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doi:10.1111/j.1741-2358.2009.00319.x Exfoliative cytology of the oral mucosa in burning mouth syndrome: a cytomorphological and cytomorphometric analysis Objective: The aim of this study was to evaluate oral epithelial cells by exfoliative cytology in burning mouth syndrome (BMS). Material and methods: Oral smears were collected from clinically normal-appearing mucosa by liquid-based exfoliative cytology in 40 individuals (20 BMS patients and 20 healthy controls matched for age and gender) and analysed for cytological and cytomorphometric techniques. Results: Mean values of nuclear area (NA) for experimental and control groups were, respectively, 67.52 and 55.64 μm2 (p

The planning of the construction and operation of Near Surface Disposal (NSD) facilities for Radioactive Waste at Serpong Nuclear Area needs a safety assessment in order to provide assurance that the radioactive waste disposal system functions properly. The purpose of the safety assessment is to provide a reasonable scientific assurance that the disposal system will provide an adequate level of safety and meet the requirements to protect human health and the environment. The NSD facility is vault type with a size of 20.2 × 20.2 m with a depth of 2.6 m from the ground surface (4.43 m above the highest ground water level). The facility was designed to accommodate waste packaged in 200 l drums and 950 l concrete shells. The total capacity of the disposal will be 1350 drums and 144 concrete shells. The waste that was considered in the study contain radionuclides which are dominantly contained in the low level packaged waste and, as reference for the safety assessment, with short half-lifes (less than or equal to about 30 years), namely Co-60 and Cs-137. This study used the PRESTO software version 4.2 to calculate the migration/exposure of radionuclides from the facility into the environment as well as individual doses. The results indicate that the predicted individual doses of the population around the facility at the Serpong site still fulfill the dose limit value of the public based on BAPETEN Chairman Regulation No. 4/2013 on the Protection and Radiation Safety in the Utilization of Nuclear Energy.

The imaging analytical capabilities of laser scanning cytometer (LSC) have been used to assess morphological features considered to be typical of the senescent phenotype. The characteristic “flattening” of senescent cells was reflected by the decline in the density of staining (intensity of maximal pixel) of DNA-associated fluorescence [4,6-diamidino-2-phenylindole (DAPI)] paralleled by an increase in nuclear size (area). The decrease in ratio of maximal pixel to nuclear area was even more sensitive senescence biomarker than the change in maximal pixel or nuclear area, each alone. The saturation cell density at plateau phase of growth recorded by LSC was found to be dramatically decreased in cultures of senescent cells, thereby also serving as an additional marker. The induction of cyclin dependent kinase inhibitors p21WAF1 and p27KIP1 and γH2AX and activation of ATM markers of DNA damage response were measured in parallel with DNA/DAPI maximal pixel and nuclear area. These biomarker indices were expressed in quantitative terms by reporting them as a fraction of the respective controls. The effect of treatment of A549 and WI-38 cells with different concentrations of mitoxantrone (Mxt) and trichostatin A for various time periods was studied to assess the degree (depth) of cell senescence. Also assessed was the effect of 2-deoxy-d-glucose, the agent attenuating metabolic cell activity, on the depth of senescence induced by Mxt. A relationship between the ability of cells to synthesize RNA (incorporate 5-ethynyluridine) that leads to growth imbalance and induction of cell senescence was also studied. The data show that morphometric analysis of cellular attributes by LSC offers an attractive tool to detect cell senescence and measure its degree particularly in assessing effects of the factors that enhance or attenuate this process. This methodology is of importance in light of the evidence that cellular senescence is not only a biological process that is fundamental for organismal aging but also impedes formation of induced-pluripotent stem cells providing the barrier for neoplastic transformation and is the major mechanism of induction of reproductive cell death during treatment of solid tumors. © 2010 International Society for Advancement of Cytometry.

Objectives: The objectives of this study were to quantitatively estimate the number of mitotic figures (MFs) and evaluate the cellular and nuclear features of various histological grades of oral squamous cell carcinoma (OSCC) using Feulgen and 1% crystal violet stains. Methods: This case-control study took place at the Dr D. Y. Patil Dental College & Hospital in Mumbai, Maharashtra, India, between June and December 2016. A total of 51 samples were retrieved from the hospital archives. Of these, 15 well-differentiated, 15 moderately-differentiated and six poorly-differentiated OSCC samples formed the case group while 15 samples of normal gingival mucosa constituted the control group. Each sample was dyed using Feulgen and 1% crystal violet stains and the mitotic count, nuclear area (NA), cellular area (CA), nuclear perimeter (NP), cellular perimeter (CP) and nuclear-to-cytoplasmic (N/C) ratio was calculated using computeraided morphometry techniques. Results: The number of MFs visible per field was significantly higher in Feulgen-stained sections as compared to those stained with crystal violet ( P = 0.050). In addition, the NA, NP, CA and CP values and N/C ratios of samples in the experimental group increased significantly in accordance with an increase in OSCC grade ( P

Background Malignant lymphomas are classified on the basis of morphology, immunohistochemistry, and genetic and molecular biological features. Morphology is considered the most important and basic feature. Lymphomas can be classified as small, medium, or large depending on the cell size, but this criterion tends to be rather subjective. The aim of this study was to investigate the usefulness of an objective approach based on quantitative measurements. Methods Twenty specimens of mantle cell lymphoma and 2 specimens of the tonsil were examined. The nuclear area of 6,401 tumor cells of mantle cell lymphoma and 743 normal mantle cells of reactive tonsils were measured by 3 authors by using a user-controlled image-analyzer. The images of the nuclei were outlined using the spline method and the i-solution software, and the data were assessed using ANOVA and Student's t-test. Results The mean nuclear areas of mantle cell lymphoma cells measured by the 3 authors were 37.9 [7.9] µm2, 37.9 [7.2] µm2, and 38.2 [7.7] µm2 and those of normal mantle cells in reactive tonsil were 28.6 [2.3] µm2, 28.8 [2.0] µm2, and 27.0 [3.0] µm2. There was no statistical difference between the 3 observations of mantle cell lymphoma (P=0.580) and normal tonsils. Conclusion For morphology, nuclear area is considered an important feature in the classification schemes of lymphoma. We showed that nuclear area measurement by using image analyzer can be used as an objective quantitative method. We think that nuclear morphometry may play a significant role in the diagnosis of lymphoma.

The effects of sodium arsenite exposure from postnatal day (PND) 1 to PND 14 on Leydig cells of Wistar rat testes were investigated at PND 15 and 21. Gross morphometric observations of testes did not reveal a significant change in the numerical density and volume of the testes in exposed animals compared to controls either at PND 15 or 21. However, there was significant decrease in testicular weight at PND 21 in treated animals. Measurement of nuclear area of Leydig cells revealed a decrease in nuclear area of these cells in exposed groups at PND 15 and 21. A significant decrease in the total number of Leydig cells was apparent at PND 21 in the treated group. The observations of the present study are indicative of adverse effects on rat testes Leydig cells following exposure to low doses of sodium arsenite during critical window periods. The persistence of these observations at PND 21 is suggestive of irreversible damage to testicular tissue.

800x600 Operation of nuclear facilities such as research reactor and it’s supporting installation in Serpong Nuclear Area may release controlled radionuclides to Cisadane River and then it would flow to Jakarta Bay. There are limited marine radioecology studies or radionuclides monitoring at Jakarta Bay. Therefore monitoring of 239/240 Pu and 137 Cs was carried out from Tanjung Pasir to Tanjung Kerawang. The ERICA Assessment Tool was used to evaluate radiological risk in the marine environment of Jakarta Bay area. The 137 Cs concentration in sea water and sea sediments were in range of 0.17 – 1.17 Bq.m -3 and 0.34 – 1.21 Bq.kg -1 , respectively. Moreover, result of 239/240 Pu measurement showed that concentration at sea water and sea sediment were range from < MDA – 0.53 mBq.m -3 and 2.64 – 55.70 mBq.kg -1 respectively. The results of risk analysis were indicated that all the total dose rates per organism were millions time lower than the screening rate (10 mGy.h -1 ). Normal 0 false false false EN-US X-NONE X-NONE MicrosoftInternetExplorer4 /* Style Definitions */ table.MsoNormalTable {mso-style-name:"Table Normal"; mso-tstyle-rowband-size:0; mso-tstyle-colband-size:0; mso-style-noshow:yes; mso-style-priority:99; mso-style-parent:""; mso-padding-alt:0in 5.4pt 0in 5.4pt; mso-para-margin:0in; mso-para-margin-bottom:.0001pt; mso-pagination:widow-orphan; font-size:10.0pt; font-family:"Times New Roman","serif";}

In order to support and complement the radioactive waste management facilities in Indonesia, BATAN will build a demonstration disposal facility in Serpong Nuclear Area (SNA). Demonstration disposal that will be built is Near Surface Disposal (NSD) type. Engineered vault for NSD is reinforced concrete. The calculations for determining the thickness of NSD concrete vault is based on the conceptual design as the result of the placement optimization of demonstration disposal that takes into account the inventory of radioactive waste and environmental geology conditions of the site at Serpong Nuclear Area. The thickness of the vault in this paper is focused on its ability to withstand radiation from stored waste so that workers or people who are around the disposal facility is safe with maximum radiation dose limit rate of 0.3 μSv / h. The calculation is performed with the aid of MicroShield 7:02 and Rad Pro Calculator Version 3:26 software. From the calculation so that the dose rate at the outer surface of the vault to be 0.3 μSv / h, required walls made of concrete with a density of 2:35 g / cm3 is 62.8 cm thickness.

With new numismatic data and the revisión of the old epigraphic and literary texts. The A. insists on the impossibility to identificate the Republican city of Segobrix with Cabeza de Griego (Cuenca), although this last settlement was surely the place of the Imperial Segobriga. The inhabitants of Segobrix were probably moved, after Sertorius time, from the nuclear área of the Meseta Norte, near Clunia, to another city in the Meseta Sur, Segobriga in Cuenca., Con nuevos datos numismáticos y revisión de los epigráficos y literarios, se insiste en la imposibilidad de que la Segobrix republicana estuviera en Cabeza del Griego (Cuenca). Sin embargo, si debió ser éste el emplazamiento de la Segobriga imperial, atestiguado por la presencia en la zona de su moneda augústea y tiberiana. Es posible que tras las guerras sertorianas se trasladase la ciudad desde la Meseta norte a Cabeza del Griego, ocasionando este nueve e importante emplazamiento el final de la ciudad cercana de Contrebia Carbica.

Background/aim Colorectal cancer is the seventh most common cancer in Egypt, constituting 3.47% of male cancers and 3% of female cancers. The aim of the current study is to evaluate the immunohistochemical expression of cytochrome C oxidase (CcO) as well as the nuclear morphometric measurements in colonic adenomas and adenocarcinomas. Materials and methods Sixty cases of colonic biopsies/specimens were involved in this study. The samples were received in the Department of Pathology of Kasr El-Aini Hospital, Cairo University, Egypt, and they were divided into three groups: 20 cases of normal control, 20 cases of colonic adenoma, and 20 cases of colonic adenocarcinoma. The expression of CcO was studied by immunohistochemistry in 60 formalin-fixed paraffin blocks. Nuclear morphometric parameters were evaluated using an image analysis system. The area percentage of the marker expression was correlated with nuclear morphometric parameters. Results Immunohistochemical study showed that 65% of control biopsies showed a significant difference with colonic adenocarcinoma cases in contrast and adenoma cases showed a nonsignificant difference with normal control biopsies. Mean areas percentages of CcO expression were significantly different between all three studied groups. The means of nuclear area, length, and perimeter showed a significant difference while means nuclear width and roundness showed a nonsignificant difference between all three studied groups. There was significant correlation between mean area percentage of CcO expression with mean nuclear area and perimeter in control, adenoma, and adenocarcinoma cases. Conclusion CcO protein is significantly expressed in the cytoplasm of normal colonic mucosa. It was significantly deficient in colonic adenocarcinoma. Nuclear morphometry can be introduced as a useful morphologic feature of patients with variable colonic lesions specially in combination with CcO.

H. Ohsaki, E. Hirakawa, K. Kagawa, M. Nakamura, H. Kiyomoto and R. Haba Value of computer-assisted quantitative nuclear morphometry for differentiation of reactive renal tubular cells from low-grade urothelial carcinoma Objective: To assess whether computer-assisted quantitative morphological parameters can be an effective tool for objectively distinguishing reactive renal tubular cells from low-grade urothelial carcinoma cells (LG-UCs) in voided urine. Methods: Nuclear morphometry was performed by a computer-assisted image analyser system on Papanicolaou-stained cytological specimens. The circumference of reactive renal tubular cells (n = 40) or LG-UC (n = 20) nuclei were manually traced, and the following nuclear morphometric parameters were analysed: (i) area, (ii) perimeter, (iii) roundness factor, (iv) maximum length, and (v) linear factor. For each nuclear measurement, we calculated the maximum, minimum, mean and standard deviation. Results: The mean nuclear area and nuclear perimeter were higher in reactive renal tubular cells compared to the LG-UCs. The mean of roundness and linear factors (reflecting a tendency for the nuclear outline to be regular and oval, respectively) were higher in LG-UCs compared with reactive renal tubular cells. Among nuclear areas, the nuclear perimeter, roundness factors and maximum length did not show any significant differences between reactive renal tubular cells and LG-UCs. On the other hand, the linear factor showed a mean higher value among LG-UCs than reactive renal tubular cells (P = 0.023). Conclusions: Of five quantitative nuclear morphological parameters, only linear factor was statistically significant in differentiating reactive renal tubular cells in renal disease from LG-UCs.

Mesoclemmys tuberculata is a turtle species that is distributed in northeastern Brazil, recorded mainly in the Caatinga and at some localities in the Atlantic Forest and Cerrado. In this paper we report the fi rst species record in an area of Cerrado of Minas Gerais state, and it is the only known state record in a specifi c location. During fi eld ampling, a specimen of M. tuberculata was collected in the municipality of Joao Pinheiro, northwest state, in a Cerrado nuclear area in the Sao Francisco river basin. The locality of this record is the southern and western limits of M. tuberculata's known distribution, as well as the most inland locality of species record in the Cerrado biome.

In the practice of forensic science, sometimes, it is not easy to understand whether skin lesion is due to electrocution and to differentiate the thermal burns and abrasion-type lesions, especially when electricity source cannot be revealed by death science investigation. Based on the causes of the lesions, cases were classified into three groups. Group 1 included 30 deaths from electrocution. Group 2 included 30 deaths with flame burns. Group 3 included 30 deaths from traffic accident cases, which had abrasions. In this study, epidermal nuclear area, perimeter, nuclear form factor, nuclear minimum axes, nuclear maximum axes, and minimum axes/maximum axes ratio were measured. As a result, we think that computerized image analysis beside light microscopic examination can be useful in the differentiation of the electrocution, flame burn, and abrasion type lesions.

This research aimed to study the effects of Myristica fragrans Houtt. (Myristicacae) oil on male reproductive functions in Wistar strain rats, the oil of Myristica fragrans at the dose levels of 0.2, 0.4, and 0.5 mL/kg body weight/day for 60 days. The oil reduced fertility to 0% by 60 days of treatment at the highest dose level. The results also revealed the oil caused significant reduction in the weight of testis, epididymis, seminal vesicle, ventral prostate and vas deferens. Sperm motility as well as sperm density were significantly reduced which resulted in the reduction of male fertility by 100% at the highest dose level tested. Leydig cell nuclear area and mature Leydig cell numbers were significantly reduced when compared with controls. The round spermatids were decreased by 72.71%. The hematological parameters show a non-significant change when compared with the control group. A recovery of 80% fertility was recorded after withdrawal of 120 days, proving that the drug is safe to use. These results...

Nucleolar and nuclear size of bone marrow plasma cells has been studied in 26 cases of myeloma, 19 of benign essential monoclonal gammapathy (BEMG) and 9 without an M-component. Bone marrow smears were Feulgen-stained to visualize nucleoli and nuclei. Nucleolar area, nuclear area and the ratio nucleolar area/nuclear area were calculated. The myeloma group differed from the other two groups in having plasma cells with larger nucleoli, larger nuclei and an increased ratio nucleolar area/nuclear area. No difference was found between the BEMG group and the cases without an M-component. In some cases--in which the absence of osteolytic lesions at X-ray examination, a low plasma cell percentage in bone marrow smears or a low M-component concentration initially made the diagnosis of myeloma uncertain--the observation of enlarged nucleoli could help to establish the diagnosis. Three cases were observed in which the onset of myeloma was preceded by a long period of essentially unchanged M-component concentration. In these cases a sudden increase of M-component concentration was accompanied by an increase of mean nucleolar size of bone marrow plasma cells.

Summary. Chromatin packing, mostly due to disulfide bond formation, is one important step in epididymal sperm maturation. In this study the in vitro effect of a reducing agent such as thioglycolate (Tg) was tested. Thioglycolate was assayed on rat sperm obtained from caput, corpus and cauda epididymides. Changes were verified via light microscopy (and area measurements) and both transmission and scanning electron microscopy using standard techniques. Due to the low molecular weight of Tg, it does not require detergent to enter sperm. The nuclear decondensation elicited by Tg and its effect on sperm surface, differ from caput to cauda, although the final nuclear area is not significantly different. The pattern of sperm nuclear decondensation suggests that it starts at the caudal segment of the head, perhaps due to the abundance of nuclear pores in this region. Under the experimental conditions described below the perforatorium was not affected. Thus, the nature and role in the rat sperm of the perforatorium deserves more detailed analysis.

Stained cytological specimens from 7 canine anal sac gland adenomas and 11 canine anal sac gland carcinomas were analyzed by computer-assisted nuclear morphometry. In each case, the nuclei of at least 100 neoplastic cells were measured, and the mean nuclear area (MNA), mean nuclear perimeter (MNP), mean nuclear diameter (MND) and nuclear roundness (NR) were calculated. The study aimed to evaluate (1) the possibility of using nuclear cytomorphometry as an auxiliary diagnostic method to differentiate between canine anal sac gland adenomas and adenocarcinomas, and (2) the prognostic value of nuclear morphometry in canine anal sac gland adenocarcinomas. The results indicated that (1) MNA, MNP, MND and NR could be used as effective auxiliary tools for differential diagnosis between canine anal sac gland adenomas and adenocarcinomas, and (2) MNA, MNP and MND are reliable prognostic indicators for canine anal sac gland adenocarcinomas.

Twenty-four feline spontaneous basal cell carcinomas (BCCs) were analyzed by computerized nuclear morphometry. The study included 15 non-recurrent and 9 recurrent tumours. In each case, the nuclei of at least 100 neoplastic cells were measured, and the mean nuclear area (MNA), mean nuclear perimeter (MNP) and mean nuclear diameter (MND) were calculated. The analysis of data of the non-recurrent BCCs and the recurrent tumours revealed statistically significant differences between those groups (p < 0.001) as well as between infiltrative and clear types of BCCs (p < 0.05). The results indicate that nuclear morphometry is able to predict recurrent tumour growth and helps to differentiate histological subtypes of BCCs in cats.

Objective This study assessed the effect of smoking crack cocaine on the nuclear area (NA), the cytoplasmic area (CA), and the nucleus-to-cytoplasm area ratio (NA/CA) of oral squamous epithelial cells. Study design Oral smears were collected from clinically normal-appearing mucosa by liquid-based exfoliative cytology of 40 individuals (20 crack users and 20 nonusers) and analyzed for quantitative techniques using an image analysis system. Results Mean values of NA for experimental and control groups were, respectively, 49.85 μm2 and 62.68 μm2 (P < .01). CA showed the following mean values: 1820.9 μm2 (experimental) and 1780.8 μm2 (control). NA/CA for the experimental group was 0.03; the control group was 0.04 (P < .01). Conclusion This study revealed that crack cocaine was able to induce significant changes on the oral epithelial cells. Since this illicit drug is normally used in association with other risk factors for oral cancer (tobacco and alcohol), crack cocaine abusers should have frequent preventive oral exams.

Objective: To evaluate the effect of manufactured cigarettes on the nuclear area (NA), cytoplasmic area (CA), and nuclear/ cytoplasmic ratio (NA/CA) in oral ...

Ionic silver has well-proven bactericidal properties, and silver-containing wound dressings are now widely used to aid in the creation of an antimicrobial environment in wounds. The effect of silver ions on bacterial ultrastructure can best be studied by viewing bacterial cells under a transmission electron microscope (TEM). Bacterial cells of Pseudomonas aeruginosa were incubated within a control dressing (e.g. a non-antimicrobial Hydrofiber dressing) (Hydrofiber is a registered trademark of E.R. Squibb and Sons, L.L.C.) and a silver-containing Hydrofiber dressing, followed by processing for TEM. Liquid cultures, with and without silver, were prepared for comparison. The addition of silver to growing bacterial cultures stopped growth of the cells very quickly. Ultrastructurally, the presence of silver was found to affect both the shape of the bacterial nucleoid and the organization of bacterial DNA. X-ray microanalysis of bacteria from liquid cultures showed the presence of silver within silver-treated cells and the absence of calcium. It is suggested that the presence of available silver ions within the Hydrofiber dressing could lead to the loss of cellular ions, vital for maintaining the structural integrity of the nuclear area.

As a historiographical category, ‘big science’ was elaborated from the point of view of advanced countries. However, some developing countries decided to invest a significant part of their rather modest science budgets in building many-million-dollar facilities. A comparative approach to the study of the first stages of the Argentine TANDAR heavy ion accelerator and the Brazilian National Laboratory Synchrotron Light (LNLS) projects may help understand specificities in patterns of organisation of big science in peripheral contexts. Oversimplification of the decision-making processes linked to authoritarian political contexts—which allowed to overcome the lack of consensus within the physics community as well as financial uncertainties—seem to have been a necessary condition for TANDAR and LNLS, which differentiated them from big science in developed countries. Additionally, the different ways in which the institutionalisation of the nuclear area took place in Argentina and Brazil seem to have been responsible for the different paths followed by experimental physics between the 1960s and 1980s: nuclear physics in Argentina and particle physics in Brazil.

Fifty-two spontaneous canine mammary gland tumours (fibroadenomas (n = 8), tubulopapillary carcinomas (n = 9), solid carcinomas (n = 6), anaplastic carcinomas (n = 7), fibrosarcomas (n = 9), liposarcomas (n = 9) and osteosarcomas (n = 4) were analysed by computer-assisted nuclear morphometry in Hemacolor-stained cytological specimens. Computerized cytomorphometry was performed and the nuclear area, nuclear perimeter and mean nuclear diameter of investigated tumours were assessed. A minimum of 100 nuclei per lesion were examined. The statistical analysis revealed statistically significant differences between benign and malignant neoplasms. The results indicated that computer-assisted nuclear morphometry could be used as an additional method for differentiation of benign from malignant canine mammary gland tumours in cytological specimens.

Mediation was introduced in Russia in 2010 but its implementation is still hampered by controversies over the use of mediation terminology (MT) borrowed from English. This study presents the results of a joint research initiated by a group of linguists and specialists developing legal standards for mediation in Russia to bring into the foreground the study of mediation terminology and to suggest directions for its systematization. The research uses structural and semantic analysis building a model of source language terminology (English) based on the terminological evidence of 2263 units to elaborate standards which could be applied to Russian MT. Besides, the results of the Nuclear-Peripheral analysis (that ensures translation equivalence by preserving nuclear area of semantics in both languages) laid the basis for MT Translation Project that suggested strategies for translating the collected terms into Russian. DOI: 10.5901/mjss.2015.v6n6p302

The aims of the study are to detect whether there are any possible effects of chronic carbon monoxide (CO) exposure on the argyrophilic nucleolar-organizing region (AgNOR)–associated protein synthesis and evaluate any possible relationship between the amount of AgNOR protein and the level of myocardial injury also and between AgNOR and histopathological evaluation methods. Adult male albino Wistar rats ( n = 18) were randomly divided into three groups (groups A, B, and C). Group A served as control, while groups B and C were rats exposed to CO gas chronically (1000 and 3000 ppm CO concentration with a flow rate of 4 L/min for 30 min/day for 7 days, respectively). Total AgNOR area/nuclear area (TAA/NA) and the mean AgNOR numbers for each myocyte nucleus were determined. There were significant differences among all groups for TAA/NA ratio. These differences were not significant for mean AgNOR numbers. According to the histopathological evaluation scores, there were significant differences between the groups. The differences were significant among the groups for loss of sarcomere pattern. A strong positive correlation between histopathological injury scores and TAA/NA ratio was found ( Rsq = 0.48; p = 0.002), however, the correlation was not significant for mean AgNOR numbers ( Rsq = 0.08; p = 0.25). In conclusion, TAA/NA ratio can be used as an indicator for obtaining information about the level of myocardial damage instead of histopathological evaluation scores.

BACKDROUND: Ovarian carcinoma is a leading cause of death in gynecological malignancy. Ovarian surface epithelial serous and mucinous tumours are classified as benign, borderline, and malignant. The identification of borderline tumours most likely to act aggressively remains an important clinical issue.AIM: This work aimed to study DNA ploidy and nuclear area in ovarian serous and mucinous; benign, borderline and malignant tumours.MATERIAL AND METHODS: This study included forty ovarian (23 serous and 17 mucinous) tumours. Paraffin blocks were sectioned; stained with haematoxylin and eosin for histopathologic and morphometric studies and with blue feulgen for DNA analysis.RESULTS: All four serous and six out of nine mucinous benign tumours were diploid. All eight serous and five mucinous malignant tumours were aneuploid. Nine of eleven (81.8%) serous and all three mucinous borderline tumours were aneuploid. There were highly significant differences in mean aneuploid cells percentage between serous benign (1.5%), borderline (45.6%) and malignant (74.5%) (p = 0.0001) and between mucinous benign (13.2%) and both borderline (63.7%) and malignant (68.4%) groups (p = 0.0001).There were significant differences in nuclear area between serous benign (26.191%), borderline (45.619%) and malignant (67.634 %) and a significant positive correlation between mean percentage aneuploid value and mean nuclear area in all serous and mucinous groups.CONCLUSION: We suggest that DNA ploidy and nuclear area combined, may be adjuncts to histopathology; in ovarian serous and mucinous benign, borderline and malignant neoplasms; identifying the aggressive borderline tumours.

An automatic method for estimating the density of cell nuclei in whole slide images (WSI's) of hepatic histological sections is presented. The method can be applied to hematoxylin-eosin (H&E) stained sections with slight histological atypism, such as early well-differentiated hepatocellular carcinoma (ewHCC). It is shown that measured nuclear density is affected by the nuclear size due to fragments of nuclei. This size-dependent problem has been solved by estimating the standard nuclear area for each image patch. The method extracts typical nuclei, which are used to automatically adjust the parameters, including the standard nuclear area. The method is robust for variations in contrast, color, and nuclear size. 40 image patches sampled from 20 WSI's of surgical sections were used for accuracy evaluation. The mean absolute percentage error of estimated nuclear densities was 8.2%. It was also confirmed that the distributions of nuclear density were successfully estimated and visualized for all 20 WSI's. The computation time for a WSI of typical surgical section (754 mm2, about 1,280,000 nuclei) was about 57 minutes on a PC.

The purpose of this study is to evaluate the association between the age and the density of anterior, posterior, and nuclear parts of lens and detect the normal lens density (LD) values in different age groups. 370 eyes of 370 voluntary healthy Turkish subjects were included. These were divided in four groups according to their age. These were Group 1 (51–65 years), Group 2 (41–50 years), Group 3 (31–40 years) and Group 4 (20–30 years). LD values of anterior, nuclear, posterior, and total (average) were measured using Pentacam HR Scheimpflug camera system. Both eyes of each subject were scanned twice in automatic release and 50-picture 3-dimensional (3D) scan modes by one experienced observer. 207 of the participants were females and 163 were males, with a mean age of 40.57 ± 13.06 years. The differences between groups in terms of nuclear area, posterior area, and average LD measurements were significant but anterior area was not. The nuclear area and average LD had high positive correlation with age (r = 0.72, p

Se presentan los resultados de los analisis realizados a micro-restos botanicos (granos de almidon) extraidos de las paredes internas y fondo de 17 queros de madera del extremo norte de Chile. Estos queros representan cinco tipos diferentes de vasijas y estan relacionados con el consumo de chicha durante el periodo Intermedio Tardio y Horizonte Inca. Los granos de almidon fueron separados por gravimetria y observados con montaje directo. Quince de estos queros dieron positivos para la presencia de granos de almidon crudo de maiz (Zea mays), algunos de los cuales muestran signos de fermentacion. Ademas de maiz se encontraron granos de almidon de frijol (Phaseolus vulgaris), pallar (Phaseolus lunatus) y yuca (Manihot esculenta) entre otras especies. Se plantea que los aspectos tecnicos y contextuales en la produccion de chicha de maiz local, no cocida, difieren de lo descrito en las cronicas para este periodo respecto de las areas nucleares del Tawantinsuyo. Esta compleja evidencia encontrada en los queros de madera analizados incluyo el consumo de diversos tipos de vegetales, probablemente procesados e incorporados en la forma de harinas. Se concluye que esto indicaria variantes regionales del protocolo de elaboracion y consumo de chicha, posiblemente asociado a aspectos nutritivos y de ritualidad funebre local. AbstractThis paper presents the results of botanical analyses undertaken on micro-remains (starch grains) collected from the internal wall and base of five different types of queros of a total of 17 vessels. These queros are associated with chicha production during the Late Intermediate and Inca Horizon periods in northern Chile. The starch grains were separated by gravimetry and observed on microscope slides. Fifteen of these queros were positive for starch grains of corn (Zea mays) and some of these also contained evidence of fermented corn. In addition, we found starches of the common bean (Phaseolus vulgaris), lima bean (Phaseolus lunatus) and yucca (Manihot esculenta), among other species. These local findings differ from the contextual and technical aspects of chicha production described by the chroniclers for the nuclear area of Tawantinsuyo. Evidence found in the wooden queros and the different plant starches suggest grinding of these products and perhaps adding them as flour to the chicha. We conclude that this indicates regional variation in the protocols of production and consumption of chicha that were more likely related to dietary intake and local mortuary rites.

Background: Whole slide images (WSIs) involve digitally capturing glass slides for microscopic computer-based viewing and these are amenable to quantitative image analysis. Bile duct (BD) brushing can show morphologic features that are categorized as indeterminate for malignancy. The study aims to evaluate quantitative morphologic features of atypical categories of BD brushing by WSI analysis for the identification of criteria predictive of malignancy. Materials and Methods: Over a 3-year period, BD brush specimens with indeterminate diagnostic categorization (atypical to suspicious) were subjected to WSI analysis. Ten well-visualized groups with morphologic atypical features were selected per case and had the quantitative analysis performed for group area, individual nuclear area, the number of nuclei per group, N: C ratio and nuclear size differential. Results: There were 28 cases identified with 17 atypical and 11 suspicious. The average nuclear area was 63.7 µm2 for atypical and 80.1 µm2 for suspicious (+difference 16.4 µm2; P = 0.002). The nuclear size differential was 69.7 µm2 for atypical and 88.4 µm2 for suspicious (+difference 18.8 µm2; P = 0.009). An average nuclear area >70 µm2 had a 3.2 risk ratio for suspicious categorization. Conclusion: The quantitative criteria findings as measured by image analysis on WSI showed that cases categorized as suspicious had more nuclear size pleomorphism (+18.8 µm2) and larger nuclei (+16.4 µm2) than those categorized as atypical. WSI with morphologic image analysis can demonstrate quantitative statistically significant differences between atypical and suspicious BD brushings and provide objective criteria that support the diagnosis of carcinoma.

Introduction: Carbon monoxide (CO) is a colorless, tasteless, odorless, nonirritant gas and CO poisoning affects all organ systems. Aim: We aimed to detect any possible effects of CO exposure on the argyrophilic nucleolar organizing region (AgNOR)-associated protein synthesis of heart cells and whether there is any relationship between AgNOR protein amount and both carboxyhemoglobin (COHb) level and histopathological evaluation methods used for the detection of damage in heart tissue after CO exposure. Materials and Methods: The rats were divided into four groups (control, 1000, 3000, and 5000 ppm), each containing six rats. After CO intoxication, COHb levels were measured and the animals were killed on the 7th day. AgNOR staining was performed in the heart tissue. One hundred nuclei per rat were evaluated, and total AgNOR area/nuclear area and mean AgNOR number were analyzed for each nucleus. The CO exposure groups had significantly higher AgNOR values than the control group ( p < 0.0001). According to cardiomyopathy (CMY) scoring methods, the differences between groups 3 and 4 and groups 1 and 2 were significant ( p < 0.05). A significant positive correlation between AgNOR values and both CMY and COHb levels were detected. Conclusion: The detection of AgNOR protein amount may give information about the CMY levels and be used to detect the CO intoxication levels instead of COHb in later periods.

A short overview of Laser Induced Breakdown Spectroscopy (LIBS) with emphasis on the new trends is presented. Nowadays, due to unique features of this technique, LIBS has found applications in a great variety of fields. Achievements in the application of LIBS in nuclear area, for hazardous materials detection and in geology were considered. Also, some results recently obtained at VINCA Institute, with LIBS system based on transversely excited atmospheric (TEA) CO2 laser, are presented. Future investigations of LIBS will be oriented toward further improvement of the analytical performance of this technique, as well as on finding new application fields. Proceedings of SPIE, International Conference on Atomic and Molecular Pulsed Lasers XII (AMPL), Sep 13-18, 2015, Tomsk, Russia

Introduction Though we are in 21(st) century with nano technology & tissue printing, there still exist many lacunae in the field of diagnosis. Not much is known about prognostic markers till now from literature to assess potentially malignant lesions. Lesions so called potentially malignant can be termed only after clinical & malignant changes have been developed and there are no means of predicting with certainty the risk of cancerous transformation. Aim Our present study was undertaken to establish the morphometric parameters of the parabasal and spinous cells of normal oral epithelium with the changes occurring in cells of Oral Leukoplakia (OL), Oral Verrucous Carcinoma (OVC) and Oral Squamous Cell Carcinoma (OSCC). Materials and methods Total 40 patients were divided into Group I which includes patients with normal oral mucosa, group II oral leukoplakia patients, group III oral verrucous carcinoma patients and group IV includes oral squamous cell carcinoma patients. Tissue sections were taken and morphometric analysis of cell area, cell diameter, nuclear area, nuclear diameter, nuclear cytoplasmic ratio was done for parabasal and spinous layer cells. Statistical analysis was done using one-way ANOVA and T-test. Results Nuclear diameter, nuclear area, cell area, nuclear cytoplasmic ratio were significantly increased in OL, OVC, OSCC patients than normal oral mucosa, which was statistically significant. Cell diameter was decreased in OL, OVC, OSCC patients than with normal oral mucosa which was statistically significant. Conclusion Cellular & nuclear parameters showed statistically significant changes in oral leukoplakia, oral verrucous carcinoma & oral squamous cell carcinoma in comparison with normal oral mucosa.

Introduction Mean 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 methods This 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. Results The age of the patients varied from 35 to 87 years. Male: female ratio was 3.5:1. The mean MNA–10 (µm 2 ) for High Grade Malignant Potential was 104.52 ±25.64 µm 2 , 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. Conclusions MNA–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 interob server variability and to increase reproducibility of grading.

The aims of this study were to use computerized morphometry in order to differentiate between the degree of dysplasia and to predict progression to invasive adenocarcinoma in Barrett's esophagus (BE). Biopsies from 97 patients with BE graded by a consensus forum of expert gastrointestinal pathologists were available for morphometrical analysis. The study group included 36 biopsies negative for dysplasia (ND), none of which progressed to carcinoma; 16 indefinite for dysplasia (IND) and 21 low-grade dysplasia (LGD), of which three progressed in each group and 24 high-grade dysplasia (HGD), of which 15 progressed to invasive carcinoma. Computerized morphometry was used for measuring indices of size, shape, texture, symmetry and architectural distribution of the epithelial nuclei. Low-grade dysplasia was best differentiated from the ND group by nuclear pseudostratification (P=0.036), pleomorphism (P

We sought to identify morphometric descriptors predictive of nondiploidy in prostatic adenocarcinoma on prostate needle biopsies using logistic regression (LR) and binary recursive partitioning (BRP) and compare the equivalence of both methods. A total of 180 prostate needle biopsies diagnosed as prostatic adenocarcinoma were selected. Deoxyribonucleic acid ploidy and morphometry were performed separately on Feulgen-stained sections from these biopsies using the CAS-200 system and Nuclear Morphometry Suite, respectively. Seven morphometric predictors were tested as predictor variables, including nuclear area, circularity, elongation, sum optical density (OD), configuration run length, coefficient of variation (CV), and angularity. Logistic regression (LR) identified a two-parameter model including sum OD and circularity that had a 93.9% overall correct prediction rate (area under curve = 0.950; 95% CI: 0.913, 0.987). A reduced model including only sum OD was equally good without any significant loss of predictive accuracy (93.3% correct overall classification rate). BRP also selected sum OD as the most predictive parameter; a sum OD cut-off of 7.73 in this model identified 93.3% of the nondiploid cases correctly. Morphometric OD can be used as a surrogate marker of nondiploidy. LR and BRP models are both equivalent in identifying and correctly classifying nondiploid cases of prostate cancer using sum OD as the predictor variable.

Forty-five patients with papillary or follicular thyroid carcinomas with tracheal infiltration were examined by clinical, histologic, and morphometric analyses. Twenty-four patients with completely encapsulated carcinomas (encapsulated cases) and 26 patients with carcinomas confining within the thyroid capsule (nonencapsulated cases) were examined as controls. Patients with tracheal infiltration were significantly more often male and older than the patients without (P less than 0.01 and P less than 0.05, respectively). The histologic grade of differentiation did not correlate with tracheal infiltration. The nuclear area of tumor cells was significantly larger in the cases with tracheal infiltration than in the cases without (P less than 0.01). In 12 recurrent cases with tracheal infiltration, the nuclear area of recurrent tumors was significantly larger than those of their own primary tumors (P less than 0.01). These results confirm that thyroid carcinomas with tracheal infiltration were more frequent in male and older patients and had more significant nuclear atypia than the tumors without tracheal infiltration.

Summary Nucleolar organizer regions (NORs) were evaluated in 11 women with adenomyosis. The material, sampled during operations, was silver-stained using the technique of Ploton et al. and verified histopathologically. A hundred cell nuclei per slide were assessed. The examined variables comprised the number of argyrophylic nucleolar organizer regions (AgNORs) in the nucleus, the surface area of a single AgNOR and the position of the AgNORs in the cell nucleus, as well as the AgNORs’ coefficient, which were all estimated under a microscope. The parameters were quantitatively assessed using computer image analysis software Multi-Scan Base V.8. Adenomyosis was found to be associated with a significant decrease (up to 60%) in the nuclear area, as well as with a marked reduction (up to 50% as compared with normal endometrium) in the number of AgNOR granules. The total area of AgNORs in the cell nucleus also decreased from 3.55 to 1.57 μm2. There were no significant differences in the number of granules per nucleus either in the control group or in the adenomyosis group. The AgNOR coefficient was found to be lower in adenomyosis compared to normal endometrium.

Background: Nuclear texture analysis is a useful method to obtain quantitative information for use in prognosis of cancer. The first-order gray level statistics of a digitized light microscopic nuclear image may be influenced by variations in the image input conditions. Therefore, we have previously standardized the nuclear gray level mean value and standard deviation. However, there is a clear relation between nuclear DNA content, area, first-order statistics, and texture. For nuclei with approximately the same DNA content, the mean gray level increases with an increasing nuclear area. The aims of the present methodical work were to study: (1) whether the prognostic value of adaptive textural features varies with nuclear area, and (2) the effect of standardizing nuclear first-order statistics. Methods: Nuclei from 134 cases of ovarian cancer were grouped into intervals according to nuclear area. Adaptive features were extracted from two different image sets, i.e., standardized and non-standardized nuclear images. Results: The prognostic value of adaptive textural features varied strongly with nuclear area. A standardization of the first-order statistics significantly reduced this prognostic information. Several single features discriminated the two classes of cancer with a correct classification rate of 70%. Conclusion: Nuclei having an area between 2000–4999 pixels contained most of the class distance information between the good and poor prognosis classes of cancer. By considering the relation between nuclear area and texture, we avoided a loss of information caused by standardizing the first-order statistics and mixing data from cells having different nuclear area.

The present study was undertaken to compare morphometric analysis of cells in 100 effusion samples, with unaided cytological diagnosis. All the cases in the form of Leishman's stained smears were subjected to computerized-interactive morphometry (CIM) by usage of Image analysis software. On routine cytological examination, 44 cases were benign, 26 were malignant, and 30 cases were placed in "atypical" category. On morphometry, values for mean nuclear area (MNA) and mean cytoplasmic area (MCA) in benign cases were found to be 58.39 +/- 12.22 microm(2) and 185.70 +/- 43.69 microm(2), while for malignant cases were 120.12 +/- 16.28 microm(2) and 274.65 +/- 61.01 microm(2), respectively (P < 0.001, P < 0.001, highly significant). Significant differences were also noted among benign, atypical, and malignant cases for MNA (P < 0.001). The differences between atypical and malignant cases for N/C (nuclear/cytoplasmic) ratio were also found to be statistically significant (P < 0.001). Finally, 46 cases were found to be benign, 39 malignant, and 15 in the category of atypical cases. CIM helps in better and quicker sorting for malignant cells. MNA and N/C ratio are the most significant parameters in differentiating various cells in effusions. MCA constitutes as another useful parameter. Salient features of CIM, including their applications in the light of current study, are discussed.

A dualistic model for ovarian serous carcinogenesis based on morphological and molecular genetic studies has recently been proposed. This model divides serous carcinoma into low- and high-grade tumors, which develop along distinct molecular pathways. In this report, we evaluated computerized morphometry to determine its utility in distinguishing low- and high-grade serous carcinoma. The mean nuclear area (MNA) and the volume percentage of epithelium (VPE) in 93 high-grade serous carcinomas was measured and compared with 16 low-grade serous carcinomas and 21 serous borderline tumors, the putative precursor of low-grade serous carcinoma. We found that both MNA and VPE were significantly higher in high-grade serous carcinoma compared with low-grade serous carcinoma and serous borderline tumors (P.001 and P = .02, respectively). There was no significant difference in MNA and VPE between low-grade carcinoma and serous borderline tumors (P.3). Among high-grade serous carcinomas, those with an MNA of 46 microm2 or higher had a poorer survival (P = .035) than those with an MNA below 46 microm2. In contrast, VPE and tumor grade (moderately versus poorly differentiated) had no significant prognostic value. The morphometry findings lend further support to the dualistic model of ovarian serous carcinogenesis and suggest that MNA is an excellent adjunctive tool for distinguishing low- from high-grade serous carcinomas. In addition, MNA is an independent prognostic factor for high-grade serous carcinoma.

It is reported that nanoparticles can be applied as carriers and anti-cancer medicines. But the interaction of nanoparticles and cells is unclear. The purpose of this study was to discuss whether inorganic crystal nanoparticles can get through cells with intact crystal. BEL7402 hepatoma cells and titanium dioxide (TiO2) nanoparticles were selected and incubated together in vitro. All specimens were prepared and observed under a transmission electron microscope (TEM). TiO2 nanoparticles were found not in the nuclear area but in the cytoplasma. TiO2 nanoparticles maintained the plate-like shape during absorbing. The result shows that hepatoma cells can endocytose the intact TiO2 crystal nanoparticles. It implies that novel nano-effect plays an important role in the biomedicinal application of inorganic crystal nanoparticles.

Objective: In 1986, the British Society for Clinical Cytology (BSCC) published quantitative criteria to assist diagnosis in a three-tier grading system of squamous cell dyskaryosis. In dyskaryotic cells, area nuclear to cytoplasmic (NC) ratios below 50%, between 50% and 66% and over 66% were defined as equating with mild, moderate and severe grades respectively. Following the Terminology Conference in 2002, however, the BSCC recommended on their website that the three-tier model should be replaced by a new two-tier system of low- and high-grade squamous abnormalities. The latter broadly equate with the two-grade Bethesda System (TBS) for reporting squamous intraepithelial lesions. The purpose of this study was to assess the accuracy and reproducibility of the BSCC three-tier quantitative definitions, to investigate if they were applicable to liquid-based cytology (LBC) and to see how they related to the proposed new two-tier BSCC system. Methods: Quantitative image analysis was undertaken on illustrations from the 1986 BSCC terminology publication and on microscope slides from external quality assessment and Cytology Training Centre teaching sets. Results: Analysis of mean NC ratios showed that mild, moderate and severe dyskaryosis exist as statistically different populations. Overlap of NC ratio ranges, however, limits their practical application in the three-tier model, although interestingly no overlap was noted between mild and severe dyskaryosis. No grade of dyskaryosis had a mean area NC ratio over 50%, indicating that the BSCC quantitative definitions are incorrect. The mean diameter NC ratios for mild, moderate and severe dyskaryosis were found to be 40%, 49% and 66% respectively. Accordingly it is possible that those reporting cervical cytology could be interpreting the BSCC NC ratios as meaning diameter rather than area. Amalgamation of the three-tier results into the proposed two-tier model shows that the resulting mean NC area and diameter ratios identify statistically different low- and high-grade populations. The reduced degree of overlap, however, of NC ratio ranges in the two-tier model implies that NC ratios could have a useful practical role in the separation of the low- and high-grade categories. The two categories were reasonably well separated by mean area and diameter NC ratios of 25% and 50% respectively. A two-tier model combining mild with moderate rather than severe dyskaryosis was found to be a statistically valid alternative but gave rise to NC ratios that would be difficult to use in practice. Except for moderate dyskaryosis, no significant differences were identified between the mean NC ratios of either conventional and LBC preparations or LBC preparations using two different commercial methodologies (SurePathTM and ThinPrep®). Differences, however, were noted in area measurements between SurePathTM and ThinPrep® and this has potential implications for classifications (such as TBS) using area comparisons as their basis. In addition, it was found that the increased NC ratio, associated with higher grades of dyskaryosis is more a consequence of progressive cytoplasmic area reduction rather than nuclear area increase. The similar NC ratios of borderline nuclear changes associated with human papilloma virus and mild dyskaryosis support the BSCC proposal that these can be combined to constitute a low-grade category. This study shows that the BSCC area NC ratio criteria of grading squamous cell dyskaryosis require amendment. In addition, this study supports the new BSCC recommendation of low- and high-grade squamous cell categories. Conclusions: The study proposes Sheffield quantitative criteria to assist the grading of squamous cell abnormalities. Quantitative diameter NC ratio measurements, however, must always be accompanied by detailed assessment of qualitative morphological features and in particular those relating to nuclear chromatin. This is equally relevant to both two- and three-tier models.

Objective: This study assesses the accuracy of published quantitative and qualitative criteria in the Bethesda System (TBS) for squamous intra-epithelial lesions. Methods: Quantitative image analysis was undertaken on illustrations from TBS publications and also from slides in Cytology Training Centre teaching sets. Comparisons were also made with the British Society for Clinical Cytology (BSCC) terminology in cervical cytology, using the illustrations in their terminology publication and amalgamating the results into their proposed new two-tier model. Results: TBS quantitatively defines low-grade squamous intra-epithelial lesions (LSIL) in both conventional and liquid-based cytology (LBC) preparations as showing nuclear enlargement more than ×3 the area of a normal intermediate squamous cell nucleus. This study found that the increase in mean nuclear area was limited to only ×2 in conventional preparations. In LBC (SurePathTM) preparations, there was only a statistically non-significant ×1.2 increase. This study identified a progressive and statistically significant reduction in mean cytoplasmic area from normal intermediate cells to LSIL and then to high-grade squamous intra-epithelial lesions (HSIL) in both conventional and LBC preparations. Furthermore, the most consistent quantitative finding in both conventional and LBC preparations was a statistically significant increase in the mean area and diameter ratios from normal intermediate cells to LSIL and then to HSIL. In all instances this varied from ×2 to just below ×3. This is in agreement with TBS, which states that the cytoplasmic area in HSIL is decreased leading to a marked increase in nuclear to cytoplasmic (NC) ratio. With the exception of an increase in mean nuclear area in conventional preparations from normal intermediate cells to LSIL, the predominant cause for this increase in NC ratios was a reduction in mean cytoplasmic area. The numerical increase in NC ratio for LSIL identified in this study was greater than implied by the ‘slightly increased’ statement in TBS. TBS comments that some HSIL cells can have the same degree of nuclear enlargement as in LSIL and that other HSIL cells may have much smaller nuclei than in LSIL. Both of these qualitative comments were supported in this study. The mean diameter NC ratios of 33% and 50% could provide useful diagnostic assistance in the distinction of normal intermediate cells and LSIL and between LSIL and HSIL, respectively. Because of overlapping individual ranges, however, additional diagnostic features such as nuclear morphology must be used in the distinction of normal intermediate cells, LSIL and HSIL. No statistical difference was identified in the mean diameter NC ratios between ASC-US and LSIL in TBS publications. In addition, the proposed new BSCC low and high grades of squamous abnormality were not statistically different from ASC-US/LSIL and HSIL, respectively. This provides support that the proposed BSCC two-tier system of squamous abnormalities is comparable to TBS. This study shows that LBC has variable but major and significant effects on nuclear and cytoplasmic morphology and that quantitative definitions in conventional preparations cannot be automatically extrapolated to LBC methodology. Conclusions: The study shows that some TBS quantitative and qualitative criteria require amendment and that an alternative quantitative approach, such as diameter NC ratio has a more valid scientific evidence base. Furthermore, use of NC ratios avoids the problems associated with the variable changes in nuclear and cytoplasmic areas, occurring between conventional and different commercial LBC preparations. By contrast, classifications based on area comparisons must be tailored to the specific conventional or commercial LBC preparation.

Background Cytologic discrimination of cellular nodules, follicular adenoma, and follicular carcinoma in the thyroid is problematic. Methods are needed to achieve a reliable diagnosis. Some sophisticated tools, such as microarrays, offer great potential but lack accompanying morphologic information. Methods One hundred twelve samples obtained from patients with lesions histopathologically diagnosed as nodular goiter, follicular adenoma, follicular carcinoma, and papillary carcinoma were used. Eight geometric features, such as nuclear area and circular form factor, were measured. The dataset was divided into six overlapping groups to represent the frequently encountered situations in routine practice. Multivariate analysis of variance, Tukey's honestly significant differences test, and discriminant analysis were performed. Statistical analysis was carried out with two conceptually different approaches. In the first, data from all measured nuclei were used. In the second, a subset of data representing the most extreme values of variables was extracted from the entire dataset to simulate the “selection procedure” performed during conventional morphologic examination. Results When the selected dataset instead of data from all measured nuclei was used, the correct classification rates in discriminant analysis improved considerably. Conclusions Morphologic examination is based primarily on selection. Using data obtained from all of the cells in morphometry may cause a dilution effect in diagnostically important features. Morphometric studies may also be planned with a proper selection “bias.” This may be particularly helpful when isolated abnormal cells carry most of the diagnostic information. © 2005 Wiley-Liss, Inc.

Objective: The aim of this study was to evaluate the ef- fects of aging on semen quality in a population of infertile couples. Methods: A cross-sectional study of semen samples ob- tained from 1,500 men randomly selected from couples who attended an infertility clinic was conducted. The anal- yses were performed using Spearman's correlation and Mann-Whitney tests. The age groups consisted of men ≤ 35 years, from 36-45 years and > 45 years of age. The semen analysis was performed according to the WHO criteria, and morphology was evaluated using the motile sperm organelle morphology examination (MSOME). The percentages of normal spermatozoa and spermatozoa with large nuclear vacuoles (LNV, occupying > 50% nuclear area) were determined. The percentages of DNA fragmen- tation were assessed using the TUNEL assay. Results: A regression analysis revealed that the per- centages of LNV spermatozoa and sperm DNA frag- mentation positively correlated with age. Conversely, a regression analysis revealed that the percentage of nor- mal sperm, sperm progressive motility and sperm vi- tality negatively correlated with age. As in the previous test, the analysis by age group showed that there was a significant reduction (P < 0.05) in the percentage of normal sperm, sperm progressive motility and sperm vitality as age increased. Conversely, the percentage of spermatozoa with LNVs and sperm DNA fragmenta- tion significantly increased (P < 0.05) as age increased. Conclusion: Semen quality seems to be influenced by ag- ing. The age-related decrease in sperm quality suggests that delaying childbearing, not only for women but also for men, may jeopardize reproductive capacity.

The entropies of nuclear arrangements as an indicator of pleomorphism are assessed using a morphometric method. Sixty astrocytic tumors (grades I, II, III and IV; 15 cases each) were reviewed and analyzed. All slides were stained with HE and MIB-1 antibody. The MIB-1 labeling index (LI) was assessed by counting nuclei under a microscope. Images of HE-stained slides were digitized and segmented using the watershed algorithm. Then, six nuclear parameters were measured automatically: (i) the number of total nuclei in the image, (ii) percentage of total nuclear area in the image, (iii) the mean area of the nucleus, (iv) the standard deviation of the area of the nucleus, (v) the entropy of nuclear arrangement (Entropysimple), and (vi) conditional entropy of nuclear arrangement (Entropyconditional). Entropysimple was defined according to the area of the nucleus and Entropyconditional was defined according to both the area of the nucleus and the area of its neighboring nuclei. Image processing and image analysis were performed with public domain software developed in the laboratory. Segmentation of the images resulted in inappropriate segmentation in a few percent of the images. The measurements obtained for each parameter were classified using discriminant analysis. The percentage of correct classification with Entropyconditional was 62%, which was the highest value among all the measurements. Classification based on the combination of all measurements resulted in a rate of correct classification of 88%. Thus, conditional entropy of nuclear arrangement is useful for grading of astrocytic tumors and it is proposed as an indicator of pleomorphism.