Foraminifera as a model of eukaryotic genome dynamism.

In contrast to the canonical view that genomes cycle only between haploid and diploid states, many eukaryotes have dynamic genomes that change content throughout an individual's life cycle. However, the few detailed studies of microeukaryotic life cycles render our understanding of eukaryotic genome dynamism incomplete. Foraminifera (Rhizaria) are an ecologically important, yet understudied, clade of microbial eukaryotes with complex life cycles that include changes in ploidy and genome organization. Here, we apply fluorescence microscopy and image analysis techniques to over 2,800 nuclei in 110 cells to characterize the life cycle of Allogromia laticollaris strain Cold Spring Harbor (CSH), one of few cultivable foraminifera species. We show that haploidy and diploidy are brief moments in the A. laticollaris life cycle and that A. laticollaris nuclei endoreplicate up to 12,000 times the haploid genome size. We find that A. laticollaris reorganizes a highly endoreplicated nucleus into thousands of haploid genomes through a non-canonical mechanism called Zerfall, in which the nuclear envelope degrades and extrudes chromatin into the cytoplasm. Based on these findings, along with changes in nuclear architecture across the life cycle, we believe that A. laticollaris uses spatio-temporal mechanisms to delineate germline and somatic DNA within a single nucleus. The analyses here extend our understanding of the genome dynamics across the eukaryotic tree of life.IMPORTANCEIn traditional depictions of eukaryotes (i.e., cells with nuclei), life cycles alternate only between haploid and diploid phases, overlooking studies of diverse microeukaryotic lineages (e.g., amoebae, ciliates, and flagellates) that show dramatic variation in DNA content throughout their life cycles. Endoreplication of genomes enables cells to grow to large sizes and perhaps to also respond to changes in their environments. Few microeukaryotic life cycles have been studied in detail, which limits our understanding of how eukaryotes regulate and transmit their DNA across generations. Here, we use microscopy to study the life cycle of Allogromia laticollaris strain CSH, an early-diverging lineage within the Foraminifera (an ancient clade of predominantly marine amoebae). We show that DNA content changes significantly throughout their life cycle and further describe an unusual process called Zerfall, by which this species reorganizes a large nucleus with up to 12,000 genome copies into hundreds of small gametic nuclei, each with a single haploid genome. Our results are consistent with the idea that all eukaryotes demarcate germline DNA to pass on to offspring amidst more flexible somatic DNA and extend the known diversity of eukaryotic life cycles.

Synthesis of 9-(2-deoxy-beta-D-ribofuranosyl)purine-2-thione.

A synthesis of 9-(2-deoxy-beta-D-ribofuranosyl)purine-2-thione was performed by desulfurization of 2'-deoxy-6-thioguanine to give 2-amino-9-(2-deoxy-beta-D-ribofuranosyl)purine, diazotization with chloride replacement to give 2-chloro-9-(2-deoxy-beta-D-ribofuranosyl)purine, and the replacement of chloride with sulfur using thiolacetic acid and deacetylation.

Factors predicting survival, changes in activity limitations, and disability in a geriatric post-stroke population.

This study is based on data for individuals with a history of stroke taken from the NIA-sponsored Longitudinal Study of Aging (LSOA), 1984-1990. It provides information on the factors predicting survival and changes in disability and activity limitations in this cohort with stroke or cerebrovascular accident over the period of two years, 1984 to 1986. The results indicate that individuals who were less than 80 years old had higher chances of survival and were likely to show reduced activity limitations and disabilities. It was also observed that the individuals who were in excellent health were more likely to survive despite a history of stroke. Individuals with severe activity limitations had a very low incidence of survival over the two-year period. Further, the results of this study support the contention that the increased use of health care resources in the form of visits to physicians, hospitals, and nursing homes results in improved survival and lower disability and activity limitations among the elderly.

Non-Hodgkin's lymphoproliferative disorders involving the spleen.

One hundred eight splenectomy specimens involved by lymphoid neoplasms were studied to assess the frequency and pattern of involvement of the various disease groups. Cases were classified by the Working Formulation as well as by the Revised European-American classification of lymphoid neoplasms. Including the more recently described disease entities, large cell/immunoblastic lymphomas were the most common neoplasm, both primarily and secondarily, to involve the spleen (33.3% of all cases). The next most common lymphoid neoplasm to involve the spleen was chronic lymphocytic leukemia/ small lymphocytic lymphoma, found in 19.4% of cases, followed by follicular center cell lymphoma (13.0%), lymphoplasmacytoid lymphoma (9.3%), splenic marginal zone lymphoma (8.3%), mantle cell lymphoma (6.5%), and hairy cell leukemia (6.5%). The remaining 3.7% of cases included T-cell proliferations and one difficult-to-classify mixed cell lymphoma. More than 95% of the cases could be placed into one of three morphologic patterns of splenic involvement, i.e., 57.4% of spleens were involved by predominantly white pulp disease, 20.4% by predominantly nodular disease, without a predilection for white or red pulp, and 17.6% by predominantly red pulp disease. Although the white pulp and nodular patterns were primarily, but not exclusively, B-cell disorders, specimens with predominantly red pulp disease included all of the cases of hairy cell leukemia, as well as cases of both B- and T-cell lymphomas.

Human T-cell lymphotropic virus-1-positive T-cell leukemia/lymphoma in a child. Report of a case and review of the literature.

Adult T-cell leukemia/lymphoma is a monoclonal T-cell neoplasm associated with human T-cell lymphotropic virus-1 (HTLV-1) that occurs almost exclusively in adults. This report concerns a Romanian girl who had recurrent skin eruptions since infancy, subcutaneous tumors in childhood, and peripheral blood lymphocytosis, which initially developed at the age of 12 years. The circulating lymphocytes were of helper T-cell immunophenotype. Serologic studies demonstrated a number of HTLV-1 antigens in the child and her mother, and molecular analyses revealed monoclonal T-cell-receptor gamma gene rearrangement and detectable HTLV-1 proviral DNA. Conventional cytogenetic studies revealed a t(3;6)(q23;q27) chromosome translocation in most of the neoplastic cells. The patient initially responded well to interferon alfa therapy and showed regression of skin lesions and diminished lymphocytosis, but 4 years later, she developed massive lymphadenopathy and leukemic infiltration of the breast. At last clinical follow-up, at the age of 17 years, the patient had stable low-level peripheral lymphocytosis and subcutaneous tumors while being continuously treated with interferon alfa. Our review of the literature revealed six additional children with HTLV-1-associated T-cell leukemia/lymphoma, including one case with a similar clinical presentation and ethnic background. To our knowledge, the t(3;6)(q23;q27) translocation identified in this patient's neoplasm has not been previously reported in adult T-cell leukemia/lymphoma cases and may explain the early onset of disease. Although adult T-cell leukemia/lymphoma is rare in Romania, the identification of healthy carriers and vertical transmission raise the possibility that Romania might be an endemic region for HTLV-1 infection.

Rosette-forming non-Hodgkin's lymphomas.

Homer Wright rosettes, typically found in neuroblastomas and consisting of neoplastic cells surrounding an eosinophilic fibrillary centre without a lumen, have been considered as an important finding in the differential diagnosis of small round cell tumours. Rosettes in a neoplasm involving lymph nodes or bone marrow traditionally excluded a diagnosis of malignant lymphoma. In this report, we describe three cases of malignant lymphoma (two small lymphocytic and one diffuse large cell) with pronounced rosette formation. One of the two cases of small lymphocytic lymphoma was observed in the bone marrow, the other small lymphocytic lymphoma and the large cell lymphoma were in lymph nodes. The rosettes consisted of neoplastic lymphoid cells, often with participation of reactive macrophages, and ultrastructurally they had a central mass of interdigitating fibrillary cytoplasmic projections. Two cases were of B-cell lineage and one was of T-cell lineage. To the best of our knowledge, this is the first report of T-cell lymphoma with rosettes. Based on these findings, it is suggested that non-Hodgkin's lymphoma be included in the differential diagnosis of rosette-forming round cell neoplasms.

Polypharmacy among nursing home geriatric Medicaid recipients.

OBJECTIVE: To determine the factors that influence the number of different drugs prescribed to geriatric Medicaid recipients residing in Louisiana's intermediate care facilities I (ICFs I). DESIGN: Observational and cross-sectional with descriptive and analytic components. PARTICIPANTS: All geriatric Medicaid recipients in Louisiana ICFs I during 1994 (n = 19932). METHODS: Relevant data on sex, age, race, geographic region of a recipient, number of prescribing physicians, number of pharmacies used, and the number of drugs prescribed to a recipient were extracted from the state Medicaid files. Frequencies for the seven study variables were calculated. Regression analysis was used to evaluate the influence of the six predictor variables on the number of drugs prescribed. RESULTS: The study population was 73.63% women, 60.07% 81 years of age and older, 70.65% white, 23.21% African-American, 6.14% other races, and 29.83% from predominantly rural north Louisiana. A total of 44.60% of the residents received prescriptions from one physician, 8.41% of the residents were single pharmacy users, and 45.65% were prescribed more than 10 drugs during the year. The regression model accounted for 20.53% of the total variation in the number of drugs prescribed to a recipient. Race, geographic region, number of prescribing physicians, and number of pharmacies used by a recipient influenced the number of drugs prescribed. CONCLUSIONS: To reduce the number of drugs prescribed and polypharmacy among geriatric Medicaid recipients, Louisiana's ICFs I should minimize the number of physicians and pharmacies used in this population.

Inappropriate drug prescribing and related outcomes for elderly medicaid beneficiaries residing in nursing homes.

Problems of drug prescribing are related to poor economic and clinical outcomes, and there is a common perspective that prescribing in nursing homes is suboptimal. This study purported to determine how prescribing of "inappropriate" drugs, health care provider-related factors, and patient demographics were related to the cost of pharmaceutical services and the mortality of elderly Medicaid beneficiaries in Louisiana's Intermediate Care Facilities I during 1994. Inappropriate drugs for this retrospective, observational, and cross-sectional study were identified using explicit criteria. Relevant data on the population of 19,932 beneficiaries were extracted from the state Medicaid files and analyzed using multiple linear and binomial logit regression procedures. Cost of pharmaceutical services for a beneficiary was positively correlated with the number of different inappropriate drugs prescribed, number of physicians and pharmacies used, and the geographic region. It was negatively correlated with the beneficiary's age. Probability of the beneficiary's mortality was positively correlated with the number of pharmacies used and negatively correlated with one geographic region. Minimizing the number of different inappropriate drugs prescribed, as well as the numbers of prescribers and pharmacies used, for an elderly beneficiary may reduce the cost of pharmaceutical services paid by Medicaid.

Follicular small lymphocytic lymphoma.

We herein provide evidence for the existence of a distinct morphologic form of small lymphocytic lymphoma (SLL) that we term follicular small lymphocytic lymphoma (FSLL). Nine specimens of FSLL from eight patients were studied. The lymphomas in this study showed a true follicular pattern that was independent of tissue planes; the cytologic composition was identical to that seen in SLL. All six of the specimens (from five patients) for which paraffin tissue was available marked as B cell phenotype and were positive for bcl-2 protein. Polymerase chain reaction studies performed on deparaffinized tissue sections showed bcl-2 major breakpoint region rearrangements in four of five cases for which study tissue was available. Clinical information was available for all eight patients. All patients presented with lymph node disease, and three patients also had extranodal involvement at the time of presentation. Three of the patients had a relapse of disease after 33-95 months, and two of these patients died soon after relapse. Another two of the eight patients never responded to chemotherapy and died of their disease after 2 and 8 months, respectively. Two patients died of causes unrelated to their lymphoma and unrelated to any lymphoma therapy. Only one patient remains disease-free, after 65 months; this patient had a relapse at 44 months. The finding of bcl-2 rearrangements suggests that the pathogenesis of FSLL is more closely related to follicular small cleaved cell lymphoma than to classic SLL.

Non-Hodgkin's lymphoma involving the breast.

Breast involvement by non-Hodgkin's lymphoma is rare. Differences between primary and secondary breast lymphoma have been reported, and a relationship between primary breast lymphoma and lymphomas of mucosa-associated lymphoid tissue has been suggested. We reviewed 61 cases of breast lymphoma (41 primary, 13 secondary, and 7 unclear) that included 28 right-sided masses at presentation, 17 left-sided, 12 bilateral, and 4 in which the side was not known. A subgroup of bilateral breast lymphomas was identified that occurred in young women, four of which were pregnant or postpartum. A high incidence of intermediate- and high-grade lymphomas were present in both cases of primary and secondary lymphomas as was a high frequency of B-cell phenotype. Additional immunohistochemical studies failed to demonstrate evidence of marginal or mantle cell differentiation in seven of eight cases studied. Lymphoepithelial lesions were identified in a majority of cases, including 67% of primary and 64% of secondary lymphomas. This study failed to demonstrate a morphologic difference between primary or secondary lymphomas of the breast and suggests that breast lymphomas differ from other extranodal lymphomas in that the latter are frequently low grade.

Stability of fluconazole in an extemporaneously prepared oral liquid.

The stability of fluconazole in an extemporaneously prepared oral liquid was studied. An aqueous liquid formulation of fluconazole was prepared by reconstituting the powder from triturated 100-mg tablets with deionized water; the nominal fluconazole concentration was 1 mg/mL. Glass vials of the liquid were stored in the dark at 4, 23, and 45 degrees C and sampled immediately and after 1, 2, 3, and 15 days. Samples were analyzed in duplicate for fluconazole concentration by high-performance liquid chromatography. The concentration of fluconazole was virtually unchanged under all the storage conditions. The results were confirmed by analysis of variance. Fluconazole 1 mg/mL in an extemporaneously prepared oral liquid was stable at 4, 23, and 45 degrees C for up to 15 days.

High-grade transformation of chronic lymphocytic leukemia and low-grade non-Hodgkin's lymphoma. Genotypic confirmation of clonal identity.

The abrupt appearance of a high-grade tumor in patients with low-grade malignant lymphoma usually is associated with an accelerated clinical disease course. The high-grade lymphoma may take a variety of histologic forms and often, but not always, represents evolution of the original low-grade disease, as shown by immunophenotypic or immunogenotypic studies. The authors describe the transformation of a variety of low-grade B-cell neoplasms to high-grade tumors in four patients. The initial diagnoses included chronic lymphocytic leukemia and mantle cell lymphoma in one patient each and low-grade follicular lymphoma in two patients. The high-grade tumors were classified as lymphoblastic lymphoma in one patient and small noncleaved cell lymphoma in two patients. The high-grade component manifests primarily in the peripheral blood as circulating blast-like cells consistent with large-cell lymphoma in the remaining patient. In each case, immunophenotypic studies showed identical monoclonal surface immunoglobulin expression on the low- and high-grade tumors. Immunoglobulin heavy chain gene and kappa light chain gene studies showed identical clonally rearranged bands in paired samples from three of the four patients, a finding indicative of clonal identity. Unexpectedly, dissimilar immunoglobulin light and heavy chain gene rearrangements were detected in the paired samples from one patient with previously diagnosed follicular lymphoma, making the relationship of the two tumors from this patient uncertain; however, additional Southern blot analysis of the bcl-2 gene showed identical rearrangements in both lesions. Furthermore, polymerase chain reaction across the t(14;18) major breakpoint region in both tumors amplified nucleotide fragments of identical size, confirming the clonal identity of the low- and high-grade lymphomas despite the divergent immunoglobulin gene studies. These studies show that low-grade malignant lymphomas of small lymphocytic, mantle cell, or follicular small cleaved cell types may assume high-grade morphologic characteristics, that this change is the result of transformation of the preexisting low-grade malignant neoplasm, and that this progression, like typical Richter's syndrome, is associated with a dramatically accelerated clinical course. In addition, these studies confirm previous reports that disparate immunoglobulin light and heavy chain gene rearrangements are not necessarily an indicator of different cellular origins, and additional genotypic studies occasionally may be required to show the clonal identity of the cell population involved in these morphologic transformations.(ABSTRACT TRUNCATED AT 400 WORDS)

Extramedullary myeloid cell tumors. An immunohistochemical and morphologic study of 28 cases.

In an attempt to correlate the morphologic and immunophenotypic findings in extramedullary myeloid cell tumors (EMT), we studied 28 cases with a large panel of antibodies using paraffin section immunohistochemistry. A previous or concurrent diagnosis of acute myelogenous leukemia or chronic myelogenous leukemia was made in 25 cases. Six EMT were morphologically classified as well differentiated (WD-EMT), 17 as poorly differentiated (PD-EMT), and five as blastic EMT. The WD-EMT were easily recognized morphologically and displayed a relatively mature myeloid phenotype, with elastase, CD15, and CD68 positivity in all cases. On the other hand, the five blastic-EMT displayed no morphologic evidence of myeloid derivation, were completely negative for CD15, and were weakly positive for elastase in only one case. The PD-EMT, with a morphologic appearance that resembles large cell non-Hodgkin's lymphoma, variably expressed CD15 and elastase. CD68 and lysozyme were present in the majority of PD-EMT, with some variability, but were negative in most blastic-EMT. CD45 (LCA) was detected in 75% of all EMT and CD34 was positive in 36%; neither antigen was significantly associated with a specific morphology. CD30 reactivity was not evident in any case, but slight positive staining was seen with CD20 (L26) in one WD-EMT. CD43 (Leu 22) was the only antibody that was positive in 100% of cases; staining was always intense and widespread. Antimyeloperoxidase (MPO) was positive in all cases but two, both with a blastic morphology. We conclude that (a) an immunohistochemical panel including CD20, CD43, CD68, and MPO can successfully identify the vast majority (96%) of EMT in paraffin sections, and (b) there is an association between morphology and phenotype in these lesions.

Reed-Sternberg-like cells in low-grade lymphomas are transformed neoplastic cells of B-cell lineage.

Multinucleated giant cells resembling Reed-Sternberg (RS) cells are occasionally observed in high-grade lymphomas of the large-cell or immunoblastic type, but much less commonly in low-grade lymphomas. This study was conducted to determine whether RS-like cells found in seven B-cell low-grade lymphomas were immunologically similar to the neoplastic cells in the lymphoma or to the true RS cells seen in Hodgkin's disease, and whether they were therefore indicative of a composite lymphoma. Immunohistochemical studies were performed on paraffin sections of the seven low-grade (one small lymphocytic, one mantle zone, and five follicular) lymphomas with a panel of antibodies reactive with leukocyte common antigen (LCA), B-cell, T-cell, and Hodgkin's disease associated antigens. The RS-like cells were reactive with LCA (four of six), L26 (seven of seven), LN1 (five of six), LN2 (two of six), and MB2 (three of six). No positive staining was seen with either Leu-M1 or Ber-H2. The RS-like cells in the mantle zone lymphoma expressed L26, Leu-22, and kappa cytoplasmic light chains. This immunophenotype is similar to that of the neoplastic small lymphocytic cells. One of the low-grade follicular lymphomas progressed to an immunoblastic lymphoma with many RS-like cells. Paraffin immunohistochemistry on both lesions revealed a similar B-cell phenotype for the RS-like cells. Immunogenetic studies revealed B-cell and bcl-2 gene rearrangements in the immunoblastic lymphoma. These results indicate that RS-like cells in low-grade lymphomas are transformed neoplastic cells of B-cell lineage. With careful morphologic examination augmented by immunohistochemical studies, these lesions can be differentiated from Hodgkin's disease and from composite lymphomas of the combined Hodgkin's and non-Hodgkin's type.

Replication of the base pair 6-thioguanine/5-methyl-2-pyrimidine with the large Klenow fragment of Escherichia coli DNA polymerase I.

The kinetics and the fidelity of replication of the base pair 6-thioguanine (Gs)/5-methyl-2-pyrimidinone (Th) have been determined by using defined oligomers with the large Klenow fragment of Escherichia coli DNA polymerase I. The insertion efficiency, Vmax/Km (min-1 microM-1), of Th opposite Gs is 1.5 and the insertion efficiency of Gs opposite Th is 0.7. By comparison, the insertion efficiencies of C opposite G and G opposite C are 0.5 and 1.5. The insertion efficiency of the next base, A opposite T, is 2 times greater after the base pair Gs/Th than after G/C. The fidelity of replication with respect to thymine and adenine has misinsertion frequencies, or ratios of the insertion efficiency of the "wrong" base to the "right" base, of 7 x 10(-4) for T opposite Gs (T/Gs), 4 x 10(-6) for T/Th, and a maximum stable misinsertion frequency of 4 x 10(-4) for A/Th. No detectable elongation occurs after an A is inserted opposite a Gs. These values are similar to the misinsertion frequencies of G and C with T and A. The maximum stable misinsertion frequencies with G and C are 4 x 10(-2) for G/Th, 3 x 10(-2) -7 x 10(-3) for Gs/C, and 2.6 x 10(-1) for C/Gs, and the misinsertion frequency is < 1 x 10(-3) for Th/G. The kinetics results and molecular modeling suggest modifications to the Gs/Th base pair that may provide higher levels of fidelity of replication with respect to C and G.