Assessment of response to therapy in multiple myeloma by multiparameter flow cytometry. Usefulness of an eight-color single tube with monoclonal antibodies in dried formulation.

OBJECTIVE: Multiparameter flow cytometry is considered the gold standard to evaluate minimal residual disease in multiple myeloma (MM) and patients in complete remission can achieve "Flow MRD-negative" status (i.e. immunophenotypically abnormal plasma cells not detectable). In the current study we report the usefulness of an eight-color flow cytometric method with a 10-5 sensitivity, using monoclonal antibodies in dried formulation. MATERIALS AND METHODS: Forty-six patients with MM were treated with bortezomib-based regimens and, when eligible, with autologous stem cell transplantation. Response to therapy was assessed according to the criteria validated by the International Myeloma Working Group. Multiparameter flow cytometry was carried out with an 8-color panel validated by the Euroflow Consortium. A commercially available single 8-color tube in dried formulation was used and almost 2,000,000 events were acquired, in order to obtain a 10-5 sensitivity. RESULT: Sixteen patients achieved stringent complete remission and another three patients achieved complete remission. In these groups of patients, the "Flow MRD-negative" status was achieved in sixteen cases. In patients who had a different degree of response (very good partial response, partial response, minimal response) immunophenotypically abnormal plasma cells were always detected. CONCLUSION: Using a single eight-color tube in dried formulation, and an acquisition strategy able to obtain a 10-5 sensitivity, not only is it possible to detect a deep response to modern therapy in patients who obtained at least complete remission, but it is also always possible to detect minimal residual disease in patients with either complete remission or stringent complete remission.

Unusual onset of a case of chronic recurrent multifocal osteomyelitis.

BACKGROUND: Chronic recurrent multifocal osteomyelitis (CRMO) is a rare condition that commonly affects the clavicle and pelvis. CASE PRESENTATION: We report here a case a 12 years old girl with CRMO arising with recurrent episodes of left supraorbital headache, followed by the appearance of a periorbital dyschromia. Magnetic resonance imaging (MRI) of the skull and orbits revealed an important subacute inflammatory process. Few months after, the child presented a painful swelling of the left clavicle; the histological examination of the related biopsy allowed to establish the diagnosis of CRMO. CONCLUSION: CRMO presenting as acute headache involving neurocranium is rare; to our knowledge this is the first recognized case in the world literature. This pathological condition is frequently misdiagnosed as infection or neoplasm and needs a deep investigation for the differential diagnosis. The physical, laboratoristic and instrumental diagnostic investigations of the patient and the treatment employed are described in detail.

The clinical, serological, pathological and immunocytochemical features of a case of primary carcinoma of the fallopian tube.

We report the case of a patient with Stage I fallopian tube carcinoma who underwent bilateral salpingo-oophorectomy, total hysterectomy, omentectomy and appendicectomy followed by cisplatin based chemotherapy. CA 125 antigen was immunocytochemically detected in more than 50% of neoplastic cells, and serum CA 125 assay correlated with the clinical course of disease. Since CA 125 levels progressively increased during chemotherapy, an immunoscintigraphy with 111 In labeled CA 125 monoclonal antibody was performed and revealed a 5 cm area of abnormal captation placed in the posterior abdominal wall next the left kidney. A CAT scan detected an enlargement of left para-aortic lymph nodes, without cleavage plane with the left psoas muscle. The present case confirms that serum CA 125 assay is a useful noninvasive tool in the management of fallopian tube carcinoma.

Comparison between serum CA 125 and CA 19-9 assays and tissular OC 125 and 1116NS 19-9 reactivity in malignant and benign ovarian tumors.

This preliminary study included 25 patients with primary epithelial ovarian cancer (EOC) (18 serous, 3 serous-mucinous, 1 endometrioid, 2 undifferentiated carcinomas and 1 malignant Brenner carcinoma); 2 patients with borderline ovarian tumors and 20 patients with benign ovarian tumors (9 benign cystic teratomas, 6 serous cystoadenomas and 5 mucinous cystoadenomas). Blood samples for the measurement of CA 125 and CA 19-9 were drawn from all patients before surgery. Serum CA 125 (Reference Value-RV = 65 U/ml) and CA 19-9 (RV = 40 U/ml) were measured with IRMAs using the monoclonal antibodies (MoAbs) OC 125 and 1116NS 19-9. The same antigens were detected on paraffin-embedded tissue sections by immunocytochemistry with the avidin-biotin complex method employing the same MoAbs used for serum IRMAs. Among the 25 patients with EOC serum CA 125 levels were elevated in 20: tissular OC 125 reactivity was observed in 15 (75%) of them. Of the 5 EOC patients with normal CA 125 levels, 4 showed OC 125 reactivity. Only 2 of the 25 EOC patients had elevated serum CA 19-9 levels: one of them had tissular 1116 NS 19-9 reactivity. Among the 23 patients with normal serum CA 19-9 levels only 5 had immunocytochemical reactivity for this antigen. The 2 patients with borderline ovarian tumors had negative serum CA 125 and CA 19-9 assay: tissular OC 125 reactivity was observed in both patients, while 1116 NS 19-9 reactivity was detected in only one.(ABSTRACT TRUNCATED AT 250 WORDS)

Immunohistochemical detection of p185 product, p21 product, and proliferating cell nuclear antigen (PCNA) in formalin-fixed, paraffin-embedded tissues from ovarian carcinomas. Preliminary data.

Immunohistochemical techniques for the detection of oncogene products and the assessment of cell kinetics can represent promising investigational tools in clinical oncology. In the present paper the immunohistochemical expression of p185, p21 and proliferating cell nuclear antigen (PCNA) was retrospectively assessed in formalin-fixed, paraffin-embedded tissue samples taken from 28 primary ovarian carcinomas at first surgery. Positive immunostaining for p185 was found in 0% of 6 Stage I and 23% of 22 Stage III-IV tumors. Positive immunostaining for p21 was observed in 0% of early and 41% of advanced carcinomas; this immunohistochemical finding correlated significantly with histologic grade (G3 vs G1-2 = 47% vs 9%, p = 0.042). Elevated PCNA immunoreactivity was detected in 33% of Stage I and 50% of Stage III-IV tumors. Among the 20 patients with advanced carcinoma who underwent cisplatin or carboplatin based chemotherapy followed by second-look laparotomy, the pathologic complete response (pCR) rate was 36% for patients with low PCNA expression and 0% for those with elevated PCNA expression. A tendency towards a higher pCR rate was also found for patients with negative immunostaining for p185 or for p21. The prognostic value of the immunohistochemical detection of p185, p21, and PCNA in ovarian carcinoma deserves to be further investigated.

Multiparameter immunophenotyping by flow cytometry as a diagnostic tool in multiple myeloma and monoclonal gammopathy of undetermined significance.

BACKGROUND: Immunophenotyping by multiparameter flow cytometry (MFC) provides relevant information about prognosis and minimal residual disease detection in multiple myeloma (MM) and might be used to distinguish MM from monoclonal gammopathies of undetermined significance (MGUS). MATERIALS AND METHODS: We evaluated a possible usage of MFC to predict the differential diagnosis between MM and MGUS. One hundred consecutive patients were studied at diagnosis and underwent conventional diagnostic procedures. We carried out a double-blind study. Immunophenotyping was performed on samples from myeloaspirates before establishing diagnosis, while the final clinical diagnosis was established independently from MFC results. A five- or six-color method was carried out by means of monoclonal antibody combinations able to identify abnormal plasma cells (CD19-) and the most relevant immunophenotypic aberrations (loss of CD27; overexpression of CD117, CD56, CD28; asynchronous expression of CD20). MFC was applied following the indications of the European Myeloma Network. When abnormal plasma cells were /= 3.1%, MGUS was predicted. RESULTS: MFC results predicted 63 cases of MM and 37 cases of MGUS. At the end of our study, 61 cases of MM and 39 cases of MGUS were diagnosed. Therefore, 4% of patients were misdiagnosed by MFC parameters alone, with sensitivity and specificity of 0.983 and 0.92, respectively. CONCLUSIONS: Only a small proportion of patients with MM and MGUS were misdiagnosed by MFC alone and a possible systematic application of MFC in all patient with MM and MGUS at diagnosis might be proposed. Novel additional criteria could be necessary to improve the diagnostic impact of MFC in monoclonal gammopathies.

Leiomyosarcoma of the parotid gland in an HIV-positive patient: therapeutic approach, clinical course and review of the literature.

Leiomyosarcoma of the parotid gland is a rare tumor with only six cases reported in the english literature. To date, the association of this rare tumor with HIV infection has never been reported. We report the first case of a 19-year-old Caribbean woman affected by leiomyosarcoma of the parotid gland and HIV infection. Surgery, radiotherapy and chemotherapy used in this patient did not provide a good result in terms of overall survival. Intercurrent disease, opportunistic infection and chemotherapy cross-reaction have not been reported during this treatment regimen. The ability to use combined modality interventions in patients with secondary malignancies and immunosuppression requires further study with focus on both tolerance and efficacy.

Thyroid papillary adenocarcinoma; lipomatous-type.

We present a multifocal tumour of the thyroid gland with unusual histological features, occurring in a 49-year-old woman. Two foci were characterized by the presence of a lipomatous stroma. This is only the second case of its type reported in the literature.

[Post-operative examination of 2930 thyroid glands: observations on primary carcinoma. Incidental carcinoma and the preoperative diagnostic assessment of thyroidectomy for cancer]. / L'esame post-operatorio di 2930 ghiandole tiroidee: osservazioni sul carcinoma primitivo. Il carcinoma incidentale e l'iter diagnostico pre-operatorio delle tiroidectomie per cancro.

The histological examinations performed in our Institute in the years 1985-1991 and concerning thyroid diseases pertain to 2930 patients. These were considered clinically benign in 2463 cases and malignant in 467. After histological control, the actually benign cases resulted to be 2335, whereas carcinomas were 588 (495 papillary, 61 follicular, 17 anaplastic, 15 medullary). Six lymphomas and 1 fibrosarcoma have been also observed. A comparison between clinical and morphological diagnosis for thyroids thought to be clinically benign has shown a diagnostic overlapping of 93.8%, with absolute agreement for Basedow's diseases, whereas thyroiditis is the disease most often not diagnosed preoperatively. Incidental carcinomas were 132, with an attendance related directly to the number of samples randomly taken from the surgical specimen. In 538 cases of carcinoma it has been possible to evaluate the intraglandular spread of the tumor: bilateral multifocal cases were 18.4%, with an incidence of latero-cervical metastases (41.4%) nearly twice as that observed in monolateral cases (24.2%). Post-surgical examination of the whole gland in 381 patients treated with a total thyroidectomy for carcinoma has discovered 20 false positive cases: 11 were operated on only according to clinical and instrumental examinations, 2 on the basis of just the cytological results, and 7 on the basis of cytologically positive evidence though refuted by intraoperative frozen sections. In the 381 cases taken into account, intraoperative frozen sections have been done 256 times: the results have been confirmed by the histological post-surgical examination of the whole gland in 94.5% of the cases. No false positive cases did occur. False negative cases were 7, due to inadequate sampling (in the operating room or in the pathology lab). Delayed diagnoses were 4, whereas truly false negative cases have been in all 3 (1.2%). The greater part of our data has been disaggregated in two different periods (1985-1988 and 1989-1991) to point out any change with time in the parameters under observation. The most relevant reflections relating to the time were the following: (1) per cent increase of bilateral multifocal carcinomas (13.9 vs. 21.8%), (2) per cent decrease of latero-cervical metastases in monolateral carcinomas (31.8 vs 17.7%), (3) frank decrease in the median value of incidental carcinomas' diameters (2.2 cm in the first vs. 0.6 cm in the last year), (4) frank decrease of false positive cases among patients operated on without prior cytological or histological verification (6/21 vs. 1/27), (5) remarkable improvement of the rate of correlation between intraoperative frozen section diagnosis and post-surgical histological examination (92.1 vs. 97.4%).(ABSTRACT TRUNCATED AT 400 WORDS)

[Chronic calculous cholecystitis: elementary and dysplastic lesions, echographic correlations and immunohistochemical approach]. / La colecistite cronica calcolosa: lesioni elementari e displastiche, correlazioni ecografiche e approccio immunoistochimico.

The morphological analysis of 223 cases of chronic calculous cholecystitis has drawn attention to the following outstanding facts: (a) elementary lesions are numerous (above all ulceration, mucosal erosion, antral and intestinal metaplasia) and put together a complex sight; (b) mild, moderate and severe dysplasias have a surprising high incidence (67%), even in the age-group under 40; (c) dysplasia is more common in the gallbladders bearing parallel hyperplastic and metaplastic mucosal changes, if compared with gallbladders where these changes are lacking; (d) thickness of gallbladder's wall and incidence of dysplasia are in direct relation; (e) echographic preoperative assessment of wall's thickness has predictive value for presence of dysplasia; (f) CEA and CA 19-9 have a progressively increasing expression in the various degrees of dysplasia.

Treatment of hepatocellular carcinoma with percutaneous ethanol injection: evaluation with contrast-enhanced MR imaging.

OBJECTIVE: The aim of this study was to investigate the usefulness of unenhanced and enhanced MR imaging in evaluating the response of hepatocellular carcinoma to percutaneous injection of ethanol. SUBJECTS AND METHODS: Thirty-one patients with 40 hepatocellular carcinomas less than 5 cm in diameter were examined with MR imaging before and after percutaneous injection of ethanol. Unenhanced T1- and T2-weighted and contrast-enhanced T1-weighted spin-echo images were obtained. CT and percutaneous biopsy were performed 1 month after the final injection of ethanol and repeated at 6-month intervals to establish the outcome of treatment: complete tumor necrosis was shown in 36 lesions and incomplete tumor necrosis was seen in four lesions. RESULTS: Of the 36 proved necrotic lesions, 31 showed homogeneously low signal intensity on T2-weighted MR images obtained after treatment, owing to coagulative necrosis of the tumor. In the remaining five necrotic lesions, hypointense and hyperintense areas coexisted; the hyperintense areas were caused by liquefactive necrotic material in two cases and by chronic inflammatory tissue along the boundary of the necrotic area in three cases. None of the 36 necrotic lesions showed contrast enhancement on T1-weighted images obtained after IV injection of gadopentetate dimeglumine. In the four lesions with incomplete necrosis, the viable portion of the tumor was identified as a hyperintense area on T2-weighted images and as an enhancing area on contrast-enhanced T1-weighted images. No correlation was found between lesion features on unenhanced T1-weighted images and outcome of therapy. CONCLUSION: Contrast-enhanced T1-weighted MR images allow a reliable evaluation of the effectiveness of treatment and are more accurate than unenhanced MR studies. Contrast-enhanced MR imaging may be considered a valuable alternative to contrast-enhanced CT in the follow-up of hepatocellular carcinomas treated with percutaneous injection of ethanol.

[Human subcutaneous dirofilariasis: 8 more cases in Italy]. / Dirofilariasi umana sottocutanea: 8 ulteriori casi in Italia.

Eight new cases of Human Subcutaneous Dirofilariasis due to Dirofilaria repens in subjects aged 14 to 83 years, 5 men and 3 women, are recorded. The patients were living in Piedmont (5 cases), Tuscany (1), Friuli-Venetia Julia (1) and Apulia (1). The nematodes were immature in 6 cases (2 males and 4 females) and adults in 2 (both females). Microfilariae in the uterus of one of them were detected.

Interference of thyroperoxidase on immuno-cytochemical determination of steroid receptors in thyroid tissue.

The presence of sexual steroid receptor proteins in thyroid tissue has been previously demonstrated by biochemical means. The aim of this study was to determine the estrogen (ER) and progesterone (PR) receptors in malignant (12 papillary and 1 follicular carcinoma) and nonmalignant (19 multinodular goiters, 1 Graves' disease, 1 Hashimoto's thyroiditis) thyroid diseases using immunocytochemical assay employing monoclonal anti-ER and anti-PR antibodies and the peroxidase-antiperoxidase technique. Positive results were obtained in 24/34 (70%) for ER (ER-ICA+) and 22/34 (64%) for PR (PR-ICA+). To evaluate the possible interference of thyroperoxidase in the immunostaining, in consecutive sections of a positive specimen, primary antibody or primary antibody plus bridging antibody or PAP complex was omitted. Using these modified procedures, staining distribution was similar to that obtained by the standard procedure: in contrast, no staining was found in the positive control, i.e. a breast cancer specimen. The inhibition of the endogenous peroxidase caused a loss of staining in both the standard and modified procedures on thyroid specimens; no staining modification was obtained in the positive control. These results suggest that the staining observed in thyroid tissue is not specific and related to the activity of thyroperoxidase on chromogen solution. The complete loss of staining after peroxidase inhibition appears to be in contrast with the results obtained by biochemical method, and different antigenicity of thyroid receptors in comparison with breast receptors may explain this discrepancy.