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Fibroblast growth factor 2 (FGF2) has been implicated in the pathogenesis of malignant melanoma (MM), but the role of other FGFs and their receptors (FGFRs) is not elucidated. To determine whether FGF1 and FGFR1 may be involved in MM growth in vivo, we have studied the expression of the FGF1 and FGFR1 genes in 77 fresh MM biopsy samples, using RT-PCR analysis. Samples of benign nevi, normal skin and carcinoma cell lines were included as controls. Using RT-PCR analysis, expression of FGF1 and FGFR1 was observed in 69/77 and 68/77 cases, respectively. Immunohistochemical detection of the FGFR1 protein was positive in reactive stromal cells and at a much lower level in neoplastic cells. In situ hybridization experiments demonstrated FGFR1 mRNA mainly located in the stromal component. Southern blot analysis of genomic DNA prepared from MM tumors did not show any structural alteration of the FGFR1 gene. There was no correlation between FGF1/FGFR1 expression and the usual clinicopathological parameters of MM. We conclude that FGF1 and FGFR1 are frequently co-expressed in MM, a situation that may contribute to aberrant autocrine and paracrine pathways. Due to the absence of correlation with clinico-pathological parameters, this expression cannot be used as a marker of prognosis in the management of MM patients.

Bcl-x is a member of the bcl-2 family of proteins which are characterized by their ability to modulate apoptosis. Alternative splicing results in two distinct bcl-x mRNAs encoding a long isoform, bcl-xL, which acts as a bcl-2 agonist; and a short isoform, bcl-xS, which inhibits bcl-2 effects. The aim of the study was to determine whether bcl-x is expressed in lymphoma tissues and to characterize the respective production of bcl-xs and bcl-xL. We investigated the expression of bcl-x mRNA in a series of 50 non-Hodgkin's lymphomas (NHL) and Hodgkin's disease (HD) cases using a RT-PCR method in order to amplify both transcripts simultaneously, and to estimate their relative abundance. The rearrangements of the bcl-2 gene were analysed by RT-PCR expression of the hybrid bcl-2-lgH mRNA. In addition, 20 PCR-positive NHL cases and three HD cases were analysed by immunohistochemistry using bcl-x polyclonal antisera. RT-PCR showed bcl-x expression in 43/45 NHLs and 5/5 HD cases. The bcl-xL transcript was predominant in all positive cases and was associated with variable amounts of bcl-xS. There was no significant correlation between the profile of bcl-xL/bcl-xS expression and the histological and immunological subtyping. Bcl-x immunodetection was positive in the neoplastic cell component in all analysed cases, but the degree of staining was highly variable between cases. Expression of the hybrid bcl-2-IgH gene was detected by RT-PCR in five cases of follicular NHL and in one case of HD, but this group of tumours did not display a particular profile of bcl-xL/bcl-xS expression. We conclude that bcl-x is commonly expressed by malignant cells in various types of malignant lymphomas, with a predominance of the bcl-xL transcript. Since the corresponding bcl-xL isoform can block the cell death machinery and potentialize bcl-2 effects, it may be involved in some pathways of lymphomagenesis.

The fusion gene NPM-ALK occurs in a subset of anaplastic large cell lymphomas (ALCLs), as a result of a chromosomal translocation, t(2;5) (p23;q35). It has been suggested that Hodgkin's disease (HD) and ALCL share a common histogenesis because of pathological and phenotypical similarities. In order to check this hypothesis, reverse transcriptase-polymerase chain reaction (RT-PCR) was performed to detect the hybrid NPM-ALK gene in 30 tumour samples, including 22 lymph node biopsies from HD and eight ALCL specimens. The threshold level of sensitivity was shown to reach at least 1/10(4) by dilution experiments using cell lines as positive and negative controls. The expected 177 bp product indicative of the NPM-ALK rearrangement was identified in Karpas 299 and SUDHL-1 cell lines and in two out of eight ALCLs. The 22 HD cases were negative, even after two successive tests. Thus, since the ALCL-specific genetic alteration was absent in our series of HD cases, the present study does not support the hypothesis that HD and ALCL are histogenetically related entities.

The authors analyzed the possible occurrence of rearrangements and/or allelic loss of the fas/APO-1 gene in a representative series of human lymphomas, including 101 cases of Hodgkin's disease (HD) and non-Hodgkin's lymphomas (NHL). The rationale for this study was double. Chromosome 10 alterations, frequently observed in lymphoma subtypes, encompass the chromosomal localization of fas/APO-1. In addition, Ipr mouse mutants, which present with a generalized lymphoproliferative disease, were shown to exhibit alterations of fas/APO-1 structure and expression. In this retrospective study, the authors performed gene dosage of fas/APO-1 by Southern blots. No fas/APO-1 alterations were observed in the 31 HD cases. Among 70 T-cell and B-cell NHL, allelic loss of fas/APO-1 was observed in three cases. Two cases with different clinical, phenotypic, and histologic presentations showed a rearrangement of fas/APO-1. A third case showed amplification. Thus fas/APO-1 alterations do occur in human lymphomas, although at a relatively low frequency.

Forty-six days after renal transplantation for inborn tubulointerstitial nephropathy, an 18-year-old man was rehospitalized for renal failure with creatinine at 200 mumol/l. There was no sign of infection and ciclosporin level was within therapeutic range. Transplant biopsy showed minor interstitial infiltration with mononucleated cells. Methylprednisolone by 10 mg/kg/d bolus did not improve renal function and OKT3 5 mg/d was substituted for ciclosporin but had to be stopped on day 8 because of a severe infectious syndrome. The patient developed fever (39 degrees C), erythemato-pultaceous pharyngitis followed by major multiple lymph node and spleen enlargement. The diagnosis of primary Epstein-Barr infection was confirmed serologically. Histology of a submaxillary lymph node reported monomorphic immunoblastic lymphoproliferation. Immunologic phenotyping showed CD19, CD20 and CD22 surface antigens characteristic of B cells and in situ Epstein-Barr hybridization was positive in 100% of the cells. Southern Blot showed an oligoclonal pattern. Ciclosporin and azathioprin were stopped and the patient was treated with corticosteroids (15 mg/d) and aciclovir given orally (3.2 g/d) for 3 months. Outcome at six months was favorable with normalization of the renal function and complete regression of the infectious syndrome. This case demonstrated the importance of molecular biology techniques for virologic and genotypic assessment of lymphomatous proliferation allowing positive aetiologic diagnosis.

Background and Design: Polymerase chain reaction (PCR) detection of circulating tumor cells from malignant melanoma (MM) was recently described, but the prognostic value of this method in the treatment of patients with MM remained unclear. In the present prospective study, blood samples (n = 193) were collected from 93 patients with MM: 10 stage I patients after primary tumor resection, 18 patients with regional lymph node metastases before node resection, 33 disease-free but high-risk patients (previously treated for node metastases), and 32 patients with distant metastases. Circulating melanocytes were detected using a reverse transcriptase PCR method that analyzes tyrosinase gene expression. All patients were kept under regular surveillance., Results: The PCR assay was always negative in normal individuals and in subjects with non-MM metastatic cancer, while it was positive in 16 of 32 patients with disseminated MM. Five of eight patients who were PCR-positive before node dissection vs one of 10 who were PCR-negative relapsed within 6 months after surgery. In high-risk but apparently disease-free patients, the risk of relapse within the next 6 months was 3.8 times higher after a positive test result. In patients with distant metastases, a positive PCR predicted rapid disease progression., Conclusions: These data suggest that PCR detection of circulating melanocytes can be considered as a marker for rapid postoperative relapse after node dissection in patients with MM with regional node metastases, for short-term relapse in high-risk disease-free patients, and for rapid and severe progression in patients with distant metastases. This test may have a crucial interest in the treatment of patients with MM.

Expression of the p53 protein can be detected by immunohistochemistry in Reed-Sternberg (RS) cells, the presumed neoplastic component of Hodgkin's disease (HD) lesions. At present, there is no clear molecular evidence that p53 positive immunostaining in HD correlates with the presence of mutations or other structural alterations of the p53 gene. To address this question, 49 cases of HD have been investigated for p53 expression by immunohistochemistry, using the DO1 monoclonal antibody on paraffin sections. Thirty-seven out of 49 cases (75 per cent) exhibited positive immunostaining restricted to RS cells and variants. Among these 37 positive cases, ten cases were selected on the basis of a rich content of RS cells showing virtually 100 per cent DO1 positivity. A PCR-mismatch strategy was chosen for the detection of p53 mutations. The threshold level of sensitivity was assessed on positive cell-line controls. A proportion of 10-15 per cent p53 mutated cells mixed in a normal population could be identified. Total genomic DNA was extracted from the ten selected HD cases and PCR amplification of exons 5-8 of the p53 gene was performed. Heteroduplex mismatch analysis revealed no structural alterations of the p53 gene in any case. In view of these findings, it appears unlikely that the sensitivity of the method by itself can fully explain the negative results, although this possibility cannot be completely ruled out. Thus, it is conceivable that p53 positive immunostaining in HD may not necessarily imply genomic alterations in the classic 'hot spot' regions and may be related to another mechanism of p53 protein stabilization.

GV104326 is the first member of a new class of antibiotics (tribactams) selected for development. It combines a particularly broad spectrum (including gram-negative and gram-positive aerobes and anaerobes) with high potency, resistance to beta-lactamases, and complete stability to dehydropeptidases. Comparative MICs were determined for GV104326 against 415 recent clinical isolates (including beta-lactamase producers), using representative antibacterial agents (imipenem, amoxicillin-clavulanic acid, cefpirome, ciprofloxacin, gentamicin, and erythromycin). GV104326 was particularly active against gram-positive bacteria; in general, its in vitro activity was equivalent to that of imipenem, equivalent to or better than that of amoxicillin-clavulanic acid, and superior to that of cefpirome, ciprofloxacin, and erythromycin. Against gram-negative bacteria, GV104326 possessed activity similar to that of imipenem and cefpirome against enterobacteria and Haemophilus spp. but its activity was superior to that of amoxicillin-clavulanic acid. GV104326 showed excellent antianaerobe activity. GV104326 was stable to all clinically relevant beta-lactamases and was rapidly lethal to susceptible bacteria. In Escherichia coli, GV104326 bound predominantly to PBPs 1a and 2 and at low concentrations osmotically stable round forms were observed. GV104326 showed an affinity for PBPs 2 and 4 of Staphylococcus aureus.

Expression of the p53 protein has been detected recently by immunohistochemistry in Hodgkin's disease (HD), but the relationship between p53 expression and the prognosis and clinicopathological heterogeneity of HD is still unclear. To address these questions we investigated 49 cases of HD for p53 expression by immunohistochemistry using the DO1 monoclonal antibody (MAb) on paraffin sections. Thirty-five cases were simultaneously tested with the 1801 MAb on frozen sections. Thirty-seven of 49 cases (75%) were DO1 positive while 14 of 35 (40%) were PAb 1801 positive. Both MAbs gave a nuclear staining restricted to Reed Sternberg cells (RSCs) and variants and distributed among the three HD subtypes analyzed (ie, nodular lymphocyte predominant, nodular sclerosing, and mixed cellularity). The percentage of positive neoplastic cells in each case was heterogeneous, ranging from almost 100% to less than 5%. In 39 patients for whom clinical data were available statistical analysis did not show any significant correlation between p53-positive immunostaining and clinical staging, B symptoms, probability of relapse, or disease-free survival. We conclude that p53 expression is a common event in HD regardless of histological subtyping, but does not bear any pejorative significance.

Hodgkin's disease is histologically characterized by the presence of Reed-Sternberg cells (RSC) and reactive cells, including numerous T lymphocytes. Some forms of B cell-non-Hodgkin's lymphoma also contain a rich T-cell population. Previous studies have suggested that the T-cell receptor repertoire of tumor-infiltrating T lymphocytes (TITL) that act specifically against tumor-related antigens, should be restricted. The authors in this study used the polymerase chain reaction to explore the possible existence of such an immunologic response of TITL against RSC or neoplastic B cells. They studied variable (v) region genes of T-cell receptor alpha and beta chains expressed by infiltrating lymphocytes in biopsy specimens from seven patients with Hodgkin's disease and three with B cell-non-Hodgkin's lymphoma. These latter samples were selected based on a rich T-cell content. Primers specific for 18 different V-alpha and 21 V-beta families were used. In every case, TITL showed an unrestricted pattern of expression similar to the repertoire observed in peripheral blood lymphocytes. Although such experiments are limited, the apparent lack of selection of a single or a limited number of T-cell subsets in the affected tissues did not support the existence of an in vivo immunologic interaction between TITL and antigens related to RSC or neoplastic B cells.

The occasional occurrence of primary extra-cutaneous malignant melanomas (MM) has led to the hypothesis that melanocytes derived from the neural crest may be arrested in their migration and may undergo an in situ malignant transformation. However, aggregates of nevus cells have only rarely been identified by histological examination in a few organs other than skin and eye. Tyrosinase is a melanin biosynthetic enzyme that is considered one of the most specific markers of melanocytic differentiation. We have attempted to detect cells committed to the melanocytic lineage, in human tissues, by means of tyrosinase gene expression. Total RNA was extracted from normal and neoplastic tissues and analyzed using a highly sensitive reverse transcription PCR assay with primers specific for the tyrosinase gene. Peripheral blood mononuclear cells (PBMC) from healthy subjects were used as negative controls. Tyrosinase transcripts were identified in a wide range of normal organs such as skin, lymph nodes, antrum, colon, kidney, lung, testis, ovary, breast, and peripheral nerve. Tyrosinase RNA was also detected in neoplastic samples including benign cutaneous nevi, lymph nodes involved by MM, breast carcinoma, liposarcoma, malignant lymphoma, and schwannoma. PBMC from patients with metastatic MM were also positive, while no positivity was detected in blood specimens from patients with other cancers. Therefore, it appears likely that cells expressing the tyrosinase gene are present in a wide range of human tissues. Although these cells still have to be accurately identified, one could propose that they might correspond to either fully differentiated melanocytes, melanocytic precursors, or Schwann cells bearing potentialities of melanocytic differentiation.(ABSTRACT TRUNCATED AT 250 WORDS)

Background: Several lines of evidence indicate that a more favorable prognosis is correlated with the degree of lymphocyte infiltration within breast carcinomas. The characterization of these T-cell infiltrates appears necessary to explore the possible existence of an in situ immunologic response of the host against neoplastic cells. Previous studies have indeed suggested that the T-cell receptor (TCR) repertoire of tumor-infiltrating T-lymphocytes (TIL) should be restricted if they act specifically against tumor-related antigens., Methods: To address this question, the expression of variable (V) region genes of the TCR-alpha and beta chains in intratumoral lymphocytes from five breast carcinoma biopsy specimens and one normal mammary gland specimen was analyzed using the polymerase chain reaction (PCR). The neoplastic samples included one medullary carcinoma and four ductal infiltrating carcinomas selected on the basis of a rich lymphoid stromal component. Primers specific for 18 different V alpha and 21 V beta families were used for PCR., Results: In every case, TIL showed an unrestricted pattern of TCR V-region gene expression, similar to the repertoire observed in peripheral blood lymphocytes and in the normal breast tissue., Conclusions: According to these nonquantitative PCR experiments, the apparent lack of selection of a limited number of T-cell specificities in the affected tissues does not favor the existence of an in vivo lymphoid recruitment based on TCR recognition of neoplastic antigenic determinants. Further studies, however, leading to an accurate quantification of immunologically relevant T-cell clones and including larger series of cases still are necessary before it will be possible to draw a final conclusion.

The hormone gastrin is mainly produced by the G cells of the antral mucosa and plays a major role in the regulation of digestive mucosal growth. Since it permits identification of cell types containing mRNA, in situ hybridization (ISH) appears to be an interesting method for studying gastrin-producing tissues. In this study, in situ detection of gastrin mRNA has been carried out on frozen sections of four human normal antral mucosa samples and of six colonic carcinomas removed from patients with high levels of plasma gastrin, using a gastrin oligonucleotidic DNA probe. We have compared the results provided respectively by the [35s] labelling and the digoxigenin labelling of the synthetic probe. Positive cells were found in each normal sample analysed with radioactive- as well as digoxigenin-labelled antisense probes. The total number of cells expressing gastrin mRNA appeared slightly higher with the [35s]-labelled probe, while the digoxigenin-labelled probe gave a better definition of positive signals. In contrast, neither radioactive nor cold probes gave positive signals in the six colonic carcinoma samples, although gastrin expression had been demonstrated in these tumours using a reverse transcriptase-PCR method. These results show that, although ISH does not seem sensitive enough to allow the detection of very low levels of gastrin expression, it would appear to be a reliable method for visualizing gastrin mRNA in human antral mucosa.

The histopathologic pattern of tissues involved by Hodgkin's disease (HD) suggests excessive activation of environmental cells by cytokines released by Hodgkin-Reed-Sternberg (HRS) cells, which are considered as the neoplastic component of HD lesions. This hypothesis has been supported by many studies performed in vitro using HD cell lines. In this study, we have tried to demonstrate the cytokine-producing cells in an environment as close as possible to the in vivo conditions, using in situ hybridization onto frozen sections of HD samples. [35S]-labelled single-stranded RNA probes were prepared by transcribing human cDNA fragments of the TNF-alpha and IL-1 alpha genes subcloned into appropriate vectors. A total of 19 specimens of HD lesions, including 7 cases of nodular sclerosing (NS) type and 12 cases of mixed cellularity (MC), were tested with both types of probes. Clinical stages included stage I (6 cases), stage II (4 cases), stage III (6 cases) and stage IV (3 cases). TNF-alpha and/or IL-1 alpha expression was observed in 12 among 19 HD cases. However, neither the histological type nor the clinical status of the patients was correlated with the profile of cytokine secretion. Most of the cytokine-producing cells could be identified as HRS cells due to their morphological appearance. In 3 cases, simultaneous analysis by immunohistochemistry and in situ hybridization showed that IL-1 alpha/TNF-alpha mRNA-producing cells simultaneously expressed the CD30 antigen, thereby confirming the HRS nature of these cells.

A 56-year-old man presented with an inguinal lymph node enlargement. Histologic study of the tumor revealed three intermingled pathologic lesions: a nodular small cell lymphoma, an angiofollicular hyperplasia of vasculohyaline type, and a vascular neoplasia closely resembling Kaposi's sarcoma. The patient was immunocompetent and denied any homosexual relationships, transfusions, or drug use. The serum was negative for the presence of human immunodeficiency virus antibody. Computed tomographic scan and ultrasound examination revealed no other lymphadenopathies. This case shows that both hyperplastic and neoplastic lymphoid proliferations can occur simultaneously with vascular neoplasia. It thereby suggests that the neoplastic populations might interact to favor the tumor growth, the sequence and the nature of the stimulating events remaining unclear.

Genotypic analyses were performed in six primary cutaneous lymphomas whose lineage could not be assessed on the basis of histologic and phenotypic data. By immunophenotyping, these neoplasms expressed leukocyte common antigen and HLA-DR but did not show consistent immunostaining for B-cell or T-cell differentiation antigens. Expression of nonspecific histiocytic markers such as lysozyme and alpha 1-antitrypsin was found in three cases. By genotyping, three cases retained a germline configuration and immunoglobulin gene rearrangement was observed in one case, T-cell receptor gene rearrangement was found in one case, and both types of rearrangements in one case. Of the three patients in whom gene rearrangements were noted, two rapidly died and the other patient, with a dual genotype, is still alive 15 years after diagnosis. The three patients without gene rearrangements are alive and well after a mean follow-up of 2.5 years. It appears that cutaneous lymphomas with an uncertain phenotype include at least some cases of authentic B-cell or T-cell lymphomas. The germline configuration that we observed in cases with a chronic course remains difficult to explain. It may be related to a low malignancy form of histiocytic lymphoma, an atypical polyclonal hyperplasia, or even a low-grade lymphoma arising from a primitive cell without established commitment to either B- or T-cell lineage.

The expression of the mos proto-oncogene was investigated in frozen sections from 15 ovarian adenocarcinomas, 1 seminoma, 1 immature teratoma and 2 benign stromal tumors by in situ RNA/RNA hybridization with a [35S] labeled mos probe. The presence of mos transcripts was detected in one germ cell tumor and three ovarian carcinomas. The putative role of the mos proto-oncogene activation is discussed.

During the last 10 years, a very large number of molecules with antibacterial activity has been developed by pharmaceutic industry. The principal objectives of pharmacological research are to obtain ever wider spectra of activity and increased antibacterial action in order to combat "emerging" agents and those resistant to traditional treatment, as well as longer acting drugs so as to improve patient compliance. These objectives might be attained by the molecules now in the phase of clinical trial, especially by the new injectable or oral cephalosporins and new fluoroquinolones.

Recent immunohistological analyses using monoclonal antibodies (MAbs) in paraffin sections have shown that Reed-Sternberg cells (RSC) sometimes express B-cell markers, thereby suggesting a lymphoid origin in at least some cases of Hodgkin's disease (HD). In order to clarify the meaning of RSC immunoreactivities in paraffin sections, 14 cases of HD selected on the basis of a minimal RSC content of 5% (6 with mixed cellularity and 8 with nodular sclerosing HD) were B5 fixed, paraffin embedded and analysed by immunophenotyping with a panel of MAbs (LCA, MB1, MB2, L26, LN2, UCHL1, MT1, MT2, LeuM1, Ber-H2). Prior to fixation, a part of each specimen was frozen and submitted to genotypic analysis using immunoglobulin (Ig), T-cell receptor (TCR) gamma and TCR beta genes probes. RSC were strongly positive with LeuM1, BerH2 and LN2 in all cases and weakly positive with L26 in 4 cases and MB2 in 1 case. Many MT1-UCHL1 positive small lymphocytes were densely distributed close to RSC in all cases. The 14 cases retained a germline configuration with all enzyme-probe combinations tested. This finding cannot only be explained by the low number of RSC, which represented at least 5% of the cell population. Thus, neither the clonal nor the lymphoid nature of our HD cases could be determined unequivocally. LN2, L26 and MB2 expression do not correlate with Ig gene rearrangements and therefore cannot be considered as an argument supporting the B-cell derivation of RSC.

Biopsy samples from five acquired immune deficiency syndrome (AIDS)-Kaposi's sarcomas and one non-AIDS-associated Kaposi's sarcoma were assayed by in situ RNA hybridization onto paraformaldehyde-fixed, paraffin-embedded skin sections for the presence of two fibroblast growth factor gene transcripts, FGFB and FGF5. FGF5 gene expression was detected in the characteristic Kaposi's sarcoma spindle-shaped cells in the five samples from human immunodeficiency-positive (HIV+) patients. FGFB transcripts were detected in Kaposi's sarcoma cells as well as in epidermis of HIV- and HIV+ patients. These results complement the observations about growth factor gene expression done on Kaposi's sarcoma-derived cell lines, which thus appear to be representative of what happens in vivo. Furthermore, they demonstrate a contrasting expression pattern of FGF5 and FGFB genes, both involved in the growth factor pathogenic cascade leading to Kaposi's sarcoma.

Glucose-6-phosphatase (G6Pase) is a multicomponent system that catalyzes G6P hydrolysis. To determine the specificity of the histochemical reaction of G6Pase, we investigated the inhibitory effect of diethyl pyrocarbonate (DEPC), a specific and very effective inhibitor of the phosphohydrolase component of the G6Pase system, in normal human liver. The inactivation of the histochemical enzymatic activity by DEPC was monitored by determining the mean brightness of the microscopic image and the histogram of light intensity distributions. The results obtained indicate that the histogram is more sensitive than the mean brightness to variations of enzymatic activities, and that the percent of pixels brighter than a convenient level is directly proportional to DEPC concentration. This study indicates that DEPC can be used as an efficient inhibitor of the histochemical reaction of G6Pase.

Sialomucin and sulphomucin-secreting cells were studied in the normal and pathologic human pancreas with the high iron diamine-alcian blue technique which allows differentiation between the two types of mucin. In six normal autopsy pancreata, only sulphomucin was found. In benign lesions of either calcifying chronic pancreatitis (seven cases) or obstructed chronic pancreatitis (six cases), sulphomucins were widely predominant. In contrast, malignant lesions (pancreatic adenocarcinoma [12 cases], cystadenocarcinoma [two cases]) or premalignant lesions (mucinous cystadenoma [one case], ductectatic mucinous cystadenoma [four cases], villous adenoma of the main pancreatic duct [two cases]) showed a predominant sialomucin secretion, except for three poorly differentiated pancreatic carcinomas that did not show mucin staining. The sialomucin positivity was not observed at distance from the malignant lesions. In one case of benign enteroid cyst, sulphomucins predominated. These findings indicate a preponderance of sialomucin secretion in malignant or premalignant pancreatic lesions.

Immunophenotyping and immunogenotyping were performed in a series of 8 large cell lymphomas exhibiting anaplastic or "histiocytic" morphology and displaying an uncertain phenotype due to a restricted number of differentiation antigens. 6 cases expressed the Ki-1 antigen. 4 cases expressed one or two B-cell markers and contained rearrangements of the immunoglobulin genes. One of them also exhibited a T-cell receptor (TCR) beta gene rearrangement. 3 cases expressed a single T-cell differentiation antigen. Among them, only 1 displayed both gamma and beta TCR gene rearrangement; 1 only contained a gamma TCR gene rearrangement and 1 completely lacked clonal rearrangements. The eight cases expressed an inconclusive immunophenotype due to an abundant population of reactive cells but showed an immunoglobulin gene rearrangement. In conclusion, 5 out of the 8 unusual lymphomas studied here could be characterized by immunogenotyping. This approach was, however, inconclusive in the 3 remaining cases, whose lineage and differentiation stage remain poorly defined.

This report deals with two cases of villous adenoma of the Wirsung duct. The two patients presented with upper abdominal pain, diarrhea, and weight loss. Duodenal intubation showed a complete failure of exocrine pancreatic function. Ultrasound scan, computed tomography, and endoscopic retrograde cholangiopancreatography disclosed marked dilatation of the head portion of the main pancreatic duct. A proximal duodenopancreatectomy was performed. Pathological examination revealed a papillary polyp expanding the pancreatic head and filling the main duct lumen. Histological pattern consisted of a villous adenoma without any feature of malignant change. Previously reported cases exhibit many similarities to our two cases. The putative likelihood of malignant change and the relationship between villous tumors of the Wirsung duct and of other origin remain unclear.

We describe an unusual case of monocytoid lymphoma displaying a rearrangement of the T-cell receptor beta-chain gene associated with a rearrangement of the immunoglobulin heavy-chain gene, and of the lambda light-chain gene. This lymphoma was morphologically similar to previous cases in the literature, and was clinically of low grade. The lymphoma expressed the lambda light chain, HLA-DR, CD21, and CD22. Though the B-cell lineage of this lymphoma seems very likely, the genotypic profile raises a cautionary note regarding the supposed high stage of differentiation of monocytoid lymphomas.

We report a case of granulocytic sarcoma of the bone with pleural involvement diagnosed upon cytologic analysis of the pleural fluid (centrifugation spots stained by May-Grunwald-Giemsa) and confirmed by more complex investigations, i.e., demonstration of granulomonocytic membrane antigens by immunohistochemical monoclonal antibody techniques on frozen sections of the tumor. This case draws attention to the value of cytologic studies in granulocytic sarcomas whose histologic features are suggestive of lymphoma.

The authors describe the case of a 67 year-old woman bearing a malignant lymphoma displaying two different histological patterns: a Lennert's lymphoma pattern in a cervical lymph node and a diffuse centroblastic lymphoma pattern in an axillary node. Immunophenotyping on frozen and paraffin sections showed a predominant positivity of T-cell markers in the cervical node, while the axillary lymphadenopathy exhibited a major positivity of B-cell markers and a restricted expression of the lambda light chain. However, genotyping provided the evidence of a single neoplastic B-cell clone proliferating in both cervical and axillary nodes. These findings suggest that histological pattern of Lennert's lymphoma is devoid of absolute specificity. Thus, genotypic analysis is a useful tool for the diagnosis of so-called "lymphoepithelioid lymphomas".

The laminin (Lam) and type IV collagen (Coll IV) and estrogen receptor (ER) immunodetections were assessed in a large series of 400 human breast carcinomas. In all the cases the patient's age, the tumor size, the histological type and grade, the presence or the absence of axillary lymph node metastasis and of vascular invasion in tumor borders, and ER tumor content were recorded. Monoclonal anti-Lam, anti-Coll IV were applied with the avidin-biotin-peroxidase complex and monoclonal anti ER with peroxidase anti-peroxidase complex, on frozen sections. A computerized system of image analysis referred to as SAMBA (TITN) with specific software for tissue sections analysis permitted a multiparametric quantitative analysis of immunostained surfaces. With this system, in each tumor, the cellularity, the percentage of Lam, Coll IV and receptor positive surfaces versus the total cell surface and versus the epithelial (keratin positive) surface, the integrated optical density IOD histograms were obtained and correlated to morphometrical and standard histological data. From this study, it was shown that: (1) Lam and Coll IV immunostained epithelial basement membranes in carcinomas were correlated to the presence of estrogen and progesterone receptor antigenic sites within the tumors, with a significant decrease of the positive staining in ER-ICA negative tumors in comparison to ER-ICA/PR-ICA positive tumors. (2) The combined densitometric and morphometric evaluation demonstrated a decrease of Lam and Coll IV immunostaining in malignant tumors, correlated to (i) the presence of peritumorous vascular invasion and (ii) keratin positive cells in bone marrow (iii) axillary lymph node involvement. It is concluded that the variations in Lam and Coll IV antigens distribution may be relevant indicators of tumor metastatic potential in breast carcinomas and that computerized image analysis enables the standardization of the evaluation antigens distribution.

A study was carried out to determine whether the minisatellite DNA probes described by Jeffreys and coworkers could be used routinely to analyze engraftment, hematopoietic chimerism, and relapse in recipients of bone marrow transplants. The probes were informative for all of the recipient/donor pairs analyzed. Their limit of sensitivity was determined in reconstruction experiments and was found to vary from 2%, in the best cases, to 10%. We were able to confirm that engraftment and hematopoietic chimerism can, indeed, be sought routinely using this simple molecular approach. Only one set of probes is required for all patients and, unlike cytogenetic analysis, this analysis can be used whether or not blood or marrow cells are dividing.

The effect of urethan on various spleen-lymphocyte functions was studied in the rat for 45 days after administration of the carcinogen. For 3 days following the last injection of urethan, the total cell number and LPS reactivity were greatly reduced. Certain reactivities, probably T-cell dependent functions, such as responsiveness to the in vitro mitogenic effect of PHA, Con A, and primary stimulation by histocompatibility alloantigens in the one-way reaction, were selectively enriched during the first 2 days. Thereafter several dissociations of these lymphocyte functions can be observed: i.e., various intervals succeed during which one function may be enriched and other(s) diminished. It seems that the kinetics of enrichment, decay and recovery of the T-cell subsets involved in the reactions investigated follow a distinctive profile for each one of them. It is suggested, as a working hypothesis, that the unbalanced situations derived from the time-course discordance of the quantitative changes in the various lymphocyte functions may allow, or even enhance, tumor development.

Ofloxacin has been evaluated in an in vitro model where microorganisms were exposed to the varying concentrations met in human serum after the oral administration of a 200 mg dose. The recently isolated strains were selected with minimum inhibitory concentrations (MICs) representative of the MICs 90% of ofloxacin against the species considered. Results showed that ofloxacin sterilized the bacterial cultures of S. aureus, P. morganii and E. coli, with 99.99% of killing still evident for P. aeruginosa after 12 hours of exposure. All the reference compounds (amoxicillin 1 g b.i.d., cefaclor 500 mg t.i.d., rifampicin 450 mg b.i.d.) showed, in the same test, worse results than ofloxacin.

A kinetic model was used to evaluate the activity of ceftazidime against recently isolated bacterial strains. The microorganisms were subjected in vitro to drug concentrations simulating the serum kinetics of ceftazidime after i.v. administration of 1 g of the drug. In these conditions ceftazidime showed a bactericidal effect lasting up to 12 h, even at concentrations below the MICs for some strains. In no case did resistant populations develop after exposure to the drug.

The present report considers the autopsic study of an homograft recipient who had been living for 18.5 years after a cardiac transplantation. The patient was treated by immunosuppressive therapy associating azathioprine and steroids. During the exceptionally long follow-up, two skin carcinomas and a lung carcinoma occurred successively. In addition, the autopsy allowed observation of a kidney adenocarcinoma associated to a polycystic disease, and a liver regenerative nodular hyperplasia containing several areas of severe dysplasia. These findings, when compared with those usually observed in immunodeficient patients, suggested the possibility that long-term immunosuppressive therapy may give rise to malignancies other than those arising after a short therapy.

The authors report an uncommon case of giant leiomyoma of the inferior vena cava with intracardiac extension and arising from the external iliac vena. This case report illustrates the diagnostical problems usually encountered with this type of tumour. Indeed, despite ultrasound and transverse CT scan, the diagnosis was only assessed by histopathological analysis following a successful surgical removal of the leiomyoma. Only 7 cases of leiomyoma of the inferior vena cava are already reported in the literature. Relationship with intravenous leiomyomatosis of the uterus is also discussed.

The pharmacokinetics and clinical efficacy of ceftazidime, a cephalosporin with activity against Pseudomonas aeruginosa, were studied in children with cystic fibrosis. Ceftazidime had high in vitro activity against P. aeruginosa strains isolated from our patients, with a mean MIC90 of 8 micrograms/ml. The first dose of 50 mg/kg IV of ceftazidime achieved serum concentrations of 40.8 +/- 19.7 micrograms/ml at one hour and 3.8 +/- 1 micrograms/ml at four hours after administration (mean values +/- SD). The MIC90 was exceeded by serum concentrations for up to three hours. The elimination of the drug from the blood followed a biexponential decay with an elimination half-life of 60.4 +/- 6.8 min and a total body clearance of 6.0 +/- 3.1 ml/min/kg. Renal clearance of the drug accounted for 75% of the total clearance; these are higher values than those reported for adults. Peak concentrations of ceftazidime in the sputum two hours after a single administration were 4.13 +/- 2.06 micrograms/ml; after seven and 14 days of treatment, sputum concentrations were significantly lower. Eleven children with acute pulmonary exacerbation were treated for 14 days with ceftazidime at a dose of 150 mg/kg/day (12 treatment courses). Significant clinical and radiologic improvements were obtained in nine of 12 patients. In six of 11 cases the P. aeruginosa count decreased by 10(4) CFU/ml. Ceftazidime was well tolerated, and no side effects appeared during treatment.

We investigated the pharmacokinetics of cefuroxime sodium, a new parenteral beta-lactam antibiotic, in 15 patients with stable chronic renal failure during intermittent peritoneal dialysis (IPD). Eight patients were administered 1 g cefuroxime as an intravenous bolus 1 h before the start of dialysis. Mean plasma levels of cefuroxime fell from 80 mcg/ml at 1 h to 40 mcg/ml at 6-8 h. At 24 h, concentrations were higher than 20 mcg/ml. In peritoneal fluid cefuroxime reached 16.7 mcg/ml at 1 h and 7.55 mcg/ml at 6 h. Seven patients received cefuroxime added to the dialysis solution at a dose of 2.5 g/10 liters. After 6 h of dialysis, cefuroxime reached plasma levels of 60 mcg/ml; after 24 h, concentrations were 37.5 mcg/ml. These results demonstrate that cefuroxime, administered by the i.v. route, easily diffuses from blood to peritoneal fluid and, from peritoneal fluid to blood when added to the dialysis solution. In both cases concentrations reached by cefuroxime are sufficient to treat peritoneal infections associated with peritoneal dialysis.

A new case of primary biliary cirrhosis associated with sarcoidosis is reported in a 38 year-old woman. The diagnosis of sarcoidosis was based on the occurrence of cutaneous nodules, mediastinal lymph node enlargement, pulmonary reticular infiltrate, parotitis and diffuse arthritic pain, all disappearing after steroid therapy. Hepatic biopsy showed epithelioid-cell granuloma without necrosis. The diagnosis of primary biliary cirrhosis was made 6 years later as based on intrahepatic cholestasis associated with high titer of antimitochondrial antibodies and chronic destructive cholangitis. This association (5 other cases have already been reported) raises the problem of a possible link between the two diseases which could represent different patterns of a single immunological disorder.

The authors report an electron microscopic study of the liver in a case of Niemann-Pick disease type C. The diagnosis was supported by clinical data, moderate increase of phospholipids, total cholesterol, leukocytes and liver sphingomyelin and sphingomyelinase activity. Intrasinusoidal liver foamy histiocytes were filled with lamellar and multivesicular inclusions similar to those observed in sphingomyelinase deficient patients. Hepatocytes contained a few non-specific lamellar inclusions occasionally associated with triglycerides and cholesterol crystals. These results are discussed with reference to the recent biochemical findings supporting the evidence of excessive accumulation of unesterified cholesterol in Niemann-Pick disease type C.

The activity of ceftazidime, amikacin and sisomicin was investigated in an in vitro model using varying concentrations of antibiotic which mimic the serum levels of patients after the intramuscular administration of a 500, 250 and 70 mg dose respectively. Using this test, during the time of the agar MIC value correlation, ceftazidime, amikacin and sisomicin proved to be active against strains sensitive to 16 micrograms/ml, 8 micrograms/ml and 4 micrograms/ml respectively. Using the above concentrations as the cut-off points in defining the sensitivity of the strains, ceftazidime revealed the same level of activity as amikacin (6 and 5 resistant strains respectively out of the 185 tested) and proved much more active than sisomicin (48 resistant strains).

Adequate drug concentration at the infection site is a very important objective of antibiotic therapy. In this study we investigated the pharmacokinetics of the new methoximine, cefuroxime, in peritoneal and pleural fluids in 16 patients. After a 1-g dosage of cefuroxime, its concentration reaches a peak in ascitic fluid within 4-5 h. With a 2-g dose the peak level is reached in 2-3 h. After 5 h, cefuroxime concentration in ascites exceeds that in serum. Cefuroxime diffuses readily into the peritoneal space, where high concentrations (8-10 mcg/ml) are maintained for at least 6 h. Data on the kinetics in pleural fluid also seem to show high antibiotic diffusion rates, even if concentrations are never as high as those in blood at the same time following 1 g i.v. These favorable kinetic properties are probably due to low serum protein binding and to slow renal excretion.