Application of the FICTION technique for the simultaneous detection of immunophenotype and chromosomal abnormalities in routinely fixed, paraffin wax embedded bone marrow trephines.

The use of interphase fluorescence in situ hybridisation (FISH) to study cytogenetic abnormalities in routinely fixed paraffin wax embedded tissue has become commonplace over the past decade. However, very few studies have applied FISH to routinely fixed bone marrow trephines (BMTs). This may be because of the acid based decalcification methods that are commonly used during the processing of BMTs, which may adversely affect the suitability of the sample for FISH analysis. For the first time, this report describes the simultaneous application of FISH and immunofluorescent staining (the FICTION technique) to formalin fixed, EDTA decalcified and paraffin wax embedded BMTs. This technique allows the direct correlation of genetic abnormalities to immunophenotype, and therefore will be particularly useful for the identification of genetic abnormalities in specific tumour cells present in BMTs. The application of this to routine clinical practice will assist diagnosis and the detection of minimal residual disease.

The generation and in vivo differentiation of murine embryonal stem cells genetically null for either N-cadherin or N- and P-cadherin.

Many mutations of the murine genome are recessive embryonic lethals precluding phenotype analysis at subsequent stages of development. This is true for embryos genetically lacking either N-cadherin or N- and P-cadherin. To circumvent this, we have generated pluripotent embryonal stem (ES) cells of the same genotype in vitro and differentiated them in vivo in the form of teratomas. All of the ES cells isolated in this study had a normal ES cell morphology in vitro and were able to generate teratomas. Histological analysis revealed that some differentiation and histogenesis had occurred within the teratomas. Epithelial formation was, for example, unaffected in all cadherin null cells. Surprisingly, however, the differentiation of cells lacking both N- and P-cadherin was, in general, even more pronounced both quantitatively and qualitatively. Tumours lacking either N- cadherin or N- and P-cadherin contained more striated muscle (apparently cardiac muscle) than heterozygote controls, and this was most strikingly conspicuous in teratomas from N- and P-cadherin null cells. This more pronounced differentiation was not seen for all tissues, however, as structures with a simple neural tube-like morphology were never found in teratomas lacking both N- and P-cadherin and organoid-like structures were rare in Ncad-/-tissue.

The significance of cancer cells contaminating postoperative seroma.

Sixth postoperative day seromae of 142 breast cancer patients were searched for the presence of malignant cells. They were found in 32 patients. These patients showed significant propensity of developing distant metastases or having an unfavourable outcome of their disease although the size of their tumours was smaller than in women whose postoperative seromae were malignant-cell free. At the same time TNM tumour staging and axillary lymph node analyses failed to prove any difference between these two groups of breast cancer patients. Finally, the analyses of the state of patients' menstrual cycles showed that cycling women whose postoperative seroma contained malignant cells are at high risk of developing metastases or dying.

Endoscopic studies of nabumetone in patients with rheumatoid arthritis. A comparative endoscopic and histologic evaluation.

Gastric tolerance to 1 g of nabumetone administered in a single nightly dose was assessed in two trials in patients with rheumatoid arthritis. Gastroscopy and histology of mucosal biopsy specimens were performed before and after the end of treatment in both trials. Trial 1 was an open study that compared the effects of 1 g of nabumetone at night with those of naproxen (dose, 500 mg twice daily) and indomethacin (dose, 50 mg three times daily) in 41 hospitalized patients. After three weeks of treatment, nabumetone was significantly better tolerated than naproxen or indomethacin. Trial 2 was a randomized trial with 24 patients per group that compared 1 g of nabumetone given at night with 250 mg of naproxen given in the morning and 500 mg given at night for a period of three months. This single-blind study revealed that the number of patients with microscopic or macroscopic mucosal lesions was significantly smaller following intake of nabumetone. Concerning efficacy, as judged clinically by a rheumatologist, treatment with nabumetone was superior as well.

Prognostic value of cytophotometric analysis of DNA in lymph node aspirates from patients with non-Hodgkin's lymphoma.

To classify cases of non-Hodgkin's lymphoma in terms of expected clinical behaviour and survival, kinetic parameters measured by cytophotometry were assessed in 62 patients between 1978 and 1987. The influence of the number of cells with increased DNA content (more than 2N) on survival was evaluated. Analyses were carried out on the small samples obtained by needle aspiration biopsy of lymph nodes before treatment, using microdensitometry and Feulgen staining. Patients whose lymphomas contained less than 6% of cells with increased DNA content had a mean survival of 81.3 months and those whose lymphomas contained 6% or more of such cells had a mean survival of 18.5 months. A significant difference in survival using the same criteria was also noticed for patients with both low grade lymphomas and those with intermediate and high grade lymphomas. It is concluded that cytophotometric analysis of lymph node aspirates is of prognostic value in the initial assessment of non-Hodgkin's lymphoma.

Jumping translocations involving 11q in a non-Hodgkin lymphoma.

This paper presents the results of a cytogenetic analysis in an 11-year-old boy with non-Hodgkin lymphoma. The investigation was performed on slides obtained from short-term culture of lymph node cells. The analyses revealed an abnormal clone with loss of Y, gain of an X chromosome, t(3;22), trisomy 11, and three cytogenetically-related subclones with jumping translocations involving 11q13 as the common breakpoint region. This region is an unusual site of chromosome breakage in jumping translocations, and has not been reported thus far. Contrary to most published reports, the jumping translocation in our patient is associated with long survival.

Embryonal rhabdomyosarcoma with 92,XXYY, +dmin and evidence of spontaneous cell fusion.

The results of a cytogenetic analysis in an embryonal rhabdomyosarcoma are presented. A tetraploid karyotype with double minute chromosomes (dmin) was identified using a direct method of tumor tissue treatment. In 5% of the examined cells, evidence of spontaneous cell fusion was observed.

Cytogenetic analysis in ataxia telangiectasia with malignant lymphoma.

We present the results of cytogenetic analysis in a brother and sister with ataxia telangiectasia (AT), one of whom had malignant T-cell lymphoma. In both children, cytogenetic analysis of phytohemagglutinin (PHA)-stimulated lymphocytes showed chromosomal instability and inv(7) in 10% of the cells examined. The malignant lymphoma was analyzed cytogenetically on slides obtained from short-term culture of the lymph node cells; 64 cells were analyzed. A heterogeneous cell population was noted. Fourteen cells (21.9%) had a normal male karyotype; t(7;14)(p14;q12) and inv(7)(p14q35) were observed in 6.3% and 3.1% of metaphases. Owing to low frequency, these cells are probably a characteristic of the basic disease and have no features of malignant cells. Forty cells (62.5%) had a pseudodiploid karyotype 46,XY,dup(1)(p22p36),del(5)(q33),del(12)(p11), without cytogenetically evident aberrations of chromosomes 7 and 14. The results of these investigations suggest that the cells with rearrangements of chromosomes 1, 5, and 12 are malignant cells and did not originate by transformation of cells with inv(7) and t(7;14).

Molecular insight into the diagnosis of lymphoma.

Malignant lymphoma may be very difficult to diagnose with routine histopathological methods because they may recapitulate benign architecture or contain benign infiltrates. The best method of diagnosis is to establish the clonal profile of the lymphocyte population, since a monoclonal proliferation is highly suggestive of neoplasia. By means of a PCR (polymerase chain reaction) method it is possible to detect the immunoglobulin heavy chain (IgH) gene rearrangement and therefore establish the lymphocyte clonality. PCR with primers complementary to relatively conserved regions called frameworks (FR1-FR3) laying among hyper variable regions (CDR1-CDR3) of IgH gene unable us to detect monoclonal versus polyclonal B-cell population. The length of the PCR product with these primers is unique if we deal with a monoclonal population. On the contrary, a polyclonal population gives PCR products of a different size. In this retrospective study we used a semi-nested PCR to analyse 37 paraffin-embedded specimens. All of them had been evaluated previously by pathohistological and immunophenotypic criteria. A number of polyclonal (PBL and tonsils from healthy donors) and monoclonal cells (PBL from CLL patients, Raji cell line) were analyzed as controls. Clonality was successfully determined in all specimens. Our results support the concept that molecular techniques such as PCR provide a helpful approach for detection of monoclonal immunoglobulin rearrangements in malignant lymphoma. This is especially true for border cases, but always in the combination with other pathohistological methods.

Electron microscopy of large cell undifferentiated and giant cell tumors.

Electron microscopy is a valuable morphologic method for the diagnostic evaluation of undifferentiated tumors composed of polygonal or oval large cells and mononuclear or multinucleated giant cells. Although few ultrastructural details are pathognomonic, electron microscopic findings may add significantly to the formulation of the final diagnosis if used in the context of other pathomorphologic and clinical data. Contributions of electron microscopy to tumor diagnosis are summarized and illustrated with appropriate examples from personal experience in a routine university hospital laboratory.

Histologic and cytologic liver changes in 120 patients with malignant lymphomas.

The purpose of this study was to determine the frequency of liver involvement in malignant lymphomas. Non-specific liver changes were also registered. Percutaneous liver biopsy was performed on 120 patients with untreated malignant lymphomas. There were 38 patients with Hodgkin's disease, 42 with histiocytic and 40 with lymphocytic lymphomas. All the biopsy specimens were histologically and cytologically analyzed. Positive liver findings (lymphomatous infiltration) were observed in 27.5% of patients with lymphocytic, 23.8% with histiocytic lymphomas, and 7.8% with Hodgkin's disease. Liver involvement in non-Hodgkin's lymphomas was significantly higher (P less than 0.025) than in Hodgkin's disease. In the whole group of patients, there were non-specific liver changes: 23 chronic persistent hepatitis, 5 aggressive hepatitis, 9 liver steatosis and 4 liver hemosiderosis. Based on these results, it can be concluded that liver involvement with lymphomatous tissue is more common in non-Hodgkin's lymphomas. Knowledge of this is relevant for clinical staging and the treatment program. These findings also confirm that percutaneous liver biopsy is a valuable diagnostic procedure in the staging of malignant lymphomas.

Intensive care of severely wounded military and civilian casualties in Zadar, Croatia.

From August 1, 1991, to May 30, 1992, 148 severely wounded military and civilian casualties with the injury severity score of 3 to 5 were treated in the intensive care unit of the Zadar General Hospital. There were 138 male and 10 female patients; their mean age was 32 years. There were 64 wounded civilians and 84 wounded soldiers. The average evacuation time was 3 hours. Twelve (8%) severely wounded persons died. The cause of death was craniocerebral injury in 7 patients (58%) and hemorrhage in 4 patients (33%). Complications following shock-like acute renal failure, gastrointestinal bleeding, coagulopathy, and hepathopathy developed in 18 wounded persons (12%).

[Castleman's disease]. / Castlemanova bolest.

In this paper we have presented five patients in whom the diagnosis of Castleman's disease was established. Although it was described for the first time in 1956, the etiology of this lymphoproliferative disorder is still unknown, and the life expectancy is uncertain. Lymph nodes and spleen were analysed with the standard histological procedures and hyaline-vascular or plasma type of Castleman's disease was diagnosed. It is known that plasma-cellular type, especially its systemic presentation, can transform into malignant lymphoma or Kaposi sarcoma. Hyaline-vascular type was considered as a localized, benign lesion until malignant proliferation of the stromal cells were found in patients with this disorder. Out of five patients plasma-cellular type was diagnosed in three of them. In two of these three, malignant lymphoma appeared. Out of two patients with hyaline-vascular type one has developed follicular dendritic cell tumor.

[Fatal "overwhelming" strongyloidiasis in an immunosuppressed patient]. / Smrtonosna "preplavljujuca" strongiloidoza u bolesnika sa suprimiranim imunitetom.

The increased use of immunosuppressants in the treatment of malignant and non-malignant diseases in today's medicine has significantly contributed to the increased interest in infections caused by opportunistic microorganisms and rare parasites. A fifty-eight-year old male patient, professor, born in Bosnia, was admitted to the Institute due to poor general condition and decompensated steroid diabetes. He had been under immunosuppressant therapy for the previous 5 weeks. Six months before, he noticed squamous and crusted changes on capilli, and afterwards on his body too. As these changes did not respond to local therapy he was admitted to the Department of Dermatovenereology, Zagreb University School of Medicine. Histologic analysis indicated pemphigus erythematosus. He was treated with immunosuppressants (methylprednisolone + azathioprine). Endoscopic examinations revealed duodenal ulcer, in addition to diabetes which could not be regulated by oral hypoglycemics. He received antiulcerative therapy for ulcer treatment. Several hours upon admission the patient became highly febrile, and vomited a sanguinolent content. In spite of intensive therapy, he became comatose and died 20 hours later. On autopsy, generalized strongyloidosis of the lungs, liver, duodenum and small intestine, and a bleeding duodenal ulcer due to strongyloidosis were found. This review should remind us that hyperinfestation with strongyloides is a rare and severe complication, and could be expected in immunocompromised patients.