bluuebunny/crossref_metadata_2025 huggingface dataset:
Stem cell leukemia protein directs hematopoietic stem cell fate
Authors: Kunisato Atsushi , Chiba Shigeru , Saito Toshiki , Kumano Keiki , Nakagami-Yamaguchi Etsuko , Yamaguchi Tomoyuki , Hirai Hisamaru
Published: January 2004

Stem cell leukemia (SCL) protein has been shown to be an essential transcription factor during hematopoietic development in the embryo. In adult hematopoiesis, however, the role for SCL has remained largely unknown, whereas it is expressed in bone marrow hematopoietic stem cells (HSCs). In this study, we performed HSC transplantation and an in vitro HSC differentiation assay using retrovirally transduced HSCs with wild-type (WT) and dominant-negative (DN) SCL. The transplantation experiments showed that SCL does not affect the long-term repopulating capacity of HSCs but that WT SCL and DN SCL increase the short-term contribution of the transduced HSCs in myeloid and lymphoid lineages, respectively. An in vitro single-cell assay using a fetal thymus organ culture system further demonstrated that WT SCL facilitates HSCs to differentiate into the myeloid lineage but that DN SCL facilitates HSCs to differentiate into the lymphoid lineage. We conclude that the up-regulation or down-regulation of SCL directs HSCs toward myeloid or lymphoid lineage, respectively, although SCL does not affect their long-term repopulating capacity. (Blood. 2004;103: 3336-3341)
Jagged1-induced Notch signaling drives proliferation of multiple myeloma cells
Authors: Jundt Franziska , Pröbsting Kristina Schulze , Anagnostopoulos Ioannis , Muehlinghaus Gwendolin , Chatterjee Manik , Mathas Stephan , Bargou Ralf C. , Manz Rudolf , Stein Harald , Dörken Bernd
Published: January 2004

Notch receptors expressed on hematopoietic stem cells interact with their ligands on bone marrow stromal cells and thereby control cell fate decisions and survival. We recently demonstrated that Notch signaling is involved in proliferation and survival of B cell-derived tumor cells of classic Hodgkin disease and described a novel mechanism for the oncogenic capacity of Notch. In this study we investigated whether Notch signaling is involved in the tight interactions between neoplastic plasma cells and their bone marrow microenvironment, which are essential for tumor cell growth in multiple myeloma (MM). Here we demonstrate that Notch receptors and their ligand Jagged1 are highly expressed in cultured and primary MM cells, whereas nonneoplastic counterparts show low to undetectable levels of Notch. Functional data indicate that ligand-induced Notch signaling is a growth factor for MM cells and suggest that these interactions contribute to myelomagenesis in vivo. (Blood. 2004;103:3511-3515)
Kaposi sarcoma-associated herpesvirus (KSHV) induces heme oxygenase-1 expression and activity in KSHV-infected endothelial cells
Authors: McAllister Shane C. , Hansen Scott G. , Ruhl Rebecca A. , Raggo Camilo M. , DeFilippis Victor R. , Greenspan Deborah , Früh Klaus , Moses Ashlee V.
Published: January 2004

Kaposi sarcoma (KS) is the most common AIDS-associated malignancy and is characterized by angiogenesis and the presence of spindle cells. Kaposi sarcoma-associated herpesvirus (KSHV) is consistently associated with all clinical forms of KS, and in vitro infection of dermal microvascular endothelial cells (DMVECs) with KSHV recapitulates many of the features of KS, including transformation, spindle cell proliferation, and angiogenesis. To study the molecular mechanisms of KSHV pathogenesis, we compared the protein expression profiles of KSHV-infected and uninfected DMVECs. This comparison revealed that heme oxygenase-1 (HO-1), the inducible enzyme responsible for the rate-limiting step in heme catabolism, was up-regulated in infected endothelial cells. Recent evidence suggests that the products of heme catabolism have important roles in endothelial cell biology, including apoptosis and angiogenesis. Here we show that HO-1 mRNA and protein are up-regulated in KSHV-infected cultures. Comparison of oral and cutaneous AIDS-KS tissues with normal tissues revealed that HO-1 mRNA and protein were also up-regulated in vivo. Increased HO-1 enzymatic activity in vitro enhanced proliferation of KSHV-infected DMVECs in the presence of free heme. Treatment with the HO-1 inhibitor chromium mesoporphyrin IX abolished heme-induced proliferation. These data suggest that HO-1 is a potential therapeutic target for KS that warrants further study. (Blood. 2004;103: 3465-3473)
Donor treatment with pegylated G-CSF augments the generation of IL-10-producing regulatory T cells and promotes transplantation tolerance
Authors: Morris Edward S. , MacDonald Kelli P. A. , Rowe Vanessa , Johnson Diana H. , Banovic Tatjana , Clouston Andrew D. , Hill Geoffrey R.
Published: January 2004

We investigated whether the protection from graft-versus-host disease (GVHD) afforded by donor treatment with granulocyte colony-stimulating factor (G-CSF) could be enhanced by dose escalation. Donor treatment with human G-CSF prevented GVHD in the B6 → B6D2F1 murine model in a dose-dependent fashion, and murine G-CSF provided equivalent protection from GVHD at 10-fold lower doses. Donor pretreatment with a single dose of pegylated G-CSF (peg-G-CSF) prevented GVHD to a significantly greater extent than standard G-CSF (survival, 75% versus 11%, P < .001). Donor T cells from peg-G-CSF-treated donors failed to proliferate to alloantigen and inhibited the responses of control T cells in an interleukin 10 (IL-10)-dependent fashion in vitro. T cells from peg-G-CSF-treated IL-10-/- donors induced lethal GVHD; T cells from peg-G-CSF-treated wild-type (wt) donors promoted long-term survival. Whereas T cells from peg-G-CSF wt donors were able to regulate GVHD induced by T cells from control-treated donors, T cells from G-CSF-treated wt donors and peg-G-CSF-treated IL-10-/- donors did not prevent mortality. Thus, peg-G-CSF is markedly superior to standard G-CSF for the prevention of GVHD following allogeneic stem cell transplantation (SCT), due to the generation of IL-10-producing regulatory T cells. These data support prospective clinical trials of peg-G-CSF-mobilized allogeneic blood SCT. (Blood. 2004;103:3573-3581)
Elevated telomerase activity and minimal telomere loss in cord blood long-term cultures with extensive stem cell replication
Authors: Gammaitoni Loretta , Weisel Katja C. , Gunetti Monica , Wu Kai-Da , Bruno Stefania , Pinelli Silvana , Bonati Antonio , Aglietta Massimo , Moore Malcolm A. S. , Piacibello Wanda
Published: January 2004

Telomerase activity, telomere length, stem/progenitor cell production, and function of CD34+ cells from cord blood (CB), bone marrow, and mobilized peripheral blood were evaluated in long-term cultures. CB cells were cultured either on OP-9 stromal cells transduced with an adenovector expressing thrombopoietin (TPO) or stimulated by a cytokine cocktail in the absence of stroma, with, in one method, CD34+ cells reisolated at monthly intervals for passage. Continuous expansion of stem cells as measured by in vitro cobblestone area and secondary colony-forming assays was noted for 18 to 20 weeks and by severe combined immunodeficiency (SCID)-repopulating cells (SRCs), capable of repopulating and serially passage in nonobese diabetic/SCID mice, for 16 weeks. Despite this extensive proliferation, telomere length initially increased and only at late stages of culture was evidence of telomere shortening noted. This telomere stabilization correlated with maintenance of high levels of telomerase activity in the CD34+ cell population for prolonged periods of culture. Cytokine-stimulated cultures of adult CD34+ cells showed CD34+ and SRC expansion (6-fold) for only 3 to 4 weeks with telomere shortening and low levels of telomerase. There is clearly a clinical value for a system that provides extensive stem cell expansion without concomitant telomere erosion. (Blood. 2004;103:4440-4448)
T-cell protein tyrosine phosphatase deletion results in progressive systemic inflammatory disease
Authors: Heinonen Krista M. , Nestel Frederick P. , Newell Evan W. , Charette Gabrielle , Seemayer Thomas A. , Tremblay Michel L. , Lapp Wayne S.
Published: January 2004

The deregulation of the immune response is a critical component in inflammatory disease. Recent in vitro data show that T-cell protein tyrosine phosphatase (TC-PTP) is a negative regulator of cytokine signaling. Furthermore, tc-ptp-/- mice display immune defects and die within 5 weeks of birth. We report here that tc-ptp-/- mice develop progressive systemic inflammatory disease as shown by chronic myocarditis, gastritis, nephritis, and sialadenitis as well as elevated serum interferon-γ. The widespread mononuclear cellular infiltrates correlate with exaggerated interferon-γ, tumor necrosis factor-α, interleukin-12, and nitric oxide production in vivo. Macrophages grown from tc-ptp-/- mice are inherently hypersensitive to lipopolysaccharide, which can also be detected in vivo as an increased susceptibility to endotoxic shock. These results identify T-cell protein tyrosine phosphatase as a key modulator of inflammatory signals and macrophage function. (Blood. 2004;103:3457-3464)
Endothelial cell apoptosis in systemic lupus erythematosus: a common pathway for abnormal vascular function and thrombosis propensity
Authors: Rajagopalan Sanjay , Somers Emily C. , Brook Robert D. , Kehrer Christine , Pfenninger Dana , Lewis Emily , Chakrabarti Anjan , Richardson Bruce C. , Shelden Eric , McCune W. Joseph , Kaplan Mariana J.
Published: January 2004

Women with systemic lupus erythematosus (SLE) are at risk for premature atherothrombosis independent of Framingham risk factors. We investigated whether endothelial cell (EC) apoptosis predicts abnormal vasomotor tone and contributes to circulating tissue factor (TF) levels in this disease. Brachial artery flow-mediated dilation (FMD) and nitroglycerin-mediated dilation were determined in women with SLE, healthy control subjects, and subjects with coronary artery disease (CAD) (n = 43/group). Quantification of circulating apoptotic ECs was performed by flow cytometry (CD146+ cells that stained for Annexin V [CD146AnnV+]) and immunofluorescent microscopy. Plasma TF was measured by enzyme-linked immunosorbent assay (ELISA). Compared with healthy control and CAD subjects, patients with SLE had higher numbers of circulating CD146AnnV+ cells (10 ± 3, 18 ± 5, and 89 ± 32 cells/mL, respectively, mean ± SEM; P < .01). Increased CD146AnnV+ cells correlated strongly with abnormal vascular function (P = .037). After adjusting for known predictors of endothelial function, CD146AnnV+ was the only variable that predicted FMD (β = –4.5, P < .001). Increased CD146AnnV+ was strongly associated with elevated levels of circulating TF (r = .46, P = .002). Circulating apoptotic ECs are elevated in young women with SLE and strongly correlate with markedly abnormal vascular function and elevated TF levels. Heightened endothelial apoptosis may represent an important mechanism for development of atherothrombosis in SLE.
SCL is required for normal function of short-term repopulating hematopoietic stem cells
Authors: Curtis David J. , Hall Mark A. , Van Stekelenburg Leonie J. , Robb Lorraine , Jane Stephen M. , Begley C. Glenn
Published: January 2004

The stem cell leukemia (SCL) gene is essential for the development of hematopoietic stem cells in the embryo. Here, we used a conditional gene targeting approach to examine the function of SCL in adult hematopoietic stem cells (HSCs). Flow cytometry of bone marrow from SCL-deleted mice demonstrated a 4-fold increase in number of Linneg c-kit+ Sca-1+ cells. Despite this increase in the number of phenotypic HSCs, competitive repopulation assays demonstrated a severe multilineage defect in repopulation capacity by SCL-deleted bone marrow cells. SCL-heterozygous cells also showed a mild repopulation defect, thus suggesting haploinsufficiency of SCL. The transplantation defect of SCL-deleted cells was observed within 4 weeks of transplantation, indicating a defect in a multipotent progenitor or short-term repopulating HSCs. Although the defect persisted in secondary transplants, it remained relatively stable, suggesting that SCL was not required for self-renewal of the HSCs. Generation of SCL-deleted cells within SCL-wild-type mice rescued the early repopulating defect. Together, our results suggest that SCL is required for the normal function of short-term repopulating HSCs. (Blood. 2004;103:3342-3348)
Induction and maintenance therapy with intermittent interleukin-2 in HIV-1 infection
Authors: Farel Claire E. , Chaitt Doreen G. , Hahn Barbara K. , Tavel Jorge A. , Kovacs Joseph A. , Polis Michael A. , Masur Henry , Follmann Dean A. , Lane H. Clifford , Davey Richard T.
Published: January 2004

Studies establishing that intermittent subcutaneous interleukin-2 (IL-2) therapy can lead to substantial CD4 cell increases in many HIV-infected patients have generally been of limited duration. We studied 77 patients participating in active longitudinal studies of subcutaneous IL-2 therapy at our center in order to determine the long-term feasibility of this approach. Following initial induction, patients in each trial were eligible to receive intermittent 5-day cycles of subcutaneous IL-2 treatment at individualized doses and frequencies capable of maintaining CD4 counts at postinduction levels. The mean duration of study participation to date is 5.9 years (range, 1.0-9.3 years). Mean baseline CD4 cell count and CD4 percent values of 0.521 × 109/L (521 cells/μL) and 27% have risen to 1.005 × 109/L (1005 cells/μL) and 38%, respectively, at 90 months. The mean number of subcutaneous IL-2 cycles required to achieve and maintain these increases was 10 cycles (range, 3-29 cycles), and the current mean interval of cycling required to maintain these elevations is 39 months (median, 35 months; range, 2-91 months). We conclude that subcutaneous IL-2 therapy is capable of maintaining CD4 cell increases for an extended period using a remarkably low frequency of intermittent cycling. These observations may contribute to patients' acceptance of subcutaneous IL-2 as a favorable long-term treatment strategy. (Blood. 2004;103:3282-3286)
Proteomics techniques and their application to hematology
Authors: Cristea Ileana M. , Gaskell Simon J. , Whetton Anthony D.
Published: January 2004

The recent sequencing of a number of genomes has raised the level of opportunities for studies on proteins. This area of research has been described with the all-embracing term, proteomics. In proteomics, the use of mass spectrometric techniques enables genomic databases to be used to establish the identity of proteins with relatively little data, compared to the era before genome sequencing. The use of related analytical techniques also offers the opportunity to gain information on regulation, via posttranslational modification, and potential new diagnostic and prognostic indicators. Relative quantification of proteins and peptides in cellular and extracellular material remains a challenge for proteomics and mass spectrometry. This review presents an analysis of the present and future impact of these proteomic technologies with emphasis on relative quantification for hematologic research giving an appraisal of their potential benefits.
SAP mediates specific cytotoxic T-cell functions in X-linked lymphoproliferative disease
Authors: Sharifi Reza , Sinclair Joanna C. , Gilmour Kimberly C. , Arkwright Peter D. , Kinnon Christine , Thrasher Adrian J. , Gaspar H. Bobby
Published: January 2004

Cytotoxic T cells (CTLs) and natural killer cells play a major role in the immune response to Epstein-Barr virus (EBV) infection. In X-linked lymphoproliferative (XLP) disease, a severe immunodeficiency, immunodysregulatory phenomena are observed following EBV infection, suggesting that defects exist in these effector populations. The gene defective in XLP is SAP (signaling lymphocytic activation molecule [SLAM]–associated protein), an adaptor protein that mediates signals through SLAM and other immunoglobulin superfamily receptors including 2B4. We generated EBV-specific T-cell lines from controls and XLP patients and examined CTL function in response to different stimuli. We show that XLP patients can generate EBV–T-cell lines that are phenotypically similar to those from controls. XLP patient EBV–T-cell lines showed a significant decrease in interferon-gamma (IFN-γ) production in response to 2B4 and autologous EBV-transformed lymphoblastoid cell line (LCL) stimulation but not in response to SLAM. Furthermore, XLP EBV–T-cell lines demonstrated markedly decreased cytotoxic activity against autologous LCLs. By retroviral gene transfer of the SAP gene into XLP EBV–T-cell lines, we show reconstitution of IFN-γ production and of cytotoxic activity confirming SAP-dependent defects. These studies demonstrate that in XLP the lack of SAP affects specific signaling pathways resulting in severe disruption of CTL function.
Bioengineering of coagulation factor VIII for improved secretion
Authors: Miao Hongzhi Z. , Sirachainan Nongnuch , Palmer Lisa , Kucab Phillip , Cunningham Michael A. , Kaufman Randal J. , Pipe Steven W.
Published: January 2004

Factor VIII (FVIII) functions as a cofactor within the intrinsic pathway of blood coagulation. Quantitative or qualitative deficiencies of FVIII result in the inherited bleeding disorder hemophilia A. Expression of FVIII (domain structure A1-A2-B-A3-C1-C2) in heterologous mammalian systems is 2 to 3 orders of magnitude less efficient compared with other proteins of similar size compromising recombinant FVIII production and gene therapy strategies. FVIII expression is limited by unstable mRNA, interaction with endoplasmic reticulum (ER) chaperones, and a requirement for facilitated ER to Golgi transport through interaction with the mannose-binding lectin LMAN1. Bioengineering strategies can overcome each of these limitations. B-domain-deleted (BDD)-FVIII yields higher mRNA levels, and targeted point mutations within the A1 domain reduce interaction with the ER chaperone immunoglobulin-binding protein. In order to increase ER to Golgi transport we engineered several asparagine-linked oligosaccharides within a short B-domain spacer within BDD-FVIII. A bioengineered FVIII incorporating all of these elements was secreted 15- to 25-fold more efficiently than full-length FVIII both in vitro and in vivo. FVIII bioengineered for improved secretion will significantly increase potential for success in gene therapy strategies for hemophilia A as well as improve recombinant FVIII production in cell culture manufacturing or transgenic animals. (Blood. 2004;103: 3412-3419)
X-linked ectodermal dysplasia and immunodeficiency caused by reversion mosaicism of NEMO reveals a critical role for NEMO in human T-cell development and/or survival
Authors: Nishikomori Ryuta , Akutagawa Hiroshi , Maruyama Kyoko , Nakata-Hizume Mami , Ohmori Katsuyuki , Mizuno Kazunori , Yachie Akihiro , Yasumi Takahiro , Kusunoki Takashi , Heike Toshio , Nakahata Tatsutoshi
Published: January 2004

X-linked ectodermal dysplasia and immunodeficiency (XL-EDA-ID) is an X-linked recessive disease caused by a mutation in the nuclear factor-κB (NF-κB) essential modulator (NEMO). Here we report an XL-EDA-ID patient with atypical features of very few naive-phenotype T cells and defective mitogen-induced proliferation of peripheral blood mononuclear cells (PBMCs). The patient's NEMO defect was diagnosed by flow cytometric analysis of intracellular NEMO staining. Specific cell lineages (monocytes and neutrophils) expressed reduced levels of NEMO, but 2 populations of T, B, and NK cells were detected with normal and reduced expression of NEMO. Genomic analysis revealed that duplication of a 4.4-kb sequence ranging from intron 3 to exon 6 caused the reduced expression of NEMO. Polymorphism analysis showed that the patient's B- and T-cell lines with reduced and normal expression of NEMO had the same X chromosome, indicating that the somatic mosaicism was not due to fetomaternal transfusion but was most likely due to postzygotic reversion. This XLEDA-ID case adds to our understanding of NEMO biology, indicating that NEMO is critical for T-cell development and/or survival in humans as well as in mice. (Blood. 2004;103:4565-4572)
Alemtuzumab is an effective therapy for chronic lymphocytic leukemia with p53 mutations and deletions
Authors: Lozanski Gerard , Heerema Nyla A. , Flinn Ian W. , Smith Lisa , Harbison Jennifer , Webb Jennifer , Moran Mollie , Lucas Margaret , Lin Thomas , Hackbarth Marcy L. , Proffitt John H. , Lucas David , Grever Michael R. , Byrd John C.
Published: January 2004

The presence of p53 mutation or deletion predicts for poor response to conventional therapy in chronic lymphocytic leukemia (CLL). We sought to determine whether the humanized anti-CD52 antibody alemtuzumab was effective in this patient group. Thirty-six patients with fludarabine-refractory CLL were treated with alemtuzumab, 15 (42%) of whom had p53 mutations or deletions. Clinical responses in patients with p53 mutations, deletions, or both were noted in 6 (40%) of 15 versus 4 (19%) of 21 of patients without. The median response duration for this subset of patients was 8 months (range, 3-17 months). These data suggest that alemtuzumab may be an effective therapy for patients with CLL with p53 mutations or deletions. (Blood. 2004;103:3278-3281)
Hematopoiesis is severely altered in mice with an induced osteoblast deficiency
Authors: Visnjic Dora , Kalajzic Zana , Rowe David W. , Katavic Vedran , Lorenzo Joseph , Aguila Hector L.
Published: January 2004

We previously reported a transgenic mouse model expressing herpesvirus thymidine kinase (TK) gene under the control of a 2.3-kilobase fragment of the rat collagen α1 type I promoter (Col2.3ΔTK). This construct confers lineage-specific expression in developing osteoblasts, allowing the conditional ablation of osteoblast lineage after treatment with ganciclovir (GCV). After GCV treatment these mice have profound alterations on bone formation leading to a progressive bone loss. In addition, treated animals also lose bone marrow cellularity. In this report we characterized hematopoietic parameters in GCV-treated Col2.3ΔTK mice, and we show that after treatment transgenic animals lose lymphoid, erythroid, and myeloid progenitors in the bone marrow, followed by decreases in the number of hematopoietic stem cells (HSCs). Together with the decrease in bone marrow hematopoiesis, active extramedullary hematopoiesis was observed in the spleen and liver, as measured by an increase in peripheral HSCs and active primary in vitro hematopoiesis. After withdrawal of GCV, osteoblasts reappeared in the bone compartment together with a recovery of medullary and decrease in extramedullary hematopoiesis. These observations directly demonstrate the role of osteoblasts in hematopoiesis and provide a model to study the interactions between the mesenchymal and hematopoietic compartments in the marrow. (Blood. 2004; 103:3258-3264)
Case report: fatal poisoning with Colchicum autumnale
Authors: Brvar Miran , Ploj Tom , Kozelj Gordana , Mozina Martin , Noc Marko , Bunc Matjaz
Published: January 2004

Introduction

Colchicum autumnale, commonly known as the autumn crocus, contains alkaloid colchicine with antimitotic properties.

Case report

A 76-year-old man with a history of alcoholic liver disease and renal insufficiency, who mistakenly ingested Colchicum autumnale instead of wild garlic (Aliium ursinum), presented with nausea, vomiting and diarrhea 12 hours after ingestion. On admission the patient had laboratory signs of dehydration. On the second day the patient became somnolent and developed respiratory insufficiency. The echocardiogram showed heart dilatation with diffuse hypokinesia with positive troponin I. The respiratory insufficiency was further deteriorated by pneumonia, confirmed by chest X-ray and later on by autopsy. Laboratory tests also revealed rhabdomyolysis, coagulopathy and deterioration of renal function and hepatic function. The toxicological analysis disclosed colchicine in the patient's urine (6 μg/l) and serum (9 μg/l) on the second day. Therapy was supportive with hydration, vasopressors, mechanical ventilation and antibiotics. On the third day the patient died due to asystolic cardiac arrest.

Discussion and conclusion

Colchicine poisoning should be considered in patients with gastroenterocolitis after a meal of wild plants. Management includes only intensive support therapy. A more severe clinical presentation should be expected in patients with pre-existing liver and renal diseases. The main reasons for death are cardiovascular collapse, respiratory failure and leukopenia with infection.
Conditional macrophage ablation in transgenic mice expressing a Fas-based suicide gene
Authors: Burnett Sandra H , Kershen Edward J , Zhang Jiayou , Zeng Li , Straley Susan C , Kaplan Alan M , Cohen Donald A
Published: January 2004

Transgenic mice expressing an inducible suicide gene, which allows systemic and reversible elimination of macrophages, were developed. A macrophage-specific c-fms promoter was used to express enhanced green fluorescent protein and a drug-inducible suicide gene that leads to Fas-mediated apoptosis in resting and cycling cells of the macrophage lineage. Transgenic mice were fertile, of normal weight, and showed no abnormal phenotype before drug exposure. The transgene was expressed constitutively in macrophages and dendritic cells (DC) but not significantly in T cells or B cells. Induction of the suicide gene led to depletion of 70–95% of macrophages and DC in nearly all tissues examined. Depletion reduced the ability to clear bacteria from the blood and led to increased bacterial growth in the liver. Depleted mice displayed several abnormalities, including splenomegaly, lymphadenopathy, thymic atrophy, extramedullary hematopoiesis, and development of peritoneal adhesions. This new, transgenic line will be useful in investigating the role of macrophages and DC.
TNFerade Biologic, an Adenovector With a Radiation-Inducible Promoter, Carrying the Human Tumor Necrosis Factor Alpha Gene: A Phase I Study in Patients With Solid Tumors
Authors: Senzer Neil , Mani Sridhar , Rosemurgy Alexander , Nemunaitis John , Cunningham Casey , Guha Chandan , Bayol Natalia , Gillen Michelle , Chu Karen , Rasmussen Camilla , Rasmussen Henrik , Kufe Donald , Weichselbaum Ralph , Hanna Nader
Published: January 2004

Purpose

TNFerade is a replication deficient adenovector that expresses human tumor necrosis factor alpha under control of the radiation-inducible Egr-1 promoter. The goals of this study were to determine the safety and toxicity of TNFerade in combination with radiation therapy.

Patients and Methods

TNFerade was administered by intratumoral administration, weekly for 6 weeks with concomitant radiation (30 to 70 Gy). Seven dose levels were studied (4 × 107particle units [pu] to 4 × 1011pu) in patients with solid tumors being treated with radiation.

Results

Thirty-six patients were assessable for toxicity and 30 for tumor response. Most frequent TNFerade-related toxicities were fever (22%), injection site pain (19%), and chills (19%). No dose-limiting toxicities were observed. Overall, 21 of 30 patients (70%) demonstrated objective tumor response (five complete responses, nine partial responses, and seven minimal responses). In four of five patients with synchronous lesions, a differential response between lesions treated with TNFerade + radiation compared with radiation only was observed.

Conclusion

This is the first human study with TNFerade and radiation. The integrated treatment was well tolerated in patients with predominantly prior treatment-refractory solid tumors. Controlled prospective clinical trials have been initiated to more fully define the therapeutic contribution of TNFerade.
Selective CD4+ Lymphopenia in Melanoma Patients Treated With Temozolomide: A Toxicity With Therapeutic Implications
Authors: Su Y.B. , Sohn Sejean , Krown Susan E. , Livingston Philip O. , Wolchok Jedd D. , Quinn Carolyn , Williams Linda , Foster Theresa , Sepkowitz Kent A. , Chapman Paul B.
Published: January 2004

Purpose

Standard schedule temozolomide (TMZ; daily for 5 days every 4 weeks) is often used in melanoma patients, but phase III data show that it is no more effective than standard dacarbazine. Extended TMZ dosing regimens may be superior by delivering the drug continuously at a higher dose over time. Using an extended dosing schedule, we noted a high incidence of lymphopenia and occasional opportunistic infections (OIs). Here we report our retrospective experience in the first 97 patients.

Materials and Methods

TMZ was administered at 75 mg/m2/d orally for 6 weeks every 8 weeks, although nine patients were treated continuously without a break. Seventeen patients were treated with TMZ alone; 73 patients received TMZ with thalidomide; seven patients received TMZ with low-dose interferon alfa.

Results

Median duration of TMZ treatment was 113 days; 29% received ≥ 24 weeks of therapy. Lymphopenia was seen in 60% of patients (absolute lymphocyte count < 800/μL) with a median of 101 days to lymphopenia. TMZ did not cause significant neutropenia or thrombocytopenia. Lymphopenia was not more common in patients treated concomitantly with thalidomide. In all patients analyzed for lymphocyte subsets, lymphopenia induced by TMZ affected the CD4+ compartment preferentially. There were two documented OIs (Pneumocystis and Aspergillus pneumonia) as well as other infections indicative of T-cell dysfunction in another 21 patients.

Conclusion

TMZ at this dose and schedule results in CD4+ lymphopenia in a majority of patients that can result in OIs. Pneumocystis pneumonia prophylaxis should be considered for patients who develop sustained lymphopenia on TMZ.
Reproductive Abnormalities in Human Insulin-Like Growth Factor-Binding Protein-1 Transgenic Male Mice
Authors: Froment Pascal , Staub Christophe , Hembert Stéphanie , Pisselet Claudine , Magistrini Michèle , Delaleu Bernadette , Seurin Danielle , Levine Jon E. , Johnson Larry , Binoux Michel , Monget Philippe
Published: January 2004

Adult transgenic mice overexpressing human insulin-like growth factor-binding protein-1 in the liver present reproductive abnormalities in both sexes. In the present work, we have investigated the mechanisms responsible for limiting breeding capacity in these transgenic male mice. Homozygous adult transgenic male mice (3–6 months old) exhibited irregular copulatory behavior and a reduction of the number of pregnancies per female as well as of litter size per pregnancy. Genital tract weight, more specifically epididymal and seminal vesicle weights, were reduced by 45% in homozygous transgenic vs. nontransgenic mice. Homozygous transgenic mice exhibited a 30% reduction of the length of seminiferous tubules (P = 0.007), a 30% decrease in daily sperm production per testis (P = 0.019), and a 50% decrease in the number of spermatozoa in testis (P = 0.037), associated with morphological abnormalities of the sperm heads leading to an approximately 50% reduction of fertilized two-cell eggs (P = 0.002) and of implanted embryos on d 5.5 after mating (P = 0.004). The round spermatids also appeared altered in their morphology. In addition, Leydig cells in homozygous transgenic mice exhibited an altered appearance, with a 1.8-fold increase in lipid droplets in their cytoplasm (P < 0.001). Moreover, the concentration of 3β-hydroxysteroid dehydrogenase was 66% lower in testis from transgenics compared with those from normal mice (P = 0.01), leading to a tendency toward lower plasma testosterone levels (P = 0.1). Interestingly, LH concentrations were increased by 40% in transgenic pituitary extracts (P = 0.02), and basal LH secretion by pituitary explants in vitro was increased by 60% in homozygous transgenic vs. normal mice (P = 0.04), suggesting an alteration of LH pulsatile secretion in vivo. In conclusion, these data suggest that the breeding impairment of human insulin-like growth factor-binding protein-1 transgenic males is due at least in part to an alteration of the process of spermatogenesis, leading to a diminution of sperm production and of its quality. Minor impairment of steroidogenesis may also contribute to the reduced reproductive capacity of these animals. Our observations are consistent with the idea that normal spermatogenesis and perhaps also steroidogenesis are dependent on the actions of sufficient concentrations of unbound IGF-I.
A Novel Missense Mutation in the Mouse Growth Hormone Gene Causes Semidominant Dwarfism, Hyperghrelinemia, and Obesity
Authors: Meyer Carola W. E. , Korthaus Dirk , Jagla Wolfgang , Cornali Emmanuelle , Grosse Johannes , Fuchs Helmut , Klingenspor Martin , Roemheld Stephanie , Tschöp Matthias , Heldmaier Gerhard , de Angelis Martin Hrabé , Nehls Michael
Published: January 2004

The SMA1-mouse is a novel ethyl-nitroso-urea (ENU)-induced mouse mutant that carries an a→g missense mutation in exon 5 of the GH gene, which translates to a D167G amino acid exchange in the mature protein. Mice carrying the mutation are characterized by dwarfism, predominantly due to the reduction (sma1/+) or absence (sma1/sma1) of the GH-mediated peripubertal growth spurt, with sma1/+ mice displaying a less pronounced phenotype. All genotypes are viable and fertile, and the mode of inheritance is in accordance with a semidominant Mendelian trait. Adult SMA1 mice accumulate excessive amounts of sc and visceral fat in the presence of elevated plasma ghrelin levels, possibly reflecting altered energy partitioning. Our results suggest impaired storage and/or secretion of pituitary GH in mutants, resulting in reduced pituitary GH and reduced GH-stimulated IGF-1 expression. Generation and identification of the SMA1 mouse exemplifies the power of the combination of random mouse mutagenesis with a highly detailed phenotype-analysis as a successful strategy for the detection and analysis of novel gene-function relationships.
Human Chorionic Gonadotropin-Dependent Regulation of 17β-Hydroxysteroid Dehydrogenase Type 4 in Preovulatory Follicles and Its Potential Role in Follicular Luteinization
Authors: Brown Kristy A. , Boerboom Derek , Bouchard Nadine , Doré Monique , Lussier Jacques G. , Sirois Jean
Published: January 2004

17β-Hydroxysteroid dehydrogenase type 4 (17βHSD4) has a unique multidomain structure, with one domain involved in 17β-estradiol inactivation. The objective of the study was to investigate the regulation of 17βHSD4 during human chorionic gonadotropin (hCG)-induced ovulation/luteinization. The equine 17βHSD4 cDNA was cloned and was shown to encode a 735-amino acid protein that is highly conserved (81–87% identity) compared with other mammalian orthologs. RT-PCR/Southern blot analyses were performed to study the regulation of 17βHSD4 transcripts in equine preovulatory follicles isolated between 0–39 h after hCG treatment. Results showed the presence of basal 17βHSD4 mRNA expression before hCG treatment, but an increase was observed in follicles obtained 24 h after hCG (P < 0.05). Analyses of isolated preparations of granulosa and theca interna cells identified basal mRNA expression in both layers, but granulosa cells appeared as the predominant site of follicular 17βHSD4 mRNA induction. A specific polyclonal antibody was raised against a fragment of the equine protein and used to study regulation of the 17βHSD4 protein. Immunoblots showed an increase in full-length 17βHSD4 protein in follicles 24 h after hCG (P < 0.05), in keeping with mRNA results. Immunohistochemical data confirmed the induction of the enzyme in follicular cells after hCG treatment. Collectively, these results demonstrate that the gonadotropin-dependent induction of follicular luteinization is accompanied by an increase in 17βHSD4 expression. Considering the estrogen-inactivating function of 17βHSD4, its regulated expression in luteinizing preovulatory follicles appears as a potential complementary mechanism to reduce circulating levels of 17β-estradiol after the LH surge.
Distinct Components of Janus Kinase/Signal Transducer and Activator of Transcription Signaling Pathway Mediate the Regulation of Systemic and Tissue Localized Renin-Angiotensin System
Authors: Guo Yueling , Mascareno Eduardo , Siddiqui M. A. Q.
Published: January 2004

In an attempt to demonstrate the linkage between the Janus kinase (Jak)/signal transducer and activator of transcription (STAT) signaling and the activity of the systemic or local renin-angiotensin system in vivo, we produced transgenic mice harboring angiotensinogen (ANG) promoter containing the wild-type or mutant STAT target site (St-domain) fused to the luciferase reporter. The ANG-promoter-driven luciferase expression was dependent upon phosphorylation of Jak2, as administration of tyrphostin AG490, a potent inhibitor of Jak2, down-regulated the ANG promoter activity and abolished the stimulated endogenous ANG mRNA level in the liver. Administration of angiotensin II peptide to the mice resulted in prominent expression of luciferase in the liver and heart of animals containing wild type St-domain, but not in transgenes with mutant St-domain. Angiotensin II-induced signaling caused activation of STAT proteins in the liver (systemic), the pattern of which was distinct from that in the heart (local). The inducible expression of ANG promoter appears to be mediated by physical association of p300 with STAT 5B in liver and STAT 3 and STAT 5A in heart. Taken together, these results point to the differences in signaling mechanisms in the circulating and localized renin-angiotensin system and identify at least two molecular steps, the tyrosyl phosphorylation of Jak2 and the STAT/St-domain interaction, as pivotal in the regulation of ANG gene transcription.
Ectopic Myf5 or MyoD prevents the neuronal differentiation program in addition to inducing skeletal muscle differentiation, in the chick neural tube
Authors: Delfini Marie-Claire , Duprez Delphine
Published: January 2004

Forced expression of the bHLH myogenic factors, Myf5 and MyoD, in various mammalian cell lines induces the full program of myogenic differentiation. However, this property has not been extensively explored in vivo. We have taken advantage of the chick model to investigate the effect of electroporation of the mouse Myf5 and MyoD genes in the embryonic neural tube. We found that misexpression of either mouse Myf5 or MyoD in the chick neural tube leads to ectopic skeletal muscle differentiation, assayed by the expression of the myosin heavy chains in the neural tube and neural crest derivatives. We also showed that the endogenous neuronal differentiation program is inhibited under the influence of either ectopic mouse Myf5 or MyoD. We used this new system to analyse, in vivo, the transcriptional regulation between the myogenic factors. We found that MyoD and Myogenin expression can be activated by ectopic mouse Myf5 or MyoD, while Myf5 expression cannot be activated either by mouse MyoD or by itself. We also analysed the transcriptional regulation between the myogenic factors and the different genes involved in myogenesis, such as Mef2c, Pax3, Paraxis, Six1, Mox1, Mox2 and FgfR4. We established the existence of an unexpected regulatory loop between MyoD and FgfR4. The consequences for myogenesis are discussed.
Moesin contributes an essential structural role inDrosophilaphotoreceptor morphogenesis
Authors: Karagiosis Sue A. , Ready Donald F.
Published: January 2004

Ezrin-Radixin-Moesin (ERM) family proteins organize heterogeneous sub-plasma membrane protein scaffolds that shape membranes and their physiology. In Drosophila oocytes and imaginal discs, epithelial organization, fundamental to development and physiology, is devastated by the loss of Moesin. Here, we show that Moesin is crucial for Drosophilaphotoreceptor morphogenesis. Beyond its requirement for retinal epithelium integrity, Moesin is essential for the proper assembly of the apical membrane skeleton that builds the photosensitive membrane, the rhabdomere. Moesin localizes to the rhabdomere base, a dynamic locus of cytoskeletal reorganization and membrane traffic. Downregulation of Moesin through RNAi or genetic loss of function profoundly disrupts the membrane cytoskeleton and apical membrane organization. We find normal levels and distribution of Moesin in photoreceptors of a Moesin mutant previously regarded as protein null, suggesting alternative interpretations for studies using this allele. Our results show an essential structural role for Moesin in photoreceptor morphology.
Hydroformylation of Mixed Octenes Using Rhodium–Bulky Phosphonite Complexes with Excellent Catalytic Activity and Stability
Authors: Jeon Jong-Ki , Park Young-Kwon , Kim Ji Man
Published: January 2004

Rhodium catalyst liganded with tetrakis-(2,4-di-t-butylphenyl)-4,4′-biphenylphosphonite exhibited an excellent catalytic activity as well as stability for the hydroformylation of mixed olefins, especially butene dimer.
Formation of Micro and Nanoscale Patterns of Monolayer Templates for Position Selective Immobilization of Oligonucleotide Using Ultraviolet and Electron Beam Lithography
Authors: Niwa Daisuke , Omichi Kaoru , Motohashi Norikazu , Homma Takayuki , Osaka Tetsuya
Published: January 2004

Formation of organosilane monolayer templates using ultraviolet and electron-beam (EB) lithography was investigated. The oligonucleotides were covalently immobilized with high selectivity only to the amino-monolayer modified regions locally formed on the template surfaces at micro and nanometer scale. By using EB lithography, patterned immobilization in nanometer scale, as small as 20 nm, was achieved.
Intramolecular Proton Transfer Inducing a Biszwitterion Structure in a N4 Schiff Base
Authors: Karamé Iyad , Tommasino M Lorraine , Faure René , Fenet Bernard , Lemaire Marc
Published: January 2004

X-ray and NMR evidences of a sulfonated and tetradentate N4 Schiff base pointed out unusual iminium groups. This produces amide groups and the protons transfer leads to an overall biszwitterion structure.
A Novel 1-D Ladder-like Coordination Polymer [Eu(dipic)1.5(H2O)4·3H2O]∞
Authors: Liu Zhiliang , Zhang Deqing , Liu Caiming , Zhu Daoben
Published: January 2004

A novel one-dimensional infinite ladder-like lanthanide (EuIII) coordination polymer was obtained by hydrothermal method and its structure and magnetic and photoemission properties were investigated.
Resolution of the Acid-base Fraction Curves of the Calixarene Derivatives with Chemometric Methods
Authors: Wang Li , Zhu Zhong-Liang , Shi Xian-Fa
Published: January 2004

The acid-base properties of several new calix[4]arene derivatives were studied with chemometric methods by measurement of their UV absorbances under different pH. Resolving the acid-base fraction curves with chemometric methods, the pKa of these derivatives were determined and the fraction curves and pure absorbing spectra of each absorbing component in the calixarene system were obtained. The proton dissociation behavior of these derivatives was also discussed in this paper.
Interconversion between syn and anti Conformations of 1,3-Bis(O-cyanomethyl)-p-tert-butylthiacalix[4]arene
Authors: Bhalla Vandana , Kumar Manoj , Kabuto Chizuko , Hattori Tetsutaro , Miyano Sotaro
Published: January 2004

1,3-Bis(O-cyanomethyl)-p-tert-butylthiacalix[4]arene (5) has been found to interconvert between syn and anti conformations in solution. The equilibrium shifts toward the anti form with increasing solvent polarity. In the solid state, it adopts a pinched cone conformation with the syn arrangement of the cyanomethyl groups. Reduction of the equilibrium mixture of 5 with LiAlH4 gives only anti stereoisomer of 1,3-bis(O-aminoethyl)-p-tert-butylthiacalix[4]arene.
An Unbridged Platinum(III) Dimer with Added Chloro Ligands in Equatorial Sites, [Pt2Cl2(phpy)4] (Hphpy = phenylpyridine), Synthesized by an Oxidation with Aurous Complex
Authors: Yamaguchi Tadashi , Kubota Osamu , Ito Tasuku
Published: January 2004

An unsupported platinum(III) dimer having added chloro ligands at equatorial sites and novel axial-equatorial chelate ligands, [Pt2Cl2(phpy)4] (Hphpy = phenylpyridine) was obtained from an oxidative addition reaction of [Pt(phpy)2] with [AuCl(SMe2)].
Short Report: False-negative primary neonatal thyroid screening: the need for clinical vigilance and secondary screening
Authors: Yunis K A , Nasr M R , Lepejian G , Najjar S , Daher R
Published: January 2004

The screening of newborn babies for congenital hypothyroidism has changed the natural history of this abnormality. We describe here a case of a female patient with congenital hypothyroidism that was missed by primary neonatal thyroid screening (using thyroid-stimulating hormone) at two days of age; it was detected only after the development and persistence of jaundice during the first three weeks of life. A normal neonatal screening result does not preclude the development of hypothyroidism later in infancy. Clinical vigilance must be maintained by practitioners. A second screening between two and six weeks of age may be useful in order to detect the few cases missed at first screening.
Original Paper: Informed consent for breast screening: what should we tell women?
Authors: Marshall T , Adab P
Published: January 2004

Objective: To illustrate visually the lifetime probabilities of the principal outcomes of the UK breast screening programme in a readily understandable format.

Methods: We derived prognostic data from a modelling exercise using published effectiveness data and routine data sources. We calculated the probability that a woman will survive to the age of 75 if she chooses to participate fully in breast screening from age 50 to 64 and if she chooses not to participate. We also calculated her probability of being referred for assessment, undergoing fine-needle biopsy and undergoing open biopsy. We present these data in two alternative decision aid formats. These alternative formats illustrate visually the outcomes for 1000 women and 100 women choosing each alternative: breast screening or no breast screening.

Results: A woman participating in breast screening from age 50 to 64 increases her chances of surviving to age 75 by 0.6%. She has a 21.8% probability of surviving to age 75 and being referred for assessment but no further investigation. She has a 5.7% probability of undergoing core biopsy and a 0.9% probability of undergoing open biopsy. This information can easily be presented visually.

Conclusions: We can provide realistic estimates of the effects of the breast screening programme on mortality in a readily understandable format. If we wish women to make an informed choice about breast screening they must be given this information.