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Zeitschrift für Zytologie und Histologie

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Volumen 4, Ausgabe 4 (2013)

Forschungsartikel

Ossicles Autograft Safety in Cholesteatoma: A Histological Study

Giuseppe Attanasio, Mammola CL, Covelli E, Cagnoni L, De Seta D, Minni A, Gaudio E and Filipo R

Objective: The aim of the study is to define, just before any sort of cleaning procedure, if there is any epithelial inclusion inside the ossicles of patients with cholesteatoma and if the findings could be correlated with surgical aspect of cholesteatoma.

Methods: The specimens used for this study were 19 malleus and 15 incuss which were intraoperatively obtained from 24 patients. Each ossicles was grouped on the basis of cholesteatoma intraoperatively aspect as follows: Stage 1: ten ossicles obtained from encapsulated cholestatoma, non-invasive, easily cleavable. Stage 2: fourteen ossicles obtained from partially encapsulated cholesteatoma, non-invasive, non-easily cleavable. Stage 3: ten ossicles obtained from not encapsulated cholesteatoma, invasive, not cleavable. Two stapes and 1 malleus have been taken from patients who underwent middle ear surgery for conductive hearing loss and they have been used as control. The ossicles were examined histopathologically after the removal.

Results: Our results do not show any epithelial inclusion inside the ossicles independently from the macroscopic aspect or growing aggressiveness of cholesteatoma. In addition there was not inflammatory cells infiltration in stage 1, but it was present in one incus (7.1%) of stage 2 and in five ossicles (50%) of stage 3. In ossicles of grade 3 it has been found up to four layers of epithelial cells on the surface of the ossicles.

Conclusion: The results of the present study reject the hypothesis that epithelial inclusions into the ossicles could cause cholesteatoma recurrences, but strongly suggest to perform safe cleaning procedure of ossi

Forschungsartikel

Identification of NKCC1 and Aquaporin 1 in Blood Vessels of Human Dura and Chronic Subdural Hematomas. A New Target for Bumetanide?

Mahlon D Johnson, Mary J O’Connell and Jason H Huang

Chronic Subdural Hematoma (CSH) may be challenging to manage due to neurological complications and frequent recurrence. The pathophysiology of fluid collection in CSHs is incompletely understood and the role of water and cation cotransporters in the movement of fluid from dural capillary/venous complexes into the dural border zone and hematomas is unknown.

In the present study, normal dura was collected at autopsy from 9 fetal, 6 neonatal or infant, 11 adult and 16 aged patients. CSHs were collected from 15 surgical specimens. Na-K-2Cl cotransporter (NKCC1) and aquaporin 1 (AQP1) were evaluated using immunohistochemistry.

In the normal dura, NKCC1 immunoreactivity was extensive in neonatal, infant and adult capillaries. NKCC1 was not detected in arterioles and large venules in the fetal and adult dura or in unlined channels at any age. AQP1 was extensive in capillaries in all fetal, infant, aged and 82% of adult dural samples. In venules, AQP1 was extensive in most immature and adults. In CSHs, NKCC1 and AQP1 were extensive in capillaries and sinusoids in the outer dural membrane of each case and exhibited an overlapping distribution.

This study shows that NKCC1 and AQP1 are present on endothelial cells in the developing and adult dura and likely participate in fluid and ion movement from dural and hematoma capillaries into CSHs. Potential pharmacological interventions using NKCC1 inhibitors such as bumetanide and AQP1 inhibitors, along with other agents inhibiting angiogenic factors, should augment current management options and reduce the morbidity and mortality of CSHs.

Fallbericht

Well-Differentiated Adenocarcinoma of the Urinary Bladder with Minimal Deviation Adenocarcinoma-Like Features

Naoki Kanomata and Muramaki M

Urinary bladder adenocarcinoma is an uncommon disease. The histological diagnosis of bladder adenocarcinoma is usually straightforward. However, bladder carcinoma rarely shows benign-mimicking features. We report on a 64-year-old woman who was diagnosed with urinary bladder adenocarcinoma, which was well differentiated with the features of minimal deviation adenocarcinoma. Most of the tumor cells were slightly atypical, and the tumor was differentiated barely from the benign disease. Careful examination was necessary for a correct diagnosis. The pathology comprised invasive foci with desmoplastic reaction in the extravesicular adipose tissue.

Rezensionsartikel

The Stem Cell Niche as the Key to Early Cancer Development

Julio Roberto Cáceres Cortés

Cancer cells are remarkable for their multiform character. It is now apparent that for normal cells to become cancerous, they shall not have reached a high degree of differentiation and must be properly nourished. Another important factor in the transformation of a normal cell is probably the alteration of the microenvironment that otherwise would hold the cell in check. Loss of niche control over normal stem cells could allow for the reversion of differentiation to a less specialized form, leading to unrestrained growth. The tumor microenvironment consists of the properties conferred by abnormal interactions between tumor and host cells. The aim of the current review is to map out the steps leading to this abnormal interaction process. If indeed the cancer stem cell niche is key to unrestrained proliferation, insights into the alteration in the microenvironment should certainly provide clues as to more effective cancer therapy.

Forschungsartikel

Digital Image Processing Assessment of the Differential in vitro Antiangiogenic Effects of Dimeric and Monomeric Beta2-Glycoprotein I

Camila Machado, Miriela Escobedo Nicot, Carolina Nigro Stella, Sara Vaz, Cassia Prado, Durvanei Augusto Maria, Francisco Palacios Fernandez and Ligia Ferreira Gomes

The β2 -glycoprotein I (β2GPI) is an endothelial cell ligand, accessible for systemic, autocrine and paracrine signaling. In vivo, β2GPI is immobilized by binding, mainly to the endothelial cell membrane heparan sulfate but also to anionic phospholipids, and functional receptors. The β2GPI was attributed antiangiogenic properties both in vitro and in vivo. This work was designed to evaluate the antiangiogenic effects of native, monomeric, and dimeric β2GPI. Monomeric as well as dimeric forms were purified from human plasma, and the native protein was obtained as a balanced mixture of both components present in human plasma. The proliferation, migration, and differentiation of Human Umbilical Vascular Endothelial Cells (HUVEC) were considered in an in vitro angiogenesis model based on tridimensional cultures and quantitative digital image processing techniques. The early events of the in vitro HUVEC growth and differentiation in the tridimensional cultures microenvironment were addressed by the morphological analysis. The morphological aspects were correlated to the cell growth, oxidative balance outcome and mitochondrial toxicity assays, leading to the evidence that non-confluent HUVEC cultures temporarily stop growing in the presence of the native protein, but remain competent to proliferate. The β2GPI monomer allowed the in vitro differentiation of the HUVECs into typical trabeculæ and incomplete capillary-like tubes, along with lowering the available proliferation fraction. In opposition, the dimer rich purification fraction exposure halted cell elongation and migration, and prevented the organization of the tubular structures in tridimensional cultures, maintaining cell growth. The morphological approach was useful to attribute to β2GPI dimerization the cell migration inhibition modulation, which potentially leads to overcome the diminished sprouting antiangiogenic effect of the monomer fraction of the native protein.

Forschungsartikel

CD44-Positive Cells and Hyaluronan are a Hallmark of a Rat Model of Aristolochic Acid Nephropathy

Anne-Emilie Decleves, Agnieszka Pozdzik, T Baudoux, Isabelle Habsch, Eric De Prez, Bruno Flamion, Joelle Nortier and Nathalie Caron

Background: Aristolochic acid (AA) Nephropathy (AAN) is characterized by tubulointerstitial injury leading to fibrosis and tubular atrophy. Defective regeneration of tubular cells was hypothesized as a mechanism leading to Proximal Tubular Epithelial Cell (PTEC) atrophy. Here, we examined the distribution of CD44, an adhesion molecule involved in differentiation, and its main ligand, Hyaluronan (HA), in an experimental AAN model.

Methods: Wistar rats were injected daily with AA or vehicle during 35 days. Tubular enzymuria and tissue expression of CD44 and HA were evaluated at days 3, 7, 10, 21 and 35. Co-expressions of CD44 and PCNA (proliferation), vimentin (mesenchymal phenotype), a-SMA (myofibroblasts), and CD133 (progenitor cells) were investigated.

Results: AA induced acute tubular damage followed by PTEC atrophy. In controls, CD44 was limited to the basolateral membrane of collecting ducts while HA expression was confined to the medullary interstitium. CD44 was overexpressed as soon as 3 days after AA exposure, spreading to the apical and basolateral membranes of dedifferentiated PTECs and appearing on numerous interstitial cells along with marked HA accumulation around necrotic tubules. Both CD44 and HA expressions increased steadily throughout the 1-month follow-up. A large amount of tubular and interstitial CD44+ cells were proliferative (PCNA+) and mesenchymal-like (vimentin+). Few CD44+ cells expressed a-SMA or CD133.

Conclusions: Our results suggest CD44 is a strong marker of dedifferentiation, and thus of regeneration, in a rat model of AAN. Concomitant HA accumulation around necrotic tubules may create a sustained crosstalk between CD44+ cells and their interstitial ligand. PTEC atrophy is accompanied by overexpression, not deficiency, of the CD44-HA axis.

Rezensionsartikel

Prophylactic Strategy for Peritoneal Recurrence after Curative Operation in Pancreatic Cancer

Kenichiro Yamamoto, Shinya Shimada, Masafumi Kuramoto, Satoshi Ikeshima, Toshiro Masuda, Kenichi Nakamura, Shinichi Yoshimatsu, Masayuki Urata and Hideo Baba

Even though curative surgery has been performed for patients with pancreatic cancer without peritoneal free cancer cells at laparotomy, many patients die of peritoneal recurrence. The reason for this fact is postulated that the surgical procedure itself could cause spread of cancer cells into peritoneal cavity. To identify the free cancer cells spilled in the peritoneal cavity during the operation, we have established real-time quantitative reverse transcription Polymerase Chain Reaction (RT-PCR) system. Moreover, we have devised Extensive Intraoperative Peritoneal Lavage (EIPL) therapy as a very simple and non-aggressive prophylactic treatment for peritoneal dissemination of gastric cancer patients with peritoneal free cancer cells. In this paper, we review the validity of EIPL therapy navigated by real-time quantitative RT-PCR detecting free cancer cells in the peritoneal cavity during operation, and the effect of this therapy on the survival after curative operations in patients with pancreatic cancer

Forschungsartikel

Expression of Molecular Markers in Tumours of Patients with Breast Cancer

D.O. Shapochka, S P Zaletok and M I Gnidyuk

The expression of transcription factor NF-κB (p50 and p65), ER, PR, Her2/neu, Ki-67, p53, Bcl-2 and E-cadherin in tumor tissue of 132 patients with breast cancer was investigated by immunohistochemistry. As a result of researches the correlation between Grade and expression of ER, PR, Bcl-2, Ki-67 and p65-subunit of NF-κB was defined. The level of proliferation was correlated with ER, PR, Bcl-2, p53 and Her2/neu expression. Also the correlation between some of investigated markers was found. Thus, high level of NF-κB nuclear expression was associated with Her2/neu hyperexpression and negative status of ER, PR and Bcl-2. These data may indicate poor prognosis and resistance to chemotherapy in breast cancer patients with activated NF-κB, but it requires further research and retrospective analysis of large number of patients.

Forschungsartikel

Prevalence and Geographical Distribution of Precancerous Lesions of the Uterine Cervix in Cameroon

Tebeu Pierre Marie, Zachari Sando, Alice Ndoumba, Isaac Sandjong, Paulette Mawech-Fauceglia and Anderson Sama Doh

Objective: The aim of this study is to determine the prevalence of cervical precancerous lesions in Cameroon.

Methods: A cross-sectional study for one year period has been conducted in 6 regions of Cameroon by the National Cancer control Committee and the Gynecologic and Pediatric Hospital in Yaoundé from 1/1/2009 to 12/30/2009.

Results: Women were diagnosed of precancerous cervical lesions (97/2485), making the national prevalence of 3.9%, with the highest rate located in Centre, Littoral and Far North regions. The age of the affected women was 27-55 years with 67% women between 34 and 49 years old. The majority of these women had at least four pregnancies (32 %).

Conclusion: The prevalence of precancerous lesions of the uterine cervix in Cameroon varies among regions depending on the existence of screening activities. Continuing effort in extending these programs in different regions will make a noticeable improvement in preventing cervical cancer.

Forschungsartikel

Does Exist A Correlation Between BMI and Gleason Patterns 4 and 5 At Prostate Biopsy?

Vincenzo Serretta, Giovanni Caruana, Francesco Sommatino, Salvatore Scurria, Giuseppe Carità, Luigi Vaccarella, Federico Torretta, Giuseppe Cicero, Marcello Daricello and Vito Franco

Background: Obesity has been related with higher Gleason grade and worse prognosis. Increasing proportion of Gleason pattern 4 or 5 is a critical factor for biochemical recurrence, progression and mortality. The endocrine activity of visceral fat could be responsible of the differentation of the prostatic malignant cell towards a more aggressive fenotype. The aim of our study was to correlate Body Mass Index with the presence of Gleason pattern 4 or higher at biopsy.

Materials and Methods: Consecutive patients with positive prostate biopsy were included. A transrectal prostate biopsy procedure with 12 cores, was performed. All tissue samples were reviewed.

Results: Out of 135 patients diagnosed with prostate cancer at biopsy, a Gleason. pattern 4 or 5 was evident in 57 (42%) patients, while it was not detected in 78 (58%). The statistical analysis did not demonstrate a correlation between the histological expression of Gleason pattern 4 or 5 in the bioptic specimens and the BMI class.


Conclusion: Although high risk prostate cancer has been reported more frequent in patients with higher BMI, in our experience, no significant correlation between BMI and Gleason patterns 4 and 5 at biopsy was detected. Other factors responsible for the worse prognosis and the more aggressive behavior of prostate tumors in obeses should be investigated.

Forschungsartikel

Expression of Toll-like Receptor 9 in Cervical Intraepithelial Neoplasia from Mexican Women

Mario Adán Moreno-Eutimio, Gustavo Acosta-Altamirano and Víctor Manuel Vargas-Hernández

Introduction: Cervical cancer is a major health problem, is the second leading cause of cancer death in women worldwide and the first in developing countries. Toll-like receptors play an important role in the immune system through the recognition of pathogen-associated molecular patterns and damage associated with endogenous molecules, so that the role of Toll-like receptors is being explored in cancer.

Objective: To evaluate the expression of Toll-like receptor 9 (TLR9) in cervical intraepithelial neoplasia grade I, II and III tissues using immunohistochemical staining.

Material and Methods: Retrospective, observational and descriptive study, the expression of TLR9 incervical intraepithelial neoplasia grade I (n=124), II (n=68) and III (n=29) tissues were evaluated using immunohistochemical staining. All samples used in this study were from Mexican women.

Results: We found the expression levels of TLR9 was higher in CIN I (p<0.001) and CIN-II (p<0.001) than in CIN-III.

Conclusion: The low expression of TLR9 may play important roles in the development and progression of cervical intraepithelial neoplasia in Mexican women.

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