Stem-like cells traffic from heart ex vivo, expand in vitro, and can be transplanted in vivo.

BACKGROUND: Cells with stem cell surface markers have been identified in heart tissue. Early indications suggest that these are cardiac progenitor cells that could contribute to cardiac repair/regeneration. Clinically relevant therapeutic strategies based on these cells will require improved methods for their isolation and characterization of determinants of their mobilization, proliferation and differentiation. METHODS: An ex vivo culture system was developed that promotes trafficking of progenitor-like cells from mouse ventricles to a culture surface. Cells that "trafficked" from cardiac tissue were phenotyped by flow cytometry and immunohistochemistry. RESULTS: Morphologically distinct cells spontaneously trafficked from mouse ventricular tissue, adhered in culture, and proliferated for up to 4 weeks in Dulbecco's minimal essential media supplemented with fetal calf serum. After 4 weeks in culture, cell number declined. Co-culture with unfractionated bone marrow restored the proliferation of these trafficked cells. A significant population of the trafficked cells expressed a phenotype consistent with that of a myogenic progenitor such as: c-kit+, Sca-1+, CD45-, CD34-, CD90.2-, MyoD1-, desmin-, muscle-specific actin-, and, infrequently, myogenin+. An expanded population of trafficked cells from ventricles of mice expressing green fluorescent protein (GFP+) and containing cardiac-derived progenitor cells were injected into the pericardial space of GFP- mice. GFP+ cells trafficked throughout the heart but retained a primitive undifferentiated morphology. However, when injected into the pericardial space of Apo-E-deficient mice with coronary vasculopathy, progenitor-like cells trafficked into myocardium, and GFP+ cells differentiated into vessel-lining endothelial cells and, rarely, smooth muscle and cardiomyocytes. CONCLUSIONS: Progenitor-like cells in the heart can be mobilized by tissue injury to spontaneously traffic from cardiac tissue and can expand in culture by co-culture with bone marrow. When re-infused by pericardiocentesis, these primitive cells traffic into heart, retain immature morphology, but are capable of undergoing injury-induced differentiation. The novel method described herein permits further characterization of cardiac-derived progenitor cells, which are a candidate for cardiac regeneration strategies.

Foreign matter salpingitis 3 years after typhlitis.

This case suggests that typhlitis may cause delayed abdominal pathology. A history of this condition should be considered in the work-up of any patient with gastrointestinal or genitourinary pathology.

Rapid autologous marrow recovery and eradication of infectious mononucleosis despite severe immunosuppression following second transplantation for aplastic anemia.

A patient with aplastic anemia failed to respond to immunosuppressive therapy and first marrow transplantation (BMT). Recovery of autologous hematopoiesis was rapid following a second stem cell transplant with a non-myeloablative preparatory regimen. The autologous immune response to infectious mononucleosis (IM) 4 weeks post-transplant was normal despite recent and ongoing severe immunosuppression.

Ki-1+ large-cell anaplastic lymphoma after Ewing sarcoma.

PURPOSE: A large cell anaplastic lymphoma that developed after treatment of a Ewing sarcoma (ES) is described. PATIENT: An 11-year-old girl with a pelvic ES developed a large cell, Ki-1+, anaplastic lymphoma in the same anatomic location 10 months after multimodal therapy. RESULTS: ES recurred in the primary site 16 months after allogeneic marrow transplantation and 3.5 years after initial diagnosis, but the patient remains in remission from her lymphoma. CONCLUSION: The occurrence of lymphoma and ES in a short time interval in the same patient is very unusual. Whether etiologic factors other than chemoradiotherapy, including genetic disposition, play a role remains to be elucidated.

Pre-B acute lymphoblastic leukemia in a 3-year-old boy with pre-acute myelogenous leukemia myelodysplastic syndrome: cytogenetic evidence of common early progenitor cell ontogeny.

PURPOSE: Myelodysplastic syndromes in children commonly evolve into acute leukemia, usually acute myelogenous leukemia (AML) and rarely acute lymphoblastic leukemia (ALL). The lineage of the leukemia can be predicted based on characteristic morphologic and cytogenetic findings of the marrow and peripheral blood. PATIENT AND METHODS: A 3-year-old boy had refractory anemia with excess blasts and abnormalities suggestive of pre-AML with highly unusual cytogenetic changes. ALL of pre-B phenotype developed. RESULTS: Leukoerythroblastic anemia, pseudo Pelger-Huet neutrophils, and dysmyelopoietic hyperplasia of the marrow suggested likely early progression to AML. Complex cytogenetic abnormalities (monosomy 17 and 20, ring chromosome 11 with deletion of bands q23, and a derivative dicentric chromosome 12) were present in both the myelodysplastic marrow and the subsequent ALL. CONCLUSION: This case presents cytogenetic evidence of common early progenitor cell ontogeny of both malignancies (refractory anemia with excess blasts and ALL).

Plasma creatinine results derived from an endpoint modification of the Jaffé method.

For values in the normal pediatric range, endpoint modifications of the Jaffé method for measuring plasma creatinine (PCr) yield higher results than other commonly used techniques. In an effort to evaluate the Olympus AU5000 endpoint method used by the large reference laboratory to which many of our patients are directed by their third-party payor, we compared results with a kinetic Jaffé technique using paired samples from the same specimens. In 46 samples, the kinetic method measured Pcr at < or =0.8 mg/dl, whereas the endpoint technique PCr was higher by 0.1 mg/dl in 6 (13%), 0.2 mg/dl in 23 (50%), and 0.3 mg/dl in 16 (35%) samples (P<0.0001). The combination of these higher values and the same reported normal range for all children ages 2-12 years (0.3-1.0 mg/dl) and 13-17 years (0.7-1.4 mg/dl) makes interpretation of Olympus AU5000 endpoint method results difficult, particularly for younger children. The results reinforce the need for each laboratory to provide comprehensive age- and sex-adjusted normal PCr ranges.

RT in situ PCR for the detection of mRNA transcripts of Fas-L in the immune-privileged placental environment.

Rapid and reproducible detection of RNA in cells and tissue sections is routinely accomplished using in-situ hybridization technique provided that the target number of mRNA copies is above a minimum number. Detection of low copy transcripts is problematic when threshold detection occurs below clear signal resolution or alternatively, when technical problems result in background noise which occludes clear signal. RT in-situ PCR methodology utilizes both the power and specificity of PCR to amplify target whose localization is subsequently detected at the cellular level. RT in-situ PCR methods routinely involve a two-step methodology. mRNA copies are initially transcribed into cDNA. This step is followed by a separate PCR step wherein amplification of the newly synthesized cDNA takes place. A simplified one-step procedure biochemically compartmentalizes these sequential steps within a single applications methodology using the enzyme rTth. This method was successfully applied to detect and localize mRNA transcripts for Fas ligand within the immune privileged placental environment and to provide verification of immunohistochemical localization of gene product.

Trophoblasts express Fas ligand: a proposed mechanism for immune privilege in placenta and maternal invasion.

Cross-linking of Fas (CD95, APO-1) and Fas ligand (FasL; CD95L) induces apoptosis of Fas-bearing cells. Recent evidence suggests that FasL. expression plays an important role in maintenance of immune privilege in murine testis and eye and in tumour escape from immune rejection in colon cancer, melanoma and hepatocellular carcinoma. Bcl-2 is a membrane protein that suppresses apoptosis in response to a variety of stimuli. In this paper we describe abundant expression of FasL protein and mRNA transcripts within the immune privileged environment of the placenta by immunohistochemistry and reverse transcription in-situ polymerase chain reaction methods. The syncytiotrophoblast layer, the main site of feto-maternal interface, and extravillous trophoblasts, demonstrated consistent immunoreactivity for FasL in term placentae. Co-occurrence of Fas and Bcl-2 were detected with a similar pattern of distribution with FasL. The TUNEL method revealed evidence of apoptosis in the placental tissues. We speculate that abundant presence of FasL in the trophoblast contributes to immune privilege in this unique environment, perhaps by fostering apoptosis of activated Fas-expressing lymphocytes of maternal origin. An apoptotic process mediated by FasL may also play a role in placental invasion during implantation and underscores similarities between the trophoblast and neoplastic cells.

Asymptomatic lymphoma associated with elevation of immunoglobulin E.

BACKGROUND: Elevation of immunoglobulin E (IgE) is rarely detected in patients with underlying malignancy. There has been only one report of such a finding in a patient with a T cell lymphoma and none to our knowledge in association with a B-cell lymphoma. OBJECTIVE: To report a patient with elevation of IgE associated with a B-cell lineage lymphoma. MATERIAL AND METHODS: Case report and review of literature. RESULTS: A 73-year-old woman was found to have elevated IgE (11,766 IU/mL) in a routine evaluation for rhinitis. No underlying allergic diathesis or disease associated with IgE elevation was noted. Because of an abnormal lymphocyte profile, an extensive malignancy evaluation was performed which revealed a B-cell lymphoma. CONCLUSION: B-cell lineage lymphoma can be associated with elevated of IgE and should be included in the differential diagnosis in a patient presenting with this finding.

Successful allogeneic bone marrow transplantation for agnogenic myeloid metaplasia in a 3-year-old boy.

PURPOSE: Agnogenic myeloid metaplasia (AMM) is a myeloproliferative disorder characterized by marrow fibrosis, extramedullary hematopoiesis, splenomegaly, and leukoerythroblastosis with abnormalities of red blood cell morphology. This is rarely encountered in children. No conventional curative therapy is known; however, allogeneic bone marrow transplantation (BMT) may eradicate the underlying stem cell abnormality with subsequent normal hematopoiesis. RESULTS: We report a 3-year-old Arab boy who had AMM and who had normal marrow hematopoiesis and markedly reduced fibrosis after high-dose chemotherapy and matched sibling BMT. CONCLUSIONS: Allogeneic BMT offers a potential cure for patients with agnogenic myeloid metaplasia. A preparatory regimen containing busulfan and cyclophosphamide appears promising for patients with the disease.

Simultaneous expression of CD2 on a B-cell, small lymphocytic neoplasm.

An unusual neoplasm, best classified as a B-cell chronic lymphocytic leukemia on the basis of cytofluorographic, immunohistologic, and gene rearrangement analysis also co-expressed the T-cell associated marker CD2 (sheep erythrocyte receptor), but without other cell markers restricted to T cells. This case was associated with a more aggressive course than typically seen with B-CLL or malignant lymphoma, small lymphocytic type, implying along with previous reports that CD2 expression may serve as a marker of small lymphocytic tumor cell behavior and the ultimate clinical course. Thus, surface phenotypic analysis of these particular hematopoietic neoplasms may serve not only for identification of the malignancy, but could also render prognostic information.

Central nervous system expression of a monoclonal paraprotein in a chronic lymphocytic leukemia patient.

An unusual complication of chronic lymphocytic leukemia (CLL) is reported. The patient, a 79-year-old man, had a long standing history of CLL, that had been complicated by the development of a Guillain-Barré-like syndrome and a peripheral biclonal gammopathy. The biclonal immunoglobulins identified in the serum were IgM lambda and IgG lambda. The patient's condition progressed and he eventually developed ophthalmologic complications. Cerebrospinal fluid (CSF) obtained during evaluation of his visual dysfunction contained numerous small, mature lymphocytes consistent with the presence of CLL cells in the central nervous system (CNS); immunoperoxidase staining of these cells revealed a monoclonal population. Protein electrophoretic evaluation of the patient's CSF showed a single monoclonal band and immunofixation electrophoresis of the CSF revealed that the immunoglobulin present was IgG lambda. No evidence for the monoclonal IgM paraprotein identified in serum could be appreciated in the CSF by immunofixation. Taken together, these findings strongly implied that there was CNS involvement by the leukemia and this process caused the patient's neurologic symptoms. Furthermore, this study demonstrates that chronic lymphocytic leukemia should also be considered as one of the hematopoietic malignancies associated with monoclonal gammopathies involving the CNS.

Nuclear grooves in cytologic preparations. A study of the utility of this feature in the diagnosis of papillary carcinoma.

The sensitivity and specificity of nuclear grooves and inclusions for papillary carcinoma was investigated in 32 touch preparations and 69 fine needle aspiration cytologic preparations of the thyroid. Ultrastructurally, these grooves and inclusions are cytoplasmic invaginations into the nucleus. Overall, 100% of the papillary carcinomas contained nuclear grooves while only 70% contained inclusions. Grooves, however, could be seen in 70% of nonpapillary neoplasms and in 56% of nonneoplastic conditions of the thyroid, albeit generally fewer in number and often not as distinct. Inclusions were present in 13% of nonpapillary neoplasms and were absent in nonneoplastic conditions. Some nuclei on cytologic preparations contain lines that are probably artifacts of chromatin alignment and do not represent true nuclear grooves. Since such lines may be indistinguishable from true grooves, grooves should be used cautiously and in conjunction with other criteria in the diagnosis of papillary carcinoma.