Vasomotor dysfunction early after exposure of normal rabbit arteries to an adenoviral vector.

We aimed to investigate whether infection of normal rabbit arteries with a recombinant adenovirus vector would result per se in alterations in contractile and endothelial functions. In one group of rabbits, right or left femoral and ear artery segments were injected in vivo with a replication-deficient adenoviral vector expressing a beta-galactosidase (beta-Gal) reporter gene (4 x 10(10) pfu/ml) to demonstrate efficient gene transfer. Contralateral arteries were injected with the same concentration of a recombinant adenoviral vector carrying no transgene (Ad.MLPnull). In another group of animals, Ad.MLPnull was injected into the lumen of femoral and ear artery segments. The contralateral arteries were used as controls with the injection of vehicle alone. Histochemical assessment of gene transfer using beta-Gal activity (group 1) or in vitro contractility and endothelial function (group 2) was performed 3 days after adenoviral infection. Gene transfer was efficient and reproducible in the endothelium and was associated with the presence of inflammatory cells in the media. In Ad.MLPnull-injected arteries, in vitro contractile response of femoral artery rings to either KCl 60 mM or phenylephrine (10 microM) was reduced to 10.5 +/- 2.3% (n = 14; p < 0.001) and 8.8 +/- 2.0% (n = 7; p < 0.001) of the control values, respectively. Furthermore, in arteries injected with Ad.MLPnull, the endothelium-dependent relaxation produced by acetylcholine (10 microM) was virtually abolished. Similarly, the relaxant effects of the alpha 2-adrenoreceptor agonist UK14304 (1 microM) or the Ca2+ ionophore A23187 (1 microM) were also abolished. By contrast, sodium nitroprusside (10 microM) was still able to relax adenovirus-infected arteries. We conclude that infection with a recombinant adenoviral vector can induce early severe vasomotor alterations in both contractile function and endothelium-mediated relaxation of normal rabbit arteries.

Thrombus generation after adenovirus-mediated gene transfer into atherosclerotic arteries.

Thrombosis represents a major issue during arterial local delivery. We evaluated the occurrence of thrombosis after adenovirus (Ad)-mediated gene transfer into normal and atherosclerotic arteries. A replication-deficient Ad vector expressing the beta-galactosidase reporter gene (Ad.RSV betagal; 4 x 10(9) PFU) was injected into normal and atherosclerotic arteries (n = 11 in both groups). The contralateral artery received either an Ad vector carrying no transgene (Ad.MLPnull) (n = 7 in both groups, 4 x 10(9) PFU) or vehicle buffer (n = 4 in normal group, n = 8 in atherosclerotic group). Animals were sacrificed 3 days following gene transfer for thrombus detection and assessment of beta-galactosidase activity. Thrombus was absent in normal arteries and in atherosclerotic arteries injected with vehicle buffer only. In contrast, nonocclusive thrombus was present in atherosclerotic arteries injected with either Ad.RSV betagal (5 of 11) or Ad.MLPnull (3 of 7). Beta-galactosidase activity was predominantly found in the endothelial layer of the transfected arteries. Gene transfer and expression occurred despite the presence of the thrombus (4 of 5), and its efficiency did not significantly differ regardless of the thrombus. We conclude that thrombus frequently occurred in atherosclerotic arteries after Ad-mediated gene transfer. Further studies are warranted to identify the mechanisms of thrombus generation after Ad-mediated gene transfer into atherosclerotic arteries.

Mycobacterium genavense infection in normal and immunodeficient mice.

Mycobacterium genavense is a recently described microorganism causing disseminated infections in AIDS patients. In this study, we investigate its pathogenicity in mice and some mechanisms of the host response to this bacterium. Following an intravenous challenge of 10(6) organisms, M. genavense grew progressively in the spleens and livers of BALB/c and CBA mice over at least an 8-month period. Granulomas were present in the spleens, livers and lungs of the animals. The numbers of bacteria recovered from the spleens and livers were higher in BALB/c (Bcg(s)) than in CBA (Bcg(r)) mice from day 30. The role of the Bcg gene, in the early phase of infection, was supported by the fact that the bacterial load, on day 15, was higher in BALB/c than in the congenic C.D2 (Bcg(r)) mice. The role of T cells in the host response was suggested by the high susceptibility of nude mice to M. genavense infection. In vivo depletion experiments in CBA mice indicated that gamma interferon and both CD4(+) and CD8(+) T cells participate in the containment of the bacterial load.

MUC1 mucin gene, transcripts, and protein in adenomas and papillary carcinomas of the thyroid.

MUC1 mucin is found in a variety of epithelial tissues and is overexpressed in several epithelial cancers. This molecule could modulate cellular adhesion and thereby influence tumor invasion and metastasis. Little is known of MUC1 gene expression in thyroid tissues. We investigated whether MUC1 mucin gene alteration and/or expression correlated with thyroid tumor progression by studying 21 fresh thyroid tissue specimens comprising 10 macrofollicular adenomas and 11 papillary carcinomas. Normal adjacent tissue from the same patients was also studied. To determine the integrity and expression of the MUC1 mucin gene, a complementary DNA (cDNA) probe was used for Southern blot analysis of DNA and Northern blot analysis of RNA. A detailed immunohistochemical analysis of MUC1 protein expression was performed with DF3 monoclonal antibody, and was compared with other tumor characteristics and clinical manifestations at diagnosis. Of the 14 tumors informative (heterozygous) with the pMUC10 polymorphic probe, 2 (14%) showed loss of heterozygosity (1 adenoma and 1 carcinoma). Overexpression of MUC1 RNA, compared with normal thyroid tissue, was observed in 6 of the 11 papillary carcinomas and in none of the 10 adenomas. Immunostaining of the corresponding formalin-fixed paraffin-embedded tissue sections detected MUC1 mucin protein at the apical domain of follicular cells. Most of the lining was thin in normal tissue and follicular adenomas, but the protein was more irregularly and less strongly expressed in adenomas. In carcinomas the epithelial mucin produced by the MUC1 gene was present irregularly as a thin and/or thick lining at the apical domain of tumor cells. In addition, 5 of the 6 samples with MUC1 mRNA overexpression showed intracytoplasmic staining. Moreover, intracytoplasmic MUC1 mucin staining was found in 75% of "high-risk" papillary thyroid carcinoma (PTC) (PTC with extrathyroid extension at initial diagnosis and/or lymph node involvement), and in only 28.5% of "low-risk" PTC (purely intrathyroidal carcinomas).

Impact of different gases and pneumoperitoneum pressures on tumor growth during laparoscopy in a rat model.

BACKGROUND: The influence on intraperitoneal tumor growth of the choice of gas and pneumoperitoneum pressure during laparoscopy is still unknown. This study compared tumor growth after laparoscopy with different gases and pneumoperitoneum pressures in an immunodeficient model. METHODS: In an initial experiment, 60 nude rats were randomly allocated to undergo laparoscopy at different pneumoperitoneum pressures (gasless, 4 mmHg, or 8 mm Hg.) In a second experiment, 23 nude rats were randomly allocated to undergo laparoscopy with different gases (carbon dioxide or helium). Surgery was carried out 7 days after intraperitoneal injection of IGR-OV1 cells. The rats were killed 7 days after surgery. Tumor growth was assessed by the weight of the omental metastasis. For statistical analysis, we used analysis of variance (ANOVA). RESULTS: Mean omental weight was similar for all groups, regardless of the pneumoperitoneum pressure (p = 0.86) or the type of gas (p = 0.80). CONCLUSION: Physical parameters of gas have a limited impact on tumor growth.

Description of two new human ovarian carcinoma models in nude rats suitable for laparoscopic experimentation.

BACKGROUND: Experimental laparoscopic trials require relevant models of ovarian carcinomatosis. METHODS: Female nude rats were inoculated intraperitoneally either with the IGR-OV1 or the NIH:OVCAR-3 human adenocarcinoma cell lines. Serial clinical checks and sacrifices were used to evaluate the rates of tumor take, survival, and patterns of tumor spread. Finally, laparoscopies with various pneumoperitoneum pressures were performed to verify the "surgical" relevancy of out models. The learning curve was measured. RESULTS: The best results were obtained when twenty-seven 106 IGR-OV1 cells and thirty-six 106 NIH:OVCAR-3 cells were injected in 28-day-old rats. The IGR-OV1 model provided a mean survival of 17.8 days (range, 13-22 days), with a high take rate (94%). The NIH:OVCAR-3 model resulted in a longer mean survival (59 days; range, 49-77) and also a high take rate (83%). The two models differed in their patterns of tumor spread: solid bulky omental metastasis having a diffuse microscopic peritoneal carcinomatosis with the IGR-OV1 line (the weight of the omental cake correlated significantly with the stage of development) and diffuse macroscopic peritoneal carcinomatosis having no large solid tumor, but visceral and paraaortic metastases, with the NIH:OVCAR-3 line. In both models, CA125 was high. Anesthesia could be performed and repeated in healthy and tumor-bearing rats. Laparoscopy was feasible, with pneumoperitoneum pressures as high as 8 mmHg lasting 1 h. Laparoscopy provided a reliable evaluation of the tumor spread into the peritoneal cavity. The plateau of the learning curve was soon obtained for take rate and survival after laparoscopy. CONCLUSION: We report two new human ovarian carcinoma xenografts in nude rats suitable for laparoscopy. The IGR-OV1 model mimics an advanced stage of the disease, and the NIH:OVCAR-3 model presents an earlier stage. These two models appear useful for experiments involving laparoscopy.

Peritoneal closure reduces port site metastases: results of an experiment in a rat ovarian cancer model.

BACKGROUND: The arm of this study was to assess the role of peritoneal closure in the prevention of port site metastasis after carbon dioxide (CO2) CO2 pneumoperitoneum. METHODS: We developed a xenograft ovarian cancer model by intraperitoneal injection of 27 106 IGR-OV1 line cells in nude rats Seven days after the inoculation, the animals underwent a CO2 pneumoperitoneum. At the end of the procedure, port sites were randomly closed either with suture of peritoneum (n = 14, group A) or without suture of peritoneum (n = 12, group B). The rats were killed 7 days after surgery and their port site scars were resected. Tumor implantation was assessed by a pathologist who was blinded to the type of wound closure. RESULTS: The animals in group B were significantly more likely to have at least one port site metastasis frequent (seven of 12, or 58.3%) than those in group A (two of 14, or (14.3%) (p = 0.037). Port sites with metastases were seen more frequently in group B (eight of 24, or (33.3%) than in group A (three of 28, or 10.7%) (p = 0.046). CONCLUSIONS: Our results shows that peritoneum closure decreases the risk of port site metastasis.

Impact of pneumoperitoneum on tumor growth.

AIM OF THE STUDY: To compare intraperitoneal tumor growth after CO2 laparoscopy (L), gasless laparoscopy (GL), midline laparotomy (ML), and general anesthesia (GA) as a control. MATERIALS AND METHODS: A prospective randomized trial was carried out in nude rats. A carcinomatosis was obtained by intraperitoneal injection of either one of the two human ovarian cancer cell lines IGR-OV1 or NIH:OVCAR-3. Rats secondly underwent randomly different kind of procedures: CO2 L (8 mmHg, 60 min), GL (traction by a balloon for 60 min), ML (bowel removed and let on a mesh for 60 min), or GA. The rats were finally killed 10 or 35 days after surgery (respectively in IGR-OV1, or NIH:OVCAR-3 models). Tumor growth was assessed by the weight of the omental metastasis and MIB1 immunostaining. Peritoneal dissemination as well as abdominal wall metastases were assessed by pathological examination. Statistical analysis used the chi-square test (or Fisher exact test) and Bonferroni method for multiple comparison between groups. RESULTS: Fifteen rats were included in each group. Mean omental weight was significantly increased after surgery (3.1 to 5.6 g), when compared to control (2.4 g), but no significant difference was recorded between the three surgical accesses. MIB1 immunostaining was poor in the PNP group (37%), whereas it was higher after midline laparotomy (51%), but the difference was not significant (p = 0.07). Similarly, no significant variation was recorded in the NIH:OVCAR-3 model for omental weight or MIB1 staining. CO2 pneumoperitoneum significantly increased right diaphragmatic dome involvement in the NIH:OVCAR-3 model. Abdominal wall metastases were significantly more frequent after surgery when compared to the control group, but no significant difference could be demonstrated between surgical groups in each model. CONCLUSION: In these solid tumor models, CO2 pneumoperitoneum had no deleterious effect on tumor growth when compared to gasless laparoscopy or midline laparotomy.

Laparoscopic pelvic lymphadenectomy in an anatomical model: results of an experimental comparative trial.

OBJECTIVES: The aim of this paper was to compare the accuracy of laparoscopic versus open pelvic lymphadenectomy in an experimental trial. STUDY DESIGN: We performed unilateral laparoscopy pelvic lymphadenectomy (LPL) in 33 non-embalmed cadavers between the external iliac vein, the obliterated umbilical artery and the obturator nerve. Then a laparotomy was performed to inspect the LPL limits, look for laparoscopic complications and finally realize a controlateral lymphadenectomy. The LPL side was randomly decided. A pathologist counted the number of lymph nodes collected with both techniques. We compared the number of retrieved lymph nodes, the completeness of the dissection and the complication rate with those two procedures. Student's t-test, chi 2-test and non-parametric tests were used when appropriate. RESULTS: No dissection had to be aborted. One hundred and twelve nodes were removed laparoscopically (mean, 3.73; S.E., 2.9) and 84 at laparotomy (mean, 2.77; S.E., 2.06). There was no significant difference in the number of nodes retrieved with both procedures. Effectiveness of laparoscopy was not significantly different in the first ten procedures, in the second ten or in the last ten LPL. Residual tissue was observed after LPL in 13.3% of the procedures whereas all open lymphadenectomies were complete. LPL sensitivity reached at least 86% in this paper. Failures were more frequent at the beginning of the study (50% among the first ten dissections), in obese subjects or in subjects with prior history of laparotomy (but the difference was not significant). Two venous injuries occurred during LPL (6.7%). Complication rates for the two techniques were not significantly different. However, the LPL complication rate was higher at the beginning of the study and increased significantly in subjects with prior history of laparotomy (P < 0.05). CONCLUSIONS: This randomized study shows that LPL and laparotomy have similar effectiveness. Incomplete dissections and complications are more frequent in obese subjects or in case of prior history of laparotomy. Fifteen procedures seems necessary to learn the technique and provide constant and safe results in routine practice.

Effect of methotrexate on tubal epithelium. A report of three cases.

BACKGROUND: Tubal effects of methotrexate injections have been poorly reported. CASES: Three fallopian tubes were examined with light microscopy 9-13 months after tubal methotrexate injection (one case) and intramuscular methotrexate injection (two cases) given for the treatment of unruptured ectopic pregnancies. No evidence of tubal damage was found. CONCLUSION: These three cases confirm previous experimental and clinical data showing the absence of a direct adverse effect of methotrexate on the fallopian tubes in the treatment of ectopic pregnancy.

[Microinvasive carcinoma uterine cervix. Which approach in 1998?]. / Cancer micro-invasif du col utérin. Quelle approche en 1998?

Numerous definitions of microinvasive carcinoma (MIC) have been proposed. The SGO takes into account the depth of stromal invasion and presence of capillary like space involvement (LVI). The Figo uses the lesion width and describes different substages according to the depth of stromal invasion. Two major prognostic factors can be identified in the literature: the depth of invasion and the presence of LVI. The lesion volume is probably more accurate than the depth of stromal invasion but cannot be measured in routine. Taking into account that a classification must be a guide for the evaluation of prognosis and treatment, the SGO definition seems more reliable. Pelvic lymph node metastasis rate and recurrence increase with these two factors. MIC with stromal invasion under 3 mm and without LVI have a little risk of parametrial and nodal involvement: with a high rate of survival. Conversely, MIC with invasion over 3.1 mm depth or LVI have a greater risk of spread beyond the cervix (1% versus 7.7%) and many authors now consider them as true invasive cancers. For lesion invading the stroma within 3 mm, the treatment can be limited to a standard hysterectomy with good results. Some authors have proposed more conservative therapy as conization. This procedure is interesting for young women willing to preserve their anatomy, fertility and sexual function. In selected cases, short term results are similar to those of hysterectomy but there is a lack of controlled studies with long term follow-up. Lesions over 3.1 mm with LVI should be treated as true invasive cancers. Intermediate cases should have a conservative therapy associated with a laparoscopic lymphadenectomy.

[Visceral Kaposi sarcoma with angioimmunoblastic lymphadenopathy and latent sicca syndrome (author's transl)]. / Sarcome de Kaposi viscéral associé à und lymphadénopathie angioimmunoblastique avec syndrome sec latent.

A case clinically and morphologically typical angio-immunoblastic lymphadenopathy in a 64-year old man is reported. A sicca syndrome was discovered with histological localization in the labial accessory salivary glands. Death occurred after a course of a few months. The autopsy showed an association of angio-immunoblastic lymphadenopathy lesions of the bone marrow and the lymph node and of typical multivisceral localizations of Kaposi's sarcoma (lymph node, liver, thyroïdd gland) without skin involvement. This association of two diseases characterized by severe dysimmunity with B lymphocytes hyperplasia is discussed, and compared to the others diseases which could be associated with Kaposi's sarcoma.

Microinvasive carcinoma of the cervix. Rationale for conservative treatment in early squamous cell carcinoma.

Numerous definitions of microinvasive carcinoma (MIC) have been proposed. Taking into account that a classification must be a guide for the evaluation of prognosis and treatment, the authors reviewed the risk of spread in MIC. Two major prognostic factors can be identified in the literature: the volume of the lesion and the presence of capillary-like space involvement (LVI). The former item is generally assessed by the depth of invasion. Two kinds of MIC can be distinguished. Those with stromal invasion under 3 mm and without LVI, and those with invasion over 3.1 mm depth or LVI. The former have little risk of parametrial and nodal involvement and a high rate of survival. The latter have a greater risk of spread beyond the cervix and many authors now consider them as true invasive cancers. The Society of Gynaecologic Oncologists (SGO) definition seems more reliable. For lesions invading the stroma within 3 mm, treatment can be limited to a standard hysterectomy with good results. Some authors have proposed more conservative therapy such as conization. This procedure could be desirable for young women who want to preserve their anatomical integrity, fertility and sexual function. In selected cases, short term results are similar to those of hysterectomy but there is a lack of controlled studies with long term follow-up.

[Significance of lymphoplasmocytic infiltration in arrhythmogenic right ventricular dysplasia. Apropos of 3 own cases and review of the literature]. / Signification des infiltrats lymphoplasmocytaires dans la dysplasie ventriculaire droite arythmogène. A propos de 3 cas personnels et d'une revue de la littérature.

Three patients with typical histological signs of arrhythmogenic right ventricular dysplasia progressed to fulminating heart failure in 1 case and unexplained sudden death in the other two. There were signs of superacute myocarditis in the first case, previous healed pericarditis in the second and chronic myocarditis in the third case. These cases suggested the presence of an inflammatory process complicating the substrate of the dysplasia. Arrhythmogenic right ventricular dysplasia seems to be a developmental defect which is complicated in 50 to 70% of patients, according to a review of 74 cases reported in the literature, by a varying degree of myocarditis suggesting particular susceptibility of these patients to infection and explaining the presence of unusual amounts of fibrous tissue in some and the so called "progressive" nature of the disease in other patients.

[Extensive aortic dissection and aneurysm of the left coronary trunk. 20-year spontaneous development]. / Dissection aortique étendue et anévrisme du tronc corconaire gauche. Vingt ans d'évolution spontanée.

A case is reported of dissection of the aorta in a lady of 35. The condition was complicated first by myocardial infarction and secondly by severe aortic incompetence which led, 20 years after the onset of the dissection, to the death of the patient from intractable heart failure. At post-mortem, an extensive dissection was found to involve the whole of the aorta and several of its branches, but did not involve the coronary arteries, which were normal except for the trunk of the left coronary; this was dilated. The natural history of dissections of the aorta is reviewed. The incidence, course and etiology of coronary aneurysms are also discussed.

[Breast lesions associated with cancer of the breast. Study of 200 excision specimens]. / Lésions mammaires associées au cancer du sein. Etude de deux cents pièces d'exérèse.

Two hundred cases of breast carcinoma were studied, with particular attention to adjacent non-malignant modifications considered by some authors as preneoplastic changes. Diverse proliferative lesions were recognized in 82% of cases. Atypical hyperplasia was present in 51.5% of cases and was more often ductal (83/200) than lobular (40/200). Microscopic cysts were noted in 72% of cases. Calcifications, easily identified by mammography were present in 20% of cases and were associated with atypical hyperplasia in 4/5 of cases. The mean of patients was 58 years. For comparative studies of histologic lesions the following age categories were evaluated: category I less than 50 years (57 patients), category II greater than or equal to 50 years (143 patients). Atypical lobular hyperplasia, adenosis, and, to a lesser degree, sclerosing adenosis and macroscopic cysts were more frequently associated with breast cancer in women younger than 50. Fibro-atrophic or lipomatous changes were more often found in the older group of women, as expected. Sclerosing adenosis was significantly more frequent in breasts with multifocal infiltrating carcinoma. Such changes must be considered as risk factors by pathologists when present in mammary specimens without carcinomatous lesions, and careful study should be undertaken to locate a possible in situ microscopic carcinoma. No differences appeared in lesions associated with breast cancer in women with or without a familial history of breast cancer.

[Carcinosarcoma of the submaxillary gland]. / Carcinosarcome de la glande sous-maxillaire.

Salivary gland carcinosarcoma is a rare neoplasm, with a poor prognosis, that should not be confused with the more common carcinoma ex-pleomorphic adenoma, in which the epithelial component alone is malignant. The authors report one case of carcinosarcoma of the submandibular gland, with no previous history of pre-existent tumor. The tumor exhibited two intermingled and very atypical cellular components, one undifferentiated and the other with a chondroid pattern. Immunostaining with epithelial markers in the undifferentiated area allowed to distinguish this tumor from salivary gland sarcomas. In addition, the positivity for EMA, vimentin, and S100 protein in the two predominant components suggested a common origin for the different tumoral cell types, and led to discuss the nomenclature and the still unclarified cellular origin of these tumors.

[Neuroendocrine carcinoma of the larynx with secretion of calcitonin: primary tumor or metastasis of the medullary thyroid carcinoma?]. / Carcinome neuroendocrine laryngé avec sécrétion de calcitonine: cancer primitif ou métastase d'un cancer médullaire de la thyroïde?

Neuroendocrine carcinomas of the larynx are rare and their links with thyroid neoplastic lesions are not specified in most reported cases. We report such a case secondary to medullary thyroid carcinoma. In a 63-year-old man, a supra glottic laryngeal carcinoma confirmed by biopsy was initially treated by chemotherapy. A second biopsy of the laryngeal lesion after incomplete remission displayed a neuroendocrine carcinoma with calcitonin in neoplastic cells. Laryngectomy, cervical bilateral neck dissection and thyroid isthmic adenoma dissection were performed. Numerous neoplastic cells from these tissues contained calcitonin revealed by immunohistochemical method. Calcitoninemia was highly increased and multiple bone metastases were discovered. Secondary total thyroidectomy was not possible and the patient died 6 months after. In this case the neuroendocrine laryngeal carcinoma was probably secondary to a latent medullary thyroid carcinoma. Medullary thyroid carcinoma is most often unknown in similar previously reported cases of neuroendocrine laryngeal carcinoma. Thus a neuroendocrine tumour of the larynx should require search for extra thyroid and over all thyroid neuroendocrine carcinoma, even limited to a small and latent lesion.

[Fatal pneumopathy linked to subcutaneous injections of liquid silicone into soft tissue]. / Pneumopathie mortelle liée à l'injection sous-cutanée de silicone liquide dans les parties molles.

The case of a 23-year old transsexual male who developed a lethal lung disease 48 hr after a 1 l subcutaneous silicone fluid injection is reported. Histologic examination showed optically empty oil red O negative vacuoles in several specimens, and particularly in the lungs; refractile particles of silicium were also found. We identified the material as silicium, and evaluated its quantity in tissues by atomic absorption and spectrometric and toxicological analysis. Experimental studies and rare human accidents suggest that silicone fluid is transported by migrating phagocytes, thereby explaining lymph node, spleen, liver, adrenal and lung deposits. In our patient's report, we added gradual blood contamination to explain the progressive development of respiratory failure with the severity of the respiratory disorder linked to the massive dose injected. Massive silicone blood embolization could be excluded because of the interval between the injection and emergence of the first clinical signs.

[Elastofibroma dorsi. Apropos of 5 cases]. / Elastofibrome dorsal. A propos de cinq observations.

Elastofibroma is a rare tumor mainly found under the scapula. Based on a recent study of five cases, we would like to stress the importance of making the diagnosis of elastofibroma, as this lesion simply consists of dysplasia of the elastic fibres and is not malignant as could be suggested by clinical examination.