Mean Nuclear Area Research Articles

Hypoxic Preconditioning Protects Cord Blood-Mesenchymal Stromal Cells from Ischemic Damage in Vitro

Objectives: We have previously shown that cord blood mesenchymal stromal cell (CBMSC)-secreted factors act anti-apoptotic and pro-mitotic on endothelial cells and cardiomyocytes to ischemia-like conditions. However CB-MSCs themselves suffer ischemic damage in the infarcted heart, hampering their potential clinical efficacy. To increase CB-MSC tolerance to ischemia, we developed a specific hypoxic preconditioning (HP) protocol and investigated the molecular mechanisms involved. Methods: Human CBMSCs were subjected to different durations (12, 24 or 36 hour) of hypoxic preconditioning (1% O2 in glucose/serum-containing medium) prior to a final phase of simulated ischemia (24 hour, 1% O2 in glucose/serum-free medium). Cell loss, metabolic activity (WST-8 conversion), apoptosis (nuclear fragmentation and shrinking) and proliferation (BrdU incorporation) were measured. The effects of HP on Akt-, ERK1/2 and STAT3-phosphorylation and Bcl-2, Bcl-XL and Bag-1 mRNA expression were determined by Western blot and quantitative real time PCR. Results: Ischemic damage of CBMSCs was lowest after 24 hour HP. Here, the percentage of remaining cells was higher (74 ± 7% versus non-HP 39 ± 11%, p < 0.05) and metabolic activity was enhanced (35 ± 2% versus non-HP 18 ± 6%, p < 0.05). Nuclear fragmentation index was lower (11.9 ± 0.2 versus non-HP 13.8 ± 0.3, p < 0.05), nuclear shrinking was less pronounced (mean nuclear area 179 ± 5 µm2 versus non-HP 155 ± 5 µm2, p < 0.05), and the proliferation index was enhanced (49 ± 1 versus non-HP 16 ± 3, p < 0.05). While phosphorylation of Akt and STAT3 was increased in preconditioned cells, ERK1/2 phosphorylation and Bcl-2 mRNA expression were downregulated in response to HP, and mRNA expression of Bcl-XL and Bag-1 was not altered. Conclusions: Hypoxic preconditioning protects CBMSCs from the deleterious impact of combined oxygen/glucose/serum-deprivation via STAT3-dependent signaling. This might by a useful strategy to increase their tolerance to ischemia and improve their therapeutic efficacy in clinical applications.

Significance of Morphometric Parameters in the Categorization of Breast Lesions on Cytology.

The aim of this study was to assess the utility of the morphometric parameters in cytological aspirates in the categorization of breast lesions. The study was carried out on 50 females presenting with a breast lump for fine needle aspiration cytology. Air-dried smears from the aspirate were stained with Leishman and Giemsa stains while alcohol-fixed preparations were stained with the PAP stain. They were subsequently categorized into benign, borderline and malignant categories on light microscopy. Morphometry was performed on the stained smears. The morphometric parameters studied were mean nuclear area, mean cytoplasmic area, perimeter and nuclear/cytoplasm ratio. Morphometric parameters revealed a progressive and statistically significant increase in values from benign to borderline to malignant cases. On comparing benign with borderline and malignant, all the four parameters were found to be statistically significant with a p-value of less than 0.05 while on comparing borderline with malignant, two of the parameters, i.e. mean nuclear area and mean cytoplasmic area, were statistically significant. This study introduces morphometry as a highly objective tool to supplement the entirely subjective fine needle aspiration cytology in the crucial differentiation of benign from malignant lesions and especially the borderline cases in the gray zone comprising of atypical ductal hyperplasia and ductal carcinoma in situ.

Cytomorphometric analysis of buccal mucosa cells in tobacco chewers

Objective: The objective of the present study was to assess the effect of tobacco chewing on buccal mucosa by using cytomorphometry. Study Design: The study group consisted of 100 subjects divided into 50 tobacco chewer subjects (Group C) and 50 control subjects (Group N) aged between 31 and 80 years. We analyzed and compared the nuclear and cell morphometric features, that is, mean nuclear area (MNA), mean nuclear perimeter (MNP), mean of maximum nuclear diameter (Max-ND), mean of minimum nuclear diameter (Min-ND), mean cell area (MCA), mean cell perimeter (MCP), mean of maximum cell diameter (Max-CD), mean of minimum cell diameter (Min-CD) and nuclear to cell parameter ratio. Buccal epithelial cells of these individuals were collected with a brush and fixed smears were stained with Papanicolaou stain and cytomorphometric analysis performed using Image J image analysis software. Results: The mean of above nuclear and cell morphometric parameters and their ratio were found to be statistically significant by Univariate analysis of variance (ANOVA) being P value < 0.001. Conclusions: There is progressive increase in nuclear parameters, decrease in cellular parameters and increase in ratio of nuclear to cellular parameters in smears from tobacco chewers, as compared with normal subjects. It is possible to conclude that these adaptive changes in the cellular and nuclear parameters tend to be a progression toward dysplastic or premalignant change.

Evaluation of the Long-term Growth Potential of Benign Non-toxic Thyroid Nodules by the AgNOR Method; an 11-year Follow-up Study

Objective: The classification of benign thyroid nodules as stable or as fast growing may have clinical and economical relevance. The proliferative activity of thyroid cells determined by argyrophilic staining of the nucleolar organizer region (AgNOR) displays significant differences between benign and malignant thyroid lesions. We tested if that parameter can predict the growth potential of benign nodules. Methods: On cytological samples of 106 benign nodules of 89 euthyroid patients, the mean AgNOR number per cell (AgNORc), the mean AgNOR area per cell (AgNORa) and the mean nuclear area were determined. The mean follow-up period was 11.9 years. The initial (V1) and the final (V2) nodule volumes and respective TSH values (TSH1 and TSH2) were determined. Results: 66 nodules (62.2%) increased in volume. The median of V2/V1 was 1.31. V2/V1 correlated significantly with AgNORa, AgNORc, nuclear area and TSH-1. There was no significant correlation between V2/V1 and either age, duration of follow-up or TSH-2. No correlation was found between V2 and any of the parameters. We found on both forward and backward stepwise analysis that AgNORc values and initial TSH levels were significant determinates of the increase in volume size, i.e. both higher AgNORc values and higher initial TSH levels carry a greater risk for nodule growth. Conclusions: The determination of AgNORc during FNA evaluation may contribute to the estimation of growth potential of thyroid nodules. Further studies using other enzyme techniques or new molecular methods are justified to achieve a better discrimination between stable and growing nodules.

Role of morphometry and proliferative parameters in grading of urothelial neoplasms

IntroductionMean nuclear area of 10 nuclei (MNA–10), mitotic activity index (MAI) and Ki–67 are highly reproducible and can be routinely used as adjuncts to histopathological grading in classifying tumors. Assays of these biomarkers are non–invasive, rapid, easy to perform, more objective and accurate, with high sensitivity and specificity, and correlate well with tumor grade.Material and methodsThis study was conducted at the Department of Pathology PGIMS, Rohtak on 50 cases, of which 25 cases were high–grade, 15 low–grade, 6 Papillary Urothelial Neoplasm of Low Malignant Potentialand 4 reactive lesions as per the 2004 ISUP/WHO classification. MNA–10, MAI and Ki–67 immunoquantitation were performed on stained sections.ResultsThe age of the patients varied from 35 to 87 years. Male: female ratio was 3.5:1. The mean MNA–10 (μm2) for High Grade Malignant Potential was 104.52 ±25.64 μm2, which was significantly higher than in PUNLMP (47.64 ±10.23) and LMP (51.57 ±15.66). MAI (/10 HPF) showed an increasing trend from reactive lesions to HMP, with a mean of (3 ±1.16)/10 HPF to (21.36 ±5.31)/10 HPF respectively. Ki–67 labelling index, a proliferative marker, revealed increasing trend lowest with reactive lesions (10 ±2.83%) and highest in high grade tumors (65.96 ±14.44). Spearman's correlation showed maximum correlation between MAI and Ki–67 and the increasing grade of tumor.ConclusionsMNA–10 in combination with Ki–67 and MAI was found to be stronger than MNA–10 alone. MAI has high reproducibility in differentiating low and high grade, with simple assessment in paraffin embedded sections allowing adequate histopathological analysis and visualization of proliferating cells simultaneously. This multivariate grading model should be applied in routine grading to overcome interobserver variability and to increase reproducibility of grading.

Diagnostic application of computerised nuclear morphometric image analysis in fine needle aspirates of breast lesions

Background: Alterations in nuclear structure are the indications of cancer diagnosis. This study has thus focused on variation in nuclear morphometry in breast lesions. Aim: The aim of the study is to compare morphometric characteristics of different types of breast lesions and evaluate its utility in differentiating benign against malignant. Study Design: Sixty fine needle aspiration cytology (FNAC) of ethanol fixed samples were obtained and stained with Papanicolou stain. All diagnoses reported were confirmed histopathologically. Nuclear morphometric parameters for free cells in smears were calculated using the ImageJ 1.47 morphometric computer software. Result: The nuclear morphometric parameters including mean nuclear area, perimeter, diameter, long axis, and short axis were found to be statistically significant (ANOVA, P < 0.0001) in differentiating benign and malignant breast aspirates. Conclusion: Nuclear morphometry has been found to be a valuable objective method in differentiating benign and malignant breast lesions, especially in the grey zones, when diagnostic dilemmas are encountered. This study results demonstrate that the size-related parameters (area, perimeter, diameter, long, and short axes) of the nucleus are the most appropriate nuclear morphometric parameters for differentiating between benign lesions and infiltrative ductal carcinoma of the breast.

Applicability of preoperative nuclear morphometry to evaluating risk for cervical lymph node metastasis in oral squamous cell carcinoma.

BackgroundWe previously reported the utility of preoperative nuclear morphometry for evaluating risk for cervical lymph node metastases in tongue squamous cell carcinoma. The risk for lymph node metastasis in oral squamous cell carcinoma, however, is known to differ depending on the anatomical site of the primary tumor, such as the tongue, gingiva, mouth floor, and buccal mucosa. In this study, we evaluated the applicability of this morphometric technique to evaluating the risk for cervical lymph node metastasis in oral squamous cell carcinoma.MethodsA digital image system was used to measure the mean nuclear area, mean nuclear perimeter, nuclear circular rate, ratio of nuclear length to width (aspect ratio), and nuclear area coefficient of variation (NACV). Relationships between these parameters and nodal status were evaluated by t-test and logistic regression analysis.ResultsEighty-eight cases of squamous cell carcinoma (52 of the tongue, 25 of the gingiva, 4 of the buccal mucosa, and 7 of the mouth floor) were included: 46 with positive node classification and 42 with negative node classification. Nuclear area and perimeter were significantly larger in node-positive cases than in node-negative cases; however, there were no significant differences in circular rate, aspect ratio, or NACV. We derived two risk models based on the results of multivariate analysis: Model 1, which identified age and mean nuclear area and Model 2, which identified age and mean nuclear perimeter. It should be noted that primary tumor site was not associated the pN-positive status. There were no significant differences in pathological nodal status by aspect ratio, NACV, or primary tumor site.ConclusionOur method of preoperative nuclear morphometry may contribute valuable information to evaluations of the risk for lymph node metastasis in oral squamous cell carcinoma.

Morphometric evaluation and clinical correlations in pediatric malignant small round cell tumors.

Aims:Nuclear size increases in malignant tumors and reflects DNA content, ploidy and proliferation index. Present study investigated if the nuclear morphometry could differentiate histomorphologically similar paediatric malignant small round cell tumors on hematoxylin and eosin stained sections for diagnostics in a resource poor setting.Settings and Design:All the consecutive malignant pediatric tumors received in Pathology Department from other faculties of King George's Medical University and also those referred directly to Pathology Department from other hospitals of city/other cities during 3 years period were recorded.Materials and Methods:Morphometric analysis was done in 22 confirmed (by higher ancillary techniques) but histomorphologically difficult to differentiate round cell tumors. All sections were analyzed by cell images from six different areas, using Leica Q win 500 images software.Results:Nuclear measurements were obtained for retinoblastoma (RB) (nine cases), neuroblastoma (five cases), Wilms tumor (WT) (three cases), rhabdomyosarcoma (three cases), malignant hemangiopericytoma (one case) and non-Hodgkin lymphoma (one case). Among the RBs, maximum mean nuclear area percent (24.93) was seen in a case with nerve involvement and metastasis, followed by cases with only nerve involvement (21.60) and smallest area (16.57) was in non-nerve involving, nonmetastatic cases. All five cases of neuroblastoma had almost similar mean nuclear area percent (18.05-18.29). WT case with metastasis had higher nuclear area (21.25) than nonmetastatic (19.47). Amongst all the tumors, minimum value (14.93) was seen in malignant hemangiopericytoma.Conclusion:Morphometric evaluation in paediatric malignant round cell tumors have generated useful data, and needs further multicentric confirmation for implementation.

Karyometric Analysis of Squamous Metaplasia, Dysplasia and Squamous Cell Carcinoma of the Lung

Summary The sequence of precursor lesions for squamous cell carcinoma may be hyperplasia-metaplasia-dysplasia-carcinoma in situ. The aim of this study was to perform a karyometric analysis of squamous metaplasia, moderate dysplasia and squamous cell lung carcinoma. Bronchoscopic biopsies of normal mucosa in chronic bronchitis patients (n=10), squamous metaplasia (n=10), moderate dysplasia (n=11), squamous cell lung carcinoma (n=48), and normal appearing mucosa surrounding carcinoma (n=11) were retrieved. Three nuclear variables were estimated using an image analysis system. The mean equivalent diameter, nuclear area and volume of equivalent sphere of squamous cell lung carcinoma were significantly larger than in moderate dysplasia, squamous metaplasia and normal bronchial mucosa. Also, the values of equivalent diameter, nuclear area and volume of equivalent sphere were significantly larger in normal appearing mucosa surrounding carcinoma compared to normal mucosa in chronic bronchitis patients. Karyometric analysis may be a helpful ancillary tool in distinguishing squamous cell lung carcinoma from dysplasia, and dysplasia from squamous metaplasia in bronchoscopic biopsy specimens.

Characterization of squamous cell carcinomas of the head and neck using methods of spatial statistics.

In the present study, 53 cases of squamous cell carcinomas of the head and neck were characterized by a quantitative histological texture analysis based on principles of spatial statistics. A planar tessellation of the epithelial tumour component was generated by a skeletonization algorithm. The size distribution of the virtual cells of this planar tessellation, and the size distribution of the profiles of the tumour cell nuclei were estimated in terms of area and boundary length. The intensity, the reduced second moment function (K-function) and the pair correlation function of the point process of the centroids of the profiles of the tumour cell nuclei were also estimated. For both purposes, it is necessary to correct for edge effects, which we consider in this paper in some detail. Specifically, the point patterns of the tumour cell nuclei were considered as realizations of a point process, where the points exist only in the epithelial tumour component (the permitted phase) and not in the stroma (the forbidden phase). The methods allow to characterize each individual tumour by a series of summary statistics. The total set of cases was then partitioned into two groups: 19 cases without lymph node metastases (pN0), and 34 nodal positive cases (pN1 or pN2). Statistical analysis showed no significant differences between the intensities, the mean K-functions and the mean pair correlation functions of the tumour cell nucleus profiles of the two groups. However, there were some significant differences between the sizes of the virtual cells and of the nucleus profiles of the nodal negative cases as compared to the nodal positive cases. In a logistic regression analysis, one of the quantitative nuclear size variables (mean nuclear area) was found to be a significant predictor of lymph node metastasis, in addition to tumour stage. The study shows the potential of methods of spatial statistics for objective quantitative grading of squamous cell carcinomas of the head and neck, and provides an example for modelling histological point patterns as realizations of planar point processes occupying a reference phase which is only a partial component of the total tissue.

Quantitative Analysis of High-Resolution Microendoscopic Images for Diagnosis of Esophageal Squamous Cell Carcinoma

High-resolution microendoscopy is an optical imaging technique with the potential to improve the accuracy of endoscopic screening for esophageal squamous neoplasia. Although these microscopic images can be interpreted readily by trained personnel, quantitative image analysis software could facilitate the use of this technology in low-resource settings. In this study, we developed and evaluated quantitative image analysis criteria for the evaluation of neoplastic and non-neoplastic squamous esophageal mucosa. We performed an image analysis of 177 patients undergoing standard upper endoscopy for screening or surveillance of esophageal squamous neoplasia, using high-resolution microendoscopy, at 2 hospitals in China and at 1 hospital in the United States from May 2010 to October 2012. Biopsy specimens were collected from imaged sites (n= 375), and a consensus diagnosis was provided by 2 expert gastrointestinal pathologists and used as the standard. Quantitative information from the high-resolution images was used to develop an algorithm to identify high-grade squamous dysplasia or invasive squamous cell cancer, based on histopathology findings. Optimal performance was obtained using the mean nuclear area as the basis for classification, resulting in sensitivities and specificities of 93% and 92% in the training set, 87% and 97% in the test set, and 84% and 95% in an independent validation set, respectively. High-resolution microendoscopy with quantitative image analysis can aid in the identification of esophageal squamous neoplasia. Use of software-based image guides may overcome issues of training and expertise in low-resource settings, allowing for widespread use of these optical biopsy technologies.

Quantitative morphology in canine cutaneous soft tissue sarcomas.

Stained cytological specimens from 24 dogs with spontaneous soft tissue sarcomas [fibrosarcoma (n = 8), liposarcoma (n = 8) and haemangiopericytoma (n = 8)], and 24 dogs with reactive connective tissue lesions [granulation tissue (n = 12) and dermal fibrosis (n = 12)] were analysed by computer-assisted nuclear morphometry. The studied morphometric parameters were: mean nuclear area (MNA; µm(2)), mean nuclear perimeter (MNP; µm), mean nuclear diameter (MND mean; µm), minimum nuclear diameter (Dmin; µm) and maximum nuclear diameter (Dmax; µm). The study aimed to evaluate (1) possibility for quantitative differentiation of soft tissue sarcomas from reactive connective tissue lesions and (2) by using cytomorphometry, to differentiate the various histopathological soft tissue sarcomas subtypes in dogs. The mean values of all nuclear cytomorphometric parameters (except for Dmax) were statistically significantly higher in reactive connective tissue processes than in soft tissue sarcomas. At the same time, however, there were no considerable differences among the different sarcoma subtypes. The results demonstrated that the quantitative differentiation of reactive connective tissue processes from soft tissue sarcomas in dogs is possible, but the same was not true for the different canine soft tissue sarcoma subtypes. Further investigations on this topic are necessary for thorough explication of the role of quantitative morphology in the diagnostics of mesenchymal neoplasms and tumour-like fibrous lesions in dogs.

Dietary Fat and Aging Modulate Apoptotic Signaling in Liver of Calorie-Restricted Mice

Imbalance between proliferation and cell death accounts for several age-linked diseases. Aging, calorie restriction (CR), and fat source are all factors that may influence apoptotic signaling in liver, an organ that plays a central metabolic role in the organism. Here, we have studied the combined effect of these factors on a number of apoptosis regulators and effectors. For this purpose, animals were fed diets containing different fat sources (lard, soybean oil, or fish oil) under CR for 6 or 18 months. An age-linked increase in the mitochondrial apoptotic pathway was detected with CR, including a decrease in Bcl-2/Bax ratio, an enhanced release of cytochrome c to the cytosol and higher caspase-9 activity. However, these changes were not fully transmitted to the effectors apoptosis-inducing factor and caspase-3. CR (which abated aging-related inflammatory responses) and dietary fat altered the activities of caspases-8, -9, and -3. Apoptotic index (DNA fragmentation) and mean nuclear area were increased in aged animals with the exception of calorie-restricted mice fed a lard-based fat source. These results suggest possible protective changes in hepatic homeostasis with aging in the calorie-restricted lard group.

Cytomorphometric evaluation of alterations in buccal mucosal cells of smokers and non-smokers

The objective of the present study was to assess the effect of smoking on buccal mucosa by using cytomorphometry. A case control study on 50 smoker subjects and 50 non-smoker controls between the age of 28 and 71 years. Buccal epithelial cells of these individuals were collected with a brush and fixed smears were stained with Papanicolaou stain and cytomorphometric analysis performed using image analysis software (Image J v 1.47). Cytomorphometric changes could be the earliest indicators of cellular alterations. There is progressive increase in nuclear parameters i.e. mean nuclear area (MNA), mean nuclear perimeter (MNP), mean of maximum nuclear diameter (Max-ND) and mean of minimum nuclear diameter(Min-ND) and decrease in ratio of cellular to nuclear parameters in smears from smokers, as compared to normal subjects. This indicates that there could be cause–effect relationship between smoking and quantitative alterations. This increase determined in nuclear parameters shows smoking-related cellular adaptation. It is possible to conclude that this adaptive change in the cell nucleus tends to be a progression towards dysplastic change.

Nuclear morphometric and morphological analysis of exfoliated buccal and tongue dorsum cells in type-1 diabetic patients

Background:Diabetes mellitus type 1 that results from immunologically mediated damage to the β-cells in the pancreas. Diabetes mellitus is characterized by recurrent or persistent hyperglycemia. Hyperglycemia can be associated with salivary gland dysfunction and alterations in the oral epithelial cells.Aim:The aim of this study was to evaluate the qualitative and quantitative changes in buccal and tongue dorsum epithelial cells using an exfoliative cytology method in type 1 diabetic patients.Materials and Methods:We performed light microscopic analysis of the buccal and tongue dorsum smears in thirty type 1 diabetic patients and thirty healthy individuals. The oral smears were stained using Papanicolaou method for cytological examination and nuclear morphometric analysis. In each case, the mean nuclear area, perimeter, length, breadth, and roundness factor were evaluated in each smear using the image analysis software (Q Win, Leica™).Results:The nuclear area, length, breadth, and perimeters were significantly higher in the diabetic group from tongue dorsum smear than that of the control group (P < 0.05). In the cytological examination, karyorrhexis-karyolysis-karyopyknosis, binucleation, nuclear membrane irregularity, cytoplasmic polymorphism, perinuclear halo were observed in oral smears with type 1 diabetic patients. Binucleation (P = 0.002) and nuclear membrane irregularity (P = 0.024) were significantly more common in buccal smears of diabetic group. Furthermore, the sensitivity of buccal mucosa was significantly higher in the diabetic group (P = 0.006).Conclusion:The light microscopic and nuclear morphometric study indicates that type 1 diabetes can produce morphological and nuclear morphometric changes in the oral mucosa that are noticeable with exfoliative cytology.

Exfoliative cytology of buccal squames: A quantitative cytomorphometric analysis of patients with diabetes.

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.

Evaluation of galectin-3, β-catenin, and nuclear morphometry in different thyroid lesions

This study aimed to evaluate the expression pattern of galectin-3 and β-catenin and assess the nuclear morphometric features of different thyroid lesions in order to investigate their diagnostic significance. Sixty specimens of benign and malignant thyroid lesions, in addition to 10 cases of normal thyroid tissue used as control, were immunohistochemically stained with antibodies against galectin-3 and β-catenin. Hematoxylin and eosin-stained slides were examined using image analyzer Proplus V software. Morphological parameters measured included mean nuclear diameter, mean nuclear perimeter, and mean nuclear area. Cytoplasmic immunoreactivity for galectin-3 was strongest in papillary thyroid carcinomas (88.9%), whereas staining was less intense in the follicular variant of papillary thyroid carcinoma and in follicular (60 and 75%, respectively) and poorly differentiated carcinomas (28.6%); it was expressed focally and weakly in follicular adenoma cases and even more weakly in the reactive follicular epithelium of nodular goiter (P<0.5). Membrane β-catenin expression was decreased in only two of 13 (15.4%) adenomas and in 21 of 38 (55.3%) carcinomas (P<0.5). Among carcinomas, reduced membrane β-catenin was associated with progressive loss of tumor differentiation and increased invasiveness (P<0.5). The mean nuclear diameter and the mean nuclear perimeter were higher in undifferentiated carcinomas when compared with other subtypes and were the least for follicular neoplasms. Galectin-3 immunostaining and analysis of β-catenin dysregulation may be useful as an adjunct to distinguish benign from malignant thyroid lesions. Quantitative estimation of nuclear features can play a role in assessment of the morphological features and thus help in the diagnosis of thyroid lesions.

Immunohistochemical analysis of chromatin remodeler DAXX in high grade urothelial carcinoma

Background/AimsThe chromatin remodeler DAXX, a predominantly nuclear protein, regulates the status of chromatin organization. The aim of this exploratory immunohistochemical study was to evaluate DAXX protein expression in high grade invasive urothelial carcinoma (UC) of the bladder as a biological regulator of aggressiveness.MethodsQuantitative analysis was made on DAXX immunostained nuclei in tissue sections from 5 cases of bladder normal urothelium (NU) and 5 cases of bladder pT1 UC. Carcinoma in situ (CIS) and high grade papillary carcinoma (HGPCa) were identified in 2 out of 5 UC cases.ResultsThe nuclei in UC show an open configuration of the chromatin composed of granules varying in size and distribution and a mean nuclear area 1.7 times greater than that in NU (UC: mean and SD 24.4 ± 11.4 square microns; NU: 14.8 6.5 square microns. The differences are statistically significant). 70% of the NU nuclei are immunostained, whereas 90% of UC nuclei are positive. The mean gray level value in UC, related to the intensity of nuclear immunostaining, is lower than in NU by a factor of 0.94 (UC: mean and SD 100 ± 15; NU: 106 ± 15. The differences are statistically significant). In particular, the value in the nuclei adjacent to the stroma in UC is slightly lower than in the intermediate cell layers by factor of 0.98, whereas in NU it is slightly greater by a factor 1.02 and 1.04 compared to the intermediate and superficial cell layers. The values in CIS and HGPCa are similar to those in UC.ConclusionsThe quantitative immunohistochemical analysis shows an altered protein expression of chromatin remodeler DAXX in UC and in its preinvasive phases, when compared to NU. DAXX evaluation, if associated with markers related to global DNA methylation and histone acetylation, could be used in clinical practice as a marker of aggressiveness.Virtual slidesThe virtual slides for this article can be found here:http://www.diagnosticpathology.diagnomx.eu/vs/1398457297102379

Cytological Study of Breast Carcinoma Before and After Oncotherapy with Special Reference to Morphometry and Proliferative Activity.

Our study was done to assess the cytological changes due to oncotherapy in breast carcinoma especially on morphometry and proliferative activity. Cytological aspirates were collected from a total of 32 cases of invasive ductal carcinoma both before and after oncotherapy. Morphometry was done on the stained cytological smears to assess the different morphological parameters of cell dimension by using the ocular morphometer and the software AutoCAD 2007. Staining was done with Ki-67 and proliferating cell nuclear antigen (PCNA) as proliferative markers. Different morphological parameters were compared before and after oncotherapy by unpaired Student's t test. Statistically significant differences were found in morphometric parameters, e.g., mean nuclear diameter, mean nuclear area, mean cell diameter, and mean cell area, and in the expression of proliferative markers (Ki-67 and PCNA). Statistical analysis was done by obtaining p values. There are statistically significant differences between morphological parameter of breast carcinoma cells before and after oncotherapy.

The effects of angiotensin-converting enzyme-inhibitory peptide LAP on the left common carotid artery remodeling in spontaneously hypertensive rats

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.