Detection of c-Ki-ras mutation by PCR/RFLP analysis and diagnosis of pancreatic adenocarcinomas.

BACKGROUND: The c-Ki-ras oncogene (also known as KRAS2) is activated by point mutations involving codon 12 in 72%-100% of primary pancreatic adenocarcinomas, but the gene is not activated in nonneoplastic tissues. Therefore, the detection of c-Ki-ras mutations can facilitate the diagnosis of pancreatic adenocarcinomas, which are not always identified with current tests. Detection is usually performed by oligonucleotide hybridization combined with polymerase chain reaction (PCR), RNAse mismatch cleavage assay, or non-isotopic mismatched PCR, methods that are not feasible for routine screening of large numbers of samples because they are time consuming and/or expensive. PURPOSE: Our purpose was to evaluate a rapid, non-radioactive method of detection of a mutation in codon 12 of the c-Ki-ras gene in pancreatic tumor samples obtained by fine-needle aspiration for diagnostic screening. METHODS: Twenty consecutive patients (15 with pancreatic adenocarcinoma, one with pancreatic cystadenocarcinoma, one with endocrine islet cell tumor, and three with chronic pancreatitis) were selected for this study. A sample of pancreatic tissue from each patient with a tumor or pancreatitis was obtained and evaluated by fine-needle aspiration biopsy under computerized tomography scan or ultrasound guidance using a two-needle coaxial technique. Pancreatic DNA from each of these samples was evaluated by PCR amplification and restriction fragment length polymorphism (RFLP) analysis with nucleotide substitution in PCR primers, creating BstNI restriction patterns that distinguished mutated from normal alleles. The accuracy of the PCR/RFLP assay was validated with DNA from SW480 and HT29 colonic carcinoma cell lines with known mutated and wild-type c-Ki-ras gene sequences. Sensitivity was tested with a series of titration experiments. RESULTS: PCR/RFLP analysis can detect a mutation present in 1% of cells. No amplification could be performed in four (20%) samples because of the absence of cells in the aspirated sample. In the 16 samples adequate for PCR/RFLP analysis, a c-Ki-ras gene mutation was detected in 11 (92%) of 12 adenocarcinomas. Overall, diagnosis was obtained by pathologic (cytomorphologic) examination alone in 13 samples (81%). The presence of malignant cells and/or mutated c-Ki-ras gene was detected in 12 of 12 adenocarcinomas but not in chronic pancreatitis or islet cell tumor. CONCLUSION: Screening of pancreatic tissue samples obtained by fine-needle aspiration for c-Ki-ras mutation using PCR/RFLP analysis combined with pathologic examination could facilitate diagnosis of pancreatic tumors.

Immunohistological study of precancerous mucus modification in human distal colonic polyps.

The M1 antigens associated with gastric fucomucins, oncofetal markers of the distal colonic mucosa, were demonstrated to be more closely associated with adenomas [92 of 139 (66%)] than with invasive adenocarcinomas [27 of 218 (12%)]. They were always expressed in tumors containing the M3 antigen normally associated with the intestinal mucus. The M1 antigens, present in 100% of hyperplastic polyps (30 of 30), were not specific for a particular histological type of adenoma but were found to be more closely associated with those showing a villous differentiation [41 of 47 (87%)] than with those having a tubular pattern [51 of 92 (55%)]. The presence of these M1 antigens depended neither on the size nor on the degree of cytological atypia of the nodular adenomas. However, M1 antigens were found in 94% of the adenomas (35 of 37) concomitant with adenocarcinomas; in contrast, only 56% of adenomas (55 of 102) observed on noncancerous mucosa contained these M1 antigens. As already demonstrated during rat colonic carcinogenesis, mucus modification characterized by the presence of M1 antigens could represent early molecular changes occurring before malignant transformation related to a chemical carcinogen. These M1 antigens might be regarded as early precancerous markers of an oncofetal type, associated with human distal colonic mucosa.

Sentinel node biopsy or limited oriented axillary dissection (LOAD) in early breast cancer.

AIMS: The value of the sentinel node biopsy technique is recognised by the majority of surgical teams as an alternative to conventional axillary lymph node dissection for the treatment of small breast cancers. Secondary procedures are necessary when lymph node invasion not detected by frozen section examination is discovered post-operatively. In order to avoid or limit these reoperations, our sentinel node biopsy technique has gradually been transformed into limited oriented axillary dissection (LOAD), which avoids secondary procedures in the majority of patients. PATIENTS AND METHODS: Three hundred and eighty two patients were operated on by the same surgeon, using the patent blue sentinel node identification technique. This technique failed in nine patients, seven of whom were obese. Only one lymph node was removed in 75 patients, two in 88 patients, 3-5 in 174 patients and more than five lymph nodes were removed in the remaining patients. Eighty-eight percent of patients had no lymph node invasion on intraoperative and post-operative examination. RESULTS: Only seven patients were reoperated by secondary conventional lymph node dissection and there was residual cancer in only one patient. CONCLUSION: The oriented limited axillary dissection technique, combined with frozen section histological examination, avoids the usually unnecessary secondary operations in small breast cancers, in which axillary lymph node invasion rarely exceeds more than two nodes. This technique requires surgeons experienced in axillary surgery and conventional sentinel lymph node biopsy. It needs to be validated on a larger scale by a multicentre randomized prospective trial comparing LOAD to conventional axillary lymph node dissection.

Sclerosing cholangitis induced by formaldehyde solution injected into the biliary tree of rats.

Sclerosing cholangitis has been reported after surgical treatment of hydatid disease of the liver and has been hypothetically attributed to the caustic effect of the parasiticide solution injected into the cyst and diffusing into the biliary tree through a cystic-biliary fistula. In this experimental study, we showed that, in rats, injection into the biliary tract of 20% hypertonic saline solution or 2% formaldehyde solution, the most commonly used scolicidal solutions, was followed by lesions of the biliary epithelium. As compared with 20% hypertonic saline solution, the 2% formaldehyde solution caused more severe lesions of the biliary epithelium and, in addition, induced the development of sclerosis. This experimental study confirms the deleterious effect of scolicidal solutions to the biliary epithelium, shows that their effect is mainly related to the causticity of the scolicidal solution, and indicates that intracystic injection of 2% formaldehyde solution should be abandoned.

Clinical and biological characteristics of breast cancers in post-menopausal women receiving hormone replacement therapy for menopause.

The paradox of an excess breast cancer incidence among current users of hormone replacement therapy (HRT) without excess in breast cancer mortality raises the question of possible differences in the clinical and biological characteristics of cancers in current HRT users compared to non-users. A consecutive series of 129 post-menopausal patients under HRT for at least 6 months, in whom an operable breast cancer was diagnosed from January 1992 to December 1996, were identified retrospectively. In most cases women had received combination HRT (estrogen and progestative) and the mean duration was 60.4 (range: 6-360) months. Breast cancers diagnosed in post-menopausal patients during 1992-1993 at the Institut Curie constituted the reference series. Cancers in patients receiving HRT were smaller: 78% versus 32% T1 and 12 mm in larger diameter versus 29.5 mm. They were also more often diagnosed radiologically (49 versus 33%). A second group of 420 post-menopausal breast cancer patients whose samples had been referred for steroid hormone receptor and flow cytometric analysis in 1992-1996 were used for comparing biological and pathological information. Cancers of patients receiving HRT tended to be more often grade I and rarely grade III in the Scarf Bloom Richardson classification. Percentage of cells in S-phase as measured by flow cytometry was considerably lower in HRT users compared to control (mean 2.4 versus 3.7, median 2.2 versus 2. 6). Lymph node invasion, ploidy, and steroid hormone receptor expression did not differ significantly between the 2 groups. This apparently more favourable phenotype of breast cancers diagnosed in post-menopausal patients receiving HRT compared to unselected non-HRT users was not confirmed when analysis was restricted to breast cancers of less than 25 mm in diameter. If, as expected, the phenotypic information bears out in terms of prognosis, this may contribute to overcome the reticence in prescribing HRT due to the increased risk of breast cancer. However, it is still not clear whether the biologically less aggressive phenotype is related to the hormone treatment or is simply due to early detection.

Neo adjuvant Tamoxifen in post menopausal patients with operable breast cancer.

AIMS: Tamoxifen is widely used as adjuvant therapy in receptor positive post menopausal breast cancer. Little is known about its efficacy as neo adjuvant therapy in terms of breast conservation and improved survival. METHODS: We analyzed the tumour response to 20-30 mg Tamoxifen for 6 months in post menopausal patients with oestrogen receptor positive tumours. Treatment included Tamoxifen for 6 months, surgical resection, and irradiation for post menopausal patients refusing initial mastectomy; aged > or =70 years; or with other factors delaying surgery. RESULTS: Between April 1994 and June 1998, 102 patients, age 73+/-87 (54-90) were studied. There were 24 T1, 56 T2, 14 T3, and 8 T4 tumours. Clinical response to Tamoxifen was observed in all patients, with a median size reduction from 31+/-15 (9-70) to 16+/-9 mm (0-50), 15 clinical and 6 complete responses. 88/102 patients were treated conservatively. Radiotherapy was given to 80 and a flash technique to 8 patients. All patients but one are still alive. CONCLUSION: Neo adjuvant Tamoxifen in operable post menopausal ER positive breast cancer is associated with a good clinical response rate and facilitates conservative surgery. Tamoxifen has a valuable role as neo-adjuvant treatment in terms of breast conservation and survival.

[Hyperplastic colorectal polyp: histological, histochemical and immunohistochemical study of 43 cases]. / Le polype hyperplasique colo-rectal: étude histologique, histochimique et immunohistochimique de 43 cas.

Forty-three solitary hyperplastic polyps removed from the colon or the rectum (HPC) were examined under light microscopy. A histochemical and immunohistochemical study was undertaken in order to evaluate semiquantitatively the nature and the distribution of epithelial mucins and the secretion of carcinoembryonic antigen (CEA). Ten HPC had a peculiar morphologic pattern (four with regenerative dysplasia and six with adenomatous foci). CEA secretion was always increased (37 per cent of cases) or highly increased (63 per cent of cases) with respect to the normal colonic mucosa. The nature and distribution of the secreted acid mucins were modified: sulfomucin was equal (25 per cent of cases) or higher (75 per cent of cases) than that in normal rectal or sigmoid colonic mucosae; sialomucin was strongly decreased in 91 per cent of cases. Some of these functional changes (CEA) are also observed in neoplastic lesions. These findings are not in accord with the hypothesis that hyperplastic polyp is a simple hyperplasia of the mucosal epithelium and suggest a disorder in cellular differentiation, particularly for the larger polyps.

[Primary histiocytic lymphoma of the liver. Study of 2 cases and review of the literature]. / Lymphome histiocytaire primitif du foie. Etude de deux cas et revue de la littérature.

The authors report two cases of primary histiocytic malignant lymphoma of the liver successfully treated by right hepatectomy. The first case, was free of secondary spread with a follow-up of ten years, although he received no adjuvant therapy. The second patient developed skin, bone and brain metastases, which disappeared after chemotherapy and radiotherapy. He has been free of recurrence for four years. Such primary liver tumors are very rare and are worth knowing because of the efficiency of treatment and of their rather good prognosis. Evolution is different from that observed in histiocytic malignant lymphomas developed in other sites. It is suggested that, in the liver, this tumor might originate and grow from Kupffer's cells. This histologic origin might explain the particular clinical and evolutive features of primary histiocytic malignant lymphomas of the liver.

[Acute alcoholic hepatitis and death of cirrhotic patients]. / Hépatite alcoolique aiguë et mort du cirrhotique.

Alcoholic hepatitis (AH) seems to be less frequent and to play a lesser role in the death of cirrhotic patients than previously acknowledged. The purpose of this work was: 1) to study the cause of death of cirrhotic patients 2) to determine the prevalence of AH among these patients and 3) to describe the clinical and laboratory features of cirrhotic patients with AH. The data were collected from a series of 107 necropsies in cirrhotic patients without hepatocellular carcinoma. The statistical analyses were carried out with an IRIS 80 computer. Severe liver failure with jaundice and encephalopathy, hemorrhage and uncontrolled infection with septic shock represented 84 p. 100 of the causes of death in patients with cirrhosis. Seventy-nine out of 107 patients (74 p. 100) had no AH (group 1), and 28 (26 p. 100) had AH (group 2): AH was mild in 15 cases and severe in 13 cases. All patients with AH died from a complication directly related to their liver disease while 21.5 p. 100 of patients without AH died from a complication not related to cirrhosis. The clinical and laboratory features of the patients without AH and cirrhosis differed from those of patients without AH by: a more frequent presence of fever (p less than 0.01), the absence of important weight loss (p less than 0.001), the total absence of abstinence (p less than 0.05), a higher value of ASAT/ALAT ratio, of serum levels of total bilirubin (p less than 0.01) and conjugated bilirubin (p less than 0.05), gamma glutamyl transpeptidase (p less than 0.001) and total cholesterol (p less than 0.05).(ABSTRACT TRUNCATED AT 250 WORDS)

[Sentinel node and operable breast cancer: utilization of blue dye injection. Pilot study]. / Ganglion sentinelle et cancer opérable du sein: utilisation du bleu patent. Etude pilote.

STUDY AIM: Sentinel node detection in breast cancer can be realized with colorimetric and isotopic procedures often associated. The aim of this study was to report results obtained with blue dye injection only. PATIENTS AND METHOD: From September 1998 to July 1999, blue dye injection was performed in 73 consecutive patients (mean age: 51 years, range: 36-71 years); 51/70 70% were post-menopausal and half of them were under substitute hormonal treatment; 70% of cancers were discovered through routine mammography. There were 12 bilateral cancers, six of them synchronous, and 84% of cancers were located in the external quadrants. Individualization of sentinel node was performed through blue dye injection into the tumor in case of preoperative diagnosis or in the tumoral site in case of discovery of the cancer through extemporaneous histological examination. RESULTS: 71 out of 73 cancers were classified pT1 and 70% measured 10 mm and over. Individualization of sentinel node failed in two obese patients. Sentinel node invasion concerned one node (n = 7), two nodes (n = 1) and three nodes (n = 1). Conservative treatment was performed in 72 patients out of 73; in case of sentinel node invasion, axillary irradiation was performed without reoperation. CONCLUSION: Blue dye injection for sentinel node individualization is an accurate technique in selected patients in case of small tumors. Reoperation can be avoided and replaced by axillary irradiation in case of N+ tumors. Duration of hospitalization was 48 hours or under in 70/73 patients. Nevertheless isotopic procedure must be recommended as a routine technique in learning centers and for most surgical teams.

Estrogen receptors evolution in neoadjuvant aromatase inhibitor (AI) therapy for breast cancer in elderly women: stability of hormonal receptor expression during treatment.

UNLABELLED: In France, 20% of breast cancers occur in women over the age of 70 and 10% in women over the age of 80. As these women are not included in screening programs, breast cancer is often diagnosed later, at the stage of a large tumor. PURPOSE: To analyze clinical response, possibilities of conservative treatment and course of hormonal receptors in patients receiving neoadjuvant aromatase inhibitor (AI) therapy for at least 6 months. PATIENTS AND METHODS: There were 75 patients, with a mean age of 75 +/- 8 years (range, 58-91 years) received AI for 6 months after the diagnosis of invasive breast cancer with positive hormonal receptors. Clinical and radiologic tumor reduction, the number of conservative treatments and the course of estrogens receptor-labeled cells were determined for each patient. RESULTS: All but 1 of these patients obtained clinical reduction of their tumor. Of these, 86% patients received conservative treatment. In the majority of patients, estrogen receptor (ER) level did not vary between the initial assay and analysis of the operative specimen. DISCUSSION AND CONCLUSION: Aromatase inhibitors are effective as neoadjuvant therapy in ER positive elderly patients with large tumors, as is tamoxifen. Changes in hormone receptor expression during treatment do not predict clinical response. In our experience, neoadjuvant AI therapy should be administered for at least 6 months to optimize clinical response before deciding upon surgery. Discrepancy observed in the literature could be explained by the duration of the treatment.

[Experimental sclerosing cholangitis following intrabiliary injection of formol in the rat]. / Cholangite sclérosante expérimentale secondaire à l'injection intra biliaire de formol chez le rat.

Our study was aimed at assessing the effect on the biliary epithelium of formol and hypertonic salt serum solutions used as parasiticide for hydatid cysts of the liver in rats. One hundred and forty six rats were administrated an intrabiliary injection of isotonic salt serum (control group), of 20% hypertonic salt serum and of 0.5% and 2% formol solutions. A histological study performed 3 months later did not show any abnormality in the rat control group (n = 11). In the hypertonic salt serum group (n = 14) and 0.5% formol group (n = 12), moderate lesions of biliary epithelium were observed. After a 2% formol injection, a periductal sclerosis was observed in 11 cases out of 16, associated with pseudocirrhosis lesions in 4 cases. A sequential histo-immunochemical study did not show any histological abnormalities in the rat control group (n = 24). In rats which were administrated a 2% formol solution (n = 36), first abnormalities were observed as soon as the 7th day with a IA antigen expression at the biliary epithelium level. Fibrosis lesions were observed after 2 months. After 9 months, the infiltration included only T lymphocytes in direct contact with biliary tracts. A cholangiographic study showed a normal aspect in the rat control group (n = 11) or in rats having received hypertonic salt serum (n = 11), and stenoses of biliary tracts, mainly intrahepatic, in 10 rats out of the 11 of the 2% formol group.(ABSTRACT TRUNCATED AT 250 WORDS)

Carcinoid tumor of the common bile duct.

The fifth case of a common bile duct carcinoid tumor is reported. The importance of endoscopic retrograde cholangiography on the one hand and specialized staining on the other for diagnosis is emphasized. The slow growing nature of these tumors favors extensive surgery whenever possible.