[RALP: comparison of the oncological and functional outcomes of the intrafascial and the interfascial approaches]. / Résultats carcinologiques et fonctionnels de l'abord interfascial de la prostatectomie robotique comparé avec l'abord intrafascial.

INTRODUCTION: Due to its technical ease and greater precision Robotic Assisted Laparoscopic radical Prostatectomy (RALP) allows a better preservation of the neurovascular bundles, thereby improving functional outcomes. The intrafascial dissection has been proposed to allow a more complete preservation of these bundles. However, this technique harbors a high rate of positive surgical margins, justifying another trend: the interfascial approach. To date, there are still few publications directly comparing these 2 techniques and our study is the first to offer a 2-year follow-up. MATERIALS AND METHODS: Our study focused on a two-hundred patients population divided into two consecutive groups. All the patients were continent preoperatively and had a satisfactory IIEF5 score: (1) Group 1 consisted of 100 patients who underwent RALP with the intrafascial approach. They had a mean age of 60.3 years (45-70). The majority of cancers were of the low or moderate risk group of d'Amico. The mean PSA was 7.43ng/ml. Seventy-five patients had a pT2, 24 a pT3 and one patient had a pT4. (2) Group 2 included 100 patients who underwent RALP with the interfascial technique. Patients had a mean age of 61.6±5.96 years (45-72), and their cancers were mostly of the low or moderate risk groups of d'Amico. The mean PSA was 6.3ng/ml. Seventy-four patients had a pT2, 22 a pT3a, and 4 had a pT3b. All patients were evaluated after one and two years of follow-up. RESULT: Rates of positive surgical margins were 45% and 19% respectively for groups 1 and 2 (P<0.0001). The rates of biochemical failure (PSA>0.2ng/ml) at 2 years were 10% and 3%, respectively for groups 1 and 2 (P=0.0447). At 2 years, 2 patients in group 1 and one patient in group 2 were using 2 or more urinary pads. Erection with or without oral medication was maintained in 65 (65%) and 31 (31%) patients respectively for groups 1 and 2 at one year. At 2 years 86 and 65 patients were having spontaneous erection, respectively in groups 1 and 2 (P=0.0006). In addition, 65 and 55 patients were also capable of sexual penetration, respectively in groups 1 and 2 (P=0.0045). CONCLUSION: The intrafascial approach exposed to a very high rate of positive surgical margins while offering only a little benefit in the erectile function preservation at 2 years compared to the interfascial variant. In our series, we did not notice any significant difference between the two techniques concerning the urinary continence. LEVEL OF EVIDENCE: 5.

[Value of stereotactic breast core biopsy in patients with atypical ductal hyperplasia]. / Valeur des macrobiopsies mammaires sous guidage stéréotaxique en cas de découverte d'une hyperplasie canalaire atypique.

PURPOSE: Based on a dual center retrospective study, the risks of under-diagnosing clusters of microcalcifications on core biopsies are analyzed. Imaging and histopathological factors affecting this risk are explored. MATERIALS AND METHODS: A total of 1,400 lesions (ACR BI-RADS 2-5) were reviewed and analyzed for size, degree of excision (%), and histology result. A total of 381 patients underwent surgery. Inter-center review of some histological slides was also performed. RESULTS: The rate of under-diagnosis was 5.9% for ductal carcinoma in-situ (DCIS) and 12.5% for atypical ductal hyperplasia (ADH). Most cases of under-diagnosis involved clusters larger than 20 mm in diameter where the percentage of excision decreased from 98% (clusters<10 mm) to 9%. Review of histological slides showed inter-observer variability that decreased in relation to experience. ADH was never under-diagnosed when 3 or less foci were present. The risk of under-diagnosis for DCIS increased when the number of biopsies containing DCIS was superior to 50% and if the grade was high. CONCLUSION: The presence of ADH on biopsy specimens requires surgical biopsy, but optimal core biopsies with cluster removal and histological analysis by an experienced observer allows identification of low risk patients that could undergo close follow-up.

[Value of vacuum assisted biopsies under sonography guidance: results from a multicentric study of 650 lesions]. / Place des macrobiopsies mammaires assistées par le vide sous guidage échographique: étude multi-centrique de 650 lésions.

OBJECTIVE: With this retrospective, multi-centric study, the authors are showing the technique of Vacuum assisted biopsies under ultrasound guidance and comparing it with the other widely used diagnostic techniques. Material and method. Six hundred and fifty biopsies were performed between May 2000 and December 2004, on 644 patients, in 3 centres, following a unique protocol. Lesions were categorized, using the classification from Stavros, between "probably benign", "indeterminate", "probably malignant" and "malignant" Histology was validated only after review of the clinical and radiological data, as well as surgical data when available. All benign cases were included in an on-going follow-up protocol. RESULTS: We have identified 471 benign lesions and 179 malignant lesions. The mean size of the lesions was 9 mm. Three cancers were diagnosed in the cases of "probably benign lesions" and in the cases of "probably malignant lesions" 18 (27%) were inflammatory disorders. In 5 cases vacuum biopsy underestimated the pathology with regard to surgery: 2 cases of atypical duct hyperplasia (HCA) were in situ ductal carcinoma (DCIS) at surgery and 3 cases of DCIS were infiltrative ductal carcinoma (DCI) at surgery. With this technique we have avoided surgery for 71% of all women who presented an "indeterminate" or "probably malignant" condition. Specificity is excellent with no cancer detected so far among the patients with benign findings, under follow-up. CONCLUSION: Ultrasound guided Vacuum assisted biopsy is a fairly recent minimally invasive technique, with short learning curve. The ability to collect a larger volume of tissue overcomes the targeting issues on small lesions and avoids underestimation of heterogeneous and larger abnormalities and some specific at-risk lesions such as papilloma. This technique thus appears indicated in such cases because it overcomes some of the limitations of core needle biopsy and should be considered as an alternative to surgical biopsy.

High levels of stromelysin-3 correlate with poor prognosis in patients with breast carcinoma.

Stromelysin 3 (ST3) is a matrix metalloprotease (MMP) expressed in fibroblast-like cells of most human invasive carcinomas. In this investigation, ST3 was measured by semiquantitative immunohistochemistry in 111 primary breast cancers. ST3 levels showed no correlation with tumor size, axillary-node status or tumor grade (Scarff-Bloom-Richardson system; SBR) but were significantly associated with higher nuclear grade (modified SBR). In addition, ST3 levels were significantly higher in ductal than in lobular cancers. Patients with high scores of ST3 staining had a shorter disease-free interval and shorter overall survival than patients with low scores. ST3 is thus one of the first MMPs to correlate with patient outcome in breast cancer. These findings are consistent with earlier clinical and experimental observations suggesting that ST3 contributes to breast-cancer progression.

Albumin as a sealant for a polyester vascular prosthesis: its impact on the healing sequence in humans.

OBJECTIVE: Although the healing characteristics of albumin impregnated vascular prostheses have been extensively studied in animal models, they have never been studied in humans. We therefore examined the healing sequence and the albumin degradation rate of this type of prosthesis harvested from humans. We also addressed the possible relationship between the implantation of cross-linked albumin and a specific inflammatory reaction. METHODS: Thirty albumin-impregnated polyester vascular prostheses were collected in our institution from January 1991 to February 1993. The mean duration of implantation of the prostheses was 8.4+/-9.7 (SD) months (range: 1 hour to 26 months). Twenty two prostheses were patent at the time of explantation and 4 had been thrombosed for less than 24 hours. In 18 cases, the prostheses were surgically removed because of a complication or a reoperation, and during an autopsy in 12 cases. Each harvested specimen was submitted to histological and immunohistochemical studies in order to demonstrate the presence of human albumin sealant, and to determine the inflammatory cell constituents. RESULTS: The albumin-impregnated prostheses were poorly infiltrated by healing tissues after 2 years of implantation. An external capsule was constantly observed after 2 months of implantation with a nonspecific chronic inflammatory reaction localized between the capsule and the polyester yarns. We observed large amounts of albumin sealant after 2 months, a gradual degradation with time, and traces after 2 years of implantation in humans. The luminal surface of the explant was mainly covered with organized fibrin. No histological signs of a specific inflammatory reaction were observed. CONCLUSIONS: The healing of the albumin impregnated prosthesis was poor and the degradation rate of the albumin sealant was significantly delayed, when compared to animal models. This difference in degradation rate could be related to interspecies differences of phagocytic cells enzymatic machinery. Finally, implantation of glutaraldehyde cross-linked albumin in humans is safe, since we observed an aspecific chronic foreign body inflammatory reaction.

Characterization of monoclonal antibodies against stromelysin-3 and their use to evaluate stromelysin-3 levels in breast carcinoma by semi-quantitative immunohistochemistry.

Stromelysin-3 (ST3) is a matrix metalloproteinase which is expressed in fibroblastic cells of most human invasive carcinomas and represents a potential new prognostic indicator. Expression of recombinant ST3 forms in Escherichia coli from cDNA constructs indicated that high levels of expression were achieved when the ST3 pro-domain was deleted. The putative mature form of ST3 thus produced and recovered from bacterial inclusion bodies was used to prepare monoclonal antibodies (MAbs) against ST3 by immunization of BALB/C mice. Ten hybridomas producing MAbs against ST3 were obtained and analyzed for their ability to detect endogenous ST3 in breast cancer and in conditioned media from human fibroblasts. One of these MAbs (5ST-4A9) was found to be suitable for the routine detection of ST3 on breast cancer tissue sections, thus opening the possibility to evaluate ST3 prognostic value in breast cancer using semi-quantitative immunohistochemistry.

A screening method to identify genes commonly overexpressed in carcinomas and the identification of a novel complementary DNA sequence.

We describe a differential screening method for cDNA libraries which used a combination of subtracted and PCR-amplified cDNA probes, and which can be applied to the selection of genes expressed in multiple tissues. This technique was used to identify genes commonly overexpressed in breast and basal cell carcinomas. These represent stromally dependent, invasive tumors with and without metastatic capacity. Thus, this screening sought to identify genes involved in the early stages of tumor progression. We identified a total of 16 genes, including c-erbB-2 and tissue inhibitor of metalloproteinases 3 whose products have been implicated in tumorigenesis or invasion. We also identified a novel sequence (D52) showing little homology with others described in any species, which maps to the human chromosomal band 8q21. In situ RNA hybridizations of breast carcinoma sections indicated that the D52 gene was expressed in cancer cells, whereas other genes identified in the differential screening were expressed in fibroblastic or inflammatory cells within the tumor stroma. Thus, the procedure developed in this study selected genes expressed in a diversity of cell types, indicating its potential usefulness in other systems.

The tissue inhibitor of metalloproteinases-3 gene in breast carcinoma: identification of multiple polyadenylation sites and a stromal pattern of expression.

BACKGROUND: Tissue inhibitor of metalloproteinases-3 (TIMP3) is the third member of the TIMP family of proteins, believed to play a significant role in controlling extracellular matrix remodeling. MATERIALS AND METHODS: Differential screening of a human breast carcinoma cDNA library using substracted and PCR-amplified cDNA probes identified a 4.6-kb TIMP3 cDNA, which was used for further cDNA library screenings, Northern blot hybridizations, and the synthesis of riboprobes for in situ RNA hybridization analyses. RESULTS: The 4.6-kb full-length TIMP3 cDNA contains 3.7 kb of 3'-untranslated sequence. Additional TIMP3 cDNAs subsequently identified were colinear with the original sequence, but revealed use of four different polyadenylation signals within the 3'-untranslated region, which accounted for the 4.6-, 2.7-, 2.5-, and 2.1-kb TIMP3 transcripts noted in this and in previous studies. In situ RNA hybridizations demonstrated that in breast carcinoma the TIMP3 gene was predominantly expressed by fibroblastic cells within the tumor stroma adjacent to cancer cells. TIMP3 transcripts were also strongly detected in fibroblastic decidual cells of pregnant endometrium. CONCLUSIONS: Modulating the length of the 3'-untranslated region may represent a mechanism by which TIMP3 gene expression is controlled in tissues. The strong expression of the TIMP3 gene by fibroblastic cells in breast carcinoma supports the importance of tumor stroma as a source of factors influencing human carcinoma growth and progression.

Membrane-type matrix metalloproteinase (MT-MMP) gene is expressed in stromal cells of human colon, breast, and head and neck carcinomas.

From breast cancer cDNA libraries, we have cloned cDNAs that proved to correspond to the membrane-type matrix metalloproteinase (MT-MMP) recently identified in human placenta and proposed to be an activator of progelatinase A [Sato, H., Takino, T., Okada, Y., Cao, J., Shinagawa, A., Yamamoto, E. & Seiki, M. (1994) Nature (London) 370, 61-65]. Using one of these cDNAs as a probe, we have detected MT-MMP gene expression in all 83 human carcinoma specimens examined by RNA in situ hybridization and have found MT-MMP transcripts in fibroblastic cells of tumor stroma but not in cancer cells. Comparison with other MMP genes expressed in fibroblastic cells of human carcinomas indicated that the expression pattern of the MT-MMP gene was more closely related to that of the gelatinase A gene than to those of the stromelysin 3 or interstitial collagenase genes. These observations are consistent with the hypothesis that MT-MMP and gelatinase A are cooperating during tumor progression and strengthen the concept that proteolytic activities originating from the stromal component of human carcinomas have a critical role in tumor progression.

Human resident peritoneal macrophages: phenotype and biology.

Macrophages represent the primary line of host defences in the peritoneal cavity. In order to study the metabolic activity and maturation stage of human resident peritoneal macrophages (PM phi). peritoneal fluid (PF) was taken by Douglas puncture from healthy hyperstimulated infertile women undergoing oocyte retrieval for in vitro fertilization. Peritoneal fluid and macrophage culture fluids were studied for different inflammatory mediators such as interleukin-1 (IL-1), tumour necrosis factor (TNF) and interleukin-6 (IL-6). The level of macrophage colony-stimulating factor (M-CSF), which represents a macrophage proliferation and differentiation factor, was determined in the PF and in the serum. Furthermore, the macrophage phenotypic profile was analysed, in particular the expression of sex steroid hormone receptors. IL-1. IL-6 and TNF were detectable in the PF and in the culture supernatants of PM phi whether stimulated or not by IFN-gamma and LPS. The mean level of M-CSF in the PF was 6.37 +/- 2.02 ng/ml as measured by RIA; this level did not correlate with the concentration of PM phi. The mean PF-M-CSF level was 1.4-fold higher than in the sera as measured by a EIA. Oestrogen and progesterone receptors could not be demonstrated on the PM phi analysed, so that a direct relationship between the ovarian steroid concentration in these women and the function of PM phi was unlikely. As compared to peripheral blood monocytes (Mo). PM phi showed a phenotypic profile, with some more mature features, e.g. increased expression of CD14, CD68, FcRII, FcRIII, CR3, CR4 and MHC class II determinants. These results indicate that resident PM phi have acquired in vivo a certain differentiation and/or activation state under micro-environmental factors where cytokines secreted by the M phi themselves or by other cells such as the mesothelium may play important roles.

Stromelysin-3 in stromal tissue as a control factor in breast cancer behavior.

BACKGROUND: It has long been proposed that secreted proteinases, including the matrix metalloproteinases, play an important part in tumor progression in mediating extracellular matrix remodeling. More recently, it has been suggested that extracellular proteinases also regulate growth factors and cytokines that may contribute to tumor progression. METHODS: RNA in situ hybridization and immunohistochemistry were used to study the expression, in breast and other types of human carcinomas, of the stromelysin-3 (ST3) gene, which encodes a putative new member of the matrix metalloproteinase family. RESULTS: The ST3 gene is overexpressed in most types of human carcinomas, including breast carcinoma where ST3 RNA was detected in 95% (99 of 104) of invasive primary tumors. Both ST3 protein and RNA are detected in fibroblastic cells immediately surrounding the cancer cells, but not in the malignant cells or in stromal cells at a distance from them. The ST3 gene also is expressed in some in situ breast carcinomas, where ST3 expression correlates with the known risk of these tumors to become invasive. CONCLUSIONS: ST3 is the paradigm of tumor proteinases that are not expressed in the malignant cells of human carcinomas but in fibroblastic cells of tumor stroma. ST3 represents a potential new prognostic parameter to identify subpopulations of aggressive tumors, particularly to evaluate the likelihood of in situ breast carcinoma progression to invasive cancer. Furthermore, the specific expression of the ST3 gene in fibroblastic cells immediately surrounding cancer cells suggests that ST3 may be involved in tumor progression and that it represents a potential target for cancer treatment.

Stromelysin-3 gene expression in human cancer: an overview.

Stromelysin-3 (ST3) belongs to the family of matrix metalloproteinases, a group of proteolytic enzymes which are believed to play a role in tumor invasion and metastasis. In the present study, we report that the ST3 gene, which was initially identified in invasive breast carcinoma, is expressed in most other invasive human carcinomas, but rarely in sarcomas and other nonepithelial tumors. In carcinomas, both ST3 RNA and protein were specifically detected in fibroblastic cells immediately surrounding the cancer cells. In agreement with this observation, the carcinomas which are known to progress without inducing a prominent tumor stroma are also those which usually do not express the ST3 gene. ST3 gene expression was also observed in noninvasive carcinomas of the breast, uterus cervix and bladder, where the probability of detecting ST3 RNA and protein positively correlated with the known risk of these lesions evolving towards invasion. Taken together, these observations further support the hypothesis that ST3 may contribute to tissue-remodeling processes associated with carcinoma progression, and may represent a new prognostic factor to define populations of aggressive tumors.

Production and characterization of monoclonal antibodies recognising defined regions of the human oestrogen receptor.

Mouse monoclonal antibodies were raised against the N-terminal (amino acids 151-165) and the very C-terminal (amino acids 578-595) regions of the human oestrogen receptor (hER). These antibodies recognise the hER by enzyme-linked immunosorbent assay, immunocytochemistry, immunoblotting, immunoprecipitation and gel retardation assays. The presence of hER is used prognostically in human breast cancer. We have tested the reactivity of our monoclonal antibodies on breast cancer sections, comparing with the commonly used Abbott rat monoclonal antibody H222. These studies show that the two monoclonal antibodies described here are highly versatile and will be useful tools for in vivo and in vitro studies of hER function. Furthermore, we show that the corresponding epitopes can be used as molecular "tags" for heterologous proteins and offer a powerful means of purifying and/or characterizing over-produced fusion proteins containing these regions.

The FMR-1 protein is cytoplasmic, most abundant in neurons and appears normal in carriers of a fragile X premutation.

Fragile X mental retardation syndrome is caused by the unstable expansion of a CGG repeat in the FMR-1 gene. In patients with a full mutation, abnormal methylation results in suppression of FMR-1 transcription. FMR-1 is expressed in many tissues but its function is unknown. We have raised monoclonal antibodies specific for the FMR-1 protein. They detect 4-5 protein bands which appear identical in cells of normal males and of males carrying a premutation, but are absent in affected males with a full mutation. Immunohistochemistry shows a cytoplasmic localization of FMR-1. The highest levels were observed in neurons, while glial cells contain very low levels. In epithelial tissues, levels of FMR-1 were higher in dividing layers. In adult testis, FMR-1 was detected only in spermatogonia. FMR-1 was not detected in dermis and cardiac muscle except under pathological conditions.

Stromelysin 3 belongs to a subgroup of proteinases expressed in breast carcinoma fibroblastic cells and possibly implicated in tumor progression.

The expression of the stromelysin 3 (ST3) gene, which encodes a putative matrix metalloproteinase, was studied during breast cancer progression. The ST3 gene is expressed in all invasive breast carcinomas, in a number of their metastases, and in some in situ carcinomas where the probability of detecting ST3 transcripts correlates with the known risk of these carcinomas to become invasive. ST3 RNA and protein were specifically detected in fibroblastic cells immediately surrounding the neoplastic cells in both primary and metastatic tumors. This expression pattern distinguishes the ST3 gene from other matrix metalloproteinase genes, most notably from the 72-kDa type IV collagenase gene, which can be expressed in fibroblastic cells distributed throughout the stroma of primary breast carcinomas. Furthermore, high levels of 72-kDa type IV collagenase, but not of ST3 transcripts, are detected in benign breast fibroadenomas. Interestingly, the urokinase and ST3 genes exhibit very similar patterns of expression in breast carcinomas, which suggests that their products may cooperate during cancer progression.

Increased stromelysin 3 gene expression is associated with increased local invasiveness in head and neck squamous cell carcinomas.

Matrix metalloproteinases are believed to play an important role in tumor invasion and metastasis. To examine the expression of the stromelysin 3 (ST3) gene, a new member of the matrix metalloproteinase gene family, 111 head and neck squamous cell carcinomas and 21 metastatic lymph nodes were analyzed by Northern blot. ST3 gene expression was observed in 106 carcinomas and 19 metastatic nodes, but in only 2 of 60 samples of corresponding normal tissue tested in parallel. ST3 RNA, by in situ hybridization, and ST3 protein, by immunohistochemical analysis, were specifically detected in fibroblastic cells immediately surrounding invasive cancer cells. This fibroblastic expression of the ST3 gene is characteristic among the matrix metalloproteinase genes known to be overexpressed in head and neck carcinomas, since stromelysin 2 transcripts were specifically detected in neoplastic cells, and type I collagenase transcripts in both neoplastic cells and stromal fibroblasts. Furthermore, there was a highly significant positive correlation (P < 0.0001) between ST3 RNA levels and local invasiveness by the cancer cells, suggesting that enhanced expression of the ST3 gene may contribute to the neoplastic phenotype in head and neck carcinomas.

[Neuroendocrine giant-cell epithelioma. Description of a clinical case and review of the literature]. / Les épithéliomas neuroendocrines à grandes cellules. Description d'un cas clinique et revue de la littérature.

Neuroendocrine tumors are rare neoplasms of the larynx and hypopharynx. This tumors are divided in paragangliomas (usually benign, although malignant case have been reported), large cell neuroendocrine carcinoma, described in this case as a primary pharyngo-laryngeal locate, and small cell neuroendocrine carcinoma (characterized by early and diffuse metastatic disease, and best treated by radiotherapy and chemotherapy). Through the international literature, the authors reviewed the characteristics of the large cell neuroendocrine tumors and their management.

Bronchopulmonary distress associated with toxic epidermal necrolysis.

We describe here a patient with severe TEN and respiratory distress and we review the subject of bronchopulmonary symptoms in TEN. Even if pseudostratified ciliated involvement is uncommon, bronchial lesions in the absence of other known causes, should be specifically related to TEN. The mechanisms of pulmonary involvement and ARDS associated with TEN are discussed.