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.

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.

Perception of dyspnea during exercise-induced bronchoconstriction.

UNLABELLED: After strenuous physical exercise, many subjects show a significant bronchoconstriction and report dyspnea. Despite this clinical condition being a commonly encountered situation during daily life, which may be responsible for substantial disability there is little information on the relationship between the perception of dyspnea and exercise-induced bronchoconstriction (EIB) after a standardized exercise challenge. For these reasons, we evaluated 200 consecutive outpatients (median age 13 years, ranging from 5 to 56 years) referred to our laboratory to perform an exercise test out of suspicion of EIB. On exercise challenge, perception of dyspnea was rated on a modified bipolar Borg scale immediately before each FEV1 measurement. Sixty-nine (35%) subjects had a positive exercise challenge, defined as a decrease of at least 20% in FEV1 from baseline. Both the onset and the decay of dyspnea preceded those of bronchoconstriction. Overall, the rating of dyspnea in the laboratory was well related with the reports of exercise-related symptoms. Similarly, 36 of 77 (47%) asthmatics with a history of exertional symptoms and 24 of 65 patients (40%) without a history had a positive challenge. Asthmatics reporting exertional symptoms perceived a greater magnitude of dyspnea after exercise independently from the degree of bronchoconstriction. Overall, dyspnea was significantly but loosely correlated to the magnitude of decrease in FEV1, being also influenced by age, gender and BMI. CONCLUSIONS: We conclude that dyspnea recorded in the laboratory after exercise test is related to exertional symptoms reported during real life, but not completely related to EIB. The rating of dyspnea is a well-suited model to study naturally occurring exercise-induced dyspnea and a useful tool to enlarge the results of an exercise challenge.

GENEBU Project. Equipment and drugs used for home nebulizer therapy in Italy.

The GENEBU Project is an open, observational survey evaluating home nebulizer practices in Italy. It consecutively included patients who were referred to one of the 27 participating chest clinics from May to December 1999 and who had been using a home nebulizer in the previous six months. The information source was a self-administered questionnaire compiled by the enrolled subjects. We collected 1257 questionnaires. The nebulizer equipment was heterogeneous, with at least 92 different models. Jet nebulizers were 90% of the total; 53% of these had a glass reservoir. Almost 80% of the patients selected the nebulizer themselves without any medical advice. In addition, most patients (> 80%) did not receive information on both the interface system and the optimal fill volume of the nebulizer. Corticosteroid nebulisation was widespread (74%), for both occasional and regular daily use, for both acute and chronic diseases from upper to lower airways. Beta 2-agonist (55%), anticholinergic (37%), mucolytic (32%) drugs were also often nebulised. More than 90% of patients mixed some active drugs. We conclude that the nebulizer equipment for home aerosol therapy was very heterogeneous and, probably, not always utilised at its best in Italy. The mixing of drugs and the widespread use of corticosteroids were peculiarities of home nebulizer therapy in Italy.

Increased oxidative stress in dimethylnitrosamine-induced liver fibrosis in the rat: effect of N-acetylcysteine and interferon-alpha.

Oxidative stress may represent a common link between chronic liver damage and hepatic fibrosis. Antioxidants and interferon seem to protect against hepatic stellate cell (HSC) activation and liver fibrosis. This study evaluated (1) the effect of the profibrotic agent dimethylnitrosamine (DMN) on the hepatic oxidative balance in the rat; (2) the role played by the antioxidant agent N-acetylcysteine (NAC); and (3) the antifibrotic effects of two different types of interferon-alpha: recombinant alpha-2b (rIFN-alpha) and leukocyte alpha (LeIFN-alpha). Five groups of rats received: (1) saline; (2) DMN; (3) DMN + NAC; (4) DMN + rIFN-alpha; and (5) DMN + LeIFN-alpha. Oxidative balance was evaluated by hepatic glutathione, TBARs, protein carbonyl, and sulfhydryl determination. Fibrosis was determined by hepatic hydroxyproline content and fibronectin (FN) staining (immunohistochemistry). DMN rats showed a diffuse FN deposition, an impaired oxidative balance, and higher hepatic hydroxyproline levels compared to that of controls. NAC administration significantly reduced FN deposition, increased hepatic glutathione, and decreased TBARs and protein carbonyls. Administration of IFN-alpha exerted different effects according to the type used. Both IFNs decreased FN deposition; however, LeIFN-alpha significantly improved histology and oxidative parameters compared to those of untreated DMN and rats treated with rIFN-alpha. This study shows the role of free radicals in this model of hepatic fibrosis; the protective effect of NAC against liver fibrosis; and the antifibrotic effect exerted by IFN-alpha (particularly LeIFN-alpha) independent of its antiviral activity.

Safety and effectiveness of lomefloxacin in patients with acute exacerbation of chronic bronchitis (AECB) chronically treated with oral theophyllines.

Lomefloxacin is a difluorinated quinolone with excellent activity against a wide range of pathogens including those responsible for acute exacerbations of chronic bronchitis (AECB). This open, cross-sectional, multicenter study has evaluated the efficacy and safety of a once-daily dosage of 400 mg lomefloxacin in patients with AECB chronically treated with theophylline. 137 patients (96 males, 41 females; mean age 66.1+/-11.2 yrs) were enrolled and 133 completed the study. 81% suffered from moderate AECB, 16% severe AECB. The clinical success rate was very high (95%), as well as the microbiological (93%). Side effects were scarce and were significant only in 3 patients, with 2 dropouts. All patients were using theophylline derivatives twice daily and continued without any variation in dosage during the lomefloxacin treatment. Theophylline plasma levels determined in 103 patients at baseline, during and at the end of the lomefloxacin treatment did not significantly change. We conclude that orally administered lomefloxacin at standard recommended dosage is well tolerated and effective in elderly patients with AECB. No dose adjustment is required even when it is co-administered with methylxanthines.

Detection of Helicobacter pylori cagA gene by polymerase chain reaction in faecal samples.

OBJECTIVE: The polymerase chain reaction (PCR) has been extensively and successfully used to detect Helicobacter pylori in gastric juice and gastric biopsies. In contrast, the results obtained using faeces as biological samples for PCR are rather conflicting. This may be due to the presence of faecal inhibitory compounds (polysaccharides) which can inhibit the amplification reaction. The aim of this study was to characterize the H. pylori genotype in faecal samples by using specific primers for the cagA gene. To overcome the problem of contamination by polysaccharides, we used a filter-based extraction technique already applied in a previous study. METHODS: Antral and body biopsies were obtained from 30 symptomatic patients undergoing upper endoscopy. PCR was used to detect the presence of H. pylori organisms in faecal samples by using primers selected for the urease gene A. In addition, H. pylori organisms were characterized both in faecal samples and paraffin-embedded biopsies by PCR with specific primers for the cagA gene. RESULTS: All patients showed a positive CLO test (rapid urease test) and evidence of H. pylori by Warthin-Starry stain. PCR detected the urease A gene in the faecal samples of all patients. The cagA gene was detected in the faecal and biopsy samples of 18 subjects (60%). Duodenal ulcer and/or antral erosions were observed in 15 of the 18 cagA-positive patients (83.3%) and in five of the 12 cagA-negative patients (41.7%). Endoscopic features of normal mucosa or gastritis were observed in three cagA-positive patients (16.7%) and in seven cagA-negative patients (56.3%). cagA-positive status was found to be significantly related to the endoscopic features of duodenal ulceration and/or antral erosions. CONCLUSIONS: Our findings prove that faeces are suitable samples for the detection of cagA status. Moreover, they confirm the existence of a significant relationship between cagA-positive status and duodenal ulcer and/or antral erosions.

Programmed cell death in colorectal carcinogenesis.

The most studied mechanism of malignant transformation has been cell proliferation. The relationship between programmed cell death (apoptosis), cell proliferation, and apoptosis regulatory genes (p53 and bcl-2), was studied in normal colonic epithelium, 26 sporadic adenomas both early and late, 25 FAP adenomas, and 34 carcinomas. We showed a decrease in programmed cell death and an increase in cell proliferation during the transition from adenoma to carcinoma. The increase of expression of p53 from early (10%) to late adenomas (87%) contrasted with the decrease of bcl-2 staining. Sixty-two per cent and 23% of carcinomas were reactive for p53 and bcl-2 respectively. Abnormal early activation of the bcl-2 gene, rather than late p53 gene mutation appears to be responsible for inhibition of apoptosis in colorectal carcinogenesis. bcl-2 was higher in FAP adenomas than in sporadic cases, and in carcinomas favouring the accumulation of long-living cells, which are more subject to mutation and thus cancerization.

[Core biopsy in the preoperative diagnosis of breast carcinoma]. / La "core biopsy" nella diagnostica preoperatoria del carcinoma mammario.

BACKGROUND: In spite of the widespread use of cytological smears for the diagnosis of breast cancer lesions, many surgeons are still reluctant to accept the cytological report as the only criterion for performing definitive surgery. Modern surgical strategy requires a preoperative planning of the surgical treatment which is possible with the use of core biopsy (CB) or FNB (fine needle biopsy) for the diagnosis of tissue specimens. METHODS: The authors report their five-year experience with this technique and evaluate its diagnostic usefulness and substitution of interoperative biopsy. The histological examination of 92 palpable breast lesions, clinically and mammographically detected, was performed with CB, and diagnosis was confirmed with the surgical sample in 80 cases. RESULTS: A definitive histological diagnosis was obtained with CB in 90% of cases. Only nine cases required confirmation with frozen section diagnosis at the time of definitive surgery. The sensibility of CB was 93%, specificity and predictive value of a positive result were 100%, and diagnostic efficiency was 95%. CONCLUSIONS: This study confirms the usefulness of the systematic use of CB for accurate preoperative diagnosis of breast cancer, and the simplicity, safety and accuracy of this method also make ultrasound-guided CB applicable to non-palpable lesions.

Core Biopsy as Alternative to Fine-Needle Aspiration Biopsy in Diagnosis of Breast Tumors.

In spite of the widespread use of cytological smears for diagnosis of breast cancer lesions, many surgeons are still reluctant to accept the cytological report as the only criterion for performing definitive surgery. Modern surgical strategy requires a preoperative planning of the surgical treatment, possible through the use of core biopsy, which provides a diagnosis based on tissue specimens, thus permitting the study of both the architectural and cytological patterns. The authors report their five-year experience with this technique and evaluate its diagnostic usefulness and ability to reduce intraoperative biopsy procedures. The histological examination of 92 palpable breast lesions, clinically and mammographically detected, was performed with core biopsy, and diagnosis was confirmed with the surgical sample in 80 cases. A definitive histological diagnosis was obtained with core biopsy in 90% of cases. Only nine cases required confirmation with frozen section diagnosis at the time of definitive surgery. The sensibility of core biopsy was 92%, specificity and predictive value of positive result were 100%, and diagnostic efficiency was 86%. This study confirms the usefulness of systematic use of core biopsy for definitive preoperative diagnosis of breast cancer; the simplicity, safety and low cost of this method also make ultrasound-guided core biopsy applicable to nonpalpable lesions.

A rare association between ovarian endometriosis and bilateral ovarian teratoma. Case report.

A 28 year woman complained for a sudden abdominal pain with some of the classical signs of the acute abdomen. Since a bilateral ovarian mass was found at ultrasound examination, clinician decided that patient underwent a urgent surgery. At histology, this mass resulted to be a bilateral triphillic mature teratoma. A focus lined by endometrioid epithelium and embedded in the modified stroma of endometrial type, was found beyond the residual cortex of the left ovary. There was some fibrotic tissue separating the endometrial and teratomatous lesions. To our knowledge, these two diseases have never been reported as associated in literature, but this association has a clinical relevance because an endometriotic pathology can reveal a silent teratoma with bilateral ovarian localization.

[The risk factors for hepatitis D viral infection in southern Italy]. / Fattori di rischio di infezione da virus dell'epatite delta nel Sud Italia.

The aims of this study were to evaluate the prevalence of hepatitis delta virus (HDV) infection and risk factors associated to it. Three hundred sixty-one HBsAg chronic carriers from southern Italy were studied and 13.8% of them resulted anti-delta positive. 80% of these subjects were less than 50 years old. When anti-delta positive subjects were compared with anti-delta negative ones, a lower number of healthy HDV carriers and a higher frequency of cirrhotics were noted among anti-delta positive. Of lower than 50 years, imprisonment, sexual contacts with drug abusers and male homosexuality were risk factors of HDV infection. No association was found with sex, household contacts with HBV or HDV carriers, number of family members and transfusion of blood products. These data confirm the high prevalence of HDV infection in southern Italy.

The immunohistochemical expression of cathepsin D in colorectal cancer.

It has been suggested that cathepsin D expression in stromal cells affects the prognosis of breast cancer. With reference to colon and breast cancer, this study verified cathepsin D immunostaining in epithelial and stromal cells of primary tumours and lymph-node metastases from 46 colorectal carcinomas. Eight of 46 cases (17%) were cathepsin D+ both in cancer and stromal cells. No staining was found either in cancer or stromal cells of the remaining cases. The results presented here suggest the possible paracrine influence of another diffusible factor produced by cancer cells which stimulates cathepsin D production in stromal and cancer cells.

The efficacy and safety of moguisteine in comparison with codeine phosphate in patients with chronic cough.

We conducted a multicentre, double-blind, parallel group study to compare the clinical efficacy of a new antitussive drug, moguisteine (100 mg t.i.d.), to that of a reference standard, codeine (15 and 30 mg, t.i.d.). Both drugs were given orally for a period of two days. A group of 119 patients (mean age 54 yrs; 61 females and 58 males) with chronic, dry or slightly productive cough, associated with various respiratory disorders (including chronic obstructive pulmonary disease, respiratory malignancies and pulmonary fibrosis) were enrolled at six participating centres. The percentage reduction in the number of morning coughs over a period of 6 h after the first administered dose compared to baseline assessment, was 21% with moguisteine (n = 39), 28% with codeine 15 mg (n = 38), and 29% with codeine 30 mg (n = 36). Differences between treatments were not significant. The percentage reduction in the number of nocturnal coughs per hour, after the last evening dose compared to baseline assessment, was 33, 46 and 52%, respectively. Subjective assessments (patients' visual analogue scale scores of cough frequency, cough intensity and sleep disturbance, and investigators' ranking of cough severity) indicated that there was a similar improvement in cough symptoms in all treatment groups. Adverse events were observed in two patients on moguisteine, three on codeine 15 mg, and five on codeine 30 mg. No event was serious, but discontinuation of treatment was required in two patients on codeine 30 mg. The results of our study suggest that moguisteine 100 mg t.i.d. is safe, and seems to have an antitussive activity similar to that of codeine 15-30 mg t.i.d.

P-170 glycoprotein expression in gastric and colorectal carcinomas and normal mucosa. An immunocytochemical study.

During the past few years it has become apparent that simultaneous resistance of tumour cells to a number of heterocyclic drugs (multidrug resistance) is often correlated with overexpression of a P-glycoprotein (P-gp or P-170). P-gp expression can be studied by molecular biology and immunohistochemical techniques. The latter provide a rapid, sensitive and specific screening method suitable for testing even a relatively small number of tumour cells like those obtained at biopsy. The aim of this study was to detect and localize the immunohistochemical expression of P-gp in normal and neoplastic gastrointestinal tissue using the Mab JSB-1 in formalin-fixed paraffin-embedded specimens. A particularly striking finding of our study was the consistent, prevalent higher expression of P-gp in the stomach than in the colon, with a higher percentage of immunostaining in normal than in neoplastic tissue. This is in agreement with the fact that not only is the prognosis known to be worse for stomach cancer, but also the response to treatment is lower. Further studies should be carried out to verify the possibility of making routine tests of this kind for the evaluation of multidrug resistance, to guide the selection of patients for treatment of cancer with chemotherapeutic drugs.

Carcinoma and synchronous hyperplastic polyps of the large bowel.

The prognostic and biological meaning of the association of colonic cancer with hyperplastic polyps (HP) is not as well known as that with adenomatous polyps (AP). In order to gain some insights into this matter, we have retrospectively studied two hundred and twelve patients with colon-rectal carcinoma in which 64 (30.18%) had synchronous AP, 24 (11.32%) had synchronous HP, 13 (6.13%) had both AP and HP and 111 had no synchronous polyps (52.36%). The 34 cases of synchronous HP, whether or not associated with AP, were located in the same colonic segments of the cancer and were found usually in the sigmoid-rectum. The AP were found throughout the colon-rectum with a similar rate of association with the cancer in each segment. The cancer associated with HP have a higher prevalence in the better prognostic stages of both Dukes and Jass-Morson systems. Conversely both AP and AP+HP associated cancers exhibit prevalences rates higher in the worst prognostic stages. Our observations suggest that separate factors might promote the growth of HP and AP and that a relationship between colonic cancer and synchronous HP might exist and differ from that demonstrated for AP and colonic cancer.

[DNA content in colorectal carcinomas: application of image analysis to archival material]. / Contenuto di DNA nei carcinomi colo-rettali: applicazione dell'analisi di immagine su materiale di archivio.

The most studied biological parameter of human tumours is the DNA nuclear content. In the present study we assessed the DNA content of 197 colorectal cancers from 180 patients using archival material (tissues routinely processed and paraffin-embedded) for image analysis. The metastatic lymph nodes were also included in the study in 54 Dukes "C" stage cases and 19/25 Dukes "D" stage cases. A total of 452 sections, 5 mu thick, were submitted to image analysis in static cytometry. DNA diploid pattern was found in 72/452 samples (15.9%) and the DNA-aneuploid pattern in 380/452 (84.1%). Two different areas for each tumour were examined but we found a concordance of diploid pattern in only 23 cases. DNA-aneuploidy, regarded as presence of at least one sample with an aneuploid peak in the same tumour, reached 88% in our study. In conclusion the image analysis of several samples obtained from different areas of the same tumour clearly revealed: a pronounced heterogeneity of ploidy patterns in colon cancer; an overall prevalence of DNA-aneuploidy in colon cancer; a prevalence of diploid pattern in tumours localized in right portion of the colon and in younger patients.

[Granulomatous hepatitis in a hospital population in southern Italy]. / Epatiti granulomatose in una popolazione ospedaliera dell'Italia meridionale.

In a southern Italy hospital, in five years 1523 liver biopsy specimens have been performed and histologically examined. Granulomas are found in specimens from 15 patient (1%). They are seven females and eight males with an average age of 57 years (range 43-71). Seven of the 15 specimens are Menghini-type percutaneous needle, five are surgical and three are laparoscopic bioptic specimens. Four patients are correlated with infectious diseases: 2 with hepatitis C virus (HCV), 1 with hepatitis B virus (HBV) and 1 with Mycobacterium Tuberculosis. In three patients the diagnosis is primary biliary cirrhosis (PBC), in two sarcoidosis, in other two pseudosarcoid reaction to abdominal tumours (a gall-bladder cancer and a non-Hodgkin lymphoma of the stomach). Finally there are 2 lipogranulomas, 1 foreign-body granuloma and 1 cholesterin granuloma. This work underlines the high prevalence in our series of PBC and sarcoidosis in the etiology of hepatic granulomas and the high frequency of patients with markers of HCV or HBV in granulomatous hepatitis.

The m1g8 monoclonal-antibody - a comparison between paraffin-embedded and frozen-sections from breast-cancer.

Till now the immunohistochemical studies on the procathepsin D expression in breast cancer have been performed using polyclonal antibodies on paraffin-embedded sections because the monoclonal ones were considered effective only on frozen sections. In this study we compared the MAb M1G8 reactivity, which recognizes the 52 Kd procathepsin D, on a series of frozen and paraffin-embedded sections of the same specimen from 22 breast cancers, using the immunohistochemical method. This methodological choice was due to the necessity to perform further studies with monoclonal antibodies using archival material to clarify the procathepsin D prognostic role. MAb M1G8 was clearly positive with a 100% concordance rate both in frozen and paraffin sections of the same specimen.

Immunohistochemical expression of p53 paraffin sections from colon-cancer specimens - comparison between 2 new monoclonal-antibodies.

The early monoclonal antibodies (MAbs) to p53 are considered reliable only on frozen sections. We examined the immunohistochemical expression of p53 in 40 paraffin-embedded samples obtained from colon carcinomas using the two new MAbs DO-1 and DO-7. Thirty-one frozen samples were obtained from the same surgical specimens and were stained using MAb PAb 1801 which has a high specificity to p53. Fifty-five percent of 40 paraffin-embedded samples was positively stained with both MAbs DO series and showed a 100% concordance rate while 52% of 31 frozen samples was positively stained with MAb PAb 1801. The concordance rate between the reactivity of MAbs DO series and that of MAb PAb 1801 was 98%. Therefore, the reactivity of MAbs DO Series in paraffin-embedded material is similar and reflects accurately the overexpression of p53 when compared with the reactivity of MAb PAb 1801. Further investigations, moreover, can be performed using the MAbs DO series on archival material for a full understanding of the p53 involvement in human cancer.