Uterine cervix metastasis from primary colon adenocarcinoma: a case report and review of the literature.

INTRODUCTION: Metastases to the female genital tract from extragenital primary tumors are unusual. We report a rare case of uterine cervix metastasis from colon adenocarcinoma and discuss diagnostic and therapeutic issues. CASE REPORT: We report a case of a 38-year-old North African Caucasian woman treated for a non-metastatic colon adenocarcinoma. She had a sigmoidectomy and incomplete adjuvant chemotherapy. Six months later, she consulted with vaginal bleeding caused by a cervical tumor, which was confirmed to be metastatic disease, and the patient underwent decompressive and hemostatic radiotherapy. CONCLUSION: Uterine cervix metastasis from primary colon adenocarcinoma is rare. The resection remains the standard protocol for the local treatment of resectable metastatic disease. Otherwise, systemic therapy is the preferable option.

Polymorphism of ftsI gene in Haemophilus influenzae and emergence of cefotaxime resistance in two Tunisian hospitals.

The decreased affinity to ß-lactams in Haemophilus influenzae is usually caused by specific alterations in penicillin-binding protein 3 due to varieties of substitutions in ftsI gene. This study aimed to characterize the polymorphism of ftsI gene in 19 H. influenzae strains, isolated between 2014 and 2016 (different resistance phenotypes to ß-lactams (n = 9) and susceptible strains (n = 10) used for comparative purposes). All strains were characterized for capsular type by PCR and agglutination tests and for ß-lactam resistance by amplification and sequencing of ftsI. Biotyping and clonality were performed by API-NH and pulsed-field gel electrophoresis, respectively. Four strains were ß-lactamase-negative ampicillin-resistant and five were ß-lactamase-positive clavulanic-acid-resistant. One strain from each group was resistant to cefotaxime. Our isolates belonged mainly to biotype IV and I and were non-typeable and genetically unrelated. According to mutation profiles of their ftsI, strains were classified as group I (n = 3), group II (n = 4), group-III-like (n = 1) and group III (n = 1). All group II strains were further classified as subgroup IIb, except for one strain, which harboured a new mutation (N422I). Ampicillin MICs of ß-lactamase-negative ampicillin-resistant strains were 6 to 12 times the MICs of susceptible strains. Only bla TEM-1 was detected in ß-lactamase-positive clavulanic-acid-resistant strains, and was responsible for high MICs for ampicillin (>256 mg/L), whatever the ftsI mutational resistance group. The emergence of cefotaxime-resistant isolates in our country is a matter of concern and requires strict surveillance and rationalization of antibiotic use to preserve these molecules.

Partial volume corrected 18F-FDG PET mean standardized uptake value correlates with prognostic factors in breast cancer.

AIM: The aim of this paper was to assess the prognostic role of pretherapy partial volume corrected (PVC) 18F-fluorodeoxyglucose mean standardized uptake value (SUV) in breast cancer (BC). METHODS: Forty oncological patients, BC diagnosed by biopsy, with breast tumor mass diameter >1 cm measured to the mammography, designed for surgical intervention, underwent a pretherapy semi-quantitative 18F-FDG positron emission tomography/computed tomography (18F-FDG PET/CT) whole-body study for tumor staging. Mean Body-Weight Standardized Uptake Value with Correction for Partial Volume effect (PVC- SUVBW-mean) was calculated in all mammary detected lesions. Excised tissues from primitive BC were sectioned and classified according to the WHO guidelines, evaluating biological features. Univariate (Mann-Withney/Kruskal-Wallis) and multivariate (linear regression, hierarchical clustering) statistical tests were performed between PVC-SUVBW-mean and biological indexes. ROC analysis was performed. PVC-SUVBW-mean thresholds were derived allowing to distinguish groups of BC patients with different biological characteristics. Specificity and Sensitivity were also calculated. RESULTS: Statistical and multiple correlations between pretherapy 18F-FDG PET PVC-SUVBW-mean and histological type, grade, ER/PgR hormone receptors and Mib-1 cellular proliferation index were found. In our samples, PVC-SUVBW-mean <≈4 g/cc was found correlated to BC patients with Invasive Lobular Carcinoma (ILC) or well differentiated Invasive Ductal Carcinoma (IDC), a positive expression of ER and PgR and a negative expression of MiB-1, while PVC-SUVBW-mean >≈7.00 is associated to BC patients with moderately and poorly differentiated IDC, negative expression of ER and PgR and a positive expression of MiB-1. CONCLUSION: Pretherapy PVC 18F-FDG PET PVC-SUVBW-mean measurement correlates with prognostic factors in BC and could be used to stratify patients before intervention.

Nonpalpable breast lesions: preoperative radiological guidance in radioguided occult lesion localisation (ROLL).

PURPOSE: This study evaluated the methods, technical aspects and impact of preoperative radiological guidance in radioguided occult lesion localisation (ROLL) for single nonpalpable breast lesions. MATERIALS AND METHODS: A total of 288 patients underwent ROLL before surgery. Human serum albumin macroaggregates labelled with 3.7-7.4 MBq of technetium(99) were injected into the lesion. In the case of ultrasonographic guidance (221/288 patients), inoculum positioning resulted in a change of echogenicity at the lesion site. In the case of mammographic guidance (67/288 patients), iodinated contrast medium was injected following the radiotracer for subsequent mammographic evaluation. Patients underwent surgery within 24 h from ROLL. A gamma-detecting probe was used to locate the lesion during surgery and guide its removal. After excision, the specimen was examined by either ultrasonography or mammography to verify complete lesion removal before histological evaluation. RESULTS: The lesion was correctly localised in 281/288 patients (97.5%). One ROLL procedure failed because surgery could not be performed within 24 h and the radioactivity decayed. Of the six incorrect localisations, 2 were due to the radiological guidance and 4 to technetium(99) dispersion. CONCLUSIONS: Radiological guidance in ROLL ensured the outcome of the procedure of localisation and removal of single, nonpalpable breast lesions in the majority of cases.

The role of DNA ploidy in postoperative management of stage I endometrial cancer.

BACKGROUND: Definition of high-risk stage I endometrial cancer (EC) patients who might benefit from adjuvant therapy (AT) is controversial. Decision is on the basis of traditional prognostic factors. We report our experience in which ploidy has found to play a role in clinical practice since 1999. PATIENTS AND METHODS: Two hundred and twenty-two patients with stage I EC with a median follow-up of 4.57 years were studied. After primary surgery, patients are chronologically divided in group A, from 1990 to 1998 (n = 141), receiving AT in IC stage and group B, from 1999 to 2003 (n = 81), receiving AT in case of DNA index >1.2 or stage IC grade 3 with unknown lymph node status. We analyzed prognostic factors, survival and relapse rate of the two groups. RESULTS: Since ploidy was introduced as a decision-making factor, only 30.6% (n = 11) of patients with stage IC received AT. Despite this considerable decrease of AT, no tumor-related deaths were reported in the group of patients with diploid IC stage who did not receive AT. Only DNA ploidy and age at diagnosis were independent predictors of overall survival. CONCLUSIONS: Our results indicate the important role of ploidy in order to identify high-risk patients who need AT and avoid overtreatment.

Identification of high risk patients with endometrial carcinoma. Prognostic assessment of endometrial cancer.

OBJECTIVES: To verify the importance of DNA ploidy on clinical outcome in endometrial carcinoma and to investigate whether the prognostic information obtained by this variable is independent from other clinical-pathologic features. MATERIALS AND METHODS: Univariate and multivariate analysis of clinical and pathologic prognostic factors obtained from 203 consecutive cases of endometrial cancer, that had been surgically treated in our hospital, were performed. RESULTS: Significant prognostic factors according to the Kaplan-Meier method were age at the time of diagnosis, grade of differentiation, peritoneal cytology, node involvement, vascular invasion, myometrial infiltration and ploidy. At multivariate analysis only DNA ploidy resulted to be an independent variable. CONCLUSIONS: In our analysis DNA content is the only parameter which preserved prognostic significance in multivariate analysis.

Adjuvant role of MIB1 index in differentiating serous ovarian tumors-preliminary report.

Atypically proliferating serous tumors (APST) account for 15% of all ovarian serous epithelial neoplasms. The differences between benign, borderline and malignant ovarian tumors is principally due to their cellular proliferative potential. By means of MIB1 expression we could recognize differences in proliferation among serous ovarian tumors, overcoming interobserver variability. Thirty-three patients with serous ovarian tumors, treated at S. Raffaele Hospital, University of Milan between November 1, 1992 and July 31, 1994 were used as study the population: 9 patients had serous cystoadenocarcinoma, 14 patients had APST and 10 patients had serous cystoadenoma. Pathological slides of all the cases were reviewed and immunohistochemical analysis was performed on formalin fixed, paraffin was embedded tissue. Pearson's Chi-square test and Fisher's exact test were performed for statistical evaluation. The percentage of MIB1 positive neoplastic cells ranged from, 0% to 2.1% (median 0.45%; mean 0.69%) in cystoadenomas, 1.3% to 7% (median 2.9%; mean 3.98%) in APSTs and 4.7% to 20.3% (median 6.95%; mean 9.51%) in cystoadenocarcinomas (p < 0.0001; F = 47.7). A High percentage expression of MIB1 in a serous tumor, initially diagnosed as APST, promoted a wider sampling of the surgical specimen confirming the presence of a carcinomatous component. MIB1 index is reported as representative of cellular proliferative potential. The analysis of MIB1 index provided valuable information in addition to that gained by conventional microscopic study in all cases where diagnostic difficulties arose in assessing APST.

Breast carcinoma detection with a combination of radiolabeled monoclonal antibodies. Promising results from immunohistochemistry studies.

BACKGROUND: Recent studies have demonstrated that the use of radiolabeled monoclonal antibodies (MoAbs) directed against tumor-associated antigens could help in the recognition of primary tumors, their extent, and their metastases by external scintigraphy (used preoperatively) or by hand-held gamma-detecting probe (GDP) (used intraoperatively). METHODS: The authors evaluated carcinoembryonic antigen (CEA), c-erb B-2 protein, and TAG-72 expression in 100 cases of breast carcinoma using F023C5 (anti-CEA), B72.3, and anti-c-erb B-2 protein MoAbs that were previously investigated for their usefulness in radioimmunoguided surgery and external scintigraphy. The goal of this study was to examine the biodistribution of each antibody in primary, multifocal, and metastatic lesions to evaluate the suitability of their simultaneous use in GDP and external scintigraphy. RESULTS: Results showed immunoreactivity for c-erb B-2 protein in 39 of 99 cases, for B72.3 in 41 of 100 cases, and for CEA in 15 of 100 cases. Multifocal lesions demonstrated positivity for c-erb B-2 protein in 37.4% of cases (6 of 16), for B72.3 in 68.8% of cases (11 of 16), and for CEA in 6.2% of cases (1 of 16). In lymph node metastases, immunoreactivity was found for c-erb B-2 protein in 36.4% of cases (12 of 33), for B72.3 in 63.7% of cases (21 of 33), and for CEA in 24.3% of cases (8 of 33). When the authors considered the immunoreactivity of all three MoAbs, the percentage of positive cases they observed was 60% in primary tumors (60 cases), 78% in lymph node metastases, and 81.2% in multifocal lesions. CONCLUSIONS: These results suggest that in vivo tumor radioimmunodetection could be improved by the use of more antibodies directed against different tumor-associated antigens.

Flow cytometric analysis of c-myc oncoprotein in non-small-cell lung carcinoma: comparison with immunohistochemical results.

c-myc oncoprotein expression was evaluated in 36 non-small-cell lung carcinomas using immunohistochemical and flow cytometric analysis. Formalin fixed and paraffin embedded material was used. The same monoclonal antibody (mouse anti-human c-myc) was employed both for immunohistochemistry and flow cytometry. For the immunohistochemical evaluation we calculated the percentage of stained cells on 200 neoplastic cells. c-myc oncoprotein was measured using flow cytometry by linking the monoclonal with a secondary FITC-conjugated antibody; for each sample 20,000 events were analysed and the percentage of cells positive for green fluorescence was calculated. DNA content was obtained by propidium iodide staining. Results showed a high correlation between immunohistochemical and cytometric data, suggesting that flow cytometry could be used as an alternative to immunohistochemistry in evaluating nuclear antigens. Moreover, using flow cytometry information on DNA content can be obtained simultaneously.

Role of DNA ploidy and ERB-B2 oncogene expression in the prognosis of endometrial carcinoma.

Seventy-three patients with endometrial carcinoma have been retrospectively evaluated in this paper. Stage, grade, depth of myometrial invasion, flow cytometric DNA ploidy and ERB-B2 oncogene expression by immunohistochemical method have been analyzed on paraffin embedded tissue. Results showed the existence of a significant correlation between stage and grade of neoplasia and between DNA ploidy and course of disease; it has been observed that patients with aneuploid tumor tend to have a shorter time before relapse of disease. No significant correlation between depth of myometrial invasion and survival was found. Besides, it has been shown that tumours with ERB-B2 oncogene hyperexpression seem to have a more aggressive evolution.

Avidin-biotin system in radioimmunoguided surgery for colorectal cancer. Advantages and limits.

PURPOSE: Radiolabeled monoclonal antibodies (MAbs) have been reported to allow tumor intraoperative detection by means of a gamma-detecting probe. The technology is called the Radioimmunoguided Surgery (RIGS) system. The main inconveniences of the method are 1) the long interval needed for clearance of unattached MAbs from the patient's body, between the injection of the MAb and surgery, and 2) the low sensitivity of current MAbs used in detecting small tumors. We describe a new method to overcome these inconveniences using biotinylated MAbs and avidin in order to obtain a rapid blood clearance of the radiolabeled MAbs both anticarcinoembryonic antigen and antitumor-associated glycoprotein-72 MAbs. METHODS: Twenty patients with primary and recurrent colorectal cancer have been enrolled in the study; 125I-biotinylated MAbs FO23C5 (anticarcinoembryonic antigen) and B72.3 (antitumor-associated glycoprotein-72) followed by cold avidin were injected in 13 patients and 7 patients, respectively. RESULTS: A decrease of 94 +/- 3 percent of circulating radioactivity was achieved in 3 to 5 days. Patients underwent surgery approximately seven days after MAb injections rather than after four weeks. Tumors were localized in 14/20 (70 percent) patients (true positive), 2 (10 percent) were false negative, and 4 (20 percent) were true negative. The overall sensitivity level in early-stage primary cancers was 37 percent when related to the presence of disease and 75 percent when related to antigenic expression. The sensitivity for more advanced cancer and for recurrences was 100 percent. Moreover, the in vivo tumor targeting of biotinylated MAb was demonstrated in frozen tumor section by direct streptoavidin-peroxidase staining. CONCLUSIONS: The avidin-biotin system may enhance applicability and effectiveness of radioimmunoguided surgery (RIGS).

[Correlations between the HPV type, DNA index and evolution of cervical intraepithelial neoplasia (CIN)]. / Correlazione tra tipo di HPV, DNA index ed evoluzione delle neoplasie intraepiteliali della cervice (CIN).

Correlation between human papillomavirus infection and DNA ploidy in the prognosis of uterine cervical intraepithelial neoplasia. Due to the increasing frequency of human papillomavirus (HPV) infection of cervical epithelium in patients with cervical intraepithelial neoplasia (CIN) of different grades, methods are needed to identify progressive lesions. HPV typing as well as quantitative DNA analysis are possible tools to identify high-risk lesions. The aim of our study was to compare the results of "in situ" hybridization and of DNA content analysis with behavior of CIN I and CIN II lesions in 14 patients with HPV infection. Three of the 4 cases with regression of the cervical lesions were diploid, with HPV 16/18 detected in 2 of the 3. The 5 cases with progression were aneuploid, and 4 of them were HPV positive. In the 3 cases with no changes a near-diploid DNA content was observed. DNA analysis was not available in 2 cases. These preliminary results suggest that progressive CIN cases are aneuploid, and that DNA ploidy could be an objective prognostic marker.

Reproducibility of FCM-DNA ploidy analysis in prostatic cancer: comparison between needle biopsy and surgical specimens.

Cytofluorometric analysis of FCM-DNA ploidy also seems to have prognostic value in prostatic carcinoma. Determination of the DNA index preoperatively from needle biopsy samples would be advantageous since it would enable the type of intervention to be established in advance. The finding of intratumoural variability in the DNA index in different solid tumours prompted us to compare the data obtained from needle biopsy and surgical specimens of prostatic cancer. Although the results obtained confirmed the heterogeneity of DNA content in prostatic carcinoma, they nonetheless indicate that preoperative DNA analysis is useful, in particular when aneuploidy is observed, since it provides the clinician with additional information on which to base treatment decisions.

Localization of biotinylated monoclonal antibodies (B72.3 and F023C5) in patients with colorectal cancer.

The recognition and treatment of neoplastic lesions in early phases are among the most important aims of research using monoclonal antibodies (MoAb). Recent studies have demonstrated that the use of radiolabelled MoAb directed against tumor associated antigens and hand-held gamma detecting probe (GDP) could help in the recognition of primary tumors and metastases. The goal of this study was to investigate the in vivo antibody reaction as shown by histologic preparations after injection of 125I biotinylated MoAb (B72.3 or F023C5) before surgery and to compare the immunohistochemical results with those obtained with GDP in colorectal cancer. We studied 15 cases of patients with primary or recurrent colorectal cancer. The biotinylated, radiolabelled antibody administered in vivo could be seen in frozen sections with streptoavidin peroxidase complex. In 14 cases there was agreement between GDP observations and detection of the in vivo injected biotinylated MoAb with direct staining using streptoavidin peroxidase conjugate. The use of MoAbs thus provides a means of correlating the intraoperative signal with the presence of the injected antibodies on the tumor.

[The use of flow cytometry in the study of parathyroid hyperplasia and adenomas]. / L'impiego della citofluorimetria nello studio delle iperplasie e degli adenomi paratiroidei.

To discriminate benign from malignant parathyroid glands lesions is often difficult, because the morphologic features lack absolute diagnostic value. The differential diagnosis between chief cell hyperplasia and parathyroid adenoma is frequently based on physical features such as increased gland weight and number of enlarged glands. A carcinoma is identified by the evidence of local invasion, metastases or recurrence. Nevertheless the lesions classified as benign for lack of histologic features of aggressiveness can show nuclear atypia, increased mitotic figures and broad fibrous bands. Since DNA aneuploidy is present in a great number of human neoplasms and DNA aneuploidy has been suggest to be a marker of malignancy, flow cytometric assessment of ploidy appeared a possible method for rapid and objective distinction between benign and malignant lesions. Flow cytometric DNA content was evaluated on 113 parathyroid glands: the parathyroids were resected from 26 patients with hyperparathyroidism and from 17 patients with adenoma. The analysis was performed on paraffin-embedded specimens according to Hedley with minor modifications. Interpretable histograms were obtained for 103 parathyroids gland (mean CV = 5.3). Aneuploidy was detected in 22.5% of glands; in 12 instances of parathyroid hyperplasia the glands of the same patient showed different DNA Indexes. Cytometric results and morphological features do not correlate as far as aneuploidy and cellular atypia are involved. Although our results fail to show any correlation between morphology of parathyroid cells and DNA content, and abnormal DNA content suggests a careful follow up of these patients.

Flow cytometric diagnosis of partial mole.

To differentiate histologically partial hydatidiform moles (PM) and complete hydatidiform moles (CM) may be difficult. Cytogenetic studies have shown that PMs often had a triploid karyotype while CMs were always diploid. We assessed the DNA content of 31 paraffin-embedded cases of trophoblastic disease with flow cytometry. Twenty-four cases were histologically diagnosed as PM, 3 cases as CM; the others as hydropic abortion (2 cases), choriocarcinoma (1 case), and persistent trophoblastic disease (1 case). Four normal term placentas were used as diploidy controls. In 9 cases the results of the cytogenetic analysis were available. All placental specimens included also maternal tissue as an internal control. Eight of the 24 histologically diagnosed PMs were triploid; there was agreement in 8 cases out of 9 (90%) between the flow cytometric analysis and the karyotypic determination of ploidy. All normal controls as well as the hydropic abortion, the CM and the persistent trophoblastic disease were diploid. Abnormal content of DNA (DI = 1.3) was observed in the choriocarcinoma. Our results show that flow cytometric analysis of DNA content is a reliable and fast method of diagnosing PM on paraffin-embedded material.

Two-step tumour targetting in ovarian cancer patients using biotinylated monoclonal antibodies and radioactive streptavidin.

A new method for intraperitoneal tumour targetting in ovarian cancer using biotinylated monoclonal antibodies (MoAb) and radioactive streptavidin is described. Fifteen patients with histologically documented ovarian carcinoma were injected intraperitoneally with 2 mg of biotinylated MoAb MOv18, followed 3-5 days later by 100-150 micrograms of indium-111 streptavidin, at the specific activity of 280-370 MBq/mg in 500 ml of normal saline. No toxicity was observed. Tumours were imaged from 2 to 48 h after radioactivity injection by recording both planar and single photon emission tomography (SPET) data. All patients underwent surgery 1-8 days later (mean 3 days) after scanning. The resected tumour and normal tissue radioactivity were measured. On the day of surgery, the tumour to normal tissue ratio was 9:1 (range 3:1-30:1) and 45:1 (range 12:1-120:1) for intra- and extraperitoneal samples, respectively. The mean tumor to blood ratio was 14:1 (range 4:1-30:1). The injected dose (i.d.) per gram of tumour was 0.112 (range 0.01-0.3) for recurrences and 0.05 for primary tumour (range 0.005-0.2). Over 24-48 h 14% i.d. (range 8-18% i.d.) was found in the urine, 14% i.d. (range 6-29% i.d.) in the blood and 63% i.d. (range 56-70% i.d.) was still in the peritoneal cavity. These preliminary clinical data suggest that this two-step strategy may be superior to the conventional approach (radiolabelled antibodies) for intraperitoneal radioimmunolocalization and radioimmunotherapy of ovarian cancer.