Evaluation of dermal collagen stained with picrosirius red and examined under polarized light microscopy

Abstract: The special picrosirius red staining highlights the natural birefringence of collagen fibers when exposed to polarized light. The results from birefringence allow to evaluate the organization of the collagen fibers in the tissues. The authors intend to elucidate all steps to obtain and capture images of histological sections stained with picrosirius red and evaluated under polarized light microscopy, as well as possible artefacts that may occur.

Diagnostic value of rosette sign under a polarized dermoscope / 中华皮肤科杂志

Objective To investigate the diagnostic value of rosette sign under a polarized dermoscope.Methods Lesions with rosette sign were selected from polarized dermoscopic image database of the Department of Dermatology of Peking University Third Hospital between September 2014 and March 2017.Then,histopathologically confirmed lesions were further chosen,and the correlations between the rosette sign and diseases were analyzed.These histopathologically confirmed lesions were divided into actinic keratosis (AK) group and non-AK group,and differences in clinical and dermoscopic features were analyzed between the 2 groups.Statistical analysis was carried out by nonparametric test for comparisons of the number of rosette sign between the AK group and non-AK group,as well as between different body sites.Results A total of 4 956 dermoscopic images of skin lesions were analyzed retrospectively,among which there were 144 (2.91％) skin lesions with rosette signs.Among the 144 skin lesions,74 were histopathologically diagnosed,37 (50.00％) of which were diagnosed as AK.Compared with the non-AK group,the AK group showed significantly higher proportions of lesions on the face (x2 =23.786,P ＜ 0.001) and at sun-exposed sites (x2 =12.921,P ＜ 0.001),and prevalence of superficial scales (x2 =7.056,P =0.008),keratotic plugs (x2 =6.167,P =0.013) and hair follicle openings surrounded by a white halo (x2 =4.893,P =0.027) under a dermoscope.Moreover,the number of rosette sign was significantly higher in facial lesions than in non-facial lesions (Z =-2.581,P =0.010),as well as in lesions at exposed sites than in those at unexposed sites (Z =-2.098,P =0.036).Conclusions The rosette sign is mainly observed in AK lesions.If lesions on the face or at sun-exposed sites are characterized by rosette sign,and superficial scales,keratotic plugs and hair follicle openings surrounded by a white halo can be observed under a dermoscope,these lesions can be diagnosed as AK with a high probability.

Effect of thermocycling to adhesive interface of enamel analyzed by polarized light microscopy / Análise do efeito da termociclagem na interface adesivo do esmalte analisada por microscopia de luz polarizada

Objetivo: A desmineralização menos agressiva do esmalte por sistemas adesivos autocondicionantes resulta em maior descoloração, infiltração marginal e falhas em suas restaurações. Deste modo, este estudo teve como objetivo avaliar a microinfiltração por meio da técnica da infiltração de nitrato de prata e formação de tags na interface esmalte/adesivo. Material e Método: Duzentos fragmentos de esmalte foram divididos aleatoriamente em 10 grupos (n = 10) de acordo com o sistema adesivo (Single Bond Adper Plus- (SB), Clearfil Tri-S Bond- (CF), ou Scotchbond Universal- (SBU)) e a superfície do esmalte (lixada (GE), não lixada- (UE), condicionada com ácido fosfórico 37%- (PHA), ou sem condicionamento): (SB-UE), (SB-GE), (CF-UE), (CFGE), (CF-ue/pha), (CF-ge/pha), (SBU-ue), (SBU-ge), (SBU-ue/pha) e (SBU-ge/pha). Metade das amostras restauradas foram submetidas a 20.000 ciclos térmicos. Quatro fatias de 1,0 mm/amostra foram obtidas para avaliar a formação de tags e infiltração de nitrato de prata. Todas as amostras foram examinadas com microscopia de luz polarizada e a percentagem de infiltração foi quantificada. Resultados: Não foram observadas interações entre os três fatores. O sistema adesivo e envelhecimento exibiram uma interação. Foram encontradas diferenças significativas somente após termociclagem: os grupos SB e SBU condicionados apresentaram menor porcentagem de infiltração comparados aos outros grupos, independente do tipo de esmalte. Quanto à analise qualitativa, o comprimento dos tags após condicionamento ácido foi maior para o GE comparado ao UE, independente do sistema adesivo. Conclusão: A aplicação dos sistemas adesivos na técnica autocondicionante mostrou uma formação de tags significativamente menor comparada à técnica convencional. O condicionamento ácido do esmalte previamente aos sistemas adesivos multi-mode é fundamental para reduzir o grau de infiltração na interface adesiva após envelhecimento.

Objective: The less aggressive demineralization of enamel by self-etching systems results in greater staining, marginal leakage, and failure in their restorations, so this study aimed to assess the silver nitrate infiltration and tag formation of the enamel/adhesive interface. Material and Methods: Two hundred enamel fragments were randomly assigned into 10 groups according to the adhesive system (Single Bond Adper Plus- (SB), Clearfil Tri-S Bond-(CF), or Scotchbond Universal-(SBU)) and enamel surface (ground- (ge), unground-(ue), phosphoric acid etching- (pha), or none) (n=10): (SB-ue), (SB-ge), (CF-ue), (CF-ge), (CF-ue/pha), (CF-ge/pha), (SBU-ue), (SBU-ge), (SBU-ue/pha), and (SBUge/pha). Half of the restored samples were submitted to thermocycling. Four slices of 1.0mm/ sample were obtained to evaluate tag formation and silver nitrate infiltration. All of the specimens were examined with Polarized Light Microscopy, and the percentage of infiltration was quantified. Results: No interactions were found among the three factors. The adhesive and aging exhibited an interaction. Significant differences were found only after thermocycling: the SB and SBU-etched groups had decreased infiltration compared with the other groups. The tag length after etching was higher for ge compared with ue, regardless of the adhesive system. Conclusion: The selfetching techniques resulted in significantly less tag formation compared with the conventional technique. The acid pre-etching of enamel with the multi-mode adhesive was fundamental for reducing the degree of infiltration of the adhesive interface after aging

Clinical factors and treatment outcomes associated with failure in the detection of urate crystal in patients with acute gouty arthritis

BACKGROUND/AIMS: To investigate the rate of detection of monosodium urate (MSU) crystals in the synovial fluid (SF) of patients with acute gouty arthritis and factors associated with false-negative results. METHODS: A total of 179 patients with acute gouty arthritis who had undergone SF crystal examination were identified from the data warehouse of two university hospitals. Clinical and laboratory data were obtained from the medical records. RESULTS: The overall rate of detection of MSU crystals was 78.8%. In univariate analyses, the only significant differences between the variables of crystal-negative and crystal-positive patients were a lower C-reactive protein level (p = 0.040) and fewer patients undergoing emergent surgery in the crystal-positive group (p = 4.5 x 10(-6)). In logistic regression analyses, MSU crystal-negative results were significantly associated with the interval from arthritis onset to crystal examination (p = 0.042), and this was the most significant risk factor for arthroscopic surgery (p = 2.1 x 10(-4)). Seventeen patients who underwent arthroscopic surgery had a significantly longer hospital stay (p = 0.007) and a significant delay in gout treatment (p = 8.74 x 10(-5)). The distribution of crystal-negative patients differed significantly between the SF samples that were evaluated by both the laboratory medicine and the rheumatology departments (p = 1.2 x 10(-14)), and the kappa value was 0.108. CONCLUSIONS: Although several clinical features were associated with detection failure, SF MSU crystal identification was critically dependent on the observer. Considering the impact on the treatment outcomes, implementation of a quality control program is essential.

Polarized dermoscopy of mammary Paget disease / Dermatoscopia da doença de Paget mamária

Mammary Paget's disease is a rare intraepithelial adenocarcinoma, located on the nipple/areola complex, highly associated with breast cancer. Although the international literature emphasizes the dermatoscopic pattern of mammary Paget's disease pigmented variant, the authors describe the dermoscopic findings of classical Paget's disease and demonstrate the presence of chrysalis-like structures, criteria recently described in the literature and not yet reported in Paget's disease. .

Doença de Paget mamária é considerada um adenocarcinoma intra-epitelial raro, localizado no complexo mamilo-aréola,com alta associação ao câncer de mama. Apesar da literatura mundial realçar o padrão dermatoscópico da doença de Paget mamária variante pigmentada os autores descrevem os achados dermatoscópicos da doença de Paget clássica realçando a presença das estruturas crisálida-símiles,critério recentemente descrito na literatura mundial e ainda não relatado na Doença de Paget. .

Análise invasiva e não invasiva do fuso meiótico de oócitos humanos obtidos de ciclos estimulados: dados preliminares / Invasive and noninvasive analysis of the meiotic spindle of human oocytes obtained from stimulate cycles: preliminary results

OBJETIVOS: Avaliar a concordância entre as técnicas de microscopia de polarização e microscopia confocal na avaliação do fuso meiótico de oócitos humanos maturados in vivo. MÉTODOS: Estudo prospectivo que avaliou oócitos com o primeiro corpúsculo polar extruído obtidos de mulheres inférteis submetidas à estimulação ovariana para realização de injeção intracitoplasmática de espermatozoide. Os oócitos com o primeiro corpúsculo polar extruído foram avaliados por meio da microscopia de polarização e, imediatamente após, foram fixados e corados para avaliação dos microtúbulos e cromatina pela microscopia confocal de alto desempenho. Foram comparadas as técnicas de microscopia de polarização e confocal, de acordo com a visualização ou não do fuso meiótico pela microscopia de polarização e a presença ou não de anomalias meióticas à análise pela microscopia confocal. Foram calculados os intervalos de confiança, o índice de Kappa e a concordância entre as metodologias, considerando a análise da microscopia de imunofluorescência como padrão-ouro para avaliação de normalidade do fuso e distribuição cromossômica oocitária. RESULTADOS: Observou-se que 72,7% dos oócitos em metáfase II com fuso celular não visível à polarização apresentaram anormalidades meióticas à análise confocal e que 55,6% dos oócitos em metáfase II com fuso celular visível à polarização apresentaram-se como oócitos anormais à análise confocal. Somente 44,4% dos oócitos com fuso celular visível à polarização apresentaram-se como normais à análise confocal. A concordância entre os métodos foi de 51,1% (Kappa: 0,11; IC95% -0,0958 - 0,319). CONCLUSÕES: A baixa concordância entre a microscopia de polarização e a confocal na avaliação do fuso meiótico oocitário sugere que a visualização do fuso meiótico de oócitos humanos em metáfase II pela microscopia de polarização tem limitado o valor preditivo de normalidade meiótica oocitária.

PURPOSE: To evaluate the concordance between polarization microscopy and confocal microscopy techniques in the evaluation of the meiotic spindle of human oocytes matured in vivo. METHODS: Prospective study that evaluated oocytes with the first polar extruded body obtained from infertile women who had undergone ovarian stimulation for intracytoplasmic sperm injection. The oocytes with the first polar extruded body were evaluated by polarization microscopy and were then immediately fixed and stained for microtubule and chromatin evaluation by high-performance confocal microscopy. We determined the correlation of polarization microscopy with confocal microscopy in the detection of meiotic oocyte anomalies, and we also evaluated the percentage of oocytes with a visible and non-visible cell spindle by polarization microscopy and with meiotic normality and abnormalities by confocal microscopy. Confidence intervals, Kappa's index and concordance between the methodologies were calculated, considering immunofluorescence microscopy analysis as the golden-standard for evaluating normal spindle and oocyte chromosome distribution. RESULTS: We observed that 72.7% of metaphase II oocytes with a nonvisible meiotic spindle by polarization microscopy showed no meiotic abnormalities by confocal analysis and 55.6% of metaphase II oocytes with a visible meiotic spindle by polarization microscopy were found to be abnormal oocytes by the confocal analysis. Only 44.4% of oocytes with a visible meiotic spindle by polarization microscopy were found to be normal by confocal analysis. Concordance between the methods was 51.1% (Kappa: 0.11; 95%CI -0.0958 - 0.319). CONCLUSIONS: The low correlation between polarization microscopy and confocal microscopy in the assessment of oocyte meiotic spindle suggests that visualization of the meiotic spindle of human oocytes at metaphase II by polarization microscopy is not a good indicator of oocyte meiotic normality.

Oócitos aparentemente maduros injetados em telófase I apresentam piores resultados de reprodução assistida / Apparently matured oocytes injected in telophase I have worse outcomes from assisted reproduction

OBJETIVO: Avaliar o estágio de maturação nuclear de oócitos com o primeiro corpúsculo polar (CP) visível de pacientes inférteis submetidas à estimulação ovariana para injeção intracitoplasmática de espermatozoide (ICSI) e comparar os resultados da injeção intracitoplasmática de espermatozoide entre os oócitos em telófase I (TI) e metáfase II (MII), e entre aqueles em metáfase II com e sem fuso celular visível. MÉTODOS: Estudo prospectivo que incluiu 106 pacientes inférteis submetidas à injeção intracitoplasmática de espermatozoide. Foram incluídas pacientes com idade menor ou igual a 38 anos, hormônio folículo estimulante (FSH) basal menor que 10 mIU/mL e índice de massa corpórea (IMC) menor que 30 kg/m². Foram excluídas pacientes com doenças sistêmicas, com qualquer infecção ativa, tabagistas ou que fizeram uso de medicações hormonais e anti-inflamatórias hormonais e não hormonais nos últimos dois meses, previamente à programação para o procedimento de reprodução assistida. Os oócitos com extrusão do primeiro corpúsculo polar foram avaliados pela microscopia de polarização, imediatamente antes da realização da injeção intracitoplasmática de espermatozoide, e caracterizados quanto ao estágio de maturação nuclear (telófase I ou metáfase II). Os oócitos em metáfase II foram avaliados de acordo com a presença ou não do fuso meiótico. Foram analisadas as taxas de fertilização, clivagem e o número de embriões de boa qualidade no segundo dia (D2) de desenvolvimento. Os dados foram analisados comparativamente através do teste exato de Fisher. Em todas as análises foi considerado o nível de significância de 5% (p<0,05). RESULTADOS: O fuso meiótico de 516 oócitos foi visualizado através da microscopia de polarização. Dezessete dos 516 oócitos avaliados estavam em telófase I (3,3%) e 499 (96,7%) em metáfase II. Os oócitos injetados em telófase I apresentaram taxas de fertilização significativamente menores do que os injetados em metáfase II (53 e 78%, respectivamente) e não produziram nenhum embrião de boa qualidade no segundo dia. Comparando-se os oócitos com e sem fuso celular visível, não foi observada diferença significativa nos resultados de injeção intracitoplasmática de espermatozoide. CONCLUSÕES: Oócitos injetados em telófase I apresentaram menores taxas de fertilização quando comparados aos em metáfase II. É possível que a análise do estágio de maturação nuclear oocitária, por meio da microscopia de polarização, possa ser utilizada como fator de predição das taxas de fertilização pós-injeção intracitoplasmática de espermatozoide.

PURPOSE: To evaluate the nuclear maturation stage and the presence of meiotic spindles of in vivo matured oocytes from infertile women undergoing stimulated cycles for intracytoplasmic sperm injection (ICSI) and compare intracytoplasmic sperm injection outcomes between oocytes in telophase I (TI) and metaphase II (MII), and the ones with and without visible meiotic spindle. METHODS: A prospective and controlled study with 106 infertile patients who underwent ovarian stimulation for intracytoplasmic sperm injection purposes. Patients aged 38 years or less, with basal follicle stimulating hormone (FSH) less than 10 mIU/mL and body mass index (BMI) less than 30 kg/m². Were included patients presenting any systemic diseases, any active infection, smokers or patients who had been using hormonal medications and hormonal and nonhormonal anti-inflammatory drugs for the past two months prior to the assisted reproduction procedure were excluded. The oocytes with the first polar body extruded (in vivo matured oocytes) were imaged by polarization microscopy immediately before intracytoplasmic sperm injection and characterized according to nuclear maturation stage (telophase I and metaphase II) and to the presence of a meiotic spindle. We analyzed the fertilization rates, cleavage, number of good quality embryos on the second day (D2) from oocytes on telophase I versus those in metaphase II, and metaphase II visible spindle versus non-visible ones. Data were analyzed comparatively by Fisher's exact test. The level of significance was set at 5% in all analyses (p<0.05). RESULTS: The meiotic spindles of 516 oocytes were imaged using polarization microscopy. From the 516 oocytes analyzed, seventeen were in telophase I (3.3%) and 499 (96.7%) in metaphase II. The oocytes injected in telophase I had significantly lower fertilization rates than those injected in metaphase II (53 and 78%, respectively) and produced no good quality embryos on day 2. When the oocytes with and without a visible meiotic spindle were compared, there was no significant difference in the intracytoplasmic sperm injection results. CONCLUSIONS: Oocytes injected in telophase I showed lower fertilization rates when compared to those in metaphase II. It is possible that the analysis of oocyte nuclear maturation by polarization microscopy can be used as a predictor of fertilization after intracytoplasmic sperm injection.