Patent blue V dye anaphylaxis: a case report and literature review / Anafilaxia por corante azul patente V: relato de caso e revisão da literatura

Abstract Background: Anaphylaxis is a constant perioperative concern due to the exposure to several agents capable of inducing hypersensitivity reactions. Patent blue V (PBV), also known as Sulfan Blue, a synthetic dye used in sentinel node research in breast surgery, is responsible for 0.6% of reported anaphylactic conditions. We present a case of a 49-year-old female patient who underwent left breast tumorectomy with sentinel lymph node staging using PBV and experienced an anaphylactic reaction. Methods: We conducted a literature search through PubMed for case reports, case series, review and systematic reviews since 2005 with the keywords "anaphylaxis" and "patent blue". We then included articles found in these publications' reference sections. Results: We found 12 relevant publications regarding this topic. The main findings are summarized, with information regarding the clinical presentation, management, and investigation protocol. Hypotension is the most common clinical manifestation. The presentation is usually delayed when compared with anaphylaxis from other agents and cutaneous manifestations are occasionally absent. Patients may have had previous exposure to the dye, used also as a food, clothes and drug colorant. Conclusion: The diagnosis of anaphylaxis in patients under sedation or general anesthesia may be difficult due to particularities of the perioperative context. According to the published literature, the presentation of the reaction is similar in most cases and a heightened clinical sense is key to address the situation appropriately. Finding the agent responsible for the allergic reaction is of paramount importance to prevent future episodes.

Resumo Introdução: A anafilaxia pode ocorrer durante o período perioperatório devido à exposição a diversos agentes capazes de induzir reações de hipersensibilidade. O corante sintético Azul Patente V (APV), também conhecido como Sulfan Blue, é usado na pesquisa de linfonodo sentinela em cirurgia de mama, e é responsável por 0,6% dos eventos anafiláticos relatados. Descrevemos o caso de uma paciente de 49 anos de idade submetida à tumorectomia de mama esquerda com estadiamento de linfonodo sentinela, em que se empregou o APV e que apresentou reação anafilática. Método: Por meio do PubMed, pesquisamos publicações que documentavam relatos de casos, séries de casos, revisões e revisões sistemáticas desde 2005, usando as palavras-chave "anaphylaxis" e "patent blue". Em seguida, incluímos artigos encontrados na lista de referências dessas publicações. Resultados: Encontramos 12 publicações relevantes sobre o tópico. Os principais achados estão resumidos, com informações do quadro clínico, tratamento e protocolo de investigação. A hipotensão foi a manifestação clínica mais frequente. De forma geral, o quadro clínico tem início tardio quando comparado à anafilaxia por outros agentes e, ocasionalmente, as manifestações cutâneas estão ausentes. Os pacientes podem ter tido exposição prévia ao APV, que também é usado como corante de alimentos, roupas e medicamentos. Conclusão: O diagnóstico de anafilaxia em pacientes sob sedação ou anestesia geral pode ser difícil devido às peculiaridades do contexto perioperatório. Segundo a literatura publicada, a apresentação da reação é semelhante na maioria dos casos e um discernimento clínico aguçado é fundamental para enfrentar o evento adequadamente. Encontrar o agente responsável pela reação alérgica é essencial para a prevenção de futuros episódios.

A prospective randomized trial comparing patent blue and methylene blue for the detection of the sentinel lymph node in breast cancer patients / Estudo randomizado prospectivo comparando o azul patente ao azul de metileno para a detecção do linfonodo sentinela em pacientes com câncer de mama

Summary Introduction: Methylene blue is more widely available and less expensive than patent blue, with an apparently lower risk of anaphylaxis. Objective: The two dyes were compared regarding detection of the sentinel lymph node (SLN). Method: A prospective, randomized trial involved 142 patients with invasive breast carcinoma. Sixty-nine (49.3%) assigned to patent blue (group A) and 71 (50.70%) to methylene blue (group B). Thirty-five patients (25.0%) were clinical stage III or IV; 55 (38.7%) had axillary lymph nodes affected; and 69 (49.3%) underwent neoadjuvant chemotherapy. Two patients were excluded because the dye type was not recorded. Results: Patients and tumor characteristics were similar in both groups. SLNs were identified in 47 women (68.1%) in group A and 43 (60.6%) in group B (p=0.35). SLNs were affected in 22 cases (51.2%) in group A and 21 (48.8%) in group B (p=0.62). The SLN was the only node affected in 12 cases (54.5%) in group A and six (33.3%) in group B (p=0.18). The time and degree of difficulty involved in identifying the SLN were similar in both groups. There were no complications or allergies. Conclusion: Methylene blue performed as well as patent blue in identifying the SLN in breast cancer patients.

Resumo Introdução: O azul de metileno é mais facilmente encontrado para comercialização e a um preço menor que o azul patente. Parece ainda haver menor risco de anafilaxia. Objetivo: Comparar a taxa de detecção do linfonodo sentinela com o azul patente e com o azul de metileno. Método: Foram incluídas, de forma randomizada e prospectiva, 142 pacientes com diagnóstico de carcinoma mamário invasor, que consentiram em participar livremente do estudo. Foram injetados 2 mL de azul patente (grupo A) em 69 (49,3%) mulheres e de azul de metileno (grupo B) em 71 (50,70%), em localização periareolar ou peritumoral, seguido de 5 minutos de massagem. Trinta e cinco (25,0%) apresentavam estadiamento clínico 3 ou 4, e 55 (38,7%) apresentavam a axila clinicamente comprometida. Sessenta e nove (49,3%) fizeram quimioterapia neoadjuvante. Duas pacientes não tinham anotação do corante utilizado e foram excluídas. Resultados: Os dois grupos apresentaram características das pacientes e dos tumores semelhantes. Foram detectados linfonodos sentinela em 47 (68,1%) mulheres no grupo A e em 43 (60,6%) no grupo B (p=0,35). Havia linfonodos sentinela comprometidos em 22 (51,2%) casos no grupo A e em 21 (48,8%) casos no grupo B (p=0,62). O linfonodo sentinela foi o único gânglio comprometido em 12 (54,5%) casos no grupo A e em seis (33,3%) casos no grupo B (p=0,18). O tempo e o grau de dificuldade para identificação do linfonodo sentinela foram semelhantes nos dois grupos. Não houve relato de complicações ou de alergia em nenhum dos grupos. Conclusão: A utilização do azul de metileno para a identificação do linfonodo sentinela em pacientes com câncer de mama apresenta resultados semelhantes aos do azul patente.

Allergic reaction to patent blue dye in breast surgery - case report / Reação alérgica ao corante azul patente em cirurgia de mama - relato de caso

Abstract We present a case of allergic reaction to patent blue in a patient who underwent excision of sentinel lymph node associated with segmental breast resection. About 20 min after the dye injection, the patient developed hypotension (BP = 70 × 30 mmHg) associated with increased heart frequency. The patient was treated successfully with decreased inspired fraction of inhaled anesthetic and fluid replacement. At the end of the procedure, she presented with bluish urticarial-like plaques on the head, neck, upper limbs, and trunk; hydrocortisone was then used. The patient recovered uneventfully and was discharged from the PACU 2 h after the end of surgery without skin changes, and was discharged from hospital on the morning after surgery. The incidence of allergic reactions with the use of patent blue is far superior to the hypersensitivity reactions seen with anesthetic and adjuvant drugs. Therefore, the anesthesiologist must be aware of cardiovascular instability associated with skin changes during the use of patent blue, for early diagnosis and appropriate treatment of this hypersensitivity reaction to this dye.

Resumo Os autores apresentam um caso de reação alérgica ao azul patente em uma paciente submetida à exérese de linfonodo em sentinela associada a uma ressecção segmentar de mama. Paciente apresentou aproximadamente pós 20 minutos da injeção do corante hipotensão (PA = 70 × 30 mmHg) associada a aumento da frequência cardíaca. Foi tratada satisfatoriamente com diminuição da fração inspirada do anestésico inalatório e reposição volêmica. No fim do procedimento apresentava placas urticariformes azuladas em cabeça, pescoço, membros superiores e tronco e foi usada hidrocortisona. Evoluiu, sem intercorrências, na sala de recuperação pós-anestésica e teve alta duas horas após o término do procedimento cirúrgico sem a presença das alterações cutâneas. Alta hospitalar na manhã seguinte à cirurgia. A incidência de reações alérgicas com o emprego do azul patente é muito superior às reações de hipersensibilidade observadas com drogas anestésicas e adjuvantes. Portanto, o anestesiologista deve ficar atento à instabilidade cardiovascular associada a alterações cutâneas quando do uso do azul patente para o diagnóstico precoce e tratamento adequado dessa reação de hipersensibilidade com o emprego do corante.

Sentinel node biopsy using blue dye and technetium99 in advanced gastric cancer: anatomical drainage and clinical application

Lymph node metastases are an independent prognosis factor in gastric carcinoma (GC) patients. Radical lymphadenectomy can improve survival but it can also increase surgical morbidity. As a principle, sentinel node (SN) navigation surgery can avoid unnecessary lymphadenectomy without compromising prognosis. In this pilot study, 24 patients with untreated GC were initially screened for SN navigation surgery, of which 12 were eligible. Five patients had T2 tumors, 5 had T3 tumors and 2 had T1 tumors. In 33% of cases, tumor diameter was greater than 5.0 cm. Three hundred and eighty-seven lymph nodes were excised with a median of 32.3 per patient. The SN navigation surgery was feasible in all patients, with a median of 4.5 SNs per patient. The detection success rate was 100%. All the SNs were located in N1 and N2 nodal level. In 70.9% of cases, the SNs were located at lymphatic chains 6 and 7. The SN sensitivity for nodal staging was 91.6%, with 8.3% of false negative. In 4 patients who were initially staged as N0, the SNs were submitted to multisection analyses and immunohistochemistry, confirming the N0 stage, without micrometastases. In one case initially staged as negative for nodal metastases based on SN analyses, metastases in lymph nodes other than SN were found, resulting in a 20% skip metastases incidence. This surgery is a reproducible procedure with 100% detection rate of SN. Tumor size, GC location and obesity were factors that imposed some limitations regarding SN identification. Results from nodal multisection histology and immunohistochemistry analysis did not change initial nodal staging.

Comparison between hemosiderin and Technetium-99 in sentinel lymph node biopsy in human breast cancer

PURPOSE: To assess the safety and potential equivalence of the use of hemosiderin compared to the Technetium-99 in sentinel lymph node biopsy in human breast cancer. METHODS: Non-random sample of 14 volunteer women diagnosed with breast cancer with primary tumors (T1/T2) and clinically tumor-free axilla were submitted to the identification of sentinel lymph node using hemosiderin obtained from autologous blood injected in the periareolar region 24h before surgery on an outpatient basis. Patients received preoperative subareolar intradermal injection of Technetium-99 in the immediate preoperative period. Patients were submitted to sentinel lymph node biopsy, with incision in the axillary fold guided by Gamma-Probe, dissection by planes until the identification of the point of maximum uptake of Technetium-99, identifying the marked nodes and their colors. All surgical specimens were sent for pathological and immunohistochemical study. RESULTS: The results showed no evidence of side effects and/or allergic and non-allergic reactions in patients submitted to SLNB with hemosiderin. The SLN identification rate per patient was 100%. SLNB identification rate per patient with hemosiderin was the same as that of Technetium, with a concordance rate of 100% between the methods. CONCLUSION: Hemosiderin is a safe dye that is equivalent to Technetium in breast sentinel lymph node biopsy.

Efficacy of fungal decolorization of a mixture of dyes belonging to different classes

Dyes are the most difficult constituents to remove by conventional biological wastewater treatment. Colored wastewater is mainly eliminated by physical and chemical procedures, which are very expensive and have drawbacks. Therefore, the advantage of using biological processes, such as the biotransformation of dyes, is that they may lead to complete mineralization or formation of less toxic products. To prove the possibility of using fungal processes for decolorization and other applications, the analysis of the toxicity of the processes' products is required. The decolorization of the mixture of two dyes from different classes - triphenylmethane brilliant green and azo Evans blue (GB - total concentration 0.08 g/L, proportion 1:1 w/w) - by Pleurotus ostreatus (BWPH and MB), Gloeophyllum odoratum (DCa), RWP17 (Polyporus picipes) and Fusarium oxysporum (G1) was studied. Zootoxicity (Daphnia magna) and phytotoxicity (Lemna minor) changes were estimated at the end of the experiment. The mixture of dyes was significantly removed by all the strains that were tested with 96 h of experimental time. However, differences among strains from the same species (P. ostreatus) were noted. Shaking improved the efficacy and rate of the dye removal. In static samples, the removal of the mixture reached more than 51.9% and in shaken samples, more than 79.2%. Tests using the dead biomass of the fungi only adsorbed up to 37% of the dye mixture (strain BWPH), which suggests that the process with the living biomass involves the biotransformation of the dyes. The best results were reached for the MB strain, which removed 90% of the tested mixture under shaking conditions. Regardless of the efficacy of the dye removal, toxicity decreased from class V to class III in tests with D. magna. Tests with L. minor control samples were classified as class IV, and samples with certain strains were non-toxic. The highest phytotoxicity decrease was noted in shaken samples where the elimination of dye mixture was the best.

State of the art in chromovitrectomy / Estado da arte da cromovitrectomia

Vitrectomy is a surgery that involves complex and delicate techniques that treat diseases such as macular hole, epiretinal membrane and diabetic macular edema. Chromovitrectomy is one of these techniques and includes the use of coloring agents such as vital dyes or crystals to enhanced visibility of transparent structures during vitrectomy. The aim of this study was to present a modern approach, based on scientific evidence, about the application and indication of vital coloring agents during vitrectomy. The use of such agents has made this surgery more predictable and has increased its post-operative prognosis. Although research on chromovitrectomy is currently expanding there is still not an established gold standard dyeing agent.

A cirurgia vitreorretiniana é uma cirurgia que envolve técnicas complexas e delicadas que tratam doenças como buraco macular, membrana epirretiniana e o edema macular diabético. A cromovitrectomia é uma dessas técnicas que incluem o uso de corantes compostos de pigmentos vitais ou cristais para melhorar a visibilização de estruturas transparentes durante a cirurgia de vitrectomia. O objetivo desse artigo foi apresentar uma abordagem atual, baseada em evidências, sobre a aplicação e indicação de corantes vitais durante a cirurgia vitreorretiniana. O emprego desses corantes possibilitou uma maior previsibilidade para a cirurgia, melhorando assim seu prognóstico pós-operatório. Apesar do campo da cromovitrectomia está em plena expansão de pesquisas, um corante gold standard para cromovitrectomia ainda não está estabelecido.

Efficacy of calcein and Coomassie Blue dyeing of shell growing-edges and micro growth-bands: Ageing juvenile of Pinctada mazatlanica (Pterioida: Pteriidae) / Eficiencia de la tinción de bandas de crecimiento con calceina y azul de coomassie, para estimar la edad de jóvenes de Pinctada mazatlanica (Pterioida: Pteriidae)

Age validation is the first step to determine shellfish species age determination. This information is vital for different inferential models used in marine ecosystem management activities. In spite that various validation techniques are used for marking carbon calcium structures, the calcein marking technique for oysters had never been used for age validation in Pinctada mazatlanica. Thus the objectives of this study included: the evaluation of calcein to mark a shell growing-edge, and the efficacy of Coomassie Blue staining on posterior shell growth, to produce visible micro growth-bands that would enable age validation of juvenile mother-of- pearl oysters. Oysters were collected and cultivated at The Perlas del Cortez S. de R. L. MI. pearl-farming opera tion, in Pichilingue, La Paz Bay, Baja California Sur, Mexico; a total of 36 oysters (shell height 11.5-36.4mm) were injected with calcein (0.125g/L), and another 50 oysters (shell height 14.8-42.7mm) were submersed in calcein (0.4 and 0.7g/L). Shell slices of calcein-marked oysters were posteriourly stained with Coomassie Blue R-25 for micro growth-band recognition. Our results showed that Calcein marking only worked by submersion and produced a concise bright lime-green florescent band along the growing-edge with clear boundaries for both concentrations. However, marks resulted better at the lower calcein concentration (0.4g/L) with more “perfect” and “good” marks on the growing-edge (p=0.0012). Commassie Blue staining technique was successful, and allowed to conclude that one micro growth-band was laid down per day, similar to other oyster species. Mean 15-d increment of shell growth height was slightly greater at the lower calcein concentration ( =0.735mm) than at the higher one ( =0.577mm) (not significant difference, p=0.198). Calcein marking of shell growing- edges and Commassie Blue staining of posterior shell growth, as a method for age validation is recommended for shellfish shell growth-band counts. This will allow back-dating for estimation of very precise colonization dates, both spatially and temporally in future work.

La validación de la edad es el primer paso para determinar las edades de las especies de moluscos, esta información es de vital importancia para los diferentes modelos de inferencia utilizados en actividades de gestión de los ecosistemas marinos. Diversas técnicas de valida- ción se utilizan para marcar estructuras de carbonato de calcio, aunque la técnica de marcado de calceína en ostras nunca se había utilizado para la validación de la edad de P. mazatlanica. Los objetivos de este estudio fueron: evaluar la calceína como marcador interno de la concha y la eficiencia del azul de Coomassie en la tinción de la matriz proteica de la concha, para facilitar la observación y conteo de micro bandas de crecimiento que permiten validar la edad de las ostras juveniles de madre perla. Las ostras fue- ron recolectadas en la costa de la empresa Perlas del Cortez S. de RL MI., en Pichilingue en Bahía de La Paz, Baja California Sur, México. Se inyectaron 36 ostras (altura de concha 11.5-36.4mm) (0.125g/L de calceína) y otras 50 ostras (altura de la concha 14.8-42.7mm) se sumergieron (0.4 y 0.7g/L de calceína). Secciones de la concha marcadas con calceína fueron teñidos con azul de Coomassie R-25 para el reconocimiento de las micro bandas de crecimiento. El marcado con calceína fue exitoso por inmersión y produjo una banda fluorescente de color verde lima brillante con- cisa a lo largo del crecimiento interno de la concha. Sin embargo, las marcas fueron mejores a una concentración de calceína inferior (0.4g/L), con mayor cantidad de marcas “buenas” y “perfectas” (p=0.0012). La técnica de tinción con azul de Commassie también fue exitosa. Se detectó un crecimiento diario por micro banda, similar a lo encontrado en otras especies de ostras. La diferencia del crecimiento medio en relación a la altura de la concha en un lapso de 15 días, fue ligeramente mayor con una concentración de calceína inferior ( =0.735mm) que con la de mayor concentración ( =0.577mm), pero no significativamente (p=0.198). El marcado de conchas con calceína y tinción de matrices proteicas con azul de Coomassie posterior a su crecimiento, es recomendando como un método para la validación de la edad facilitando el conteo de micro bandas de crecimiento internas de la concha. Además, permitirá estimar edades con el fin de predecir fechas de colonización y ubicación de bancos naturales.

Bioremediation of dyes by fungi isolated from contaminated dye effluent sites for bio-usability

Biodegradation and detoxification of dyes, Malachite green, Nigrosin and Basic fuchsin have been carried out using two fungal isolates Aspergillus niger, and Phanerochaete chrysosporium, isolated from dye effluent soil. Three methods were selected for biodegradation, viz. agar overlay and liquid media methods; stationary and shaking conditions at 25 °C. Aspergillus niger recorded maximum decolorization of the dye Basic fuchsin (81.85%) followed by Nigrosin (77.47%), Malachite green (72.77%) and dye mixture (33.08%) under shaking condition. Whereas, P. chrysosporium recorded decolorization to the maximum with the Nigrosin (90.15%) followed by Basic fuchsin (89.8%), Malachite green (83.25%) and mixture (78.4%). The selected fungal strains performed better under shaking conditions compared to stationary method; moreover the inoculation of fungus also brought the pH of the dye solutions to neutral from acidic. Seed germination bioassay study exhibited that when inoculated dye solutions were used, seed showed germination while uninoculated dyes inhibited germination even after four days of observation. Similarly, microbial growth was also inhibited by uninoculated dyes. The excellent performance of A. niger and P. chrysporium in the biodegradation of textile dyes of different chemical structures suggests and reinforces the potential of these fungi for environmental decontamination.

Patent blue and air as an alternative for resection of nonpalpable breast lesions: a case series / Azul patente e ar como uma alternativa para a resseccao de lesoes nao palpaveis de mama: serie de casos

CONTEXT AND OBJECTIVE: Use of mammography for breast cancer screening has resulted in a significantly increased number of patients with nonpalpable radiological findings that need histopathological study for better management. The present study evaluated an alternative to excision of nonpalpable breast lesions, using injection of patent blue (CAS 3536-49-0) dye and air. DESIGN AND SETTING: Cohort study of 64 consecutive patients at a private clinic in the city of Teresina (Piauí), between January 2009 and December 2010. METHODS : The patients had received mammographic diagnoses of nonpalpable breast lesions classified as BI-RADS 3, 4 and 5, with indication of histopathological study. They underwent stereotaxy and/or ultrasound-guided injection of patent blue, for marking and subsequent excision of the lesion. RESULTS : The patients' mean age was 47.7 years. Nodes accounted for 53.1% of the breast abnormalities; microcalcifications, 37.5%; and complex cysts, 9.4%. In 89.1% of cases, the lesions were BI-RADS 4; 7.8% were BI-RADS 5 and 3.1% were BI-RADS 3. The histopathological findings were benign in 70.3% of the cases; atypical hyperplasia, 9.4%; and malignant, 20.3%. Among the malignant cases, 53.8% were carcinoma in situ and 46.2%, invasive carcinoma. The percentage of malignancy was 0% in BI-RADS 3 lesions; 14.3% in BI-RADS 4 and 100% in BI-RADS 5. In the cases of malignancy, the margins were clear in 92.3%. Reoperation to widen the margins was required in one patient. CONCLUSION: Excision of nonpalpable breast lesions marked with patent blue and air was possible in all cases. .

CONTEXTO E OBJETIVOS: A utilização da mamografia no rastreamento para o câncer de mama tem apresentado significativo aumento do número de pacientes com achados radiológicos impalpáveis que, para melhor orientação terapêutica, necessitam de estudo histopatológico. O presente estudo avaliou uma alternativa à ressecção de lesões impalpáveis de mama utilizando injeção de corante azul patente e ar. TIPO DE ESTUDO E LOCAL: Coorte de 64 casos consecutivos de uma clínica particular da cidade Teresina (PI), no período de janeiro de 2009 a dezembro de 2010. MÉTODOS: As pacientes receberam o diagnóstico mamográfico de lesões impalpáveis de mama classes BI-RADS 3, 4 e 5 com indicação para estudo histopatológico. Elas foram submetidas à injeção de azul patente, orientada por estereotaxia e/ou ultrassonografia para marcação e posterior ressecção da lesão. RESULTADOS: A média de idade das pacientes foi 47,7 anos. Os nódulos representaram 53,1% das anormalidades; as microcalcificações, 37,5% e os cistos complexos, 9,4%. Em 89,1% dos casos, as lesões eram BI-RADS 4; 7,8%, BI-RADS 5 e 3,1%, BI-RADS 3. Os achados histopatológicos foram benignos em 70,3% dos casos; com hiperplasia atípica em 9,4% e malignos em 20,3%. Dos casos de malignidade, 53,8% eram carcinoma in situ e 46,2%, carcinoma invasor. O percentual de câncer foi 0% nas lesões BI-RADS 3;14,3% nas BI-RADS 4 e 100% nas BI-RADS 5. Nos casos de neoplasia maligna, as margens estavam livres em 92,3%, sendo necessária reoperação para ampliação de margens em uma paciente. CONCLUSÃO: A ressecção de lesões impalpáveis de mama marcadas com azul patente e ar foi possível em todos os casos. .

Rapid detection of multidrug-resistant Mycobacterium tuberculosis using the malachite green decolourisation assay

Early detection of drug resistance in Mycobacterium tuberculosis isolates allows for earlier and more effective treatment of patients. The aim of this study was to investigate the performance of the malachite green decolourisation assay (MGDA) in detecting isoniazid (INH) and rifampicin (RIF) resistance in M. tuberculosis clinical isolates. Fifty M. tuberculosis isolates, including 19 multidrug-resistant, eight INH-resistant and 23 INH and RIF-susceptible samples, were tested. The sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV) and agreement of the assay for INH were 92.5%, 91.3%, 92.5%, 91.3% and 92%, respectively. Similarly, the sensitivity, specificity, PPV, NPV and agreement of the assay for RIF were 94.7%, 100%, 100%, 96.8% and 98%, respectively. There was a major discrepancy in the tests of two isolates, as they were sensitive to INH by the MGDA test, but resistant by the reference method. There was a minor discrepancy in the tests of two additional isolates, as they were sensitive to INH by the reference method, but resistant by the MGDA test. The drug susceptibility test results were obtained within eight-nine days. In conclusion, the MGDA test is a reliable and accurate method for the rapid detection of INH and RIF resistance compared with the reference method and the MGDA test additionally requires less time to obtain results.

Experimental canine model for sentinel lymph node biopsy in the vulva using technetium and patent blue dye / Modelo canino experimental para a pesquisa do linfonodo sentinela na vulva usando tecnécio e azul patente

PURPOSE: This paper aims to study and define the experimental model of sentinel lymph node biopsy of the vulva in bitches. METHODS: 0.2 ml of 99mTc phytate was injected intradermally, using a fine gauge insulin needle in the anterior commissure of the vulva. Thirty minutes after 99mTc injection, the inguinal mapping was performed using a gamma probe. After this, 0.5 ml of blue dye (bleu patenté V Guerbet 2.5 percent) was injected in the same place. After 15 minutes, a 3 cm long inguinal incision was made at point maximum uptake followed by careful dissection, guided by visualization of a bluish afferent lymphatic system that points to the sentinel lymph node (SLN). RESULTS: It was observed that 88 percent of SLN were identified. It wasn´t found a significant difference among the presence or not of sentinel lymph node in the sides, which is an indication of a good consistency. It was observed a high (88 percent) and significant (χ2=12.89 and p=0.0003) intercession between both methods (blue dye and radiation). CONCLUSION: The experimental model adopted is feasible, becoming advantageous in applying the association of Patent blue and 99mTc.

OBJETIVO: Definir um modelo experimental para a pesquisa do linfonodo sentinela na vulva de cadelas. MÉTODOS: 0,2 ml de tecnécio diluído em fitato (99mTc) foi injetado por via intradérmica na comissura anterior da vulva. Trinta minutos após a injeção de 99mTc, o mapeamento inguinal foi realizada utilizando uma sonda gama. Após isto, 0,5 ml de corante azul (bleu Patente Guerbet V 2,5 por cento) foram injetados no mesmo lugar. Após 15 minutos, uma incisão inguinal de três centímetros foi feita no ponto de captação máxima seguido de dissecção cuidadosa, guiada através da visualização de um sistema linfático aferente corado em azul que apontava para o linfonodo sentinela (LS). RESULTADOS: Foi observado que em 88 por cento das vezes o LS foi identificado. Não foi encontrada diferença significativa entre a presença ou não de linfonodo sentinela nos lados direito e esquerdo, o que é uma indicação de uma boa consistência do método. Foi observada uma alta (88 por cento) e significativa (χ2 = 12,89 e p = 0,0003) intercessão entre os dois métodos (corante azul e radiação). CONCLUSÃO: O modelo experimental adotado é viável, tornando-se vantajoso na aplicação da associação de Azul Patente e 99mTc.

Therapeutic potential of Brazilian fluoride varnishes: an in vivo study

The aim of this study was to assess in vivo the therapeutic effect of three fluoride varnishes available in the Brazilian market on the performance of white spot lesions (WSL). The sample included 36 children aged 7 to 13 years old, with a total of 67 active WSL in permanent anterior teeth. The children were randomly divided into 3 groups, according to fluoride varnish used: FL- Fluorniz (n=24), DUO - Duofluorid XII (n=22) and DF - Durafluor (n=21). Maximum WSL dimensions (mesiodistal and incisogingival) were measured in millimeters by a previously calibrated single examiner using a periodontal probe. WSL were also assessed regarding lesion activity. Initial and final S-OHI (Simplified Oral Hygiene Index) scores were recorded. Pearson's chi-square test revealed no statistically significant differences (p>0.05) in the performance of the varnishes. At the end of the 5th week, FL had 6 active and 18 inactive WSL; DUO had 7 active and 15 inactive WSL; and DL had 6 active and 15 inactive WSL. Taking into account all lesions, there was a 45.7 percent reduction in WSL dimensions. Paired Student's t-test revealed a statistically significant difference (p<0.05) between the initial size (1.88) and final size (1.02). After four applications, all varnishes obtained similar clinical results.

O propósito do presente estudo foi avaliar in vivo o efeito terapêutico de 3 vernizes fluoretados disponíveis no mercado brasileiro em lesões de mancha branca (MB). A amostra foi composta de 36 crianças entre 7 e 13 anos, portadoras de 67 MB ativas em elementos dentários permanentes anteriores. As crianças foram divididas aleatoriamente em 3 grupos FL - Fluorniz (n=24), DUO - Duofluorid XII (n=22) e DF - Durafluor (n=21). A dimensão máxima da MB (mésio-distal e cérvico-incisal) foi mensurada em milímetros por um único examinador previamente calibrado através de uma sonda periodontal. Em adição, foi verificada a atividade da MB. O IHO-S (Índice de Higiene Oral Simplificado) inicial e final foi registrado. O teste do qui-quadrado de Pearson mostrou não existir diferença estatisticamente significante (p>0,05) no comportamento dos vernizes estudados. No final da 5ª semana o FL apresentava 6 MB ativas e 18 inativas, DUO 7 MB ativas e 15 inativas e DF 6 MB ativas e 15 inativas. Levando em consideração o grupo total, houve uma redução de 45,7 por cento da dimensão das manchas brancas. O teste T pareado revelou uma diferença estatisticamente significante (p<0,05) entre o tamanho inicial (1,88) e final (1,02). Após 4 aplicações, todos os vernizes apresentaram resultados clínicos semelhantes.

Growth kinetics of mycobacterium tuberculosis measured by quantitative resazurin reduction assay: a tool for fitness studies

We standardized a method to evaluate the growth kinetics of Mycobacterium tuberculosis by measuring quantitatively the reduction of resazurin by spectrophotometry. Growth curves and the rate of growth of twenty-one M. tuberculosis clinical isolates were determined. The method showed technical simplicity and is inexpensive to assess the fitness of each isolate.

National equipment of intraoperatory gamma detection in the identification of sentinel lymph node in animal model / Equipamento nacional de detecção gama intra-operatória na identificação de linfonodo sentinela

PURPOSE: To investigate a national equipment of intraoperatory gamma detection in the identification of sentinel lymph node. METHODS: Thirty young adult male rats were used. After anesthetized, animals were divided into two groups of 15 animals each. Animals from group A received dextram 500 - Tc99 radiopharmaceutical and patent blue V and those from group B received only patent blue V to map the lymphatic drainage. The presence of radiation in the background area, in the area of injection and of the ex vivo sentinel lymph node of group A were measured. After the exeresis, each lymph node in group A and in group B was mixed forming a new random sequence and the radioactive reading of each lymph node was carried out, using both pieces of equipment. RESULTS: The hottest sentinel lymph node was identified by the national equipment when radiation was measured in the area of limphatic drainage after the Dextran 500 was injected. Also, the ex vivo sentinel lymph node. The national equipment has also detected radiation in the lymph nodes that had not received radiopharmaceutical, leading to false positive, checked by the application of Mann-Whitney tests and Student's paired t-tests. The Cronbach alpha has shown high internal consistency of data 0,9416. CONCLUSIONS: The national equipment of intraoperatory gamma detection identifies the LS and showed false positives LS and needs improvement.

OBJETIVO: Investigar o equipamento nacional de detecção gama intra-operatória na identificação de linfonodo sentinela. MÉTODOS: Foram utilizados 30 ratos machos, adultos jovens. Depois de anestesiados, os animais foram distribuidos em dois grupos de 15 animais cada. O grupo A recebeu radiofármaco dextran 500 - Tc99 e azul patente V e o grupo B, somente azul patente V para realização do mapeamento linfático. Foi realizada a medição da captação radioativa da região de fundo, do sítio de injeção e do linfonodo sentinela ex vivo do grupo A. Após a exérese, cada linfonodo do grupo A e do grupo B foram misturados formando uma nova seqüência aleatória e procedeu a leitura da radioatividade de cada linfonodo com os dois equipamentos. RESULTADOS: O linfonodo sentinela hipercaptante foi identificado pelo equipamento nacional durante as medições da captação radioativa na região do sítio de injeção e linfonodo sentinela ex vivo. O equipamento nacional detectou radiação mesmo nos linfonodos que não receberam o radiofármaco, causando falso positivo, verificado na aplicação dos testes de Mann-Whitney e t pareado de Student. O alfa de Cronbach mostrou alta consistência interna dos dados (0,9416). CONCLUSÕES: O equipamento nacional de detecção gama intra-operatória identifica o linfonodo sentinela e mostra falsos positivos e necessita de aprimoramento.

Validación y uso de la técnica del linfonodo centinela axilar con colorante exclusivo en pacientes con cáncer de mama / Exclusive use of blue dye to detect sentinel lymph nodes in breast cancer

Background: The use of a dye and radiocolloid to detect sentinel lymph nodes in breast cáncer increases the detection rates. However the use of either method alone does not modify the false negative rate. Therefore there is no formal contraindication for the exclusive use of dye to detect nodes. Aim: To repon a prospective analysis of the exclusive blue dye technique for sentinel node biopsy in patients with early breast cáncer Patients and methods: We analyzed the first 100 women with pathologically proven breast cáncer who met the inclusión criteria. Patent blue dye was used as colorant. In the first 25 cases sentinel node was identified using radiocolloid and blue dye an then an axillary dissection perfomed. In the next 25 women, blue dye was used exclusively for detection and an axillary dissection was perfomed. In the next 50 cases, blue dye was used and only isolated sentinel node biopsy was perfomed. Results: In 92 of the 100 women a sentinel node was successfully detected. In the first 50 women, the false negative rate of sentinel lymph node detection was 6.9 percent. No complications occurred. During follow-up, lasting three to 29 months, no axillary relapse was observed. Conclusions: Sentinel node biopsy in patients with early breast cáncer using exclusively blue dye is feasible and safe.

Morphological regional differences of epithelial cells along the midgut in Diatraea saccharalis Fabricius (Lepidoptera: Crambidae) larvae / Diferenças morfológicas regionais das células epiteliais ao longo do intestino médio de larvas de Diatraea saccharalis Fabricius (Lepidoptera: Crambidae)

The sugarcane borer, Diatraea saccharalis Fabricius, is a pest to sugarcane and many other crops. This work aims to characterize morphological variability in the epithelial cells (columnar, goblet and regenerative) along the midgut of D. saccharalis larvae. Fragments of the midgut (anterior, middle and posterior regions) were fixed and processed by light and scanning electron microscopy. There are both cytochemical and ultrastructural differences in the morphology of the epithelial cells, depending on their localization along the midgut. The apical surface of columnar cells shows an increase in both number and size of the apical protrusions from the anterior to the posterior midgut regions. There is an increase in the amount of PAS-positive (Periodic Acid-Schiff Reaction) granules detected in the cytoplasm of both the columnar and regenerative cells, from the anterior to the posterior region. The goblet cell apical surface is narrow in the anterior region, and enlarged in the posterior midgut; the chamber's cytoplasm extrusion are small and thin at the apical cavity surface, being thicker, longer and more numerous at the basal portion of the cavity. Our results suggest that the sugarcane borer midgut has two morphologically different regions, the anterior and the posterior; the middle region is a transitional region.

A broca da cana, Diatraea saccharalis Fabricius, é uma praga da cana-de-açúcar e outras plantações. O objetivo deste trabalho foi caracterizar variações morfológicas nas células epiteliais (colunares, caliciformes e regenerativas) ao longo do intestino médio de larvas de D. saccharalis. Fragmentos do intestino médio (anterior, mediano e posterior) foram fixados e processados para microscopia de luz e eletrônica de varredura. Existem diferenças morfológicas citoquímicas e ultra-estruturais nas células epiteliais, dependendo da sua localização no intestino médio. A superfície apical de algumas células colunares exibe projeções citoplasmáticas que aumentam em número e volume da região anterior para a posterior do intestino médio. Existe aumento dos grânulos PAS-positivos (Reação de Schiff) no citoplasma apical das células colunares e regenerativas, da região anterior para a posterior. A câmara das células caliciformes, na região anterior do intestino médio, mostra seu ápice estreito, enquanto que na posterior essa porção da câmara é alargada; as evaginações citoplasmáticas da câmara são pequenas e finas no ápice, sendo numerosas, longas e mais espessas na porção basal. Os resultados sugerem que o intestino médio da broca da cana apresenta duas regiões morfologicamente distintas, a anterior e posterior; a região mediana é uma região de transição.

Influence of thermal stress on marginal integrity of restorative materials

The aim of this study was to evaluate the influence of thermal stress on the marginal integrity of restorative materials with different adhesive and thermal properties. Three hundred and sixty Class V cavities were prepared in buccal and lingual surfaces of 180 bovine incisors. Cervical and incisal walls were located in dentin and enamel, respectively. Specimens were restored with resin composite (RC); glass ionomer (GI) or amalgam (AM), and randomly assigned to 18 groups (n=20) according to the material, number of cycles (500 or 1,000 cycles) and dwell time (30 s or 60 s). Dry and wet specimens served as controls Specimens were immersed in 1 percent basic fuchsine solution (24 h), sectioned, and microleakage was evaluated under x40 magnification. Data were analyzed by Kruskal-Wallis and Mann-Whitney tests: Thermal cycling regimens increased leakage in all AM restorations (p<0.05) and its effect on RC and GI restorations was only significant when a 60-s dwell time was used (p<0.05). Marginal integrity was more affected in AM restorations under thermal cycling stress, whereas RC and GI ionomer restoration margins were only significantly affected only under longer dwell times.

Comparação do padrão de infiltração apical mostrado por dois diferentes métodos / Comparison of apical leakage patterns shown by two different

Objetivo: Comparar a infiltração apical usando penetração decorante e métodos eletroquímicos, usados nos mesmos espécimes.Método: Foram selecionados 31 dentes humanos unirradiculares,anteriores, com canais retos. Os espécimes foram divididosaleatoriamente, respectivamente em: grupo experimental, doiscontroles positivos e dois controles negativos. Os canaisradiculares do grupo experimental e do grupo controle negativoforam obturados e o controle positivo foi deixado sempreenchimento. A superfície externa de cada dente foi cobertacom duas camadas de esmalte de unha, com exceção de 2mmapicais e a porção coronária dos canais. A infiltração em cadadente foi medida usando-se o teste eletroquímico. Dois fios decobre foram usados como elétrodos e a solução salina foi usadacomo meio eletrocondutivo. Em seguida a porção coronária dasraízes foi obturada com cimento de Oxido de Zinco e Eugenol ecoberta com duas camadas de esmalte de unha. Os dentes foramimersos em fucsina básica a 2 por cento durante 48 horas e foram lavadosem água corrente. As raízes foram então longitudinalmenteseccionadas e foi avaliada a infiltração de corante em cada dente.Resultados: Nenhuma correlação foi enconrtada entre osresultados obtidos com os dois métodos utilizados (P =.297).Conclusão: Estudos adicionais são necessários antes da avaliaçãoda infiltração marginal apical.

Diagnóstico precoz de metástasis de melanoma mediante biopsia de linfonodo centinela: informe preliminar / Early diagnosis of melanoma metastasis by sentinel lymph node biopsy

Se presenta un estudio de linfonodo centinela y la experiencia inicial en el diagnóstico de micro metástasis linfática en melanoma maligno de piel, en un período de dos años. Se identificó en el pre operatorio el sitio del linfonodo centinela mediante albúmina marcada con Tc99 y, en el intra operatorio, con tinción vital con azul de isosulfán para seleccionar a los pacientes con micro metástasis que se pueden beneficiar con tratamiento quirúrgico linfático, descartar la cirugía ganglionar en los pacientes con linfonodo centinela negativo y reducir la morbilidad asociada al tratamiento quirúrgico. El criterio de inclusión incluyó pacientes con tumores Clark III o superior. Se incluyeron 6 pacientes; 2 Clark III, 3 Clark IV y un paciente Clark V. El diagnóstico de micro metástasis se realizó mediante tinción hematoxilina-eosina y estudio histoquímico de antígenos Melan A, S 100 y HMB 45. En dos pacientes Clark III y uno Clark IV, el estudio con H-E e inmunohistoquímica no identificó micro metástasis. Un paciente Clark IV no evidenció metástasis en el primer estudio con H-E; sin embargo, la inmunohistoquímica mostró micro metástasis con los tres marcadores tumorales. La revisión de las placas H-E confirmó el hallazgo. Un paciente con micro etapa IV y uno con micro etapa V mostraron metástasis a la tinción H_E y se confirmó la presencia de anticuerpos para los marcadores Melan A, S 100 y HMB 45. En este estudio el uso de Melan A, S 100 y HMB 45 mejoró la sensibilidad del diagnóstico histológico de metástasis linfáticas en un 20 por ciento y redujo a la mitad la indicación de disección ganglionar regional.