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BackgroundRenal cell carcinoma (RCC) comprises of a spectrum of clinico-pathologically distinct entities thereby making it difficult to accurately predict the clinical outcome. Though many predictive factors have been described in literature, tumor stage and nuclear grade have been established to consistently correlate with the tumor behaviour. However, tumors in the same stage have shown to behave differently. Similarly subjectivity and lack of reproducibility in nuclear grade mandates use of more objective parameters such as digital nuclear morphometry which could provide consistent and more reliable results in predicting prognosis. The study was conducted with the main objective of comparing the histological grade and the nuclear morphometric variables in RCC for predicting the clinical outcome.Material and methodsA total of 219 cases of renal tumors in adults were retrieved retrospectively from the archives of pathology department in Sanjay Gandhi Postgraduate Institute of Medical Sciences, Lucknow and their clinical, gross and microscopic features were noted. Nuclear grading was done in 181 cases of clear cell and papillary RCC of which computer-assisted morphometry for various nuclear parameters was done in 100 cases where a follow-up data of at least 3 years was available. Nuclear grade and morphometric parameters were correlated statistically with the clinical outcome of the patients.ResultsHistological nuclear grade did not show statistically significant correlation with progression free survival (PFS). Higher values of mean nuclear area, mean nuclear circumference, mean nuclear major diameter and mean nuclear minor diameter were significant predictors of PFS with a strong inverse correlation.ConclusionNuclear morphometry is a more reliable predictor of clinical outcome in patients of RCC when compared to histological grade and should be included in predictive model with other clinical and pathological parameters to accurately determine tumor behaviour.

Background Although evaluation of nuclear morphology plays a crucial role in the diagnosis and categorisation of breast lesions, the criteria used to assess nuclear atypia rely on the subjective evaluation of several features that may result in inter- and intra-observer variation. This study aims to refine the definitions of cytonuclear features in various breast lesions. Methods ImageJ was used to assess the nuclear morphological features including nuclear diameter, axis length, perimeter, area, circularity, and roundness in 160 breast lesions comprising ductal carcinoma in situ (DCIS), invasive breast carcinoma of no special type (IBC-NST), tubular carcinoma, usual ductal hyperplasia (UDH), columnar cell change (CCC) and flat epithelial atypia (FEA). Reference cells included normal epithelial cells, red blood cells (RBCs) and lymphocytes. Results Reference cells showed size differences not only between normal epithelial cells and RBCs but also between RBCs in varied-sized blood vessels. Nottingham grade nuclear pleomorphism scores 1 and 3 cut-offs in IBC, compared to normal epithelial cells, were 1.4x that of mean maximum Feret's diameter and 2.4x that of mean nuclear area, respectively. Nuclear morphometrics were significantly different in low-grade IBC-NST vs. tubular carcinoma, low-grade DCIS vs. UDH, and in CCC vs. FEA. No differences in the nuclear features between grade matched DCIS and IBC were identified. Conclusion This study provides a guide for the assessment of nuclear atypia in breast lesions, refines the comparison with reference cells and highlights the potential diagnostic value of image analysis tools in the era of digital pathology.

This study was undertaken to compare Fuhrman grading with World Health Organization/International Society of Urological Pathology (WHO/ISUP) grading and stereologically measured nuclear area in patients with Clear Cell Renal Cell Carcinoma (ccRCC) or Papillary Renal Cell Carcinoma (PRCC) and to evaluate the independent predictive value of Fuhrman, WHO/ISUP and stereologically measured nuclear area combined with necrosis in a series of patients with ccRCC in relation to cancer-specific survival. In all, 124 cases of ccRCC and PRCC were included. All slides were blindly scored by two trained pathologists according to the Fuhrman and WHO/ISUP grading systems. Nuclear measurements were performed on digitally scanned slides in Visiopharm® and correlated to survival. Analysis of ccRCC and PRCC cases showed that application of WHO/ISUP grading resulted in a significant downgrading of cases from G2 to G1, when comparing with Fuhrman grading. Neither of these patients experienced progression. Cancer specific survival estimates in 101 ccRCC patients showed that WHO/ISUP grading was slightly superior in predicting cancer-specific survival. Novel models included WHO/ISUP grading and mean nuclear area (MNA) each of which combined with necrosis. Both demonstrated an increased ability to predict cancer-specific survival. The study demonstrates that WHO/ISUP grading provides superior prognostic information compared to Fuhrman grading and stereologically measured nuclear area. Necrosis in combination with either WHO/ISUP grading or MNA adds additional prognostic information.

Introduction: Incidence of oral epithelial dysplasia and oral squamous cell carcinoma (SCC) is very high in south Asian countries as compared to western population owing to a greater use of tobacco in these regions. While visual examination and exfoliative cytology are the most common screening and diagnostic modalities at present, it is a subjective analysis. Quantitative analyses such as nuclear size, cell size, and nuclear-to-cytoplasmic ratio may provide an accurate diagnosis and improve reproducibility. The aim of the study was to evaluate the role of morphometry as a diagnostic adjunct to exfoliative cytology and to derive a significant cutoff to identify the population at risk for development of SCC among chronic tobacco users. Material and Methods: This was an outpatient-based prospective study done in a tertiary hospital over a period of 2 years. Hundred and fifty cases with a history of chronic tobacco use for a minimum period of 5 years were evaluated. Visual inspection using acetic acid was done. Oral scrapes were taken for cytological and morphometric analysis followed by incision biopsy for histopathological evaluation, wherever possible. Results: On morphometrical analysis, mean nuclear area and nuclear:cytoplasmic (N:C) ratio increased, while the cytoplasmic area decreased from smears with normal cytology to oral dysplasia to SCC. Analysis of variance and post hoc Tukey’s honest significant difference test showed a statistically significant difference among the 3 groups (p value Conclusion: In high-risk cases, morphometry can be a useful adjunct to exfoliative cytology and visual examination for an early and accurate diagnosis and timely intervention in oral potentially malignant and malignant lesions, thereby improving the prognosis.

Breast carcinoma ranks one among the leading causes of cancer related morbidity and mortality in Indian women closely following carcinoma cervix in order of incidence. Fine needle aspiration cytology has emerged as one of the preliminary investigations besides mammography and clinical examination for the initial assessment of breast masses. Nuclear morphometry can be used as a quantitative adjuvant to FNAC in diagnosing breast lesions, especially the ones in the “gray zone”. Objectives: The aim of this study was to assess the utility of nuclear morphometry as an adjuvant in the cytological diagnosis, categorization of breast lesions and grading of malignant lesions. Materials and Methods: Nuclear parameters were assessed in fine needle aspirates of 49 cases, for which histopathological correlation was available. Morphometric parameters analyzed were nuclear area, nuclear perimeter, minimal nuclear diameter, maximum nuclear diameter, axis ratio, nuclear shape factor and nuclear compactness. Results: Morphometric parameters showed progressive and significant increase in nuclear size parameters from benign to malignant lesions. Nuclear shape parameters also showed a significant increase from grade I to grade III malignant lesions. However these shape factors could not be used to differentiate benign and malignant lesions in our study. Conclusion: Nuclear morphometry is a valuable tool in diagnosis of breast masses. It provides data in a more objective and reproducible form unlike the conventional cytological analysis. Keywords: Breast lesions, FNAC, Nuclear morphometry.

Objectives. Tobacco use is one of the most critical risk factors for different oral diseases. The aim of this study is to demonstrate the effect of tobacco on oral mucosa by cytomorphometric analysis of cells with the help of exfoliative cytology and to find out the improvement in diagnostic sensitivity of exfoliative cytology in the detection of dysplastic changes and early oral malignancy. Methods. The nuclear area (NA) and cytoplasmic area (CA) of cells were measured within cytological smear obtained from leukoplakia lesions of buccal mucosa of 90 tobacco users, 30 smokers (TS), 30 chewers (TC) and 30 with combined habit of smoking and chewing (TSC)] and from normal buccal mucosa of 30 non users (NU) of tobacco. Each habit group consisted of 30 tobacco users with oral leukoplakia lesion with mild epithelial dysplasia only. The 30 non-users of tobacco served as controls. The mean values of the CA and NA were obtained for each case, and the nuclear/cytoplasmic area (NA/CA) ratio was calculated. Results. The results showed a statistically significant increase (P

Background: Tobacco consumption is the major cause of oral cancer. Early detection of cytomorphometrical changes in the buccal mucosa of smoker by the use of exfoliative cytology could help in picking early premalignant changes and thereby reduce morbidity in oral cancer patients. Aim: To assess and compare the cytomorphometrical changes in the buccal mucosa cells among smoker and nonsmoker group and assess these findings in smokers with a duration of exposure to smoking by dividing them as per pack year groups. Materials and Methods: This study was carried out on 51 male cases (Cigarette or Bidi smoking) and 51 controls as per the inclusion and exclusion criteria. For finding the effect of smoking exposure severity, smokers were divided into three groups based on pack year like Group 1 with pack year 10. Sample was taken from the buccal mucosa and stained with Papnicoalaou stain. Images were analyzed by Image J software and nuclear area (NA), cytoplasmic area, and nuclear/cellular ratio (N/C ratio) data collected and analyzed. Results: There was a significant difference for the mean values of NA and N/C ratio in the buccal mucosa of smokers. Significantly increased NA and N/C ratio were found with increased pack year. Conclusion: Cytomorphometrical findings such as NA and N/C ratio are observed in increased severity in premalignant conditions such as leukoplakia. Hence, it is possible to pick up these findings earlier by noninvasive method such as exfoliative cytology, and it can be used as an adjunct tool for mass screening.

Background. Urine cytology is deemed a sensitive method in detection of high-grade urothelial carcinoma. In contrast, detection of low-grade urothelial carcinoma (LGUC) and its differentiation from reactive lesions is difficult with urinary cytology. Objective. Our study aims to determine the effectiveness of morphometric parameters in differentiating reactive urothelial cells from LGUC by cytological examination of urine specimens. Methods. Voided urine samples were used for the study, while the cases were randomized into two groups: those diagnosed with LGUC (first group; N=10) and those which were not diagnosed with LGUC (second group; N=10). The morphometric parameters of major nuclear diameter (MaND), minor nuclear diameter (MiND), mean nuclear area (MNA), cell diameter (CD), mean cell area (MCA), as well as MaND/CD, MiND/CD, MiND/MaND and MNA/MCA ratios were measured on 100 urothelial cells for each case through ScopeImage® 9.0 software. Results. A statistically significant difference was found between the mean values of MiND/CD (p=0.017) and MNA/MCA (p=0.002) ratios of groups. The mean value of both parameters in the first group constituted 0.2 and higher, and below 0.2 in the second group. Conclusion. The ratios of MiND/CD and MNA/MCA in urothelial cells proved significantly higher in patients with LGUC than benign/reactive cases. The reliability of these findings in differentiating LGUC from benign/reactive lesions needs to be verified through studies examining a large number of cases. These parameters can be assessed much faster through a special software enabling an automatic measurement and thus can be used in routine cytological examination., Актуальность. Цитологическое исследование мочи считается чувствительным методом выявления уротелиальной карциномы высокой степени злокачественности. В отличие от этого, выявление уротелиальной карциномы низкой степени злокачественности (УКНС) и ее дифференциация от реактивных поражений затруднено с помощью цитологического исследования мочи. Цель. Наше исследование направлено на определение эффективности морфометрических параметров в дифференцировке реактивных уротелиальных клеток от УКНС путем цитологического исследования образцов мочи. Методы. Для исследования использовались образцы мочи с мочеиспусканием, а пациенты были рандомизированы на две группы: те, у которых был диагностирован УКНС (первая группа), и те, которым не был диагностирован УКНС (вторая группа). Морфометрические параметры: большой диаметр ядра, малый диаметр ядра, средняя площадь ядра, диаметр клетки, средняя площадь клетки, а также их соотношения измеряли на 100 уротелиальных клетках для каждого случая с помощью программного обеспечения ScopeImage® 9.0. Результаты. Статистически значимая разница была обнаружена между средними значениями отношений малый диаметр ядра / диаметр клетки (p=0,017) и средняя площадь ядра / средняя площадь клетки (p=0,002) для групп. Среднее значение обоих показателей в первой группе составило 0,2 и выше, во второй - ниже 0,2. Заключение. Соотношения малый диаметр ядра / диаметр клетки и средняя площадь ядра / средняя площадь клетки в уротелиальных клетках оказались значительно выше у пациентов с УКНС, чем у доброкачественных/реактивных случаев. Надежность этих результатов в дифференциации УКНС от доброкачественных/реактивных поражений должна быть подтверждена исследованиями, изучающими большое количество случаев. Эти параметры могут быть оценены намного быстрее с помощью специального программного обеспечения, обеспечивающего автоматическое измерение, и, таким образом, могут использоваться при рутинном цитологическом исследовании., Актуальність. Цитологічне дослідження сечі вважається чутливим методом виявлення уротеліальної карциноми високого ступеня злоякісності. На відміну від цього, виявлення уротеліальної карциноми низького ступеня злоякісності (УКНС) і її диференціація від реактивних уражень утруднено за допомогою цитологічного дослідження сечі. Мета. Наше дослідження спрямоване на визначення ефективності морфометричних параметрів у диференціюванні реактивних уротеліальних клітин від УКНС шляхом цитологічного дослідження зразків сечі. Методи. Для дослідження використовувалися зразки сечі з сечовипусканням, а пацієнти були рандомізовані на дві групи: ті, у яких був діагностований УКНС (перша група), і ті, яким не був діагностований УКНС (друга група). Морфометричні параметри: великий діаметр ядра, малий діаметр ядра, середня площа ядра, діаметр клітини, середня площа клітини, а також їх співвідношення вимірювали на 100 уротеліальних клітинах для кожного випадку за допомогою програмного забезпечення ScopeImage® 9.0. Результати. Статистично значуща різниця була виявлена між середніми значеннями відносин малий діаметр ядра / діаметр клітини (p = 0,017) і середня площа ядра / середня площа клітини (p = 0,002) для груп. Середнє значення обох показників в першій групі склало 0,2 і вище, у другій - нижче 0,2. Підсумок. Співвідношення малий діаметр ядра / діаметр клітини і середня площа ядра / середня площа клітини в уротеліальних клітинах виявилися значно вище у пацієнтів з УКНС, ніж у доброякісних/реактивних випадків. Надійність цих результатів в диференціації УКНС від доброякісних/реактивних уражень повинна бути підтверджена дослідженнями, що вивчають велику кількість випадків. Ці параметри можуть бути оцінені набагато швидше за допомогою спеціального програмного забезпечення, що забезпечує автоматичне вимірювання, і, таким чином, можуть використовуватися при рутинному цитологічному дослідженні.

Background Bronchoalveolar lavage (BAL) is one of the fundamental examinations for the differential diagnosis of interstitial lung diseases (ILDs), and lymphocytosis strongly indicates alternative diagnoses rather than idiopathic pulmonary fibrosis. However, the BALF lymphocytosis is observed in several ILDs. We considered that quantitative evaluation of the BALF lymphocyte nuclear morphology would be useful in the differential diagnosis of ILDs with increased BALF lymphocyte fraction. Results One hundred and twenty-one patients with ILDs having increased BALF lymphocyte fraction were recruited (68 in the development cohort and 53 in the validation cohort). In the development cohort, BALF lymphocyte nuclei in sarcoidosis patients showed significantly smaller areas, shorter perimeters, lower radius ratios, and increased roundness than those of other ILD patients (p Conclusion This study is the first to demonstrate the usefulness of quantitative parameters of BALF lymphocyte nuclear morphology as novel biomarkers for sarcoidosis.

The 2014 Bethesda System diagnostic criteria for atypical glandular cells (AGC) aid in the classification of atypical cells in cervical cytology. Anyway, AGC diagnosis remains challenging, due to low frequencies of this finding (approximately 0.5%-1% of Pap test results), abundance of AGC mimics, and significant interobserver variability. We developed an algorithm based on nuclear areas parameter that can help to differentiate AGC from Normal and Reactive glandular cells. Nuclear areas and perimeters were measured on 16 Pap smears with AGC and 18 with Reactive glandular cells of women aged between 30 and 77. Glandular cells from nonpathological Pap smears were used as controls. For each case, the means, medians, standard deviations, and the minimum and maximum values of both nuclear areas and perimeters of the cells of interest were calculated. The nuclear area analysis showed a 100% specificity in discriminating Normal from Altered cells (either Reactive or AGC), whereas the nuclear perimeter analysis showed a lower specificity (87.5%). Both nuclear area and perimeter variability analysis resulted in high specificity values in distinguishing Reactive cells from AGC. Therefore, a stepwise two-step algorithm using nuclear areas to discriminate Normal from Altered cells, and nuclear area variability to distinguish Reactive from AGC, allowed us to reliably classify the cells into these three categories. The morphometric analysis of nuclear area is a valuable and reliable aid in AGC diagnosis and standardization, easily integrable into common automatic algorithms.

Background of Study: Diabetes is the most common endocrine-metabolic disease affecting worldwide. Diagnosis of type 2 diabetes with the help of oral exfoliative cytology is a simple, noninvasive technique. Evaluation of cytomorphometric changes in exfoliated buccal cells in diabetes can help in the early detection and examination of diabetes. Aim: The aim is to study and compare the cytomorphometric features of buccal mucosal cells of normal individuals and type 2 diabetes patients and to evaluate the role of phase-contrast microscopy for visualizing cytomorphological changes in unstained smears. Materials and Methods: Fifty known type 2 diabetic patients and fifty healthy individuals were taken as study group and control group, respectively. Smears were prepared from the buccal mucosa of both study and control groups. Fixed, unstained smears were viewed under phase-contrast microscope, and cytomorphometric analysis was done using image analysis software for evaluating nuclear area (NA), cytoplasmic area (CyA), and nuclear/cytoplasmic ratio. Results: A statistically significant increase in NA(P = 0.000), CyA (P = 0.000), and N/Cy ratio (P = 0.000) was obtained from the smears of the study group when compared with the control group. Conclusions: Cytomorphometric analysis of exfoliated cells can aid in predicting systemic disease such as type 2 diabetes. With the use of phase-contrast microscope, the cellular details can be easily and quickly evaluated and it is a cost-effective method in cytomorphometric studies.

Introduction: Liquid-based cytology (LBC) is increasingly used for nongynecologic applications. However, the cytological preparation of LBC specimens is influenced by the processing technique and the preservative used. In this study, the influence of the processing techniques and preservatives on cell morphology was examined mathematically and statistically. Methods: Cytological specimens were prepared using the ThinPrep (TP), SurePath (SP), and AutoSmear methods, with 5 different preservative solutions. The cytoplasmic and nuclear areas of Papanicolaou-stained specimens were measured for all samples. Results: The cytoplasmic and nuclear areas were smaller in cells prepared using the 2 LBC methods, compared to that prepared using the AutoSmear method, irrespective of the preservative used. The cytoplasmic and nuclear areas of cells prepared using the SP method were smaller than those of cells prepared using the TP method, irrespective of the preservative used. There were fewer differences among the cytoplasmic areas of cells prepared with different preservative solutions using the TP method; however, the cytoplasmic areas of cells prepared using the SP method changed with the preservative solution used. Conclusions: The most significant difference affecting the cytoplasmic and nuclear areas was the processing technique. The TP method increased the cytoplasmic and nuclear areas, while the methanol-based PreservCyt solution enabled the highest enlargement of the cell. LBC is a superior preparation technique for standardization of the specimens. Our results offer a better understanding of methods suitable for specimen preparation for developing precision AI-based diagnosis in cytology.

A precise diagnostic of precursor dysplastic urothelial lesions is critical for patients but it can be a challenge for pathologists. Multiple immunohistologic markers (panel) improve ambiguous diagnostics but results are subjective, with a high degree of observational variability. Our research objective was to evaluate how a classification algorithm may help morphology diagnostic. Data coming from 45 unequivocal cases of flat urothelial lesions (“training set”: 20 carcinomas in situ, 8 dysplastic and 17 reactive lesions) were used as ground truth in training a random tree classification algorithm. 50 “atypia of unknown significance” diagnostics (diagnostic set) were digitally re-classified based on morphological and immunohistochemical features as possible carcinoma in situ (20), dysplastic (17) and reactive atypia cases (13). The main sorting criterium was morphologic (nuclear area). A four-markers panel was used for a precise classification (74% correctly classified, 93% accuracy, 76% precision, averaged ROC=0.828). 3 cases were “false negative”. The performance of the immunohistologic panel was evaluated based on a stain index, calculated for CD20, p53, Ki67 and observed for CD44. Within training set, the immunohistologic performance was high. In the diagnostic set both the percentage of high stain index for each marker and the percentage of cases with 2-3 strong markers were low, explaining the initial high number of equivocal cases. In conclusion, digital analysis of morphologic and immunohistologic features may bring clarification in classification of equivocal urothelial lesions. Computational pathology supports diagnostic process as it can measure features and handle data in a precise, reproducible and objective way. In our proof of concept study, a low number of cases and the (deliberate) absence of clinical data were main limitations. Validation of the method on a high number of cases, use of genomics and clinical data are essential for improving the reliability of machine learning classification

Objective Inhalational drugs used in treating asthma have several side effects including those on oral tissues. We therefore designed a study to analyze the effects of inhalational drugs on the buccal mucosal cells of the oral cavity. Methods Smears were obtained from clinically normal buccal mucosa of 20 randomly selected asthmatic patients who had been under inhalational therapy for at least 6 months. The Papanicolaou-stained smears were then analyzed for average nuclear area, average cytoplasmic area, and average nuclear area:cytoplasmic area ratio for each patient, and the values were compared with those of 10 healthy controls. Results A statistically significant decrease in cytoplasmic area (P

Several interstitial lung diseases (ILDs) including hypersensitivity pneumonia (HP), nonspecific interstitial pneumonia (NSIP), cryptogenic organizing pneumonia (COP), collagen vascular disease-related ILD (CVD-ILD), drug-induced ILD (DILD) and sarcoidosis shows increase of lymphocyte fraction in bronchoalveolar lavage fluid (BALF). We performed the morphometric analysis in order to differentiate the BALF lymphocyte nuclear shape between sarcoidosis and the other ILDs. We recruited 68 patients with ILDs (16 of CHP, 5 of NSIP, 5 of COP, 10 of CVD-ILD, 14 of DILD and 18 of sarcoidosis) who showed 20% or more of lymphocyte fraction in BALF and measured the area, perimeter, radius ratio and roundness of the BALF lymphocyte nuclear by using the commercially available image analyzing software. As a result, among those with sarcoidosis, lymphocyte nuclear area was significantly smaller (p Based on these findings, we consider that the fraction of srLYM in BALF can be useful for the differential diagnosis of ILDs.

Background: The nuclei of most cancer cells in histopathologic preparations differ from normal nuclei and vary individually in size, shape, and chromatin pattern. Although the cytologic characteristics of squamous cell carcinoma (SCC) of the lung have been described, quantification of the cytologic features has not been established. Methods: Cytologic investigations were performed on bronchial brushings or washings, or fine-needle aspirates. We analyzed the nuclear area (NA) of 50 tumor cells in 32 patients with SCC of the lung and 50 bronchial epithelial cells in 20 patients with no evidence of malignancy including inflammatory lesions. Results: The NA of tumor cells (102.4 ± 26.2 μm2) was significantly larger than that of bronchial epithelial cells (64.1 ± 16.9 μm2) (P = 0.001). The receiver operating characteristic (ROC) curve analysis showed that an NA cutoff level of 86 μm2 had a sensitivity of 75% and specificity of 88% to detect malignant components. Conclusion: We conducted quantitative analyses of NA in SCC using cytologic specimen, NA was a useful parameter for evaluation of differential diagnosis between SCC and non-malignancies even in cytologic specimens.

Background:Radiography forms an important as well as an integral part of diagnosis which is used in various fields of medical and dental services to give an appropriate diagnosis. But, diagnostic radiations also induce some amount of damage to the cell at cytogenetic levels, depending on the dosage of the radiation administered. Objective:The objective of the present study was to observe the cytomorphometric changes in the buccal mucosa of patients who were exposed to computed tomography.Materials and methods:Study sample consisted of 15 patients who were exposed to Computed Tomography prior to treatment planning for placement of dental implants. Buccal mucosal smears were obtained pre and 10 days post radiation exposure by using a wooden tongue depressor and the contents were smeared on to clean glass slides and fixed using Biofix spray. Post fixation, these smears were stained using rapid PAP stain and viewed under compound microscope. Cytomorphometric analysis was performed using image ‘J’ software for 50 cells. Statistical analysis was done using SPSS software.Results: Cytomorphometric analysis of the buccal mucosal cells, did show variations in the nuclear and cellular parameters, but statistical significant results were obtained in case of nuclear diameter and nuclear area only suggesting genotoxic alterations following exposure to computed tomography.Conclusion: Oral buccal mucosal cells showed significant cytomorphometric changes in the nuclear parameters after exposure to X-rays during Computed Tomography. A longitudinal study consisting of a bigger sample size is required to evaluate the long-term effects of radiation on these buccal mucosa cells.Keywords: buccal mucosa; computed tomography; cytomorphometry; radiation.

Core needle biopsy (CNB) has been used as an alternative or a complementary method for diagnosis of thyroid nodules. However, morphological analysis of the nuclear features of papillary thyroid carcinoma (PTC) cells obtained via CNB remains unclear. Hence, we examined the differences between the PTC nuclear features in CNB and thyroidectomy specimens.Ten PTC patients, who underwent both CNB and thyroidectomy, were selected. Microscopic photographs of three representative areas of the PTC and adjacent parenchyma were taken. Ten cells per photograph were chosen, and 1200 cells were evaluated (300 PTC and 300 follicular cells in the CNB and thyroidectomy specimens, respectively). The area, circumference, major axis, and minor axis were measured using an image analyzer. Detailed nuclear features (size and shape, membrane irregularity, chromatin characteristics) were scored using a 3-point scale.The mean nuclear area, circumference, major axis, and minor axis of PTC cells in the CNB specimen were 1.76, 1.34, 1.34, and 1.29 times larger than those of the follicular cells (p0.001); similar results were seen in the thyroidectomy specimens (2.04, 1.41, 1.37, and 1.37: p0.001). Comparative analysis revealed that these parameters were significantly smaller in the CNB specimens than those in the thyroidectomy specimens (p0.001). Nuclear grades were also lower in the former owing to poor chromatin characteristics (clearing and margination) (p0.01).Considering that the PTC nuclei in CNB specimens are smaller with fewer irregularities and less clear than those in thyroidectomy specimens, we need to emphasize caution when using CNB specimens for diagnosis.

Objective To evaluate if dental students with the habit of smoking exhibit alterations in the epithelial cells of the cheek mucosa on the nuclear area (NA), cytoplasmic area (CA) or in the nucleus/cytoplasm ratio (NA/CA), when compared to non-smokers. Methods This is a descriptive, experimental study using a control group. The sample comprised 40 dental undergraduates (20 smokers and 20 non-smokers). Individuals who smoked at least 5 or 6 cigarettes a day were deemed to be smokers, while non-smokers were those who reported having no experience of any form of smoking. The samples of epithelial cells were obtained from the oral mucosa using the liquid-based exfoliative cytology technique. The slides were processed in the laboratory, stained using the Papanicolau technique and were analyzed via an image analysis system (analySIS getIT). A structured questionnaire was applied with the aim of measuring independent variables related to personal characteristics, health and lifestyle behavior. Results The values obtained in the group of smokers were as follows: NA (169.4 µm² ± 17.0), CA (1240.1 µm² ± 128.9) and NA/CA ratio (0.1375 ± 0.01743). The results did not evidence significant differences when compared to the control group: NA (170.7 µm² ± 20.0), CA (1255.3 µm² ± 174.5) and NA/CA (0.1380 ± 0.01436). Conclusion It was not possible to observe significant cell alterations in the NA, CA or the NA/CA ratio of epithelial cells of the oral mucosa. Other risk factors associated with tobacco should be studied, principally the length of exposure to the properties of the causative agent. RESUMO Objetivo Avaliar se os estudantes de odontologia que possuem hábitos tabagistas apresentam alterações celulares nas células epiteliais da mucosa jugal sobre as áreas nucleares (AN), citoplasmáticas (AC) e na relação núcleo/citoplasma (AN/AC) quando comparados a não fumantes. Métodos Trata-se de um estudo experimental, descritivo com grupo controle. A amostra foi composta por 40 estudantes de graduação em odontologia (20 fumantes e 20 não fumantes). Consideraram-se fumantes aqueles indivíduos que fumavam diariamente, no mínimo, cinco a seis cigarros do tipo industrializado; e não fumantes aqueles indivíduos que relataram a não experiência com qualquer tipo de fumo. As amostras de células epiteliais foram obtidas da mucosa oral pela técnica de citologia esfoliativa em meio líquido. As lâminas foram processadas em laboratório, coradas pela técnica de Papanicolau e analisadas através de um sistema de análise de imagens (Analysis getlt). Um questionário estruturado foi aplicado com o intuito de mensurar variáveis independentes referentes às características pessoais, comportamentos em saúde e estilo de vida. Resultados Os valores obtidos no grupo dos fumantes para a AN (169,4 µm² ± 17,0), AC (1240,1 µm² ± 128,9) e na relação AN/AC (0,1375 ± 0,01743) não apresentaram resultados com diferenças expressivas quando comparados ao grupo controle AN (170,7 µm² ± 20,0), AC (1255,3 µm² ± 174,5) e na AN/AC (0,1380 ± 0,01436). Conclusão Não foi possível observar alterações celulares significantes nas AN, AC e na relação AN/AC de células epiteliais da mucosa oral. Outros fatores de risco associados ao tabaco devem ser estudados, principalmente o tempo de exposição às propriedades do agente causador.

We determined cytologic features of histologically confirmed subcentimeter canine mammary tumors (CMTs) to determine reasonable criteria for an accurate cytologic diagnosis. Fifty-three CMTs from 28 bitches were included. All cytologic samples were collected by ultrasound-guided fine-needle aspiration biopsy, stained with May-Grünwald/Giemsa, and retrospectively evaluated using a scoring system established for 18 cytologic features. Mean nuclear area (MNA) was also measured for each sample by a computer-assisted program. Based on the histologic diagnosis, CMTs were divided into 2 groups: malignant tumors (25) and benign lesions (15). Data were statistically analyzed using Fisher and Mann–Whitney tests. Chromatin pattern ( p < 0.05) and macrophage infiltration ( p < 0.05) were significantly different between the groups. Median MNA was significantly ( p < 0.05) larger in malignant tumors. The evaluation of these cytologic features in subcentimeter CMTs may increase the sensitivity of cytology.

Intraductal carcinoma of the prostate gland (IDCP), which is now categorised as a distinct entity by WHO 2016, includes two biologically distinct diseases. IDCP associated with invasive carcinoma (IDCP-inv) generally represents a growth pattern of invasive prostatic adenocarcinoma while the rarely encountered pure IDCP is a precursor of prostate cancer. This review highlights issues that require further discussion and clarification. The diagnostic criterion “nuclear size at least 6 times normal” is ambiguous as “size” could refer to either nuclear area or diameter. If area, then this criterion could be re-defined as nuclear diameter at least three times normal as it is difficult to visually compare area of nuclei. It is also unclear whether IDCP could also include tumours with ductal morphology. There is no consensus whether pure IDCP in needle biopsies should be managed with re-biopsy or radical therapy. A pragmatic approach would be to recommend radical therapy only for extensive pure IDCP that is morphologically unequivocal for high-grade prostate cancer. Active surveillance is not appropriate when low-grade invasive cancer is associated with IDCP, as such patients usually have unsampled high-grade prostatic adenocarcinoma. It is generally recommended that IDCP component of IDCP-inv should be included in tumour extent but not grade. However, there are good arguments in favour of grading IDCP associated with invasive cancer. All historical as well as contemporary Gleason outcome data are based on morphology and would have included an associated IDCP component in the tumour grade. WHO 2016 recommends that IDCP should not be graded, but it is unclear whether this applies to both pure IDCP and IDCP-inv.

Purpose: Histological grade is an important prognostic factor in patients with non-muscle–invasive bladder cancer (NMIBC). However, interobserver variability is high. Previous studies have suggested that quantification of histological features is useful to objectify grading. We evaluated whether quantification of the mean nuclear area of the 10 largest nuclei (MNA-10), degree of aneuploidy (DNA index or DI) and mitotic activity index (MAI) are of diagnostic value for NMIBC grade. Additionally, prognostic value of the 3 measures was assessed. Material and methods: A consensus grade was determined by 3 uropathologists in 310 NMIBC tissues according to the World Health Organization (WHO) 1973 and the WHO2004. Logistic regression with forward selection was used to determine the optimal combination of measures (MNA-10, DI, and MAI) to diagnose grade 3 (G3) or high-grade (HG) NMIBC (WHO1973 and WHO2004, respectively). Results: In 310 tumors of 215 patients at least 1 of the measures (MNA-10, DI, or MAI) had been determined. The combination of MNA-10 and MAI was selected as the most diagnostic combination and resulted in a sensitivity of 94% (95% confidence interval [CI]: 87–100) at a specificity of 72% (95% CI: 66–78) for G3 tumors. For the diagnosis of HG tumors sensitivity was 92% (95% CI: 86–97) at a specificity of 76% (95% CI: 70–93). Conclusions: Determination of MNA-10 and MAI is promising for diagnosing G3 and HG bladder tumors. These findings warrant further studies on the diagnostic and prognostic value of proliferative and quantitative features in bladder cancer patients.

Aim: To examine and estimate cytomorphometric changes by using parameters such as cell area (CA), nuclear area (NA), cell diameter (CD), nuclear diameter (ND), and nuclear-cytoplasmic ratio (N/C) in exfoliated buccal mucosal cells in smokers, patients with hypertension, and healthy individuals by using Papanicolaou stain. Materials and Methods: This quantitative analysis was performed on totally 120 individuals who were randomly selected and were divided into four groups. Group 1: 30 healthy individuals free of hypertension and without any smoking habit comprised the control group; Group 2: 30 individuals with smoking habit and without hypertension; Group 3: 30 individuals with smoking habit and with hypertension; and group 4: 30 individuals with hypertension and without smoking habit. Buccal smears were taken and stained by Papanicolaou stain. Image analysis of 50 cells was done at 40× magnification with a digital image capture analysis software system (ProgRes capture pro, version 2.8.8). The obtained parameters were statistically compared among the groups by one-way ANOVA test and Kruskal–Wallis test. Results: Statistically significant increase in NA, ND, and N/C ratio values were found in smokers and smokers with hypertension, and minimal change was observed in patients with hypertension alone when compared with normal mucosa. Conclusion: Smoking habit alone and patients with hypertension along with smoking habit show significant cytomorphometric changes. The combined use of exfoliative cytology and cytomorphometric analysis gives an added advantage in monitoring clinically suspect lesions and early detection of malignancy in high-risk individuals such as smokers.

BACKGROUND Only a small number of studies on computerized cytomorphometry have been performed for thyroid FNAC. The present study aimed to determine the usefulness of computerized cytomorphometry methods to further classify thyroid lesions as benign or malignant and to compare the practicability and value of using Papanicolaou (Pap) and Giemsa stains in thyroid FNAC by evaluating their association to various cytologic nuclear parameters. METHODS Fifty-eight thyroid lesions diagnosed by FNAC and categorized according to the Bethesda system for reporting thyroid cytopathology were evaluated in terms of various cytologic nuclear parameters, including nuclear area (NA), nuclear perimeter (NP), nuclear density (ND), long nuclear diameter (LND), and short nuclear diameter (SND). The Pap- and Giemsa-stained slides were examined separately. RESULTS In the malignant cases, NA, NP, LND, and SND were higher than in the benign cases for both the Pap and Giemsa stains. NA, NP, LND, and SND were higher in Giemsa than Pap for both the benign and malignant groups. Statistically significant differences were detected between the benign and malignant cases in the AUS category. CONCLUSIONS Computerized cytomorphometry is useful in distinguishing between benign and malignant lesions in thyroid FNAC. The measurement of cytologic nuclear parameters in cases suggestive of AUS may be useful for the probable classification of cases as benign or malignant. Although further studies are needed, in nuclear morphometric assessment of thyroid FNAC, Giemsa staining may be more useful and valuable than the Pap stain because of its association with various cytologic nuclear parameters. Diagn. Cytopathol. 2016;44:902-911. © 2016 Wiley Periodicals, Inc.

WOS: 000378300700009 PubMed: 27546367 BACKGROUND: Ischemia/reperfusion (I/R) injury is a complex event frequently observed in vascular surgery and can cause functional and structural cell damage. Nucleolar-organizing regions (NORs) are sites of the ribosomal genes located on chromosomes and can be stained with silver when they are active. Thus these proteins are named as argyrophilic-NOR (AgNOR)-associated proteins. We aimed to investigate any possible effects of renal I/R injury on the NOR protein synthesis and association between the AgNOR proteins amount and histopathological injuring score. METHODS: Nine female wistar-albino rats with weight of 200-250g were included into the study. The animals were randomly divided in two groups, a Control Group and an I/R Group. In I/R group, rats were subjected to 45 minutes of renal pedicle occlusion followed by 24 hours of reperfusion. In the control group no drug injections or ischemia reperfusion were performed in animals. Then histopathological injury score, mean AgNOR number and total AgNOR area/nuclear area (TAA/NA) were detected for each rat. RESULTS: The differences between control and I/R groups were significant for histopathological injury scores (p = 0.016). Also the differences between control group and I/R group were significant for mean AgNOR number (p = 0.000) and TAA/NA ratio (p = 0.000). Additionally, there was a positive correlation between TAA/NA ratio and histopathological injury score (r = 0.728; p = 0.026) and between mean AgNOR number and histopathological injury score (r = 0.670; p = 0.048). CONCLUSION: The detection of AgNOR proteins amount may be used as an indicator to obtain information about the cellular behaviour (self-protective mechanism of tubular epithelial cells) against I/R injury and cellular damage levels (Tab. 2, Fig. 4, Ref. 24). Text in PDF www.elis.sk.

Yogyakarta Nuclear Area (YNA) is one of BATAN’s nuclear facility that has many nuclear facilities such as nuclear reactor, accelerator, irradiator, and waste management system. Due to nuclear accident is potentially occur, YNA has nuclear emergency response program and there is a preparedness system to support this program. Radiation monitoring system is one of utilization to acquire and record nuclear radiation exposure in the facility and its environment. An Internet of Thing based radiation area monitor was developed to improve the system not only to acquire the radiation exposure, but also to distribute the acquired data to the cloud server through the internet network. The distributed radiation data can be utilized to analyze nuclear emergence potential in the nuclear emergency response and preparedness system. The area monitor system was designed as sensor network that consists of Geiger Muller detector and high voltage power supply, signal conditioning system, and Arduino as counter and data processor. Data is acquired by a detector and it is transmitted to the server through wireless network using the node MCU communication module. The receiver station system was developed to acquire data from database server to display the radiation exposure in the environment and to identify radiological emergency status. Based on the chi square test stability method, the developed device has good stability which the probability of x 2 was obtained 0.75. The validation testing also was done using Sr-90 and x-ray spectrometer as radiation source, and the developed device has been compared using a standard survey-meter. The mean square error of this validation testing was 0.37 and 0.24 respectively for Sr-90 at two difference source distance, while for x-ray spectrometer source the mean square error value was 2.48. For further development, the system will be combined with a smart meteorological system to build the integrated data acquisition system for Nuclear and Radiological Emergency Preparedness System in Yogyakarta Nuclear Area.

BACKGROUND Urinary cytology is sensitive and specific for diagnosing and screening high-grade urothelial carcinomas (HGUC). The Paris System (TPS) for urinary cytology was introduced in 2016 to standardize reporting. According to TPS diagnostic categories of HGUC and suspicious for HGUC (SHGUC), the average nuclear-to-cytoplasm (N:C) ratio of atypical cells should be ≥0.7. The objective of the current study was to measure the N:C ratio of urine cytology specimens with HGUC and SHGUC diagnoses and biopsy-proven HGUC follow-up. METHODS A cohort of 64 cases (HGUC, 49 cases; SHGUC, 15 cases) from 57 patients was constructed. Urine cytology slides were scanned into whole-slide digital images. The nuclear and cytoplasmic areas were enumerated by digital image analysis (DIA), and the N:C ratios were measured. RESULTS In total, 640 cells were analyzed by DIA (HGUC, 490 cells; SHGUC, 150 cells). For HGUC and SHGUC, the average N:C ratios were 0.57 and 0.53, respectively. The maximum average N:C ratio was 0.73 for HGUC and 0.68 for SHGUC. HGUC had higher average N:C ratio (P < .001), higher average nuclear area (P < .001), higher average maximum N:C ratio (P = .005), and higher average maximum nuclear area (P = .006) compared with SHGUC. CONCLUSIONS The N:C ratios for the HGUC (0.57) and SHGUC (0.53) categories are lower than those previously suggested in TPS. The authors advocate reducing the N:C ratio below the current threshold of 0.7.

Medulloblastomas are the most common central nervous system tumors in childhood. Treatment and prognosis strongly depend on histology and transcriptomic profiling. However, the proliferative potential also has prognostical value. Our study aimed to investigate correlations between histogram profiling of diffusion-weighted images and further microarchitectural features. Seven patients (age median 14.6 years, minimum 2 years, maximum 20 years; 5 male, 2 female) were included in this retrospective study. Using a Matlab-based analysis tool, histogram analysis of whole apparent diffusion coefficient (ADC) volumes was performed. ADC entropy revealed a strong inverse correlation with the expression of the proliferation marker Ki67 (r = − 0.962, p = 0.009) and with total nuclear area (r = − 0.888, p = 0.044). Furthermore, ADC percentiles, most of all ADCp90, showed significant correlations with Ki67 expression (r = 0.902, p = 0.036). Diffusion histogram profiling of medulloblastomas provides valuable in vivo information which potentially can be used for risk stratification and prognostication. First of all, entropy revealed to be the most promising imaging biomarker. However, further studies are warranted.

Background Different cancers subtypes can often be effectively treated with similar Rx classes, i.e., with PARPi in the case of BRCA mutations or with anti-PD1/PDL1 inhibitors for patients with mismatch repair deficiencies. Using the Epic Sciences Functional Cell Profiling (FCP) platform utilizing high resolution digital pathology and machine learning, we analyzed ∼100,000 single CTCs from multiple indications to index metastatic cancers, with the longer-term hypothesis that sub-classification of CTCs may help define therapeutic decisions across cancer types. Methods 92,300 CTCs underwent FCP analysis (single cell digital pathology features of cellular and sub-cellular morphometrics) were collected from cancer patients comprising Prostate (1641 pts, 70,747 CTCs), Breast (268 pts, 8,718 CTCs), NSCLC (110 pts, 1884 CTCs), SCLC (141 pts, 8,872 CTCs) and Bladder (65 pts, 2079 CTCs). An equal number of CTCs from each indication was sampled to create a balanced cohort for training. K-means clustering was applied on the training set and an optimized number of clusters were determined using the elbow approach. After defining the clusters on the training set, the cluster centers were extracted from k-means, and used to train a k-Nearest Neighbor (k-NN) classifier to predict the cluster assignment for the remaining CTCs designated as the test set. Results The optimized number of clusters was 9. Breast cancer CTCs were more enriched with higher CK expressing CTCs (c1), while SCLC and some of mCRPC shared similar small cell like features (c5). Cluster subtypes and characteristics are listed. Table . 1984PD Cluster Characteristics Bladder Breast NSCLC SCLC Prostate c1 Highest CK 8% 24% 1% 5% 9% c2 Highest N/C ratio 13% 15% 7% 35% 21% c3 Large, Elongated 1% 3% 12% 1% 3% c4 Low N/C, Low CK 33% 24% 25% 8% 12% c5 Small, Higher N/C ratio, More Round 11% 12% 11% 37% 33% c6 Low CK, Elongated 8% 4% 8% 2% 4% c7 Highest Nucleoli Count 0% 0% 17% 1% 4% c8 Large Nuclear Area, Highest Nuclear Entropy 21% 14% 10% 11% 10% c9 Most Elongated Cytoplasm 4% 3% 9% 2% 4% Conclusions Heterogeneous CTC phenotypic subtypes were observed across multiple indications. Each indication harbored subtype heterogeneity and shared clusters with other cancers. Patient cluster subtype analysis for prognosis and therapy benefit as well as linkage of CTC subtype genotypes to patient survival by cancer type is ongoing. Legal entity responsible for the study The authors. Funding Epic Sciences. Disclosure A. Jendrisak: Full / Part-time employment: Epic Sciences. R. Sutton: Full / Part-time employment: Epic Sciences. S. Orr: Full / Part-time employment: Epic Sciences. D. Lu: Full / Part-time employment: Epic Sciences. J. Schonhoft: Full / Part-time employment: Epic Sciences. Y. Wang: Full / Part-time employment: Epic Sciences.

The prostatic atrophy is a lesion with small glands that can mimic adenocarcinoma, commonly diagnosed in the elderly and whose preneoplastic role is controversial. Over time several classifications have been developed for this lesions according to its architectural and cellular aspects. Materials and Methods: Aim of the study is morphometrically assessment of epithelial component nuclei in prostatic atrophy. We evaluated parameters represented by nuclear area (NA), nuclear perimeter P (μm) Elongation factor (E), average diameter (DEQ), maximum diameter (Dmax), Minim diameter (Dmin). The control group was represented by adenomatous hyperplasia component of benign hyperplasia of the prostate. Results: Morphometric analysis revealed significantly higher values of nuclear parameters in atrophy compared with adenomatous component in controls (p

Morphological alteration of cells and matrices is critical for tumor initiation and progression. Monitoring these alterations during tumor progression is vitally important for making real-time histological diagnoses of tumor staging. In this study, 20 pairs of normal and cancerous human breast tissues were imaged by multiphoton microscopy (MPM), and nuclear area and collagen density were quantified by LSM 5 software (version 3.2). Comparison of MPM images from normal breast tissue with low- and high-grade invasive ductal carcinoma (IDC) lesions clearly showed changes in both cellular features and extracellular matrix architecture during IDC development. Moreover, analysis of nuclear area and collagen density established a quantitative link between these two morphological features and progression of IDC. Present results demonstrated that MPM can provide both qualitative and quantitative evaluations of tumor progression. With additional development, this technique has the potential to make real-time histological diagnoses of tumor staging and guide development of efficacious clinical therapies.

Some technical aspects of two Spanish cooperation projects, funded by DPI and Innpacto Programs of the R&D National Plan, are discussed. The objective is to analyze the common belief about than the ultrasonic testing in MHz range is not a tool utilizable to detect internal flaws in highly attenuating pieces made of coarse-grained steel. In fact high-strength steels, used in some safe industrial infrastructures of energy & transport sectors, are difficult to be inspected using the conventional “state of the art” in ultrasonic technology, due to their internal microstructures are very attenuating and coarse-grained. It is studied if this inspection difficulty could be overcome by finding intense interrogating pulses and advanced signal processing of the acquired echoes. A possible solution would depend on drastically improving signal-to-noise-ratios, by applying new advances on: ultrasonic transduction, HV electronics for intense pulsed driving of the testing probes, and an “ad-hoc” digital processing or focusing of the received noisy signals, in function of each material to be inspected. To attain this challenging aim on robust steel pieces would open the possibility of obtaining improvements in inspecting critical industrial components made of highly attenuating & dispersive materials, as new composites in aeronautic and motorway bridges, or new metallic alloys in nuclear area, where additional testing limitations often appear. The Spanish I+D National Plan (MINECO), by funding the Projects: Fundamental Research / DPI2011 – 22438, and INNPACTO Program / IMAAD - IPT-020000-2010-0004.

Introduction: We have reported the most common underlying disease of SPAP was myelodysplastic syndrome (MDS) and its SPAP was poor prognosis. However, the cause of the dysfunction of alveolar macropghage (AM) in SPAP has been unclear. Objective: To investigate the pathogenesis of SPAP by analyzing the cell form of AM in SPAP. Methods: Bronchoalveolar lavage fluid (BALF) specimens in twenty cases of SPAP were retrospectively evaluated by microscope. Underlying disease of SPAP were all MDS. The cell area, nuclear area, and nuclear⁄cell area ratio (N⁄C ratio) of AM were measured in SPAP, compared with APAP (n=10) and healthy volunteer(n=5). These data were analyzed by hybrid cell count using digital microscope (BZ-9000), KEYENCE cooperation, Japan. Results: The median of AM cell area was 107 µm2 in SPAP and this size was significant smaller than it in APAP. The median of AM nuclear area also was significant small in SPAP (37µm2). On the other hands, median N⁄C ratio of AM in SPAP (29%) was significant larger than it in APAP(17%). AM cells with high N⁄C ratio (>40%) were remarkable in SPAP(Fig1), whereas the proportion of foamy AM cell in SPAP was less than it in APAP. Conclusion: The form of AM was significant different from between SPAP and APAP cases. It may be concerned with the variation for the phagocytosis of surfactant in each SPAP and APAP.

The aim of this retrospective pilot study was to evaluate the Aperio nuclear V9 algorithm as an image analysis tool to observe the histopathological changes of ocular surface squamous neoplasia (OSSN). A histopathological assessment, including the Ki-67 proliferative index (PI) of immunohistochemically-stained tumor conjunctiva (TC) and healthy conjunctiva (HC) tissues, was performed in six cases of OSSN. The Aperio V9 algorithm was applied to digital images of the tissue specimens to count the Ki-67 PI and to measure the nuclear area indices. This digital algorithm was validated using stereological and visual analysis methods. The visual scoring of Ki-67 PI ranged from 22 to 60% (mean, 38.5%), and from 5 to 20% (mean 9.5%) in TC and HC tissue, respectively. The computer-aided analysis, using the Aperio nuclear V9 algorithm, revealed that the Ki-67 PI ranged from 21.5 to 43.5% (mean, 33.6%), and from 1.9 to 21.0% (mean, 11.8%) in the TC and HC tissue, respectively. The stereological method demonstrated that the Ki-67 PI ranged from 30.1 to 51.5% (mean, 41.0%), and from 3.2 to 30.1% (mean, 15.1%) in the TC and HC tissues, respectively. The strongest association in the collinearity of regression analysis was observed between the Aperio nuclear V9 algorithm/stereological models in the TC tissue (r2=0.7; P=0.04) and the HC tissue (r2=0.7; P=0.03), and the visual/stereological models in the TC tissue (r2=0.7; P=0.04) and the visual/Aperio nuclear V9 algorithm models in the HC tissue (r2=0.7; P=0.04). A weak and statistically insignificant association was identified between the visual/Aperio nuclear V9 algorithm analysis in the TC tissue (r2=0.4; P=0.2) and the visual/stereological models in the HC tissue (r2=0.5; P=0.13). No significant difference was observed between the nuclear area of the TC (mean, 36.5 μm2) and HC (mean, 35.7 μm2; P=0.88) tissues. It was concluded that the Aperio nuclear V9 algorithm is a useful tool for the reliable analysis of histopathological changes of OSSN. The results of this computer-aided algorithm correlate strongly with the stereological method when assessing the Ki-67 PI.

Leukocyte cells identification is one of the most frequently performed blood tests and plays an important role in the diagnosis of diseases. The quantitative observation of leukocyte cells is often complemented by morphological analysis in both research and clinical condition. Different from the traditional leukocyte cells morphometry methods, a molecular hyperspectral imaging system based on acousto-optic tunable filter (AOTF) was developed and used to observe the blood smears. A combined spatial and spectral algorithm is proposed to identify the cytoplasm and the nucleus of leukocyte cells by integrating the fuzzy C-means (FCM) with the spatial K-means algorithm. Then the morphological parameters such as the cytoplasm area, the nuclear area, the perimeter, the nuclear ratio, the form factor, and the solidity were calculated and evaluated. Experimental results show that the proposed algorithm has better performance than the spectral based algorithm as the new algorithm can jointly use the spatial and spectral information of leukocyte cells.

Background: Diabetes is a third leading cause of mortality and morbidity in the world. Diabetes is one of the most common endocrine metabolic disorders and its prevalence has been increasing worldwide. Oral exfoliative cytology may be a more appropriate adjunctive diagnostic tool in conditions like diabetes mellitus, where the invasive techniques lose viability. Aims: The purpose of this study is to analyze the cytomorphometric changes in the exfoliated cells of the oral mucosa, as an adjunct to the diagnosis of diabetes. Materials and Methods: Smears were taken from the buccal mucosa of 30 diabetes patients (study group) and 30 healthy individuals (control group). All the smears were stained with rapid Papanicolaou stain (PAP). In the PAP smears, the nuclear area (NA), cytoplasmic area (CA), and cytoplasmic-to-nuclear ratio (CNR) were evaluated for 50 cells in each smear, using the Image Analysis Software (Magnus Pro™) and research microscope (Lawrence and Mayo™). Results: The results showed that the mean NA was significantly higher (P < 0.001) in the study group, whereas, the mean CA did not exhibit a statistically significant difference (P > 0.001). The mean CNR was significantly lower in the study group (P < 0.001). Interpretation and Conclusion: The results associated with the clinical observations suggest that diabetes can produce morphological and functional alterations in the oral epithelial cells, detectable by microscopic and cytomorphometric analysis using exfoliative cytology, which can be used in the diagnosis of the disease.

Objectives: The study aims to establish an etiological association between diabetes and precancerous lesions of the oral cavity by cytomorphometric analysis of the oral epithelium. Study Design: Smears were obtained from three distinct oral sites – buccal mucosa, dorsum of the tongue and the floor of the mouth in ten controls and ten patients previously diagnosed with type II diabetes. The oral smears were stained with Papanicolaou SA-36 solution. An eye – piece graticule was used to obtain the cytoplasm and nuclear dimension; where larger dimension was denoted as “D” and the smaller dimension was denoted as “d”. The nuclear area (NA), nuclear diameter (ND), cytoplasmic area (CA) and the cytoplasmic / nuclear ratio (C/N) were evaluated from 50 cells predominant in each oral site. Statistical Analysis: The cytomorphometric data obtained was compared between the group of diabetic patients and the control groups using the student’s t- test (SPSS version 11.0). Results: Results showed that the nuclear area and the nuclear diameter of oral epithelial cells were increased in diabetic patients, as compared to non- diabetics, while the non- diabetic patients demonstrated an increase in nuclear ratio. Conclusions: The results from this study suggest that diabetes mellitus can cause alterations in oral epithelial cells that are detectable with exfoliative cytology. Key words: Cytomorphometry, diabetes mellitus, oral exfoliative cytology

To investigate the protective effect of angiotensin-converting enzyme (ACE)-inhibitory peptide LAP on the left common carotid artery remodeling in spontaneously hypertensive rats (SHRs). A cohort of male SHRs were randomly divided into three groups (n = 10 for each group): pseudo-experimental group, enalapril-treated group as a positive control group, ACE-inhibitory peptide LAP-treated group. After the experiment, the left common carotid artery from each rat was removed for morphological evaluation. It was observed that the vascular medial thickness, media thickness/lumen diameter, medial cross-sectional area and mean nuclear area of smooth muscle cells of the left common carotid artery in the LAP group or enalapril group were significantly lower than those in the pseudo-experimental group, while there was no significant difference in these parameters observed between the LAP group and enalapril group. Additionally, the vascular area percentage of collagen fibers of the left common carotid artery in the LAP group and enalapril group was significantly lower than that of the pseudo-experimental group. The protective vessel remodeling effect in SHRs was observed with ACE-inhibitory peptide LAP in SHRs by decreasing blood pressure, inhibiting smooth muscle cell hypertrophy and reducing the proliferation of collagen fibers.

We compared the cytoarchitectural features used for the cytologic diagnosis of endometrial adenocarcinoma (EC) using ThinPrep® (TPS = ThinPrep Sample) and BD SurePath™ (SPS = SurePath Sample) preparations. In 20 patients, a direct endometrial sample using the Uterobrush was obtained. Nineteen cases of EA and one case of carcinosarcoma were studied. TPS and SPS were performed according to the manufacturer's recommendations. Moreover, after the TPS preparation, the residual material was also used to prepare an SPS sample (TP-SPS = ThinPrep-Surepath sample). The following points were investigated in both preparations: (1) number of cell clumps; SPS had a significantly higher (20.9) than TPS (1.7) and TP-SPS (10.3); (2) long axis of clumps; SPS had a significantly higher (215.4) than TPS (146.0); (3) rate of cell clumps with longer axes than 200 μm; SPS had a significantly higher (36.7) than TPS (15.2) and TP-SPS (24.2). TP-SPS showed higher values than TPS; (4) nuclear area; TPS had a significant higher (61.2) than SPS (40.8) and TP-SPS (38.6); (5) degree of overlapping nuclei; SPS (3.4) had a significantly higher number of overlapping nuclei than TPS (0.7) and TP-SPS (2.1); (6) nuclear chromatin pattern; no significant differences for the nuclear chromatin pattern were found in the three different methods. The poor performance of TPS versus SPS and TP-SPS was explained with the heavy blood contamination of the samples, and the absence of adhesive coating in the slides is used for TPS. Further investigation of technical differences in liquid-based cytology methodologies is needed.

Argyrophilic nucleolar organising region associated proteins (AgNORs) are of interest in a variety of diseases including thyroid disorders. We have investigated the cut-off values for AgNOR count and with a new approach, Total AgNOR area/nuclear area (TNORa/Na) proportions to discriminate thyrocytes obtained from benign thyroid nodules and normal thyroid tissue. Thirty patients whose fine needle aspiration (FNA) materials were compatible with a benign lesion (named as goiter group) and 30 controls (subjects with normal thyroid tissue) were included. In the control group, biopsy material was obtained from histologically normal thyroid gland operated on because FNA material was compatible with papillary thyroid carcinoma (PTC). These samples were stained for AgNOR and 100 nuclei per individual were examined. Both AgNOR values for individual cells and cut-off values were detected for each group. Patients with goiter had significantly (P

Background: Invasive lobular breast cancer (ILC) is the second most frequent form of breast cancer. While cancer cells are regularly investigated, tumor stroma represents a highly unexplored field. Aims: The aim of this study is to perform a detailed investigation of clinical, immunohistochemical, and morphometric characteristics of desmoplastic (D) and nondesmoplastic (ND) ILC. Materials and Methods: This study included twenty cases of ILC that were divided into two groups designated as D and ND groups. Medical histories and diagnosis data were obtained from the archives of the Center of Pathology, Clinical center Nis (Serbia). Morphometric analysis of hematoxylin and eosin stained slides was performed using ImageJ software, and the obtained data were further statistical processed. Results: Statistical analyses of the data revealed that no significant differences between D and ND groups when patient age, estrogen receptor (ER), and progesterone receptor (PR) expressions and morphometrical parameters (such as the distance between groups of cancer cells and nucleocytoplasmic ratio) were compared. However, D and ND groups statistically, significantly differed in the occurrence of axillary lymph node metastasis, and when the ER and PR data were included, in certain nuclear parameters (cell/nucleus area, perimeter, Feret's diameter, and circularity). Conclusions: Desmoplastic stroma was observed more frequently in patients without axillary lymph node metastases, whereas the expression of ER and PR had no influence on its development. According to the measured morphometric parameters larger cells/nuclei belonged to ND group.

Objectives To derive objective values for the diagnosis of Hirschsprung disease (HSCR) from a comparison of the morphometric profile of large intestinal neuronal plexuses in normal perinatal autopsies and surgical specimens of HSCR. Methods A cross-sectional comparative study with 40 subjects each in (i) non-HSCR perinatal group encompassing neonates and stillborn babies beyond 30 weeks of gestation on whom autopsies were conducted and (ii) HSCR group comprising all patients clinicoradiologically diagnosed as HSCR. The morphometric assessment was done on hematoxylin-and-eosin-stained sections. Key Results The morphometric profile in terms of average number of ganglia/linear mm of colon, interganglion distance, number of ganglion cells/ganglion, average ganglion cell length, ganglion cell nuclear area, ganglion cell nuclear diameter, nerve trunk thickness, and density has been outlined. On comparison with the neuroanatomically normal zone of HSCR, the cut-offs to identify hypertrophic nerve trunks (nerve trunk thickness of >37.85 μm) and reduced number of ganglia (number of ganglia/linear mm of colon 229 μm) were derived. Conclusions & Inferences The determined objective values, after testing on diagnostic rectal biopsies, may serve to formulate a diagnostic algorithm along with immunostaining for diagnosis of HSCR in colorectal specimens.

Resumo: Esse estudo objetivou avaliar o efeito da ooforectomia sob os parâmetros histomorfométricos dos cardiomiócitos e quantificação de colágeno em ratas Wistar. Vinte ratas foram utilizadas e separadas em dois grupos: GI - ooforectomizada e GII - sham-ooforectomizada. Após a recuperação anestésica os animais foram acomodados em gaiolas e separados nas condições de ciclo claro/escuro 12/12h por um período de 6 meses. Passado o período experimental, os animais foram anestesiados, o coração retirado e mergulhado em formaldeído a 10%. Os fragmentos dos ventrículos foram submetidos ao processamento histológico e corados com hematoxilina e eosina. As análises histomorfométricas (área celular e área e volume nuclear) foram realizadas pelo microscópio de luz e software ImageJ versão 1.44. Os dados foram submetidos à análise ANOVA e quando significantes, complementados pelo teste t de student (p

WOS: 000373634600001, PubMed: 27088074, Background. The aim of this study was to compare the cytomorphometric characteristics of the buccal cells of Behccet's disease patients with those of healthy controls. Methods. This case-control study compared a group of 30 patients with Behcet's disease with an age-and gender-matched control group of 30 healthy individuals. The buccal mucosal smears were stained using the Papanicolaou technique for cytomorphometric analyses. The nuclear and cytoplasmic areas were evaluated using digital image analysis; the ratio of nuclear to cytoplasmic areas and nuclear roundness are presented. Results. The nuclear and cytoplasmic areas of the BD patients' cells were significantly smaller than those of the healthy controls' cells, while the nucleus-to-cytoplasm ratio and neutrophil infiltration rate did not differ significantly between the groups. However, the nuclear area, cytoplasmic area, nucleus-to-cytoplasm ratio, and nuclear roundness factor were significantly higher in patients without aphthae. The neutrophil infiltration rate did not differ significantly in patients with or without aphthae. Conclusion. Behcet's disease can produce cytomorphometric changes in buccal cells that are detectable by exfoliative cytology and cytomorphometric analysis techniques.

The size of nuclei in histological preparations from excised breast tumors is predictive of patient outcome (large nuclei indicate poor outcome). Pathologists take into account nuclear size when performing breast cancer grading. In addition, the mean nuclear area (MNA) has been shown to have independent prognostic value. The straightforward approach to measuring nuclear size is by performing nuclei segmentation. We hypothesize that given an image of a tumor region with known nuclei locations, the area of the individual nuclei and region statistics such as the MNA can be reliably computed directly from the image data by employing a machine learning model, without the intermediate step of nuclei segmentation. Towards this goal, we train a deep convolutional neural network model that is applied locally at each nucleus location, and can reliably measure the area of the individual nuclei and the MNA. Furthermore, we show how such an approach can be extended to perform combined nuclei detection and measurement, which is reminiscent of granulometry.

This chapter considers some key aspects of Iran’s nuclear programme in technical terms and examines the implications of the particular approach that Iran has adopted. Themes addressed include the coherence of the civil rationale in light of particular technical developments, the extent and significance of nuclear opacity, and the relationship between the nuclear programme and the military sphere. The period of study for this chapter is 2002 to 2013 because it was over this timeframe that the international spotlight on Iran gained ever more intensity, and technical indicators continually emerged that raised questions over Tehran’s designs in the nuclear area. It is shown that, while these indicators did not provide ‘smoking gun’ evidence to support allegations that Iran’s civil programme disguised a nuclear weapons programme, it was evident that Tehran had moved far beyond what it strictly needed for its civil nuclear programme.

We evaluated the differences in cytologic findings between conventional and thin-layer preparations in endometrial cytology to introduce the thin-layer method into routine cytology. Eighty patients who had undergone endometrial cytology and biopsy on the same day were selected and we compared the cytological findings between conventional- and thin-layer preparations (TLP) in endometrial cytology. The numbers of neutrophils and cell clusters in the thin-layer method were lower than those in the conventional smear (CSS) method. The average number of neutrophils in endometrioid adenocarcinoma was significantly higher than that in normal morphology endometrium and endometrial hyperplasia. Regarding the shape of the cell clusters, ball-like patterns and round-edged cell clusters were not identified in CSS. The average number of clusters in CSS was significantly greater than that using the TLP. The average of the nuclear area in CSS was significantly larger than that using the TLP, indicating that the nuclear areas in CSS were more uneven than that using the TLP. In the future, it is expected that liquid-based cytology will be applied to the cytological diagnosis of a variety of lesions. The influence on cells due to fixation is considerable in liquid-based preparations. Therefore, if we strive to pick up the differences between CSS and TLP of endometrial samples, the diagnostic accuracy of the latter could be improved. Diagn. Cytopathol. 2013. © 2012 Wiley Periodicals, Inc.

Symptomatic disc degeneration is a complex pathological condition that involves a cascade of events and is not totally understood. In this context, animal models gain an important role, allowing for better understanding of the degenerative process and therapeutic interventions. There are several models with different methods of evaluation of experimental disc degeneration (EDD), including imaging, biochemical, genetics, and histological approaches, but no real gold standard has been set. The authors aim to evaluate EDD by means of automated morphometric analysis and to determine values for differentiating normal and degenerated discs by this method. The criteria mean and total cellular area, mean and total cytoplasm area, and total nuclear area of cells in the nucleus pulposus were able to differentiate the condition of degeneration (p

Stained cytological specimens from eight canine ceruminous adenomas and eight canine ceruminous carcinomas were analysed by computer-assisted nuclear morphometry. Three carcinomas had metastases in regional lymph nodes at the time of the diagnosis. The morphometric parameters evaluated in this study were mean nuclear area (MNA, µm(2) ), mean nuclear perimeter (MNP, µm), mean nuclear diameter (D mean, µm), minimum nuclear diameter (D min, µm) and maximum nuclear diameter (D max, µm). The study aimed to evaluate (1) the possibility of using nuclear cytomorphometry as an auxiliary diagnostic method to differentiate between canine ceruminous adenomas and adenocarcinomas, and (2) the prognostic value of nuclear morphometry in canine ceruminous carcinomas. The results indicated that (1) MNA, MNP, D mean, D min and D max could be used as effective auxiliary tools for differential diagnosis between canine ceruminous adenomas and adenocarcinomas, and (2) MNA, MNP, D mean and D max are reliable prognostic indicators for canine ceruminous adenocarcinomas.