Are biopsy specimens predictive of HER2 status in gastric cancer patients?

BACKGROUND: Trastuzumab has been recently proposed as a treatment for patients with HER2-positive advanced/metastatic gastric cancer (GC). Since most patients have inoperable disease at diagnosis, accurate assessment of HER2 status on biopsy specimens is essential to select the patients who may benefit from therapy. AIM: The aim of this study is to establish whether HER2 status assessed on biopsy material could be reliable for treatment decisions using anti-HER2 agents. METHODS: The HER2 status was evaluated in 61 consecutive pairs of biopsy and surgical GCs samples by immunohistochemistry and chromogenic in situ hybridization. RESULTS: The overall concordance of HER2 status between biopsy and surgical specimens was 91.8 % with a predictive positive value of 71.4 % and a negative predictive value of 94.4 %. Of five discordant cases, there were three negative and two positive false biopsy results. All the false negative cases showed heterogeneous expression of HER2 protein in surgical samples. Two cases displayed overexpression of the receptors without corresponding gene amplification. CONCLUSIONS: HER2 status as evaluated on biopsy samples is a fairly good predictor of HER2 status of surgically-excised GCs. The most important influence for discordant results is tumor heterogeneity. However, HER2 overexpression, especially without coexisting gene amplification, may only be a temporary change in a tumor population. This may explain those cases with positive HER2 evaluation on biopsy material and a negative result on corresponding surgical specimen.

One stage percutaneous transhepatic biliary stenting for malignant jaundice: a safe, quick and economical option of treatment.

OBJECTIVE: Patients with proximal malignant jaundices are often diagnosed in an advanced stage and need biliary decompression treatments, such as percutaneous transhepatic biliary drainage (PTBD) and bare metal stenting (BMS), to improve the hepatic function. Whether it is better to perform those two procedures together or in a separate time, it is not well understood. The aim of this study was to investigate the effectiveness and cost-benefit of a combined "one-stage" PTBD/BMS procedure in patients with malignant jaundices. PATIENTS AND METHODS: Forty-five patients with malignant jaundice treated with "one-stage" PTBD/BMS were retrospectively enrolled to evaluate technical success, complications, survival, and length of hospitalization. RESULTS: A full technical success of the procedures was reported for all patients, with only one major complication among 45 treated patients. A better performance in terms of hospitalization rate was achieved by the one-stage procedure compared to the two-stage, also resulting in global saving of costs. A high survival rate was observed at the 3rd and 6th month (97.7% and 86.6%, respectively), with a median overall survival time of 271,58 days. CONCLUSIONS: Our study shows that performing PTBD/BMS as a "one-stage" procedure is useful, safe, and cost-effective with a high percentage of technical success and a similar occurrence of complications compared to the two-stage procedure.

TNFα deficiency results in increased IL-1ß in an early onset of spontaneous murine colitis.

Inflammatory bowel disease (Crohn's disease (CD) and ulcerative colitis (UC)) is a multifactorial disease resulting from immune dysregulation in the gut. The underlying colitis is characterized by high levels of inflammatory cytokines, including TNFα. Biological intervention for IBD patients using anti-TNFα antibodies is often an effective therapeutic solution. However, TNFα neutralization fails to induce remission in a subgroup of IBD patients, primarily in UC patients. There is a dearth of suitable animal models representing TNFα non-responders. Here we have combined one of the best UC models currently available, namely Winnie and the TNFαKO mouse to generate a TNFα-deficient Winnie to study early onset colitis. The induced TNFα deficiency with underlying colitis does not influence general health (viability and body weight) or clinical parameters (colon weight, colon length and histological colitis) when compared with the Winnie genotype alone. The molecular characterization resulted in identification of Il1ß as the major elevated cytokine during early phases of colitis. Further, in vitro functional assay using bone marrow-derived dendritic cells confirmed IL-1ß as the major cytokine released in the absence of TNFα. This study has generated a successful model of colitis that remains TNFα non-responsive and has demonstrated that IL-1ß expression is a major pathway for the progression of colitis in this system. These data also suggest that IL-1ß can be a potential target for clinical intervention of UC patients who fail to respond to TNFα neutralization.

Immunohistochemical and serological 90K/Mac-2BP detection in hepatocellular carcinoma patients: different behaviour of two monoclonal antibodies.

To clarify the biological role of the 90K/Mac-2BP glycoprotein, we evaluated the ability of two MAbs SP-2 and 1A4.22, to reveal this glycoprotein in both serum and tissue from hepatocellular carcinoma (HCC) patients. Tissue expression of 90K was detected by the immunohistochemical method in 20 HCC patients, while the 90K serum level was assessed by the ELISA assay in 13 HCC patients. MAb SP-2 was reactive only in serum, with a mean value of 12.8+/- 6.7 microg/ml . On the contrary, MAb 1A4.22 revealed immunoreactivity both in 92% of sera and in 60% of neoplastic samples. Positive staining was seen only in the epithelial cells and was cytoplasmic and granular in all instances. The mean 90K serum level assayed with MAb 1A4.22 was 29.4 +/- 13.7 microg/ml. Patients with a 90K serum level 30 microg/ml. Moreover, a possible poor prognostic role was observed for negative 90K in tissue. Our results suggest that only MAb 1A4.22 could demonstrate 90K glycoprotein expression in paraffin-embedded tissue and that this MAb could have a diagnostic and prognostic role in both sera and tissues from HCC patients.

Nine novel APC mutations in Italian FAP patients.

Familial adenomatous polyposis (FAP) is a common hereditary syndrome characterized by early development of colorectal cancer consequent to extensive adenomatous polyps of the colon. In addition to the colonic manifestations the syndrome presents several extracolonic features including polyps of the upper gastrointestinal tract, congenital hypertrophy of the retinal pigment, jaw cysts, osteomata and desmoid tumors. In this study the entire APC coding region has been analysed for mutation in a panel of one Turcot and 33 unrelated Italian FAP patients using SSCP analysis, PTT and DNA sequencing. We detected APC mutations in 23 of them and identified nine which, to our knowledge were not previously reported. All of these novel mutations are in exon 15, including two nonsense mutations, 6 deletions or insertions leading to premature termination of the protein and one missense mutation (7697G>A). This last mutation occurs in the EB1-binding domain of the APC protein and segregates in four relatives of the patient with three of them presenting 2-3 adenomatous polyps.

Main characteristics of hepatocellular carcinoma and cirrhosis and therapeutic approaches.

Between 1995 and 1997 we studied 100 patients with hepatocarcinoma (HCC) and cirrhosis. Of these 74 were males and 26 females with a mean age of 66 years. 13% patients were only HbsAg positive, 75% only anti-HCV positive, 6% HbsAg and anti-HCV and the etiology in 6% of cases was alcoholic. Alpha-foetoprotein was >400 ng/ml in only 18% of cases and portal thrombosis was present in 12%. Mononodular HCC was observed in 63% of cases (small HCC in only 38%) and in 79% was localized to the right lobe. Of the mononodular types, 70% were shown by echography to be hypoechoic, 6% hysoechoic, 6% hyperechoic and 17% mixed patterns. Histologically, 49% were well-differentiated, 45% moderately-differentiated and 6% poorly-differentiated. No correlation was found between histologic pattern and number of nodules. Well-differentiated HCC was found in 51% of mononodular types and in 46% of multinodular types. Moderately-differentiated HCC was detected in 46% and 43% respectively and poorly-differentiated HCC in 3% and 11% respectively. No correlation was found between number of nodules and the degree of Edmonson.

Oestrogen receptors and microsatellite instability in colorectal carcinoma patients.

About 10-15% of sporadic colorectal cancers show microsatellite instability (MIN), a mutator phenotype of mismatch repair genes. It seems that oestrogens may inhibit the pathway to colorectal carcinoma which involves a mismatch repair deficiency. Oestrogen receptorial status was evaluated in the neoplastic tissue and uninvolved surrounding mucosa of 17 MIN-positive and 33 MIN-negative tumours using an immunoenzymatic assay. MIN status was examined using the polymerase chain reaction and specific microsatellite markers. MIN was significantly associated with very low levels of oestrogen receptor in tumour tissue. Our findings suggest that MIN-positive tumours might lose a possible oestrogenic modulation mechanism.

COACH syndrome: report of two brothers with congenital hepatic fibrosis, cerebellar vermis hypoplasia, oligophrenia, ataxia, and mental retardation.

Congenital hepatic fibrosis (CHF) is probably the most common cause of non-icteric hepatosplenomegaly and is encountered mainly in children and young adults. We describe here two brothers from healthy, non-consanguineous parents. The patients showed early hepatosplenomegaly, portal hypertension, and no apparent kidney involvement. Clinical and laboratory findings were similar in both patients. Liver biopsies showed the presence of broad septa of fibrous tissue containing abundant bile ducts, portal tracts enlarged by fibrosis, and preserved lobular architecture. The histological findings were suggestive of CHF. Ophthalmological assessment demonstrated visual impairment with mild exotropia, nystagmus, and oculomotor apraxia. Neurological examination showed moderate mental retardation and cerebellar ataxia. Brain MRI confirmed cerebellar malformation with inferior vermis hypoplasia. This pattern of defects is consistent with COACH syndrome (Cerebellar vermis hypoplasia, Oligophrenia, congenital Ataxia, Coloboma, Hepatic fibrocirrhosis) which has previously been reported in five other cases. Our report may contribute to a better delineation of the COACH syndrome phenotype in the spectrum of oculo-encephalohepato-renal disorders.

Helicobacter pylori strains and histologically-related lesions affect the outcome of triple eradication therapy: a study from southern Italy.

BACKGROUND: Certain evidence suggests that Helicobacter pylori strains expressing genes for cytotoxin production show a higher sensitivity than non-cytotoxic organisms to eradication treatment. No data are available on the involvement of bacterium-related lesions in different therapeutic outcomes. AIMS: (i) To investigate whether differences in eradication rates may be related to the different expression of virulent strains (cagA, vacA, iceA) in patients undergoing proton pump inhibitor-based triple therapy, and (ii) to evaluate whether therapeutic outcome may be affected by bacterium-induced gastric lesions. METHODS: One hundred and ten H. pylori-positive subjects were enrolled. H. pylori was genotyped by polymerase chain reaction. Treatment consisted of lansoprazole-amoxicillin-clarithromycin, twice daily for 1 week. Eradication was checked by urea breath test. RESULTS: The eradication rate was 70%, and the absence of cagA was associated with unsuccessful treatment. No difference between the groups with successful and unsuccessful eradication was found with regard to vacA and iceA. Lympho-epithelial lesions and fibrosis were associated with unsuccessful treatment. CONCLUSIONS: The present data confirm the importance of cagA (but not vacA and iceA) as a predictor of successful eradication. When fibrosis and lympho-epithelial lesions are present, therapy appears to be less effective. Therefore, these histological features may be involved in an unsuccessful therapeutic outcome.

Gallbladder motility in vitro in men with gallstones following Billroth II gastric resection.

Gastric surgery induces an increased incidence of gallstones. To investigate the changes in gallbladder kinetics after gastric resection, 20 male patients were studied: ten patients undergoing cholecystectomy for gallstones developed after Billroth II gastric resection and ten patients undergoing cholecystectomy for cholelithiasis without previous abdominal surgery. Longitudinal strips from the gallbladder wall were suspended in an organ bath and the isometric tension recorded. Dose-response curves to cholecystokinin-octapeptide and carbachol were obtained. Half the maximal response to cholecysto-kinin-octapeptide was 0.50 +/- 0.11 x 10(-7) M in the first group and 1.36 +/- 0.37 x 10(-7) M in the second group (P < 0.05). The ED50 to carbachol was 24.33 +/- 2.69 x 10(-7) M in the gastrectomy group and 40.39 +/- 5.01 x 10(-7) M in the control group (P < 0.01). There was no significant difference in the maximal contractile response either to cholecystokinin-octa-peptide or carbachol in the two groups. Our study shows an increased gallbladder sensitivity to cholecystokinin-octapeptide and carbachol in patients with gallstones developed after Billroth II gastric resection.

Effect of somatostatin on human gallbladder motility: an in vitro study.

In vivo studies have demonstrated that somatostatin induces human gallbladder relaxation. To determine whether this polypeptide acts directly on the gallbladder muscle, its effect on strips of human gallbladder was studied in vitro. Strips of gallbladder were set up isometrically in an organ bath containing oxygenated Krebs' solution. Dose-response curves to cholecystokinin-octapeptide and carbachol were first established. The ability of somatostatin to cause relaxation under basal conditions and during 50% maximal stimulation by cholecystokinin-octapeptide (7.2 x 10(-8) M) and carbachol (3.5 x 10(-6) M) was assessed in 32 strips at 4.3 x 10(-6) M concentration which mimics the plasma concentrations found in patients with somatostatinoma and in 12 additional strips at 4.3 x 10(-8) M concentration. Somatostatin action on the intrinsic innervation by using electrical field stimulation (EFS) (200 mA 5 msec in duration, 30 Hz; 400 mA, 1 msec in duration, 10 Hz) was also evaluated in 39 strips. Somatostatin had no effect on the basal or carbachol-generated tensions. On the contrary, somatostatin (4.3 x 10(-6) M) reduced cholecystokinin-octapeptide-generated tensions by 8% (P < 0.001) and reduced EFS-generated tensions at 30 Hz by 7.7% (P < 0.01) and those at 10 Hz by 41.2% (P < 0.01). All responses to cholecystokinin-octapeptide and carbachol were abolished by dibutyryl-guanosine 3', 5'-cyclic monophosphate (5 x 10(-3) M) and atropine (10(-5) M), respectively (P < 0.0002 and P < 0.0002). All responses to electrical field stimulation were reduced or abolished by tetrodotoxin (2 x 10(-6) M) (P < 0.001 and P < 0.0001, respectively). Our findings show that somatostatin exerts its inhibitory action on the response to cholecystokinin-octapeptide and on the intrinsic innervation of the gallbladder smooth muscle. The probable neurotransmitter is the acetylcholine.

Immunohistochemical p53 overexpression correlated to c-erbB-2 and cathepsin D proteins in colorectal cancer.

The immunohistochemical overexpression of p53 protein in 42 large bowel cancers was correlated to c-erbB-2, cathepsin D (CD) proteins and other clinical and prognostic parameters. p53 overexpression (found in 60% of specimens) was positively associated with cathepsin D staining in stromal cells from older patients and better differentiated colorectal carcinomas (G1 + G2). Cytoplasmatic staining of c-erbB-2 protein was found in 58% of cases. No staining was observed at the cell membrane level. Our findings suggest that lower p53 expression in G3 carcinomas may be due to a high genomic instability, with the loss of both alleles of the gene. Therefore, these carcinomas were immunohistochemically silent. Although our series was small, the association between p53 nuclear neoplastic cells and CD stromal cells is interesting as regards the possible implications of these markers in colorectal cancer.

Localization of p53 protein and human papillomavirus in laryngeal squamous lesions.

The p53 tumour suppressor protein can be ineffective because of mutations in the p53 gene or interactions with proteins synthesized by specific subtypes of HPV. We investigated the localization of p53 protein in association with HPV in paraffin sections of 10 dysplastic and 12 malignant laryngeal squamous epithelium specimens by using immunohistochemical and in situ hybridization techniques. Viral HPV type 16 or 18 related sequences were identified only in a squamous cell carcinoma (SCC) specimen. p53 was detected in 64% of cases studied. All p53+ specimens showed no HPVrelated sequences; the only HPV+ case was p53 negative. In our study, the increased p53 expression in the process from dysplastic to invasive SCC indicates that p53 overexpression is an early event in laryngeal carcinogenesis. Moreover, the systemic susceptibility to HPV infection suggests the need for an accurate evaluation of SCC risk not only in the genital tract in female patients shown to be positive for transforming HPV types (16 or 18).

Gastric Malt lymphoma: a clinicopathological study.

The detection of HP infection by means of non invasive methods such as breath test and urease test, which are adopted alternatively to histological examination, can delay the diagnosis of early lymphomas. On the other hand the proved regression of gastric low grade Malt lymphomas after a successful HP eradication strongly suggests the need for early diagnosis. The aim of this study was to report the results of our experience with regard to clinical, endoscopic, histopathologic, and immunohistochemical features of gastric MALT lymphomas. We studied twenty-seven consecutive cases of gastric Malt lymphoma. HP presence in gastric mucosa was detected in all cases but endoscopy only in 40.7% of cases giving a diagnosis of suspected malignancy. In conclusion, it is very important to sample gastric mucosa in HP positive patients because the histological examination represents the most effective tool to detect lesions at the earliest and curable stage.

Immunohistochemical detection of p53 protein in anogenital lesions and its relationship with HPV status.

The p53 tumour suppressor protein can be rendered ineffective by mutations in the p53 gene or by interactions with proteins of DNA-transforming viruses, including Human Papillomaviruses (HPVs). Our aim was to determine whether the inactivation of p53, caused by a mutation of gene itself or by HPV is involved in anogenital carcinogenesis. We studied p53 overexpression by immunohistochemical methods, and HPV/DNA by non isotopic in situ hybridization method in 137 anogenital lesions. Immunoreactivity for p53 was seen in 5% of condylomata acuminata, in 12% of low-grade CINs, in 3.5% of high-grade CINs, and in 17% of invasive cervical carcinomas. Two (67%) of three condylomata acuminate p53+ harboured HPV/DNA. The concomitant presence of p53 and HPV was not detected in intraepithelial and invasive cervical lesions. Our findings suggest that p53 inactivation does not seem to play an important role in anogenital carcinogenesis. Further investigation of the concomitant presence of p53 and HPV in condylomata acuminate and its role in recurrences or progression of these lesions is needed.

Multiple synchronous colorectal carcinomas: a ploidy study by image analysis.

Cytometry could represent an ancillary technique to morphology in order to understand whether multiple synchronous colorectal carcinomas arose independently. Twenty-eight multiple synchronous tumours, assessed by means of a computerized image analysis system for DNA ploidy, were categorized as diploid (4) or not diploid (24). The ploidy classes were: DNA-diploid, DNA-tetraploid, and DNA-aneuploid. The DNA Index (DI) ranged from 0,90 to 2,66. The overall concordance rate for ploidy and DI class with synchronous tumours was 69% and 31%, respectively. The high concordance rate in ploidic categories suggests the metastatic origin of our multiple synchronous tumours. Moreover, they showed site, Dukes' classification, degree of differentiation, percentage of ploidy class, and DI distribution comparable to the single colorectal carcinomas. In conclusion, image analysis is a reliable technique to determine the independent clonality or the common origin of multiple colorectal tumours when the evaluation based on the simple histopathological criteria is not satisfactory.

Clinical pathological and ploidic patterns of juvenile colon cancers.

Details of 197 colorectal carcinomas from 180 patients were retrieved from our files, with the aim of analyzing the clinical histories and pathological parameters of all patients up to 40 years, and to support these data with the results of the DNA content analysis of tumoral cells by static cytometry. The incidence of juvenile carcinomas in our series was 7%, with a high percentage (38%) of cases associated with preneoplastic conditions. Our findings show a higher frequency of mucinous (21.5%), poorly differentiated (55%) and diploid (29%) neoplasias with a higher incidence of carcinomas localized in the right colon (50%) in young patients than found in elderly one. However, juvenile carcinomas did not differ significantly from neoplasias in the elderly population, because some pathological parameters considered as unfavourable prognostic factors in the formers (e.g. increased incidence of mucinous and poorly differentiated carcinomas) are "balanced" by features suggesting a good outcome (e.g. higher frequency of diploid neoplasias).

Overexpression of p53 in a large series of patients with hepatocellular carcinoma: a clinicopathological correlation.

p53 mutant protein has been found in a variety of human malignancies. In order to assess the controversial role of p53 protein in hepatocellular carcinoma (HCC) we studied its immunohistochemical expression in a series of 193 HCC specimens. Positive immunostaining for p53 was detected in the nuclei of neoplastic cells of 29 (15%) HCCs. There was no immunohistochemical evidence of mutant p53 expression either in normal or cirrhotic tissue surrounding neoplastic tissue. Higher alphafetoprotein serum levels were significantly associated with p53 overexpression. A prevalence of p53+ HCC specimens was seen in HCV negative patients (36% vs 13%, p < 0.05). No statistically significant correlations between p53 overexpression, age, sex, and HBV infection status were found. As regards histological grading, p53 was detected more frequently in tumours with poor cellular differentiation, although this finding does not reach statistical significance. The p53+ HCC rate was comparable to that attributed to the low incidence areas for HCC, in epidemiological studies. Moreover, p53 mutation seems to be related to the reactivation of alphafetoprotein gene to a more aggressive phenotype and to a later stage of liver carcinogenesis.

Helicobacter pylori in animals affecting the human habitat through the food chain.

Helicobacter pylori (HP) is the causative agent of many gastrointestinal diseases. Horses, calves, pigs, rabbits, and chickens were evaluated for HP presence, and the pathogenetic effect on their gastric mucosa. The large-sized animals all resulted positive. No positive cases were observed in rabbits and chickens. Chronic inflammatory response to the infection with the development of acquired lymphoid tissue associated to the mucosa was revealed. The recognition of HP in animals living near the human habitat such as animals for slaughter and for technical zootechnic and alimentary use, before the witnessing of the transmission of this infection such as a zoonosis or an anthropozoonosis, can contribute to research on a common source for human and animals as reservoir. It is possible to consider that the intraspecies transmission of infection occurs by vomit, the mucus acting as a vector, while the interspecies one is due to the faecal contamination of the food chain.

Different human papillomavirus genotypes in ano-genital lesions.

In order to verify the frequency and physical state of some viral strains of Human Papillomavirus (HPV) in anogenital lesions, two-hundred and four specimens were studied. HPV/DNA was detected by using non isotopic in situ hybridization method, HPV-DNA was found in 25 lesions. The integrated pattern of HPV types 16/18 was found only in invasive carcinomas, the episomic one in all high risk lesions, never in invasive carcinomas. The low oncogenic risk genotypes 6/11 were detected only in condylomata acuminata, the high oncogenic risk genotypes 16/18 were found not only in cervical intraepithelial and invasive lesions, but also in a condyloma acuminatum. Our findings confirm the importance of the viral genotypes in the evaluation of the risk for malignancy. Therefore, the detection of a high risk viral genotype, independently of its physical state, can evoke the ghost of the malignancy also in a low risk cervical lesion.