Identification of biomarkers in laryngeal cancer by weighted gene co-expression network analysis / 中南大学学报(医学版)

OBJECTIVES@#Laryngeal cancer (LC) is a globally prevalent and highly lethal tumor. Despite extensive efforts, the underlying mechanisms of LC remain inadequately understood. This study aims to conduct an innovative bioinformatic analysis to identify hub genes that could potentially serve as biomarkers or therapeutic targets in LC.@*METHODS@#We acquired a dataset consisting of 117 LC patient samples, 16 746 LC gene RNA sequencing data points, and 9 clinical features from the Cancer Genome Atlas (TCGA) database in the United States. We employed weighted gene co-expression network analysis (WGCNA) to construct multiple co-expression gene modules. Subsequently, we assessed the correlations between these co-expression modules and clinical features to validate their associations. We also explored the interplay between modules to identify pivotal genes within disease pathways. Finally, we used the Kaplan-Meier plotter to validate the correlation between enriched genes and LC prognosis.@*RESULTS@#WGCNA analysis led to the creation of a total of 16 co-expression gene modules related to LC. Four of these modules (designated as the yellow, magenta, black, and brown modules) exhibited significant correlations with 3 clinical features: The age of initial pathological diagnosis, cancer status, and pathological N stage. Specifically, the yellow and magenta gene modules displayed negative correlations with the age of pathological diagnosis (r=-0.23, P<0.05; r=-0.33, P<0.05), while the black and brown gene modules demonstrated negative associations with cancer status (r=-0.39, P<0.05; r=-0.50, P<0.05). The brown gene module displayed a positive correlation with pathological N stage. Gene Ontology (GO) enrichment analysis identified 77 items, and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis identified 30 related signaling pathways, including the calcium signaling pathway, cytokine-cytokine receptor interaction, neuro active ligand-receptor interaction, and regulation of lipolysis in adipocytes, etc. Consequently, central genes within these modules that were significantly linked to the overall survival rate of LC patients were identified. Central genes included CHRNB4, FOXL2, KCNG1, LOC440173, ADAMTS15, BMP2, FAP, and KIAA1644.@*CONCLUSIONS@#This study, utilizing WGCNA and subsequent validation, pinpointed 8 genes with potential as gene biomarkers for LC. These findings offer valuable references for the clinical diagnosis, prognosis, and treatment of LC.

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.

Comparison of the Microleakage and Shear Bond Strength to Dentine of Different Tricalcium Silicate-based Pulp Capping Materials / 대한소아치과학회지

This study evaluated the microleakage of three restorative materials and three tricalcium silicate-based pulp capping agents. The restorative materials were composite resin (CR), resin-reinforced glass ionomer cement (RMGI), and traditional glass ionomer cement (GIC) and the pulp capping agents were TheraCal LC® (TLC), Biodentine® (BD), and ProRoot® white MTA (WMTA). Additionally, shear bond strengths between the pulp-capping agents and dentine were compared.Class V cavities were made in bovine incisors and classified into nine groups according to the type of pulp-capping agent and final restoration. After immersion in 0.5% fuchsin solution, each specimen was observed with a stereoscopic microscope to score microleakage level. The crowns of the bovine incisors were implanted into acrylic resin, cut horizontally, and divided into three groups. TLC, BD and WMTA blocks were applied on dentine, and the shear bond strengths were measured using a universal testing machine.The microleakage was lowest in TLC + GIC, TLC + RMGI, TLC + CR, and BD + GIC groups and highest in WMTA + RMGI and WMTA + CR groups. The shear bond strength of BD group was the highest and that of WMTA group was significantly lower than the others.

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.

P2X7 Receptor Antagonism Attenuates the Intermittent Hypoxia-induced Spatial Deficits in a Murine Model of Sleep Apnea Via Inhibiting Neuroinflammation and Oxidative Stress / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>The mechanism of the neural injury caused by chronic intermittent hypoxia (CIH) that characterizes obstructive sleep apnea syndrome (OSAS) is not clearly known. The purpose of this study was to investigate whether P2X7 receptor (P2X7R) is responsible for the CIH-induced neural injury and the possible pathway it involves.</p><p><b>METHODS</b>Eight-week-old male C57BL/6 mice were used. For each exposure time point, eight mice divided in room air (RA) and IH group were assigned to the study of P2X7R expression. Whereas in the 21 days-Brilliant Blue G (BBG, a selective P2X7R antagonist) study, 48 mice were randomly divided into CIH group, BBG-treated CIH group, RA group and BBG-treated RA group. The hippocampus P2X7R expression was determined by Western blotting and real-time polymerase chain reaction (PCR). The spatial learning was analyzed by Morris water maze. The nuclear factor kappa B (NFκB) and NADPH oxidase 2 (NOX2) expressions were analyzed by Western blotting. The expressions of tumor necrosis factor α, interleukin 1β (IL-β), IL-18, and IL-6 were measured by real-time PCR. The malondialdehyde and superoxide dismutase levels were detected by colorimetric method. Cell damage was evaluated by Hematoxylin and Eosin staining and Terminal Transferase dUTP Nick-end Labeling method.</p><p><b>RESULTS</b>The P2X7R mRNA was elevated and sustained after 3-day IH exposure and the P2X7R protein was elevated and sustained after 7-day IH exposure. In the BBG study, the CIH mice showed severer neuronal cell damage and poorer performance in the behavior test. The increased NFκB and NOX2 expressions along with the inflammation injury and oxidative stress were also observed in the CIH group. BBG alleviated CIH-induced neural injury and consequent functional deficits.</p><p><b>CONCLUSIONS</b>The P2X7R antagonism attenuates the CIH-induced neuroinflammation, oxidative stress, and spatial deficits, demonstrating that the P2X7R is an important therapeutic target in the cognition deficits accompanied OSAS.</p>

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.

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.

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.

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. .

Effect of 38% carbamide peroxide on the microleakage of silorane-based versus methacrylate-based composite restorations

OBJECTIVES: This study aimed to assess the effect of 38% carbamide peroxide on the microleakage of class V cavities restored with either a silorane-based composite or two methacrylate-based composites. MATERIALS AND METHODS: A total of 96 class V cavities were prepared on the buccal surface of extracted human teeth with both enamel and dentin margins and were randomly assigned into three groups of Filtek P90 (3M-ESPE) + P90 system adhesive (3M-ESPE)(group A), Filtek Z250 (3M-ESPE) + Adper Prompt L-Pop (3M-ESPE)(group B) and Filtek Z350XT (3M-ESPE) + Adper Prompt L-Pop (group C). Half of the teeth were randomly underwent bleaching (38% carbamide peroxide, Day White, Discus Dental, applying for 15 min, twice a day for 14 day) while the remaining half (control) were not bleached. Dye penetration was measured following immersion in basic fuchsine. Data were statistically analyzed using Kruskal-Wallis and Mann-Whitney U tests at a level of 0.05. RESULTS: No significant differences were found between composites in the control groups in enamel (p = 0.171) or dentin (p = 0.094) margins. After bleaching, microleakage of Z250 (in enamel [p = 0.867] or dentin [p = 0.590] margins) and Z350 (in enamel [p = 0.445] or dentin [p = 0.591] margins) did not change significantly, but the microleakage of P90 significantly increased in both enamel (p = 0.042) and dentin (p = 0.002) margins. CONCLUSIONS: No significant differences were noted between the bleached and control subgroups of two methacrylate-based composites in enamel or dentin margins. Microleakage of silorane-based composite significantly increased after bleaching.

Enamel pretreatment with Er:YAG laser: effects on the microleakage of fissure sealant in fluorosed teeth

OBJECTIVES: The purpose of this in vitro study was to evaluate the microleakage and penetration of fissure sealant in permanent molar teeth with fluorosis after pretreatment of the occlusal surface. MATERIALS AND METHODS: A total of 120 third molars with mild dental fluorosis were randomly divided into 6 groups (n = 20). The tooth surfaces were sealed with an unfilled resin fissure sealant (FS) material. The experimental groups included: 1) phosphoric acid etching (AE) + FS (control); 2) AE + One-Step Plus (OS, Bisco) + FS; 3) bur + AE + FS; 4) bur + AE + OS + FS; 5) Er:YAG laser + AE + FS; and 6) Er:YAG laser + AE + OS + FS. After thermocycling, the teeth were immersed in 0.5% fuchsin and sectioned. Proportions of mircoleakage (PM) and unfilled area (PUA) were measured by digital microscope. RESULTS: Overall, there were significant differences among all groups in the PM (p = 0.00). Group 3 showed the greatest PM, and was significantly different from groups 2 to 6 (p < 0.05). Group 6 showed the lowest PM. Pretreatment with Er:YAG with or without adhesive led to less PM than bur pretreatment. There were no significant differences among groups in PUA. CONCLUSIONS: Conventional acid etching provided a similar degree of occlusal seal in teeth with fluorosis compared to those pretreated with a bur or Er:YAG laser. Pretreatment of pits and fissures with Er:YAG in teeth with fluorosis may be an alternative method before fissure sealant application.

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.

Heat fixed but unstained slide smears are infectious to laboratory staff.

Background and Aim: In a clinical microbiology laboratory, heat fixed slide smears are commonly transported from one place to another for staining with different stains and also for onsite proficiency testing of laboratory technicians for accreditation of the laboratories. These smears are frequently handled without gloves by the staff in developing countries. Therefore, this study was conducted to check the survivability of tubercle bacilli on smears after physical and chemical treatments. Methods: A total of 196 AFB positive smears were analyzed. Of these, 116 were stained with Ziehl Neelsen (ZN), 60 with cold Kinyoun and 10 were unstained but heat fixed and 10 were neither stained nor heat fixed. The last 20 smears served as controls. The ZN and Kinyoun stained smears were 0-1.5-year-old and stored at room temperature in slide boxes, while control smears were freshly prepared. All smears were prepared from sputum samples positive for acid fast bacilli. All four sets were subjected to slide culture to see if mycobacteria could survive and grow in any. For slide culture, a new and safe device was used, which is designed for three in one purpose: cell cultivation, direct observation of the growth under microscope and cell harvesting inside the closed tube. The slide smears were directly dipped into this tube that contained liquid culture medium. The tubes were incubated at 370C for four weeks. The growth, if any, was confirmed by MPT-64 rapid test and subculture on LJ slants. Results: No growth was observed in ZN and Kinyoun stained slide smears. However, significant growth was observed in both control sets; the unstained non heat fixed as well as heat fixed slide smears. Conclusions: The results of our study indicate that tubercle bacilli remain viable even after heat fixation and carry risk of infection by contact. However, stained smears are safe for handling and storage.

A preliminary study of Heavy Brilliant Blue G for internal limiting membrane staining in macular hole surgery.

Context: Surgical outcomes of vitrectomy for idiopathic macular hole using a “heavy” Brilliant Blue G (HBBG) solution for staining and removal of the internal limiting membrane (ILM). Settings and Design: Prospective interventional case series conducted in a tertiary eye care hospital. Materials and Methods: Nineteen patients (20 eyes) with idiopathic macular hole were enrolled to undergo vitrectomy with ILM peeling using HBBG. BBG dye was made heavy by mixing with 10% dextrose normal saline (DNS) solution in 2:1 ratio. The adequacy of ILM staining was noted intraoperatively. The closure rates of macular hole and visual improvement were recorded. Patients were followed up postoperatively on day 1, week 1, and subsequently at 1, 3, and 6 months, and every 6th month thereafter. Statistical Analysis: Wilcoxon signed-rank test was used; P < 0.05 was considered significant. Results: Preoperative best-corrected visual acuity (BCVA) ranged from 20/1000 to 20/63 (median: 20/100). Intraoperatively, the ILM stained very well in all eyes, and was easily removed. All macular holes closed postoperatively. The mean follow-up was 6.15 ± 2 months (range: 4-10; median: 6 months). Final BCVA ranged from 20/20 to 20/80 (median: 20/40), amounting to a significant visual improvement (P = 0.0001). BCVA improved by 1-8 Snellen lines in 19 eyes (95%); 16 eyes (80%) improved by ≥2 lines; 13 eyes (65%) achieved a final BCVA of 20/40 or better. Conclusions: Addition of 10% DNS to BBG dye allowed good ILM staining with less dye during macular hole surgery, and provided excellent anatomic and visual outcomes.

In vitro study of microleakage of different techniques of surface preparation used in pits and fissures.

Objective : The purpose of this in vitro study was to evaluate the effect of different techniques of surface preparation on the microleakage of a sealant applied with traditional acid etching and self-etched bonding agent. Study Design : A total of 60 extracted third molars were randomly assigned into six groups (n = 10/each). The occlusal surfaces were sealed with a sealant (Clinpro) after one of the following pretreatments: (1) phosphoric acid etching; (2) Prompt L-Pop; (3) laser + etching; (4) laser + Prompt L-Pop; (5) air abrasion + etching; (6) air abrasion + Prompt L-Pop. The specimens were immersed in a 0.5% basic fuchsin solution. Buccolingual cuts parallel to the long axis of the tooth were made. The surfaces were scored 0--2 for extent of microleakage using a microscope and the data were analyzed statistically. Results : The poorest results were obtained with laser + Prompt L-Pop which showed a greater number of specimens with microleakage (80%). Air abrasion surface preparation + phosphoric acid etching showed less microleakage than the other groups (40%). Kruskal--Wallis and t-tests revealed no significant difference in microleakage between six groups. Conclusion : The self-etching adhesive studied seems an attractive alternative to the acid-etch technique for sealant application in young children where simplifications in the clinical procedure are warranted. No significant difference was noted between the different types of enamel preparation before fissure sealant.