Papanicolaou test at the recommended frequency; 58.2% incorrectly defined the test, and 69.5% did not know about the risk factors for the development of cervical cancer; the knowledge about the test showed statistically significant association with schooling and family income of the studied population. Conclusions: women present deficiencies on the proper practice of the Papanicolaou test, on knowledge about the test, risk factors and prevention methods. Therefore, it is necessary to develop primary health actions for the most vulnerable population.

Results of screening for cervical cancer among pregnant and non-pregnant women in Brazil

Objective: To compare cervical cytology test results among pregnant and non-pregnant women, and to assess associations with age, screening history, and onset of sexual intercourse. Methods: A retrospective analysis was conducted of cervical smears obtained from women aged 18-34 years in the Campinas region of Brazil between January 2000 and December 2009. Eligible participants had not undergone cytological screening within the previous year and had no history of precursor lesions or cervical cancer. Multinomial logistic regression was performed for different age groups, with high-grade squamous intraepithelial lesions (HSILs) as the endpoint. Results: Overall, 3072 (0.4%) of 861 353 non-pregnant women and 135 (0.4%) of 37 568 pregnant women had HSILs. Odds of HSIL among pregnant and non-pregnant women did not differ in any age group. An increased age at first sexual intercourse among pregnant women reduced odds of HSILs in all age groups (odds ratio 0.9 [95% confidence interval 0.8-0.9] for all). Among women aged 21-24 years, 25-29 years, and 30-34 years, some associations were identified between an interval of less than 5 years since previous screening and reduced odds of HSILs. Conclusion: Mandatory cervical cytology screening does not seem to be necessary for pregnant women; protocols in place for non-pregnant women should be followed. (C) 2015 Published by Elsevier Ireland Ltd. on behalf of International Federation of Gynecology and Obstetrics.

A conjugate of the lytic peptide Hecate and gallic acid: structure, activity against cervical cancer, and toxicity

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Synergistic effect of photodynamic therapy and cisplatin: a novel approach for cervical cancer

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Association between concurrent genital bleeding and cervical cancer: a cross-sectional study

Genital bleeding may be a common symptom among women with cervical cancer. Cross-sectional study evaluating whether the prevalence of cervical smear results is different in women with and without clinical information about concurrent genital bleeding. The sample consisted of 2 324 836 smears; of these, 0.4% had clinical information on genital bleeding. When stratified by age group, women with genital bleeding had a higher chance of a cytological result of a high-grade squamous intraepithelial lesion [30-49 years odds ratio (OR) 2.38; 95% confidence interval (CI) 1.60-3.53 and ≥50 years OR 6.30; 95%CI 3.72-10.67), of squamous cell carcinoma (SCC) (30-49 years OR 24.70; 95%CI 11.96-51.03 and ≥50 years OR 48.91; 95%CI 31.28-76.47) and of atypical glandular cells (30-49 years OR 5.72; 95%CI 3.30-9.93 and ≥50 years OR 11.56; 95%CI 5.96-22.45); there was also a higher chance of adenocarcinoma for women ≥50 years (OR 53.13; 95%CI 28.08-100.51). The sensitivity of genital bleeding for women aged 18-29 years was 0.4% for high-grade squamous intraepithelial lesion (HSIL); for women 30-49 years old the rate was 0.9% for HSIL, 8.6% for SCC and 2.1% for atypical glandular cells of undetermined significance (AGUS), while for women aged from 50 years or more the rates were 2.0% for HSIL, 13.7% for SCC, 3.6% for AGUS and 14.7% for adenocarcinoma. Women ≥30 years old with genital bleeding should be referred for colposcopy to rule out the possibility of cervical cancer.

A biópsia de linfonodo sentinela não deve ser recomendada para pacientes portadores de melanoma espesso

Objective: To ascertain whether there is any relationship between the state of the sentinel lymph node histopathology, recurrence and mortality from thick melanoma in patients undergoing SLNB over a long follow-up. Methods: Eighty-six patients with thick melanoma undergoing SLNB were selected from a prospective database. Lymphoscintigraphy, lymphatic mapping and intraoperative gamma probe detection were performed in all patients. The sentinel lymph node (SLN) was analyzed by HE and immunohistochemistry. Complete lymphadenectomy was indicated for patients with positive sentinel node. The histopathological SLN status was related to the rate of recurrence and mortality from melanoma. Results: One hundred and sixty-six SLNs were taken from the 86 patients. Ages ranged from 18 to 73 years. There were 47 women and 39 men. Micrometastases were found in 44 patients. Forty-two patients underwent complete lymphadenectomy. Seven other patients had positive lymph node. Among the 44 patients with positive sentinel node, there were 20 recurrences and 15 deaths. There were 18 recurrences and 12 deaths in the group with negative SLN. The Breslow thickness was not correlated with the histopathological SLN status. The histopathological SLN status did not affect the rates of recurrence and mortality (Fisher test, p = 1.00). The median follow-up was 69 months. Conclusion: Considering the lack of evidence of benefit, SLNB should not be indicated for patients with thick melanoma outside of clinical studies.

Polymorphisms of Lewis and Secretor genes are related to breast cancer and metastasis in axillary lymph nodes

ABH and Lewis antigen expression has been associated with cancer development and prognosis, tumor differentiation, and metastasis. Considering that invasive ductal breast carcinoma (IDC) presents multiple molecular alterations, the aim of the present study was to determine whether the polymorphism of ABO, Lewis, and Secretor genes, as well as ABO phenotyping, could be associated with tumor differentiation and lymph nodes metastasis. Seventy-six women with IDC and 78 healthy female blood donors were submitted to ABO phenotyping/genotyping and Lewis and Secretor genotyping. Phenotyping was performed by hemagglutination and genotyping by the polymerase chain reaction with sequence-specific primers. ABO, Lewis, and Secretor genes were classified by individual single nucleotide polymorphism at sites 59, 1067, 202, and 314 of the Lewis gene, 428 of the Secretor gene, and 261 (O1 allele), 526 (O2 and B allele), and 703 (B allele). No association was found between breast cancer and ABO antigen expression (P = 0.9323) or genotype (P = 0.9356). Lewis-negative genotype was associated with IDC (P = 0.0126) but not with anatomoclinical parameters. Nonsecretor genotype was associated with axillary lymph node metastasis (P = 0.0149). In conclusion, Lewis and Secretor genotyping could be useful to predict respectively breast cancer susceptibility and axillary lymph nodes metastasis.

Effects of aspirin on mesenteric lymph nodes of rabbits as basis for its use on lymph nodes metastases

OBJETIVO: Avaliar os efeitos do ácido acetilsalicílico a 10% e 20% em linfonodos mesentéricos de coelhos para posterior embasamento e uso em metástases linfonodais. MÉTODOS: Um total de 20 linfonodos de 20 coelhos (divididos aleatoriamente em quatro grupos) foi avaliado. As soluções de aspirina a 10% (grupos A e C) e 20% (grupos B e D) foram injetadas em linfonodos mesentéricos de coelhos sadios e seus efeitos macroscópicos e histológicos foram avaliados em 24 horas (grupos A e B) e em sete dias (grupos C e D). RESULTADOS: Nos grupos avaliados em 24 horas (A e B) foi verificada intensa necrose e hemorragia, aumento importante de apoptose em todo o linfonodo, com alargamento dos seios medulares e aumento dos centros germinativos. Nos grupos avaliados em sete dias (C e D) também houve aumento da apoptose, com maior elevação de histiócitos e diminuição importante da necrose; a hemorragia foi ausente e aumento de células gigantes foi visualizado, conferindo processo inflamatório crônico do tipo corpo estranho. Não houve diferença entre as concentrações utilizadas (10 e 20%) em nenhuma das comparações. CONCLUSÕES: A injeção de aspirina em linfonodos causou necrose e um aumento de apoptose após 24 horas e após sete dias de tratamento, houve regeneração dos gânglios linfáticos, com diminuição intensa de necrose e grande aumento de apoptose. Uma vez que o aumento de apoptose é um dos pilares dos tratamentos antineoplásicos, estes resultados experimentais embasam eventual aplicação clínica da aspirina no tratamento de metástases linfonodais.