[Exclusive intra-operative radiation therapy (IORT) for early stage breast cancer: pilot study of feasibility]. / Radioterapia intraoperatoria (IORT) esclusiva nel carcinoma iniziale della mammella: studio pilota di fattibilità.

AIM: To evaluate feasibility, tolerability and cosmetic outcome of intra-operative radiation therapy (IORT) as an exclusive post-surgery treatment of early stage breast cancer. PATIENTS AND METHODS: From October 2008 to October 2009 30 patients underwent wide breast cancer excision or quadrantectomy followed by IORT on tumor bed with accelerated electrons at the dose of 21Gy. The characteristics of the patients were: ductal breast cancer or invasive lobular cT1, cT2 ≤ 2,5 cm, cN0, G1-2, age over 35 years, M0. RESULTS: The average age was 51.7 (range 38 - 75) with an average follow up of 11.7 months (range 6 - 18). The pathologic stage of the lesions resulted pT1 in 29 cases (96,6%), in particular: one case pT1a (3,3%), 21 cases pT1b (70,0%) and 7 cases pT1c (23,3%). One case (3,3%) was pT2 with a diameter of 2.5 cm. The grading was G2 in 20 cases (66,6%) and G1 in 10 cases (33,3%). The toxicity, evaluated according to the EORTC-RTOG criteria, was G0 (33.3%) in 10 cases, G1 (63,3%) in 19 cases, G2 in one case (3,4%); there was no G3 toxicity. The time needed for a complete healing of the wound was less than 10 days in 96,7% of the cases, with one case of limphocele (3,3%). There were no infections of the surgical wound nor any mastitis, neither in the treated quadrant nor in the other ones. We observed a light fibrosis in 5 cases (16,6%), moderate in 2 cases (6,6%) but never severe. Cosmetics, evaluated in four levels, according to Danoff et al., was excellent in 3 cases (43,3%), good in 15 cases (50%), sufficient in 2 cases (6,7%), never insufficient. As regards local control, there was no local relapse. The global survival was 100%. CONCLUSIONS: The IORT in early breast cancer, at the doses used in this study, proved itself as a secure technique, repeatable, with limited complications. The advantages of its use are the possibility of a direct control, by the surgeon and the radiotherapist, of the structures to treat and those to protect; the absence of time needed for cellular repopulation between surgery and radiotherapy; a good cosmetic outcome; and logistic advantages. It is necessary to have a long term follow up to evaluate the efficacy in terms of long term cosmetic and local control.

IL-1-induced murine osteoblast IL-6 production is mediated by the type 1 IL-1 receptor and is increased by 1,25 dihydroxyvitamin D3.

IL-1-induced osteoblast IL-6 production represents one possible mechanism by which IL-1 augments bone resorption. In this report, we show that the murine osteoblastic cell line (MC3T3-E1) expresses type 1 IL-1 receptors based on 125I-HrIL1 alpha binding, blocked by type 1 IL-1R antibodies (35F5), and analysis of MC3T3 RNA by reverse transcription (RT)-DNA amplification and Northern analysis. MC3T3 cells do not express detectable type 2 IL-1R mRNA by RT-DNA amplification. IL-1 induces (IL-1 ED50, 0.1 pM) IL-6 production through the type 1 IL-1R as 35F5 antibodies block IL-1-stimulated IL-6 production. Vitamin D3 increases IL-1R expression dose- and metabolite-dependently, with 1,25-(OH)2D3 having the greatest potency, and also enhances IL-1's capacity to stimulate IL-6 production at low IL-1 levels. Both IL-1 and 1,25-(OH)2D3 induce type 1 IL-1R and not type 2 IL-1R upregulation based on ligand binding and RT-DNA amplification. Increased IL-1R expression requires a 5-7-h treatment and is protein/RNA synthesis dependent. These observations imply that IL-1-induced IL-6 production in osteoblasts is mediated by type 1 IL-1Rs and that increased IL-1R expression could play a role in mediating IL-1-induced skeletal responses.

Chromatin structure of the c-myc gene in HL-60 cells during alterations of transcriptional activity.

HL-60 cells have an elevated level of cellular myc RNA due to an amplified c-myc gene. Subsequent to chemically induced differentiation of HL-60 cells, both cellular myc RNA levels (Grosso, L. E., and Pitot, H. C., Biochem. Biophys. Res. Commun., 119: 473, 1984) and myc-specific transcription decrease (Grosso, L. E., and Pitot, H. C., Cancer Res., 45: 847-850, 1985). We have compared the primary DNA structure, DNA methylation, and S1 nuclease sensitivity of the myc protooncogene in HL-60 cells before and after chemically induced differentiation. We find no change in the structure or methylation of the c-myc gene. The protooncogene is hypomethylated at CCGG sequences in the 5' region but is extensively methylated at sites detected by sequences homologous to the 3' exon or 3' flanking sequences. Four S1 nuclease-sensitive sites are detected prior to differentiation. After the induction of either myeloid or monocytic differentiation, three of the S1 nuclease-sensitive sites are present. The presence of the fourth S1 nuclease-sensitive site correlates with the transcriptional activity of this gene.

Transcriptional regulation of c-myc during chemically induced differentiation of HL-60 cultures.

Previous studies have demonstrated both an elevated level of cellular myc-homologous RNA in HL-60 cultures and a decrease in this messenger RNA following the chemically induced differentiation of HL-60 cells. A nuclear transcription system isolated from HL-60 cells was used to investigate whether an alteration in the rate of transcription of the c-myc gene was associated with this decrease in myc RNA. Five days after the addition of either 180 mM dimethyl sulfoxide or 60 nM 12-O-tetradecanoylphorbol-13-acetate to HL-60 cultures, transcription of the c-myc gene was markedly reduced when compared with control cultures. This specific decrease was not accompanied by an alteration in either the bulk rate of transcription of the c-myc copy number. This suggests that the decreased cellular myc RNA levels are due to decreased transcription of the myc protooncogene.

Hemophagocytic Lymphohistiocytosis in a Patient with Classical Hodgkin Lymphoma.

Introduction. Hemophagocytic lymphohistiocytosis (HLH) is a rare hyperinflammatory syndrome that can be associated with inherited genetic mutations, malignancy, autoimmune disorders, and viral infections. Though the pathogenesis is not fully known, HLH is understood to be a reactive process in the setting of uncontrolled activation of macrophages, CD8+ cytotoxic lymphocytes, and other immune cells. Hallmark clinicopathological features of HLH include fevers, cytopenias, hepatosplenomegaly, and hemophagocytosis in the bone marrow. Case Presentation. A previously healthy 28-year-old Caucasian male presented with a one-month history of persistent fever, night sweats, and unintentional weight loss. He was diagnosed with classical Hodgkin Lymphoma (HL) by core-needle biopsy of an axillary lymph node. Both bone marrow involvement by HL and hemophagocytosis were seen on subsequent bone marrow biopsy. Other findings included pancytopenia, splenomegaly, and elevated serum ferritin. Extensive work-up for autoimmune and infectious etiologies was unremarkable. The patient had a complete response after chemotherapy with Adriamycin, bleomycin, vincristine, and dacarbazine. Conclusion. This report documents the exceedingly uncommon association between HLH and HL. HLH is a hyperinflammatory syndrome with high mortality, so it is imperative to identify and treat the underlying cause for secondary HLH. Malignancy-associated HLH should be considered in the differential diagnosis for cancer patients who present with fever, cytopenias, and splenomegaly.

Performance of U.S. citizen-foreign medical graduates on certifying examinations in internal medicine.

Increasing numbers of U.S. citizens are studying medicine abroad and returning for graduate medical education and practice. The numbers and performance of U.S. citizen and all other graduates of foreign medical schools on the certifying examination of the American Board of Internal Medicine were compared with those of graduates of U.S. medical schools for 1975-1980. Numbers of first-taker U.S. citizen foreign medical school graduates increased whereas numbers of all other foreign medical school graduates decreased during this period. In 1980, U.S. citizens represented 19 percent of all first-taker foreign medical school graduates. During the study period, annual examinations were passed by 79 to 82 percent of U.S. medical school graduates, 15 to 38 percent of U.S. citizen foreign medical school graduates, and 27 to 45 percent of all other foreign medical school graduates taking an examination for the first time. On each examination, U.S. citizen foreign medical school graduates did not do as well as all other foreign medical school graduates. Members of all groups met the same postdoctoral training requirements.

The expression of the myc proto-oncogene in a dimethylsulfoxide resistant HL-60 cell line.

A cell line partially resistant to dimethylsulfoxide (DMSO) was derived from the HL-60 cell line. Cycloleucine, theophylline, sodium butyrate and elevated levels of DMSO were found to induce myeloid differentiation in this cell line while TPA induced monocytic differentiation. The levels of myc RNA and the genomic structure of the c-myc proto-oncogene were found to be the same in HL-60 and the resistant cell line. Differentiation of the DMSO resistant line was accompanied by a reduction in myc RNA levels.

Stromal osteoclast-like giant cells in an adenosquamous carcinoma of the gallbladder.

We report a case an adenosquamous carcinoma of the gallbladder that extended to the proximal transverse colon. Metastatic tumor was present in regional lymph nodes and the liver. Microscopically, the tumor was composed of malignant epithelial cells that were cytokeratin-, epithelial membrane antigen-, and carcinoembryonic antigen-positive. The adjacent desmoplastic stroma of the primary tumor, as well as the metastasis, contained giant cells that morphologically resembled osteoclasts. Immunohistochemical studies showed that the giant cells were cytokeratin-, epithelial membrane antigen-, and carcinoembryonic antigen-negative but weakly alpha 1-antichymotrypsin-positive. While tumors containing osteoclast-like giant cells have been described in the breast, lung, liver, and thyroid, this is the first report of a tumor with this morphology originating in the gallbladder. The presence of the giant cells adjacent to both the primary and metastatic tumor and not at any other location suggests that the tumor cells are producing a substance that induces the formation of the nontumoral giant cells.

Primary Ki-1 (anaplastic large cell) lymphoma of the brain and spinal cord.

The authors report a case of primary Ki-1 lymphoma of the brain. The patient was a 4 1/2-year-old black girl who presented with a 4- and 5-day history of headaches, nausea, vomiting, neck stiffness, and difficulty in walking. Computed tomography (CT) scan of the brain showed two discrete densities in the left occipital lobe and in the brain stem. Magnetic resonance imaging (MRI) showed multiple densities scattered over the brain surface and brain stem. Microscopically, the tumor was an anaplastic neoplasm that diffusely infiltrated brain parenchyma. The neoplastic cells were large with amphophilic cytoplasm, large nuclei with irregular nuclear contours and prominent nucleoli. A high mitotic rate including atypical mitotic figures was noted. Immunohistochemical stains showed diffuse strong positivity for CD30 and moderate focal staining for epithelial membrane antigen. Leukocyte common antigen, cytokeratin, neuron specific enolase, monocyte/macrophage and B- and T-marker stains were negative. The histology was characteristic for Ki-1 large cell lymphoma. Cytologic examination of cerebrospinal fluid (CSF) demonstrated similar neoplastic cells. This is one of the first reports of this variant in the pediatric population.

Quantitation of human immunodeficiency virus type 1 RNA in cell free seminal plasma: comparison of NASBA with Amplicor reverse transcription-PCR amplification and correlation with quantitative culture.

Human immunodeficiency virus type 1 (HIV-1) is transmitted by infected males in semen. However, the inoculum required for infection is unknown. The ability to collect such information will rely on the availability of reliable quantitative assays of HIV-1 in semen. We examined the comparative performance of NASBA and Amplicor Monitor RT-PCR in quantifying HIV-1 RNA in cell free seminal plasma from seropositive men and correlated the results obtained with viral titres measured by a seminal cell quantitative microculture (QMC) assay. Of samples analysed, 68% and 56% by both NASBA and RT-PCR contained measurable HIV-1 RNA, respectively. Amplification inhibition frequently affected RT-PCR but not NASBA. Excluding samples with complete RT-PCR inhibition, there was 90% qualitative concordance and a strong positive correlation (r = 0.86) of RNA levels measured by the two methods. Comparison of the concentration of HIV-1 RNA in seminal plasma samples, as measured by NASBA, with QMC viral titres indicated that RNA levels probably reflect the infectiousness of whole semen. NASBA is a reliable technique for quantitating HIV-1 RNA in seminal plasma and should become a valuable tool in the study of factors that influence the sexual transmission of HIV.

Blastic variant of mantle cell lymphoma following interfollicular Hodgkin's lymphoma.

Infrequently, patients are diagnosed with Hodgkin's lymphoma and a morphologically distinct lymphoma. While specific subtypes of lymphomas (including Hodgkin's lymphoma) may present diagnostic difficulties, fine needle aspiration biopsy (FNAB) is sometimes useful in the evaluation and classification of these lymphoproliferative processes. We report a case of the blastic variant of mantle cell lymphoma following Hodgkin's lymphoma, interfollicular variant. A 66-year-old woman with a history of Hodgkin's lymphoma presented with increasing contralateral cervical adenopathy three years after receiving chemotherapy. FNAB with ancillary immunophenotypic characterization identified mantle cell lymphoma, blastic variant. Subsequent excisional biopsy confirmed this diagnosis and also aided in the exclusion of recurrent Hodgkin's lymphoma. In addition to identifying the previously unreported combination of blastic variant of mantle cell lymphoma and Hodgkin's lymphoma, this case emphasizes the utility of FNAB in evaluation of new masses in patient's with a previous diagnosis of Hodgkin's lymphoma.

CD7 and CD56-positive primary effusion lymphoma in a human immunodeficiency virus-negative host.

Primary effusion lymphoma is an entity with distinctive features. The majority of cases are diagnosed in patients infected with human immunodeficiency virus. We report a case of pleural-based primary effusion lymphoma in an elderly patient negative for human immunodeficiency virus. By flow cytometry, lymphoma cells expressed CD7, CD38, CD45, CD56, HLA-DR, and kappa surface light chains. A monoclonal rearrangement of the immunoglobulin heavy chain and the presence of human herpesvirus 8 genome were detected. Our case lacked CD30 or CD138 with expression of surface light chains. There was strong expression of CD7 and CD56. These findings are unusual or unique in primary effusion lymphoma. Our report suggests that aberrant expression of T cell and natural killer cell markers can be seen in primary effusion lymphoma.

Does feedback on examination performance help directors of internal medicine residencies evaluate the medical knowledge of their residents against national norms?

BACKGROUND: As part of the admission process to the American Board of Internal Medicine's certifying examination in internal medicine, training program directors evaluate residents in several components of clinical competence, including medical knowledge. Research suggested that these ratings had different meanings across programs. A report comparing certifying examination performance and ratings of medical knowledge at the program and national levels was developed and sent to program directors after the 1988 through 1992 examinations. The present study investigated whether feedback helped program directors identify where their residents ranked nationally. METHOD: Subjects were first-time takers of the 1986 through 1992 certifying examinations in internal medicine who took the examination in the year they completed training and who received ratings of 4 through 9 in medical knowledge. All subjects were from programs contributing examinees in all seven study years and that received feedback in 1988 through 1991. Year-by-year distributions of program mean percentages of examinees receiving each rating of medical knowledge (4 through 9) were generated. Program means for equated examination scores and ratings of medical knowledge were computed for each year. Correlations between program mean scores and ratings were also computed. RESULTS: The distributions of the ratings were stable across the study years. Mean scores declined while mean ratings were unchanged. At the same time, correlations between scores and ratings increased. The biggest one-year change was from 1989 to 1990 (.49 to .57). CONCLUSION: Since equated scores are directly comparable, declining mean scores but unchanged mean ratings suggest that the standards applied by program directors drifted downward. The increasing correlations suggest that program directors improved in their abilities to evaluate residents relative to a common standard. It is not clear what effect the feedback had on program directors' evaluations. It is encouraging, however, to see a higher level of agreement among program directors on the meaning of the ratings.

Efficient recovery of DNA from peripheral blood for diagnostic analysis with a vacuum manifold.

BACKGROUND: The increasing clinical use of diagnostic DNA mutation analysis requires efficient isolation of DNA from peripheral blood. METHODS AND RESULTS: The use of a vacuum manifold to isolate DNA was evaluated and compared with a similar centrifugation-based DNA isolation technique. In PCR-based assays of five-point mutations, identical results were obtained with DNA isolated from peripheral blood using either centrifugation or a vacuum system. Minor modifications to PCR procedures were encountered. CONCLUSIONS: In the clinical setting, this vacuum-driven method of DNA isolation provides an efficient, useful alternative to conventional centrifugation-based DNA isolation from peripheral-blood specimens. Providing sufficient, stable DNA for multiple assays, it is easily implemented without highly specialized, expensive equipment and decreases the time spent isolating DNA from multiple samples. In addition, the potential for specimen contamination is reduced because there are fewer transfer steps.

T-cell rich, Ki-1-positive post-transplant lymphoproliferative disorder: a previously undescribed variant following liver transplant.

Post-transplant lymphoproliferative disorders (PTLD) are a consequence of the immunosuppressive therapy following organ transplant. We describe a patient who developed PTLD seven years after liver transplant and while receiving cyclosporine and prednisone. Magnetic resonance imaging demonstrated a paraspinal mass extending from T11 to L1. Microscopically, this was composed of a diffuse infiltrate of small to intermediate sized T-lymphocytes with clusters of large anaplastic tumor cells with amphophilic cytoplasm, large irregular nuclei and prominent nucleoli. A high mitotic rate and atypical mitotic figures were noted in the clusters of large cells. Flow cytometric and immunohistochemical analysis failed identify either a monoclonal B-cell population or a T-cell population with aberrant expression of the T-cell surface markers. Strong positivity for CD30 and focal staining for epithelial membrane antigen (EMA) of the large cells was seen. Leukocyte common antigen (LCA), cytokeratin, vimentin, monocyte/macrophage and B- and T-markers were negative. The small lymphoid cells were positive for CD3, MT-1 and UCHL-1. Based on the immunophenotypic and morphological evaluation, this was characterized as a T-cell rich PTLD. PCR analysis identified a monoclonal population of B-cells. This unusual case emphasizes the morphological and immunophenotypic diversity of PTLD. The utility of PCR analysis in the evaluation of PTLD is also demonstrated.

Clinical furcation diagnoses and interradicular bone defects.

The purpose of the present study was to assess associations between clinical depth of involved furcations and their bony defect depth. Twelve patients with moderate to advanced periodontitis in molars were recruited for clinical evaluation of furcation involvement by 6 dentists. Two groups of 3 dentists were assigned to the right or left half of the dentition. All dentists assessed the clinical depth of involvement of the furcations using the Ramfjord index (2 mm) in patients 1 through 6, and using the Hamp index (3 mm) in patients 7 through 12. Diagnoses were made with calibrated and uncalibrated Nabers 2 probes. After the clinical assessments the patients received full mouth scaling and root planing. After reevaluation the molars were surgically exposed. During surgery the depth of the bony furcation defects was assessed using horizontal probing and impressions. Clinically assessed depth of furcation involvement was then compared with the surgical measurements. A total of 1,180 clinical furcation diagnoses were available, of which 426 could be surgically evaluated using both the straight probe and the impressions. These evaluations were done in a total of 72 furcations using probe and impressions. For the Ramfjord index, 5% of the clinical degree 1, 40% of the degree 2, and 43% of the degree 3 readings were overestimations. For the Hamp index, 7% of degree 1, 24% of degree 2, and 0% of degree 3 readings were overestimations. These clinical diagnoses were overestimations. Forty-three percent of surgical degree 3 involvements were not recognized when using the Ramfjord index, and 27% when using the Hamp Index. These results suggest that furcation diagnosis is of limited validity.

Protease profiles of cells isolated from regenerative membranes are associated with clinical outcomes.

Guided tissue regeneration (GTR) is a clinical procedure used to restore the attachment apparatus of periodontally diseased teeth. Guided bone regeneration (GBR) is a similar procedure used to augment bone of edentulous ridges. Both therapies enhance the ability of regenerative cells to repopulate wounds by using expanded polytetrafluoroethylene (ePTFE) membranes to exclude gingival fibroblasts and keratinocytes from the healing site. Cells were isolated from 12 membranes used in each procedure and screened for the ability to form mineralized nodules in vitro, a property of cells with osteogenic potential. Using zymography and reverse zymography, low-passage isolates of cells which formed nodules were examined for the expression of gelatinolytic and caseinolytic proteases as well as for proteinase inhibitors. These molecular data were then compared with clinical outcomes for each procedure. Cells isolated from regenerative membranes exhibited variable expression of 72 kDa gelatinase, fibroblast collagenase, stromelysin, tissue inhibitor of metalloproteinases (TIMP-1), and other unidentified proteases. The greatest proportion of clinical failures was associated with GTR therapy. Cells from GTR membranes which did not exhibit gains in clinical attachment often exhibited aberrant proteinase profiles. When compared with cells from GBR procedures, most cells from GTR procedures also secreted lower amounts of TIMP-1. The study shows that cells isolated from regenerative procedures produce degradative enzymes in vitro which may be related to the success or failure of the regenerative process in vivo. Generally, cells from unsuccessful GTR procedures produced low molecular weight gelatinases not associated with cells from successful cases.

Low-grade B-cell lymphoma and concomitant extensive sarcoidlike granulomas: a case report and review of the literature.

Sarcoidlike granulomas may occur in association with Hodgkin lymphoma and non-Hodgkin lymphoma. The granulomas may be concomitant and so extensive that they obscure the malignant process. In addition, a sarcoidosis-lymphoma syndrome has been described in which there appears to be a relationship between sarcoidosis and the development of a lymphoproliferative disorder. We report a case of a low-grade B-cell lymphoma with concomitant extensive sarcoidlike granulomas. The patient had no diagnostic clinical evidence of sarcoidosis, although she had an elevated serum calcium level and increased serum angiotensin converting enzyme activity. Increased serum calcium and serum angiotensin-converting enzyme activity have been associated with clinical sarcoidosis but have also occasionally been described in association with Hodgkin lymphoma and non-Hodgkin lymphoma without evidence of sarcoidosis. We describe our findings and illustrate the usefulness of immunoperoxidase immunophenotyping techniques in such a case.

Incidental early detection of a splenic marginal zone lymphoma by polymerase chain reaction analysis of paraffin-embedded tissue.

Splenic marginal zone lymphoma (SMZL) most commonly presents with splenomegaly and stage IV disease. To our knowledge, there have been only two reported cases of SMZL without associated splenomegaly; one was detected incidentally after a bicycle accident. This previously reported case represented an early-stage SMZL with monoclonality confirmed by immunohistochemistry. We report a case of early-stage SMZL detected incidentally following a motor vehicle accident; monoclonality was confirmed by polymerase chain reaction analysis of paraffin-embedded tissue owing to the inability of confirmation by immunohistochemical techniques. We describe our findings and emphasize the importance of recognizing early stages of SMZL in incidental splenectomies by polymerase chain reaction analysis of paraffin-embedded tissue. Frozen tissue is not generally available in such cases.

Leukemic phase of mantle cell lymphoma presenting as anemia: diagnosis by combining flow cytometry and cytomorphology.

We report a case of mantle cell lymphoma in leukemic phase, which was diagnosed by a bone marrow biopsy performed as part of a workup for chronic anemia in a patient without lymphadenopathy. The patient, a 79-year-old man with diabetes mellitus, hypertension, chronic renal failure, congestive heart failure, and atherosclerosis, presented with claudication. On admission, he also had an 8-month history of anemia, during which time he experienced a 18-kg weight loss. On presentation, the patient had normal vital signs, anemia, leukocytosis (as well as an absolute lymphocytosis), and splenomegaly; as mentioned, lymphadenopathy was absent. A bone marrow biopsy showed an increase in small to intermediate-sized, slightly irregular lymphocytes in interstitial nodules. Flow cytometric immunophenotyping of the bone marrow identified a monoclonal population of cells, representing 25% of cells within the bone marrow, with expression of CD19, CD20, immunoglobulin M/D, lambda light chain, HLA-DR, and CD5; reactions for CD10 and CD23 were absent. Based on morphologic and immunophenotypic analysis of the bone marrow, as well as morphologic review of the peripheral blood smear, a diagnosis of mantle cell lymphoma involving the bone marrow and in leukemic phase was made. Subsequent polymerase chain reaction analysis of DNA from peripheral blood identified a population of cells with the bcl-1 rearrangement. This case is unique in that the diagnosis of mantle cell lymphoma was made without lymph node or spleen analysis and the patient, although exhibiting bone marrow and peripheral blood involvement by mantle cell lymphoma at presentation, did not have lymphadenopathy.