Cytomorphometric study of epithelial cells in normal and cataractous human lenses in relation with hyperglycemia.

The aim of the study is to evaluate and correlate the morphology and cell density of epithelial cells adhering to lens capsule surgically removed from the anterior central region with lens clarity and type of cataract present in patients with or without type 2 diabetes. Capsulorhexis specimens were obtained from patients who had undergone phacoemulsification cataract surgery. All the samples were centrifuged and stained by the aid of Papanicolaou technique and were observed under light microscope. We determinated the mean cell density, the degree of epithelial damage, and morphological indicators of cells such as cell area and the nucleus-plasma ratio. Patients with cataract demonstrated a statistical significant decrease in cell density and an heterogeneous cell picture in which enlarged cells dominated. In addition, type 2 diabetics with cataract had a significantly even lower mean epithelial cell density by the presence of larger cell area with smaller nucleus-plasma ratio. More pronounced alterations in the lens epithelium were correlated not only with the presence of cortical cataract, increased fasting blood sugar, and increased HbA1c but also with the prolonged duration of diabetes and the co-existence of diabetic retinopathy. It seems that density and morphology of the anterior lens epithelial cells determine the lens epithelium damage which is more profound in hyperglycemia and in cortical cataracts. The changes in lens epithelium seem to play an important role in cataractogenesis.

Evaluation of ER, PR, MIB-1, pS2, and nuclear grade in FNA specimens of cT1 breast carcinomas: clinicopathological correlation.

Objectives were to determine oestrogen (ER), progesterone receptors (PR), and antigen related to ER (pS2) and to characterize their relationship with the cellular proliferation marker MIB-1 and the nuclear grade (NG) of the cancer cells, using fine-needle aspirates (FNA), as well as the evaluation of their clinical usefulness. The expression of ER, PR, pS2, and MIB-1 was preoperatively detected by immunocytochemistry in FNAs of 70 patients with breast adenocarcinoma and clinical tumor size up to 2 cm. The NG of the tumor cells was also assessed in these samples. We analyzed whether there was any correlation between these biocytologic markers and the invasion of ipsillateral axillary lymph nodes (LN), which were histologically identified after standard surgical treatment in each case. Of the 70 patients 50, 42.85, 50, and 41.42% were positive for ER, PR, pS2, and MIB-1, respectively. Only NG alone was strongly related to the invasion of the LN (P < 0.001). All the patients with NG1 (100%) tumors presented free LN, whereas the majority of those with NG3 (72.72%) had invaded LN (P < 0.001). Patients (14.28%) with NG1 expressed MIB-1, 85.71% ER or PR, and 71.42% pS2. Among the MIB-1-positive tumors a high proportion of NG3 (65.51%) was observed. This finding underlined a relationship between MIB-1 and NG (P < 0.05), identifying an aggressive cancer type. Remarkably 93.33% of the patients with positive MIB-1 and invaded axilla had NG3, whereas 66.66% of them expressed ER or PR and 40% pS2. The findings of the present prospective, multivariate study indicate that NG of the tumor cells, obtained from the preoperative FNAs of breast cancer patients, is a strong predictive marker for the axillary status and in parallel with MIB-1 expression can with sufficient accuracy be of clinical utility. ER, PR, or pS2 on the other hand did not show any relation to the LN status and were not dependent to NG or MIB-1.

Expression of DNA mismatch repair gene MSH2 in cytological material from lung cancer patients.

Mismatch repair genes encode for proteins responsible for the correction of bases incorrectly paired in the DNA. Loss of DNA mismatch repair activity has been associated with various cancers including tumors of the lung. In the present study, we have analyzed by immunocytochemistry the expression of MSH2 DNA repair gene in cytological material obtained by fine needle aspiration from a panel of 42 primary lung cancer patients. Specimens included 13 adenocarcinomas, 11 small cell carcinomas and 18 squamous cell carcinomas. Loss of expression or low expression was detected in 6 out of 13 (46%) adenocarcinomas and in 7 out of 18 (39%) of squamous cell carcinomas, although all 11 small cell carcinomas expressed MSH2. Our results suggest that loss of MSH2 expression is frequent in nonsmall cell carcinomas of the lung (P < 0.01, chi2 test). Evaluation of MSH2 expression can be applied for the screening of cytological material from fine needle aspirations from the lung.

Immunocytochemical panel for distinguishing between carcinoma and reactive mesothelial cells in body cavity fluids.

The morphological evaluation of cytological specimens from body cavity fluids presents difficulties in the differential diagnosis between benign reactive mesothelial (RM) cells and adenocarcinoma (AC) or malignant mesothelioma (MM). The aim of our study was to investigate whether a panel of five different antibodies can offer reliable markers in the differential diagnosis of RM, AC, and MM in serous effusions. A total of 134 cytological specimens of serous effusions from 80 ACs, 50 RMs, and 4 MMs, previously stained with Papanicolaou stain, were selected retrospectively from our files and stained with anti-human mesothelial cell (HBME-1), calretinin, epithelial specific antigen (MOC-31), Ber-EP4, and BG8. Statistical significance was found with HBME-1, calretinin, MOC-31, anti-human epithelial antigen (Ber-EP4), and blood group related antigen (BG8) when comparing AC vs. any type of mesothelial proliferation (MM or RM). The sensitivity of HBME-1 and calretinin for mesothelial cells was 98 and 100%, respectively, and the specificity was 71 and 80%, respectively. Both antibodies stained reactive mesothelial as well as MM cells, with calretinin showing a stronger intensity of immunostaining. The sensitivity of the stain for AC was 86.25% for MOC-31, 77.5% for Ber-EP4, and 67.5% for BG8, and, when combined, the sensitivity was 100%. Our data suggest that immunocytochemical studies performed on Papanicolaou-stained cytological smears with HBME-1, calretinin, MOC-31, Ber-EP4, and BG8 proved to be useful in the differentiation between metastatic AC and mesothelial proliferation. Probably, calretinin is a more preferred marker for mesothelial cells as evidenced by a more intense staining reaction.

Expression of growth hormone-releasing hormone in human primary endometrial carcinomas.

BACKGROUND: Hypothalamic GH-releasing hormone (GHRH) regulates GH release from the pituitary, but an ectopic production of GHRH has been detected in various non-hypothalamic tissues, especially cancers. OBJECTIVE: To investigate whether endometrial tumors produce GHRH. METHODS: Twenty-four endometrioid, three serous papillary (SP), three mixed type endometrioid/serous papillary adenocarcinomas and one malignant mixed Müllerian tumor (MMMT) were assessed for GHRH immunoreactivity by the polyclonal anti-rabbit antibody SV95 and for the expression of GHRH mRNA by in situ hybridization using an oligonucleotide probe. RESULTS: Increased GHRH immunoreactivity was detected in 15 out of 24 (63%) of the endometrioid tumors, including two out of three (66%) of the mixed type endometrioid/serous adenocarcinomas but not in the three SP or the MMMT tumor. Cytoplasmic staining was detected in all positive cases, while in three of them strong nuclear localization of GHRH was also revealed. In situ hybridization indicated the presence of GHRH mRNA in six cases, all characterized as positive for GHRH immunoreactivity. CONCLUSION: GHRH is expressed in a subset of endometrial tumors, of the endometrioid type in particular. A paracrine/autocrine role for GHRH in the development of the disease should be considered.

A comparative study of the int-2 gene product in primary and secondary parathyroid lesions.

OBJECTIVE: The family of fibroblast growth factors stimulates proliferation of cells of mesenchymal, epithelial and neuroectodermal origin. One of the members of this family, the product of proto-oncogene int-2, fibroblast growth factor-3, has been found to stimulate mitosis of parathyroid cells in culture. Primary and secondary hyperparathyroidism have no clear differences with regard to the histopathological features of the diseased parathyroid glands. DESIGN: This study was undertaken in order to determine whether int-2 protein is immunohistochemically expressed in normal and abnormal parathyroid glands and to investigate whether there is a differential expression of the int-2 gene product between primary and secondary parathyroid disease. METHODS: A sheep anti-human int-2 antibody was applied to tissue sections from 37 samples of primary parathyroid disease (12 sporadic adenomas, 25 hyperplastic glands), from 30 samples of renal hyperparathyroidism, and from seven normal controls. Int-2 immunostaining was evaluated semi-quantitatively. RESULTS: None of the normal parathyroid glands stained positively. Int-2 immunopositive expression was more frequently detected in specimens of uraemic patients than in those of patients with primary parathyroid growth processes (P=0.029). The reason for this differential expression appears to be the higher proportion of oxyphilic cells growing in hyperplastic glands of patients with secondary hyperparathyroidism; the latter cells were specifically found to be int-2 immunoreactive. CONCLUSION: The int-2 gene product is likely to participate in the proliferation of this parathyroid cell subpopulation.

Immunohistochemical detection of GHRH and its receptor splice variant 1 in primary human breast cancers.

OBJECTIVE: GHRH is secreted by the hypothalamus and, upon binding to specific GHRH receptors in the pituitary, stimulates growth hormone (GH) production and release from the pituitary. In addition to this neuroendocrine action, accumulated evidence implies additional roles for GHRH in carcinogenesis in non-pituitary tissues. In vitro and in vivo studies have shown that splice variant 1 (SV1) of the GHRH receptor, which is widely expressed in non-pituitary tissues and cancers, can mediate the proliferative effects of GHRH. The aim of the present study was to investigate the operation of an autocrine stimulatory loop between GHRH and SV1 in primary breast tumors. DESIGN: Fifty-three primary breast tumors were evaluated for GHRH and SV1 expression. METHODS: Expression of GHRH and SV1 was assessed by immunohistochemistry using anti-GHRH SV95 and anti-SV1 2317/5 polyclonal antibodies. RESULTS: About 40% of the specimens tested express GHRH and/or SV1 (approx. 25% each), while in 35% of these positive specimens co-expression of these antigens was detected (P<0.01). Furthermore, a correlation of GHRH, but not SV1, expression was detected in lobular compared with ductal carcinomas. CONCLUSIONS: These results constitute the first demonstration for the expression of GHRH and SV1 in primary breast cancers, and provide evidence for the operation of an autocrine stimulatory loop between GHRH and SV1 in primary cancers. Our findings indicate that GHRH analogs could have diagnostic and therapeutic applications for the management of breast cancer.

50 years after the death of George Nicholas Papanicolaou (1883-1962): evaluation of his scientific work.

The purpose of this review article is to summarise the scientific work of George Nicholas Papanicolaou, one of the most eminent figures in the 20th century history of clinical cytology and medicine. Fifty years after his death, his work still remains invaluable, from the early steps in biology and zoology to the application of the Pap test as the most important advancement in the prevention of cervical cancer. The publication of his Atlas was the first important step for the foundation of a new branch in medicine, that of exfoliative cytology. His contribution to cytology undoubtedly earned him the title of the "father of exfoliative cytology" and saved the lives of many women worldwide.

Fine-needle aspiration (FNA) biopsy: historical aspects.

This study aims to present the origins and the historical evolution of fine-needle aspiration biopsy and to also underline its importance in the history of modern cytology. The article focuses on the advances made in the 20th century that have led to the modern techniques associated with the procedure. The authors conducted a thorough review of early reports on needle biopsy, particularly those published during 19th and 20th century, examining in brief also the origins of the needle biopsy. The first report on the use of needle puncture is referred in early writings of Arab medicine. In the early 20th century, Martin and Ellis are considered to be the founders of modern needle aspiration techniques. The German doctor Mannheim was the first to publish reports suggesting the use of fine needles with a small gauge. The establishment and world-wide expansion of FNA should be attributed to the representatives of the Swedish School of Cytopathology. The school embraced FNA in the second half of the 20th century while serving as a training ground for doctors around the world. The history of needle biopsy spans ten centuries. However, the development and establishment of the technique in its modern form took place primarily during the twentieth century. Today, FNA is considered an important cytologic technique with sufficient diagnostic accuracy, especially when applied in cases of lung and prostate cancer.

Survivin expression as a strong indicator of recurrence in urothelial bladder cancer. Predictive value of nuclear versus cytoplasmic staining.

BACKGROUND: Since its discovery in 1997, survivin has garnered significant interest due to its putative role as an inhibitor of apoptosis. Few studies have investigated the immunohistochemical status of survivin in urothelial bladder carcinoma. The subcellular localization of survivin (nuclear, cytoplasmic) and its differential predictive value is a parameter that previous studies have almost ignored. The aim of this study was to investigate the expression pattern of survivin in order to determine its potential prognostic significance. MATERIALS AND METHODS: Archival tumor tissue from 80 patients with urothelial carcinoma were analysed by immunohistochemistry. RESULTS: Nuclear and cytoplasmic positive scores of 61.25% (49/80) and 22.5% (18/80), respectively were found. Nuclear positive staining correlated strongly with increased grade (p=0.001), stage (p=0.039) and the probability of tumor recurrence (p=0.029). No relationship was found between the cytoplasmic survivin level and the clinicopathological parameters. Nuclear expression was identified as a significant independent predictor of relapse-free survival (p=0.016). CONCLUSION: Nuclear expression of survivin reflects an adverse disease outcome.

Prognostic value of computer-assisted morphological and morphometrical analysis for detecting the recurrence tendency of basal cell carcinoma.

BACKGROUND: Nuclear morphometry may provide useful diagnostic and prognostic information about basal cell carcinomas (BCCs) of the skin. MATERIAL/METHODS: A morphometric analysis was performed on histological sections of 52 primary BCCs which recurred and of 52 cases of BCC which did not recur. Eighteen different morphometric parameters were considered, e.g. nuclear area, perimeter, elongation, convexivity, and gray level of the nucleus. The demographics of these patients and the histological-morphological characteristics of their tumors were also considered. Statistical analysis was performed to evaluate the prognostic and predictive value of the morphometric variables for the recurrence of BCCs. RESULTS: Increased patient age, multiple localization of BCCs at the time of diagnosis, and a low degree of peripheral palisading at the histological sections of BCCs were associated with BCC recurrence and consequently worse disease-free survival (DFS). 'Darker' values of the maximum nuclear gray level as well as greater variance of nuclear gray level values also strongly related to BCC recurrence and worse DFS. The analysis of the morphometry according to the histological types of BCC revealed that nodular BCCs consist of larger cells with statistically significant increased perimeter, minimum exterior axis, nuclear area, surface and perimeter of convexivity, and equivalent circle diameter. Infiltrating, sclerosing, and superficial BCCs, which tend to relapse, showed to consist of smaller cells with greater intercellular distance. CONCLUSIONS: Nuclear morphometry evaluated with computer-assisted image analysis may contribute to a better knowledge and outcome prediction of BCC.

Parity is associated with lower cervical E-cadherin expression in postmenopausal women.

AIM: Epithelial cadherin (E-cadherin), a transmembrane glycoprotein involved in calcium-dependent homophilic cell-cell adhesion, is expressed aberrantly during cervical carcinogenesis. E-cadherin expression and putatively implicated predictors in healthy women remain a rather under-investigated area. The objective of this study is to evaluate the possible associations between E-cadherin expression and reproductive/lifestyle factors in cervical epithelial cells from postmenopausal women. METHODS: A total of 105 healthy postmenopausal women (aged 45-68 years old) attending a university menopause clinic were enrolled in this cross-sectional study. Pap smears were derived and E-cadherin immunostaining was evaluated in squamous, glandular and squamous metaplastic cells, using a semi-quantitative method (rating scale: 0-3). Reproductive and lifestyle factors were obtained from patients' chart review. RESULTS: In squamous cells, women with a history of 0-1 deliveries presented with a higher score vs women with 2-4 deliveries (P = 0.003). Social drinkers and women drinking alcohol daily exhibited a higher E-cadherin immunostaining score in squamous cells vs non-drinkers (0.96 +/- 0.72 vs 0.56 +/- 0.65, P = 0.004). A higher dietary calcium intake was marginally correlated with a lower staining score in squamous cells (0.94 +/- 0.78 for low, 0.71 +/- 0.70 for average, 0.45 +/- 0.52 for high consumption, P = 0.073). CONCLUSIONS: E-cadherin expression seems to be associated with reproductive history and lifestyle habits in squamous cervical cells from healthy postmenopausal women. E-cadherin might participate in the molecular mechanisms underlying the role of parity as a risk factor for cervical cancer.

Expression of ERp29, an endoplasmic reticulum secretion factor in basal-cell carcinoma.

The role of endoplasmic reticulum (ER)-stress proteins in the pathogenesis of neoplasia remains obscure. ERp29 encodes for an ER protein that is thought to facilitate the transport of secretory proteins in the early secretory pathway. ERp29 is expressed at varying levels in virtually every tissue tested, yet its precise role remains obscure. To test if ERp29 is associated with the pathogenesis of skin cancer, in the present study we have assessed the expression of ERp29 in basal-cell carcinoma of the skin. A bank of 104 basal skin carcinoma, including 50 nodular, 29 infiltrating, 15 superficial, 7 sclerosing, 2 fibroepithelial, and 1 pigmented cell carcinoma, were assessed by immunohistochemistry for ERp29 expression. Thirty-nine (37.5%) of the samples tested expressed ERp29 with the infiltrating carcinomas displaying more intense (++,+++) immunoreactivity (6/29, P < 0.05) and the superficial carcinomas exhibiting the less intense anti-ERp29 staining (1/15, P < 0.05). Collectively our results suggest that ERp29 is expressed in a subset of basal-cell carcinoma of the skin with the infiltrating carcinomas exhibiting the highest incidence of immunopositivity. The role of ERp29 in the pathogenesis of the disease and its potential diagnostic value should be explored in future investigations.

Isolated talus metastasis from breast carcinoma: a case report and review of the literature.

BACKGROUND: Acrometastases are very rare and have been identified in only a few cases on the foot. At the onset, they might be misdiagnosed as arthritis. CASE REPORT: A 59-year-old woman with isolated metastasis to the talus, originating from breast carcinoma was treated by radiotherapy, letrazole, and intravenous bisphosphonates. RESULTS: The review of the literature revealed that this is the first case of an isolated metastasis to the bone of talus from a breast carcinoma, while there are a few cases originating from other organs. The differential diagnosis of acrometastases may be difficult. CONCLUSION: Pain in the foot or hand of a patient with a known history of malignancy should be considered as potential metastasis.

Comparison between morphometry and immunostaining of malignant cells in non-small cell lung cancer.

OBJECTIVE: To investigate the morphometric properties of tumor cells in combination with the expression of 5 immunomarkers, quantitatively evaluated by image analysis, in a series of 60 non-small cell lung carcinomas (NSCLCs) and to assess their prognostic significance. STUDY DESIGN: Tissue sections from 60 NSCLCs were assessed for the expression of p53, Ki-67, bcl-2, bax and FasL using immunohistochemistry and antibodies. Correlations between morphometric features and clinicopathologic variables, including overall survival and the expression of the above markers, were statistically investigated. RESULTS: Major and minor axis nuclear values, as well as nuclear area and perimeter values, were positively interrelated. No statistically significant correlations were observed between nuclear morphometry and any of the clinicopathologic parameters. p53 Accumulation, when quantitatively estimated, displayed a positive correlation with major axis (P = .008), minor axis (P = .06), nuclear perimeter (P = .006) and mainly nuclear area values (P = .003). Bcl-2- and bax-quantified immunostaining demonstrated a weak negative correlation with shape factor values (P = .039 and P = .025, respectively). Univariate and multivariate analysis showed that stage was the only significant predictor of survival in all patients. In univariate statistical analysis there was a trend between worse survival and shape factor values < 0.8 (P = .0791); this trend almost reached statistical significance in the subgroup of squamous cell carcinomas (P = .0570). CONCLUSION: p53 Accumulation, when quantified, appears to be positively linked with nuclear morphometric values. The prognostic significance of shape factor in NSCLC warrants further investigation.

Morphometric analysis of AgNORs in thin-layer, liquid-based liver specimens.

OBJECTIVE: To detect argyrophilic nucleolar organizer regions (AgNORs) in ThinPrep (Cytyc Corp., Boxborough, Massachusetts, U.S.A.) liver fine needle aspiration (FNA) specimens and to define the diagnostic value of their quantitative analysis in the evaluation of hepatic lesions. STUDY DESIGN: ThinPrep liquid-based FNA biopsy specimens from 49 malignant and benign liver lesions were resampled, fixed in 95% ethanol and stained with the AgNOR technique in accordance with the 1-step colloid method. The specimens included 11 benign and 38 malignant lesions (23 poorly differentiated hepatocellular carcinomas [HCCs] and 15 poorly differentiated metastatic adenocarcinomas [MCs]). Morphometric analysis was performed using a Zeiss Axiolab microscope (Carl Zeiss GmbH, Jena, Germany) with a mechanical stage fitted with a Sony-iris CCD videocamera (Tokyo, Japan). The videocamera was connected to a Pentium III P/C (Intel Corp., Santa Clara, California, U.S.A.) loaded with the appropriate image analysis software. The measurements were performed with ImageScan software (Jandel Scientific, Erkrath, Germany). The number of AgNORs per nucleus (NN) and the total area per nucleus occupied by AgNORs (AR) were calculated semiautomatically. Statistical analysis was performed using the SPSS software package (Chicago, Illinois, U.S.A.). RESULTS: The least significant deviance test for multiple comparisons revealed that NN differed significantly between the 3 groups of samples examined (P < .0001). The mean NN values in HCCs and MCs were significantly different (P < .0001). Logistic regression model demonstrated that as NN increased, the probability of a MC diagnosis decreased (<4%). AR values were different at a statistically significant level only between benign and malignant specimens (P = .00006), not between HCCs and MCs (P = .933). CONCLUSION: Quantitative analysis of AgNORs in ThinPrep specimens could be a diagnostically useful method in liver disease.

Ligand-dependent and -independent effects of splice variant 1 of growth hormone-releasing hormone receptor.

Existing evidence indicates that, in addition to its neuroendocrine action, growth hormone-releasing hormone (GHRH) acts directly on several nonpituitary tissues, especially neoplasms, and stimulates cell proliferation. We have recently reported that a splice variant of the receptor (SV1) is expressed in various normal tissues and particularly in tumor tissues, producing mitogenic effects on GHRH binding. By using HEC-1A human endometrial carcinoma cells, which express endogenous SV1, we show that, in addition to its ability to mediate the mitogenic effects of GHRH, SV1 also possesses relatively high intrinsic, ligand-independent activity. By using an antisense RNA-based approach we found that SV1 ablation reduces the efficacy of colony formation and the rate of cell proliferation of HEC-1A cells in the absence of exogenous GHRH, and decreases their sensitivity to GHRH when the neurohormone is added to the culture media. This ligand-independent stimulation of cell proliferation appears to be a characteristic property of the truncated form of the receptor, because the expression of SV1 and not of the full-length GHRH receptor stimulated the proliferation of 3T3 fibroblasts in the absence of exogenous GHRH, whereas both forms mediated the proliferative effects of GHRH. Evaluation of 21 specimens of human primary endometrial carcinoma for expression of SV1 by immunohistochemistry indicated that in contrast to the GHRH receptor, which is absent, SV1 is expressed in approximately 43% of the specimens. These findings indicate that SV1 can operate in a ligand-independent as well as a ligand-dependent manner. The overexpression of this form of GHRH receptor may be associated with carcinogenesis.