Histomorphometric analysis of liver biopsies of treated patients with Gaucher disease type 1

Gaucher disease (GD) is an autosomal recessive lysosomal disorder caused by a disturbance in the metabolism of glucocerebroside in the macrophages. Most of its manifestations - hepatosplenomegaly, anemia, thrombocytopenia, and bone pain - are amenable to a macrophage-target therapy such as enzyme replacement. However, there is increasing evidence that abnormalities of the liver persist despite the specific GD treatment. In this work, we adapted histomorphometry techniques to the study of hepatocytes in GD using liver tissue of treated patients, developing the first morphometrical method for canalicular quantification in immunohistochemistry-stained liver biopsies, and exploring histomorphometric characteristics of GD. This is the first histomorphometric technique developed for canalicular analysis on histological liver biopsy samples.

Morphometry of the occipital condyle and basilar process of Nigerian skulls

Background: Effective surgical approaches and conclusive forensic investigations will require a well-documented comparative data for specific regions. This study was carried out to generate Nigerian specific morphometric data for the occipital condyle and basilar process and evaluate its application in forensic and clinical anatomy using macerated skull bones. Methods: A total of 141 adult Nigerian skull bones comprising of 126 males and 15 females were used for this study. Side specific (left [L] and right [R]) morphometric measurements were taken for the occipital condyle [OC] (length, maximum and minimum width [Mx OC & MnOC width], post and anterior intercondyle distance [AICD and PICD]). OC morphology was determined by shape and analysis was carried out using the Statistical package for Social Sciences (SPSS IBM® version 23) t-test was used in assessing sex differences in the measured parameters with confidence level set at 95% and P<0.05 was considered significant. Results: The result obtained showed an equal distribution of oval and oblong shaped OC (27% for both), with less frequent crescentic shaped OC (19.9%). The mean (± S.D) of studied parameters were; L-OC Length = 20.98 ± 1.88mm, R-OC length = 20.52 ± 1.93mm, L-MxOC width = 15.11 ± 1.53mm, R-MxOC width = 14.26 ± 1.59mm, L-MnOC width = 10.85 ± 1.23mm, R-MnOC width = 10.76 ± 1.30mm, AICD = 18.09 ± 2.15mm, PICD = 30.85 ± 2.98mm and BP length = 29.37 ± 3.85mm. The paired sample t-test showed that only the MnOC width was bilateral, while OClength and MxOC width were significantly asymmetrical (P<0.001).The t-test showed that all variables except for OC length and MnOC width were significantly larger in males (P<0.05). When the dimensions of the Nigerian skull were compared with foreign values, high racial variability was observed and we had average OC length; not very short neither long, which was relatively safe for OC resection during surgical intervention at that region. Conclusion: It could be concluded that sex, side and racial differences are observable features of the occipital condyle and its morphometric relationship to other structures are useful to forensic experts and surgeons

Identification of Epstein-Barr Virus in the Human Placenta and Its Pathologic Characteristics

Epstein-Barr virus (EBV), a common pathogen in humans, is suspected as the cause of multiple pregnancy-related pathologies including depression, preeclampsia, and stillbirth. Moreover, transmission of EBV through the placenta has been reported. However, the focus of EBV infection within the placenta has remained unknown to date. In this study, we proved the expression of latent EBV genes in the endometrial glandular epithelial cells of the placenta and investigated the cytological characteristics of these cells. Sixty-eight placentas were obtained from pregnant women. Tissue microarray was constructed. EBV latent genes including EBV-encoding RNA-1 (EBER1), Epstein-Barr virus nuclear antigen 1 (EBNA1), late membrane antigen (LMP1), and RPMS1 were detected with silver in situ hybridization and/or mRNA in situ hybridization. Nuclear features of EBV-positive cells in EBV-infected placenta were compared with those of EBV-negative cells via image analysis. Sixteen placentas (23.5%) showed positive expression of all 4 EBV latent genes; only the glandular epithelial cells of the decidua showed EBV gene expression. EBV infection status was not significantly correlated with maternal, fetal, or placental factors. The nuclei of EBV-positive cells were significantly larger, longer, and round-shaped than those of EBV-negative cells regardless of EBV-infection status of the placenta. For the first time, evidence of EBV gene expression has been shown in placental tissues. Furthermore, we have characterized its cytological features, allowing screening of EBV infection through microscopic examination.

Ambiente web para citomorfometria usando imagens 3D / Web environment for histomorphometry using 3D images

Softwares de bioimage são utilizados para a análise de imagens microscópicas e auxiliam os usuários em suas tomadas de decisões, a usabilidade pode limitar o seu uso. OBJETIVOS: Desenvolver um software de bioimage que pode ser acessado em um navegador web. Auxiliar patologistas e outros usuários na tomada de decisão, minimizando a subjetividade de suas avaliações. MÉTODOS: Foram realizados estudos nos softwares de bioimage open sources, o qual foram identificadas as características positivas e negativas, permitindo a escolha das tecnologias apropriadas para desenvolver o ambiente. RESULTADOS: O trabalho proveu um software de bioimage para análise citomorfométrica usando imagens tridimensionais que pode ser acessado por meio de um navegador web. CONCLUSÃO: O ambiente proposto é capaz de subsidiar os usuários com informações sobre as estruturas das células para tomada de decisão, fornecendo dados quantitativos e permitindo a exploração por meio de uma cena tridimensional.

Bioimage softwares are used for the analysis of microscopic images and assist users in their decision making,usability can limit its use. PURPOSES: Develop a bioimage software that can be accessed through a web browser. Assist pathologists and other users in decision making, minimizing the subjectivity of its evaluations. METHODS: Studies were carried out in the bioimage open sources softwares, which have been identified the positive and negative characteristics, allowing the choice of the appropriate technologies to developing the environment. RESULTS: The work provided abioimage software for histomorphometric analysis using three-dimensional images that can be accessed through a webbrowser. CONCLUSION: The proposed environment can subsidize the users with information on the structures of cells to decision making, providing quantitative data and allowing the exploration of a three-dimensional scene.

Biomarkers for screening of lung cancer and pre-neoplastic lesions in a high risk Chilean population

BACKGROUND: The mortality of lung cancer (LC), increases each year in the world, in spite of any advances, in development of new drugs to advance stages of LC. The high incidence of LC has been associated with smoking habit, genetic diversity and environmental pollution. Antofagasta region has been reported to have the highest LC mortality rate in Chile and its inhabitants were exposed to arsenic in their drinking water in concentrations as high as 870 µg/L. Non-invasive techniques such as biomarkers (Automatic Quantitative Cytometry: AQC and DR70) and Auto Fluorescence Bronchoscopy (AFB) might be potentially useful as a supplementary diagnostic approach and early detection. Early detection is one of the most important factors to intervene and prevent cancer progression in LC. This is a work of an ongoing prospective bimodality cancer surveillance study in high risk LC volunteers. Enrolment was done in subjects from Antofagasta and Metropolitan regions. In addition, we enrolled subjects who were suspected of having lung cancer. AQC, DR70 and AFB were used as tools in the detection of pre-neoplastic (PNL) and neoplastic lesions (NL). RESULTS: Half of the samples, classified as suspicious by AFB, were confirmed as metaplasia or dysplasia by histopathology. For LC, DR70 showed a higher sensitivity (95.8%) and specificity (91.9%) than AQC. However, for PNL AQC showed a higher sensitivity (91.9%) than DR70 (27.3%), although both with low PPV values. As a pre screener, both biomarkers might be employed as complementary tools to detect LC, especially as serially combined tests, with a sensitivity of 60% and a PPV of 65.2%. Additionally, the use of parallel combined tests might support the detection of PNL (sensitivity 91.2%; PPV 49.1%). CONCLUSION: This work adds information on cellular and molecular biomarkers to complement imaging techniques for early detection of LC in Latin America that might contribute to formulate policies concerning screening of LC. Supported by INNOVA-CORFO, Chile.

Análise morfométrica do colágeno dérmico a partir da segmentação por conglomerados (clusters) de cor / Morphometric analysis of dermal collagen by color clusters segmentation

Análise morfométrica do colágeno dérmico pode fornecer subsídio quantitativo para a pesquisa em dermatologia. Os autores demonstram uma técnica de análise de imagem digital que permite a identificação de estruturas microscópicas, a partir da segmentação por conglomerados (clusters), de cor aplicada à estimativa da intensidade e densidade das fibras colágenas da derme.

Morphometric analysis of dermal collagen can provide quantitative support to dermatologic research. The authors of this article disclose a technique of digital image analysis which allows the identification of microscopic structures by color cluster segmentation regarding the estimate intensity and density of dermal collagen fibers.

The application of fluorescence in situ hybridization and automated image cytometry in the diagnosis of urothelial carcinoma of bladder / 中华外科杂志

<p><b>OBJECTIVE</b>To discuss the application of automated DNA image cytometry (ICM) and fluorescence in situ hybridization (FISH) in the diagnosis of urothelial carcinoma of bladder.</p><p><b>METHODS</b>From August 2008 to March 2009, 60 volunteers with informed consent were divided into two groups, 40 patients proven as urothelial carcinoma of bladder by pathology and 20 healthy individuals as control. Urine was collected and tested by cytology, ICM and FISH.</p><p><b>RESULTS</b>Overall sensitivity of FISH was significantly higher in detection of malignancy than that of ICM (82.5% vs 62.5%, P < 0.05) and that of urine cytology (82.5% vs 25.0%, P < 0.05), while ICM was more sensitive to diagnose urothelial carcinoma of bladder than urine cytology (62.5% vs 25.0%, P < 0.05). Specificities of urine cytology, ICM and FISH were 100% in diagnosis of urothelial carcinoma of bladder (P > 0.05). Sensitivities of urine cytology, ICM and FISH have no correlation with pathological stage (P > 0.05), but have significant correlation with grade (P < 0.05).</p><p><b>CONCLUSIONS</b>ICM and FISH have the same specificity as urine cytology in diagnosis of urothelial carcinoma of bladder, but they have significantly higher sensitivity than urine cytology. FISH has the highest sensitivity among three diagnostic methods. Therefore, FISH may become a newly non-invasive technique for the diagnosis and surveillance of urothelial carcinoma of bladder.</p>

[Developmental study of the cervicothoracic [stellate] ganglion in ovine fetus]

Sheep as an experimental animal has been used in many veterinary researches. The developmental study of the ovine cervicothoracic ganglion in the fetal stages will reveal the anatomy of the sympathetic nervous system in the adult sheep and morphological and morphometrical relation between this ganglion and surrounding structures. The aim of this research was to follow morphological and morphometrical development of cervicothoracic ganglion in sheep during fetal stages and its relation to the body sides and sex. This study was performed on 18 ovine fetuses. The approximate age of fetnses were 11 to 16 weeks and were divided in 3 groups. Coalescence of the caudal cervical ganglion and few primary ganglia of the thoracic part of sympathetic trunk were morphologically studied. In Morphometrical studies, the length and width of the ganglia were measured. The results were statistically analyzed using 1-way ANOVA. The caudal cervical and first thoracic ganglia were the most frequent cervicothoracic ganglion. Variations in the length and width of ganglia between 11 to 16 weeks were not significant [p > 0.05], but with increasing of age interval become significant [p < 0.05]. There were not any correlation between length of ganglia and side of body and sex [p> 0.05]. The results of this study can be useful for evaluation of developmental and cell changes in neuropeptides of the sympathetic ganglia especially cervicothoracic ganglion

Morphometric analysis of the vertebral artery groove of the first cervical vertebra [atlas]

Variations in the morphometry of the vertebral artery groove which presents on the superior surface of surgery. This necessitates preoperative information about the vertebral artery groove. The present study aimed at assessment of the quantitative and qualitative anatomy of the vertebral artery groove of the atlas on 76 dry specimens with comprehensive analysis. This included the study of different linear parameters of the vertebral artery groove such as the distance from the midline, the thickness, the depth of the lateral and medial entrances, the depth of the transverse foramen and the width of the transverse foramen. In addition, the different forms of posterior and lateral bridging over the groove and their percentages were assessed. It was found that the minimum distance from the midline to the medial most edge of the vertebral artery groove in the inner and outer cortex of the posterior arch were 5 and 15 mm respectively. These data suggested that dissection of the posterior aspect of the posterior arch should remain 5 and 15 mm on the inner and outer cortex from the midline. It was also found that 44 [57.96%] of the examined specimens presented with a bridge formation which projects over the vertebral artery groove. From these 44 atlas presented with a bridge formation, 42 [55.26%] presented with partial bridges and 2 [2.63%] presented with complete posterior bridges. These bridges may interfere with the normal function of vertebral artery. It was concluded that before any craniovertebral intervention is performed, collection of the morphometric data of the vertebral artery groove must be carried out

Análise da DNA-ploidia em carcinoma lobular de mama / DNA content analysis of breast lobular invasive carcinoma

O carcinoma mamário apresenta elevados números relacionados à incidência e prevalência nas mulheres em vários países. No Brasil, é a neoplasia com um dos maiores índices de mortalidade. O tipo de carcinoma mamário mais freqüente é o ductal invasor seguido do carcinoma lobular. Embora essas neoplasias tenham características próprias, há ainda muitas questões a serem dirimidas. Sabe-se que os carcinomas lobulares apresentam alto risco de desenvolvimento bilateral e recorrência; estão relacionadas à expressão hormonal e têm desfecho lento. Sabe-se que o carcinoma lobular não tem positividade para a proteína E-caderina, cuja ausência de expressão imuno-histoquímica tem sido usada para diferenciá-la do carcinoma ductal. O padrão do conteúdo de DNA no carcinoma lobular tem sido objeto de controvérsias, com trabalhos descrevendo-o como predominantemente diplóide e outros, ao contrário, aneuplóide. O objetivo deste estudo foi analisar os padrões da DNA-ploidia em carcinomas lobulares invasivos de mama, E-caderina negativos, e correlacioná-los com fatores prognósticos de notória importância: expressão da proteína p53, Cerb-B2, receptores de estrógeno, tamanho dos tumores, comprometimento linfonodal, metástases a distância e pós-cirúrgica. Para isso foi realizado um estudo retrospectivo, com 50 casos examinados no Departamento de Anatomia Patológica do Hospital do Câncer A. C. Camargo, São Paulo-SP. Essas mulheres foram tratadas com cirurgia e as que apresentaram comprometimento axilar, receberam terapia adjuvante com quimioterapia, radioterapia e hormônioterapia. As medianas dos tempos de seguimento, com início no tratamento inicial (cirurgia) até o evento foi: grupo que foram a óbito 50 ± 27,6 meses; grupo com perda de seguimento 33 ± 56,0 meses; grupo de censura por interrupção do tempo para análise 79 ± 45,3 meses. A idade média foi de 54 anos (variando de 34 a 80 anos). A análise do conteúdo de DNA mostrou-se predominantemente aneuplóide (63,16 % dos casos). Somente o comprometimento linfonodal mostrou-se significativo (p=0,043) em relação a análise de DNA-ploidia, os demais parâmetros não mostraram associação significativa. Concluímos que a análise do conteúdo de DNA não é um parâmetro relevante para avaliar a agressividade do carcinoma lobular.

Análise da DNA-ploidia em carcinoma lobular de mama

O carcinoma mamário apresenta elevados números relacionados à incidência e prevalência nas mulheres em vários países. No Brasil, é a neoplasia com um dos maiores índices de mortalidade. O tipo de carcinoma mamário mais freqüente é o ductal invasor seguido do carcinoma lobular. Embora essas neoplasias tenham características próprias, há ainda muitas questões a serem dirimidas. Sabe-se que os carcinomas lobulares apresentam alto risco de desenvolvimento bilateral e recorrência; estão relacionadas à expressão hormonal e têm desfecho lento. Sabe-se que o carcinoma lobular não tem positividade para a proteína E-caderina, cuja ausência de expressão imuno-histoquímica tem sido usada para diferenciá-la do carcinoma ductal. O padrão do conteúdo de DNA no carcinoma lobular tem sido objeto de controvérsias, com trabalhos descrevendo-o como predominantemente diplóide e outros, ao contrário, aneuplóide. O objetivo deste estudo foi analisar os padrões da DNA-ploidia em carcinomas lobulares invasivos de mama, E-caderina negativos, e correlacioná-los com fatores prognósticos de notória importância: expressão da proteína p53, Cerb-B2, receptores de estrógeno, tamanho dos tumores, comprometimento linfonodal, metástases a distância e pós-cirúrgica. Para isso foi realizado um estudo retrospectivo, com 50 casos examinados no Departamento de Anatomia Patológica do Hospital do Câncer A. C. Camargo, São Paulo-SP. Essas mulheres foram tratadas com cirurgia e as que apresentaram comprometimento axilar, receberam terapia adjuvante com quimioterapia, radioterapia e hormônioterapia. As medianas dos tempos de seguimento, com início no tratamento inicial (cirurgia) até o evento foi: grupo que foram a óbito 50 ± 27,6 meses; grupo com perda de seguimento 33 ± 56,0 meses; grupo de censura por interrupção do tempo para análise 79 ± 45,3 meses. A idade média foi de 54 anos (variando de 34 a 80 anos). A análise ...

A test of screening to predict lung cancer among dust-exposed tin miners with sputum imaging cytometry / 中华劳动卫生职业病杂志

<p><b>OBJECTIVE</b>To evaluate efficacy of sputum imaging cytometry in early diagnosis on lung cancer among tin miners exposed to dust and analyze possible risk factors related to lung cancer among tin miners.</p><p><b>METHODS</b>From a cohort of tin miners in Guangxi Province, a total of 345 male tin miners higher than 45 years old and with high exposure to crystalline silica dust were randomly selected as the objects. Imaging cytometry was used to implement the sputum analysis for the screening on lung cancer according to the experience diagnostic standard. All objects were then followed up to the end of 2006. Clinical diagnosis of lung cancer was used as the golden standard to evaluate the efficacy of screening.</p><p><b>RESULTS</b>From 1998 to 2006, 11 new cases were diagnosed as clinical lung cancer. Except of age and exposure to occupational hazards, smoking status (P = 0.0384) and mean smoking dose (P = 0.0078) were significantly associated with lung cancer, and the adjusted odds ratio of high level to the low was 18.21 (2.15 approximately 154.39). The sensitivity, specificity and Youden's index of the sputum imaging cytometry for the experience diagnosis were 27.3%, 83.9% and 11.2% respectively. According to the ROC curve analysis, area under ROC Curve (AUC) of C2.5 (the percentage when the DNA index ranged from 1.25 to 2.50) was 0.647 (0.525 approximately 0.768), with the optimal operating point (OOP) of 1.70%. Sensitivity, specificity, agreement rate, positive predictive value, negative predictive value and Youden's index for predicting lung cancers in high-exposure tin miners were found to be 72.7%, 62.3%, 62.6%, 6.0%, 98.6% and 35.0% respectively.</p><p><b>CONCLUSION</b>Smoking is confirmed as an important risk factor of lung cancer in tin miners. The diagnostic efficiency can be improved if the diagnostic point of C2.5 is adjusted to 1.70%.</p>

[ correlation of urine and serum P53 protein in patients with transitional cell carcinoma of urinary bladder]

P53 is a tumor suppressor gene which is mutated in 50% of human cancers. The purpose of this study was to evaluate the association between the protein status in the serum / urine and tumors of bladder cancer. This descriptive study was done on 38 patients with transitional cell carcinoma of bladder who had no history of chemo- or radiotherapy or immune system disease. The sera and urine of these patients were analyzed for P53 protein by Enzyme Linked Immunosorbent Assay [ELISA] and tissue P53 by Immunohistochemical Technique. Individul and laboratory data were collected in questionnaire and analyzed by descriptive statistics and frequency distribution tables. Tissue P53 was detected in 29 of 39 [74.4%] patients, serum and urine P53 protein were detected in the serum and urine of 20 of 39 [51.3%] and 27 of 39 [69.2%] patients, respectively. The mean serum and urine P53 level in positive Tissue P53 patients were 1.45 and 2.27 U/ml respectively, which was significantly higher when compared with mean serum P53 level in negative tissue P53 patients [p< 0.01]. In patients with positive serum P53 [n=20] tissue P53 was positive in 18 [90%] patients and negative in 2 [10%], which was statiscally unsignificant. In negative serum P53 patients [n=19], tissue P53 was negative in 8[42%] patients and positive in 11 [58%]. In patients with positive urine P53 [n=27], tissue P53 was positive in 25 [93%] patients and negative in 2 [%7], which was statiscally unsignificant. Statiscally significant correlation was observed between tissue P53 with pathologic grade of tumor [p= 0.05], but for serum and urine P53 such correlation was not seen. This study shew a strong relationship between tissue P53 protein overexpression and level of P53 protein in serum and urine of TCC patients. Therefore both serum and urine of patients with TCC were found to have significant clinical accuracy for determination of P53 gene status in patients with TCC of bladder

Correlation between the conventional, routine histological grading of transitional cell carcinoma of the urinary bladder and morphometric analysis

Transitional cell carcinoma of the urinary bladder is one of the important malignancies in both sex groups. It is considered as a heterogenous neoplasm with different biological behavior, in which the majority are early non invasive with tendency for recurrence and some may progress to invasive tumor. Important clinicopathological features are, the tumor stage and histological grade which are used as prognostic parameters of the tumor and play an important role in therapy. Due to the subjectivity of the histological grading, the reproducibility was low. Many studies showed the value of quantitative analysis of the tumor as an important method in determining the recurrence of the tumor and muscular invasion, some other studies showed the value of nuclear measurement as a prognostic tool for bladder carcinoma. To evaluate the benefit of nuclear image analysis as an objective method for grading of transitional cell carcinoma of the urinary bladder and compare it with the subjective routine histological grading. Sixty two cases of transitional cell carcinoma of the urinary bladder, histologically diagnosed and graded according to WHO grading system, were selected. In each case 8-10 HPF were examined and 10-20 consecutive cells were studied, also we measured the largest 10 nuclei for each case separately by a computerized image analysis system at x400 magnification. Nuclear area and roundness were determined. Statistical analysis was performed using the analysis of variance and Tukey's test [HSD]. There was no statistical difference in the mean value of nuclear roundness between the three grades [P<0.05], while there was a statistical difference between grade I and III of their mean nuclear area [MNA] and MNA of the largest 10 nuclei [P<0.05]. No such difference was found between grade I and II or grade II and III [P<0.05]. Morphometric analysis should be based on the selection of special areas and not by random measurement as done on routine histological grading

Herbal activation of mammary gland; a comparative morphometrical and biochemical analysis

Seventy five adult virgin female Norway rats [60 experimental and 15 controls] were used to evaluate the effect of seeds of three herbs [Fennel, Cumin and Garden cress] on their mammary glands. Experimental animals were fed with these herbs [each type of herb seeds was given to twenty experimental rats] for fourteen days. Rats were sacrificed and mammary gland sections were obtained, stained then morphometrically assessed. Serum prolactin level was performed too. Results revealed that Garden cress seeds are the strongest lactogenic agent among the three. Both Fennel and Cumin seeds were shown to be very weak galactagogues

Aflatoxin B[1]-induced kidney damage in rats

Aflatoxin B[1] [AFB[1]] is a widely distributed mycotoxin in nature. Several investigations have shown its biological effects on different organs and in different animal species. However, the effects of AFB[1] on the rat kidney have not been much elucidated histologically. This study aims to demonstrate the effects of AFB[1] contaminated diet on the rat kidney from the histological and morphometric aspects. Twelve mature albino rats were divided equally into a control group fed with usual diet and a treated group which was daily fed with diet contaminated with 20 mg AFB[1]/kg of body weight for 30 days. Semithin sections from renal cortex were stained with methylene blue and examined by light microscopy. Corpuscular changes were also detected morphometrically in terms of the ratio between the area of Bowman's capsule and the area of its contained glomerulus [B/G ratio]. The treated group showed a marked increase in body weight. Histologically, there was evidence of acute tubular necrosis and increase in urinary space. Morphometrically, there was a diffuse significant increase in the B/G ratio compared to the control. Gain in weight can be attributed to fluid retention that accompanies the ensuing renal damage. The dietary dose of AFB[1] [20 mg/kg of body weight] for 30 days was sufficient to produce acute tubular necrosis. The corpuscular changes indicated by the increase in the B/G ratio can be attributed to compensatory hypertrophy

Immunohistochemistry quantification by a digital computer-assisted method compared to semiquantitative analysis

PURPOSE: To compare immunostaining quantification obtained by a digital computer-assisted method with the well-established semiquantitative analysis. METHODS: Cytoplasmic staining of galectin-3 was obtained by standard immunohistochemical reactions in 25 cases of well-differentiated thyroid carcinoma. The expression index that associates the conventional area fraction of labeled cells with the immunostaining intensity score based on visual qualitative observation was used as the semiquantitative analysis. A digital computer-assisted method is described based on the use of an image processing program (ImageLab®). Three parameters were obtained: (1) percentage of labeled cells; (2) digital immunostaining intensity, and (3) digital expression index. The proposed method allows numerical analysis of the immunostaining intensity. RESULTS: There was a strong correlation between the immunostaining intensity obtained by the two methods (Pearson correlation coefficient, r = 0.71, P = 0.0001). The same was observed between expression indexes (Pearson correlation coefficient, r = 0.66, P = 0.0001). CONCLUSION: Results obtained with our proposed digital computer-assisted method for immunoexpression analysis were concordant with the semiquantitative analysis. In addition, digital values can also resolve disagreement among different observers about the quality of staining intensity because the digital method does not classify the results into groups, but rather provides a numerical value for each individual case; thus, it increases the diagnostic and, more importantly, the prognostic sensitivity of the immunohistochemical analysis.

OBJETIVO: Comparar a quantificação da imunomarcação através de um método digital assistido por computador à bem estabelecida análise semiquantitativa. MÉTODO: A marcação citoplasmática de galectina-3 foi obtida por reações imunohistoquímicas em 25 casos de carcinoma bem-diferenciado da glândula tireóide. Determinou-se o índice de expressão da análise semiquantitativa que associa a convencional fração de área de células marcadas com os escores de intensidade da imunoexpressão, com base na observação visual qualitativa. O método digital assistido por computador foi desenvolvido com o uso de um programa de análise de imagem (ImageLab®). Três parâmetros foram obtidos: (1) porcentagem de células marcadas; (2) intensidade de imunoexpressão digital e (3) índice de expressão digital. O método proposto resulta na análise numérica da intensidade de imunoexpressão. RESULTADOS: Houve importante correlação entre as intensidades de imunoexpressão obtidas pelos dois métodos (coeficiente de correlação de Pearson, r=0,71, p=0,0001). O mesmo foi observado entre os índices de expressão (coeficiente de correlação de Pearson, r=0,66, p=0,0001). DISCUSSÃO: Os resultados de intensidade de imunoexpressão obtidos com o emprego do método digital assistido por computador foram concordantes com os escores da análise semiquantitativa. Entretanto, os resultados alcançados com o emprego do método digital podem resolver a discordância entre diferentes observadores com relação a esta intensidade de imunomarcação. Além disso, o método proposto não categoriza os resultados em grupos, o que torna a análise imunohistoquímica numericamente mensurável individualmente, aumentando seu poder diagnóstico e, sobretudo, prognóstico.

Estudo da ploidia do DNA em adenomas colorretais / Study of DNA ploidy in colorectal adenomas

Objetivo: Analisar a ploidia do DNA em adenomas colorretais a fim de identificar a presença de aneuploidia como sendo um marcador de malignidade. Métodos: Trata-se de estudo transversal, prospectivo, realizado no Serviço de Endoscopia Digestiva do Hospital Geral da Universidade de Caxias do Sul, durante o período de junho de 2002 a 2004. Foram incluídos os pólipos adenomatosos de 22 pacientes submetidos a exames colonoscópicos. As variáveis estudadas foram: sexo, idade, tamanho do pólipo, tipo histopatológico e grau de displasia. Resultados: Alterações na ploidia do DNAforam vistas em 13 casos (59,09%). Todos os pacientes com displasia acentuada (n = 3) tinham estudo da ploidia de seu DNAanormal (p = 0,01). Em77,77% dos pólipos com displasia moderada (n = 7) verificou-se aneuploidia (p < 0,05). Emrelação ao diâmetro dos adenomas, verificou-se correlação positiva com a ocorrência de aneuploidia (p = 0,001). A comparação entre tipo histológico e aneuploidia não foi significativa.Conclusão: Adeterminação da aneuploidia tem sua utilidade como marcador biológico do potencial oncogênico dos adenomas colorretais. No presente estudo, em concordância com a literatura, a relação entre grau de displasia, diâmetro do adenoma e aneuploidia foi estatisticamente significativa.

Assessment of cervical intraepithelial neoplasia (CIN) lesions by DNA image cytometry / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To compare the value of conventional cytology and DNA image cytometry (DNA-ICM) assisted cytology in detection and prognostic assessment of cervical CIN lesions.</p><p><b>METHODS</b>87 women were enrolled in this study. Cervical samples were collected employing cervix brushes which were then washed in Sedfix. After preparing single cell suspensions by mechanical procedure, cell monolayers were prepared by cyto-spinning the cells onto microscope slides. Two slides were prepared from each case: one slide was stained by Papanicolou staining for conventional cytology, another was stained by Feulgen-Thionin method for measurements of the amount of DNA in the cell nuclei using an automated DNA imaging cytometer. Biopsies from the cervical lesions were also taken for histopathology and Ki-67 immunohistochemistry.</p><p><b>RESULTS</b>Of the total of 20 ASCUS cases called by conventional cytology, no CIN, nor greater lesions were found. Among the 20 cases, 7 cases did not show any cells with DNA amount greater than 5c, while CIN2 lesions were found in 11 of other 13 cases that had some aneuploid cells with DNA amount greater than 5c. Of 30 LSIL cases called by conventional cytology, CIN2 lesions were detected in 3 out of 7 cases that did not contain any aneuploid cells with DNA greater than 5c, but in 22 out of the other 23 cases that contained aneuploid cells with DNA amount greater than > 5c. Of the remaining 7 cases called HSIL by conventional cytology, all case contained aneuploid cells containing DNA greater than 5c. If cytology was used to refer all cases of LSIL and HSIL to colposcopy procedure to detect potential CIN2 or greater lesions, the sensitivity, specificity, positive predictive value and negative predictive value were 58.2%, 84.4%, 86.5% and 54.0%, respectively. If DNA-ICM were used and all cases having 3 or more cells with a DNA amount greater than 5c were assessed to be referred to pathology to detect potential CIN2 or greater lesions, the sensitivity, specificity, positive predictive value and negative predictive were 72.7% , 87.5%, 90.9% and 65.1%, respectively. We also compared Ki67 positive cells in these samples and found that DNA-ICM results were comparable to this biomarker method.</p><p><b>CONCLUSION</b>The study demonstrated that DNA-ICM approach can be successfully used to detect significant (i.e. CIN2 or greater) lesions, and also provide a prognostic assessment of CIN lesions.</p>

Quantitative pathology : historical background, clinical research and application of nuclear morphometry and dna image cytometry

Quantitative analysis of histo- and cytochemical components such as DNA, RNA or chromatin pattern on one hand (cytometry) and the quantitative analysis of geometric non-chemical cell and tissue components (morphometry and sterology) on the other, have developed somewhat independently. Today, many different techniques, such as morphometry, sterology, and static image and flow cytometry are well established and routinely used in diagnostic quantitative pathology. The potential significance of these techniques in the individualization of care in cancer patients include the objective distinction between benign, borderline and malignant lesions, objective grading of invasive tumours, prediction of prognosis, and therapy response