bluuebunny/crossref_metadata_2025 huggingface dataset:
Graft-versus-leukemia effect after suicide-gene–mediated control of graft-versus-host disease
Authors: Litvinova Elena , Maury Sébastien , Boyer Olivier , Bruel Sylvie , Benard Laurent , Boisserie Gilbert , Klatzmann David , Cohen José L.
Published: August 2002

Clinical data indicate that after allogeneic hematopoietic stem cell transplantation (HSCT) for hematological malignancies, the graft-versus-leukemia (GVL) effect is in large part mediated by the graft-versus-host reaction (GVHR), which also often leads to graft-versus-host disease (GVHD). Controlling alloreactivity to prevent GVHD while retaining GVL poses a true dilemma for the successful treatment of such malignancies. We reasoned that suicide gene therapy, which kills dividing cells expressing the thymidine kinase (TK) “suicide” gene using time-controlled administration of ganciclovir (GCV), might solve this dilemma. We have previously shown that after infusion of allogeneic TK T cells along with HSCT to an irradiated recipient, an early and short GCV treatment efficiently prevents GVHD by selectively eliminating alloreactive T cells while sparing nonalloreactive T cells, which can then contribute to immune reconstitution. Nevertheless, it remained to be established that this therapeutic strategy retained the desired GVL effect. Hypothesizing that a contained GVHR would be essential, we evaluated the GVL effect using different protocols of GCV administration. We were able to show that when the GCV treatment is initiated at, or close to, the time of grafting, GVHD is controlled but GVL is lost. In contrast, when the onset of GCV administration is delayed until day 6, a potent GVL effect is retained while GVHD is still controlled. These data emphasize that, by a time-optimized scheduling of the administration of GCV, this TK/GCV strategy can be tuned to efficiently treat malignant hemopathies.
CD8 T-cell responses to Wilms tumor gene product WT1 and proteinase 3 in patients with acute myeloid leukemia
Authors: Scheibenbogen Carmen , Letsch Anne , Thiel Eckhard , Schmittel Alexander , Mailaender Volker , Baerwolf Steffi , Nagorsen Dirk , Keilholz Ulrich
Published: August 2002

Wilms tumor gene product WT1 and proteinase 3 are overexpressed antigens in acute myeloid leukemia (AML), against which cytotoxic T lymphocytes can be elicited in vitro and in murine models. We performed this study to investigate whether WT1- and proteinase 3-specific CD8 T cells spontaneously occur in AML patients. T cells recognizing HLA-A2.1-binding epitopes from WT1 or proteinase 3 could be detected ex vivo in 5 of 15 HLA-A2–positive AML patients by interferon-γ (IFN-γ) ELISPOT assay and flow cytometry for intracellular IFN-γ and in 3 additional patients by flow cytometry only. T cells producing IFN-γ in response to proteinase 3 were further characterized in one patient by 4-color flow cytometry, identifying them as CD3+CD8+CD45RA+ CCR7−T cells, resembling cytotoxic effector T cells. In line with this phenotype, most of the WT1- and proteinase-reactive T cells were granzyme B+. These results provide for the first time evidence for spontaneous T-cell reactivity against defined antigens in AML patients. These data therefore support the immunogenicity of WT1 and proteinase 3 in acute leukemia patients and the potential usefulness of these antigens for leukemia vaccines.
High-level allogeneic chimerism achieved by prenatal tolerance induction and postnatal nonmyeloablative bone marrow transplantation
Authors: Peranteau William H. , Hayashi Satoshi , Hsieh Michael , Shaaban Aimen F. , Flake Alan W.
Published: August 2002

Clinical application of allogeneic bone marrow transplantation (BMT) has been limited by toxicity related to cytoreductive conditioning and immune response. In utero hematopoietic stem cell transplantation (IUHSCT) is a nonablative approach that achieves mixed chimerism and donor-specific tolerance but has been limited by minimal engraftment. We hypothesized that mixed chimerism achieved by IUHSCT could be enhanced after birth by nonmyeloablative total body irradiation (TBI) followed by same-donor BMT. To test this hypothesis, mixed chimerism was created by IUHSCT in a major histocompatibility complex-mismatched strain combination. After birth, chimeric animals received nonmyeloablative TBI followed by transplantation of donor congenic bone marrow cells. Our results show that: (1) low-level chimerism after IUHSCT can be enhanced to high-level chimerism by this strategy; (2) enhancement of chimerism is dependent on dose of TBI; (3) the mechanism of TBI enhancement is via a transient competitive advantage for nonirradiated hematopoietic stem cells; (4) engraftment observed in the tolerant, fully allogeneic IUHSC transplant recipient is equivalent to a congenic recipient; and (5) host-reactive donor lymphocytes are deleted with no evidence of graft-versus-host disease. This study supports the concept of prenatal tolerance induction to facilitate nonmyeloablative postnatal strategies for cellular therapy. If clinically applicable, such an approach could dramatically expand the application of IUHSCT.
Development of virus vectors for gene therapy of β chain hemoglobinopathies: flanking with a chromatin insulator reduces γ-globin gene silencing in vivo
Authors: Emery David W. , Yannaki Evangelia , Tubb Julie , Nishino Tamon , Li Qiliang , Stamatoyannopoulos George
Published: August 2002

We have previously described the development of oncoretrovirus vectors for human γ-globin using a truncated β-globin promoter, modified γ-globin cassette, and α-globin enhancer. However, one of these vectors is genetically unstable, and both vectors exhibit variable expression patterns in cultured cells, common characteristics of oncoretrovirus vectors for globin genes. To address these problems, we identified and removed the vector sequences responsible for genetic instability and flanked the resultant vector with the chicken β-globin HS4 chromatin insulator to protect expression from chromosomal position effects. After determining that flanking with the cHS4 element allowed higher, more uniform levels of γ-globin expression in MEL cell lines, we tested these vectors using a mouse bone marrow transduction and transplantation model. When present, the γ-globin cassettes from the uninsulated vectors were expressed in only 2% to 5% of red blood cells (RBCs) long term, indicating they are highly sensitive to epigenetic silencing. In contrast, when present the γ-globin cassette from the insulated vector was expressed in 49% ± 20% of RBCs long term. RNase protection analysis indicated that the insulated γ-globin cassette was expressed at 23% ± 16% per copy of mouse α-globin in transduced RBCs. These results demonstrate that flanking a globin vector with the cHS4 insulator increases the likelihood of expression nearly 10-fold, which in turn allows for γ-globin expression approaching the therapeutic range for sickle cell anemia and β thalassemia.
CCL19 induces rapid dendritic extension of murine dendritic cells
Authors: Yanagawa Yoshiki , Onoé Kazunori
Published: August 2002

Dendritic cells (DCs) possess numerous dendrites that may be of great advantage to interaction with T cells. However, it has been poorly understood how the dendritic morphology of a DC is controlled. In the present study, using a murine spleen-derived DC line, we analyzed effects of CCR7 ligands, CCL19 and CCL21, on dendritic morphology. Mature DCs, but not immature DCs, showed vigorous migration to either CCL19 or CCL21. CCL19 also rapidly (within 30 minutes) induced marked extension of dendrites of mature DCs that was maintained at least for 24 hours. On the other hand, CCL21 failed to induce rapid dendritic extension, even though a modest dendritic extension of mature DCs, compared to that by CCL19, was induced 8 or 24 hours after treatment with CCL21. In addition, pretreatment with a high concentration of CCL21 significantly inhibited the rapid dendritic extension induced by CCL19. Thus, it is suggested that CCL19 and CCL21 exert agonistic and antagonistic influences on the initiation of dendritic extension of mature DCs. The CCL19-induced morphologic changes were completely blocked by Clostridium difficiletoxin B that inhibits Rho guanosine triphosphatase proteins such as Rho, Rac, and Cdc42, but not by Y-27632, a specific inhibitor for Rho-associated kinase. These findings suggest that Rac or Cdc42 (or both), but not Rho, are involved in the CCL19-induced dendritic extension of mature DCs.
Natural killer cell receptors: new biology and insights into the graft-versus-leukemia effect
Authors: Farag Sherif S. , Fehniger Todd A. , Ruggeri Loredana , Velardi Andrea , Caligiuri Michael A.
Published: August 2002

Natural killer (NK) cells have held great promise for the immunotherapy of cancer for more than 3 decades. However, to date only modest clinical success has been achieved manipulating the NK cell compartment in patients with malignant disease. Progress in the field of NK cell receptors has revolutionized our concept of how NK cells selectively recognize and lyse tumor and virally infected cells while sparing normal cells. Major families of cell surface receptors that inhibit and activate NK cells to lyse target cells have been characterized, including killer cell immunoglobulinlike receptors (KIRs), C-type lectins, and natural cytotoxicity receptors (NCRs). Further, identification of NK receptor ligands and their expression on normal and transformed cells completes the information needed to begin development of rational clinical approaches to manipulating receptor/ligand interactions for clinical benefit. Indeed, clinical data suggest that mismatch of NK receptors and ligands during allogeneic bone marrow transplantation may be used to prevent leukemia relapse. Here, we review how NK cell receptors control natural cytotoxicity and novel approaches to manipulating NK receptor-ligand interactions for the potential benefit of patients with cancer.
2-Methoxyestradiol overcomes drug resistance in multiple myeloma cells
Authors: Chauhan Dharminder , Catley Laurence , Hideshima Teru , Li Guilan , Leblanc Richard , Gupta Deepak , Sattler Martin , Richardson Paul , Schlossman Robert L. , Podar Klaus , Weller Edie , Munshi Nikhil , Anderson Kenneth C.
Published: August 2002

2-Methoxyestradiol (2ME2) an estrogen derivative, induces growth arrest and apoptosis in leukemic cells and is also antiangiogenic. In this study, we demonstrate that 2ME2 inhibits growth and induces apoptosis in multiple myeloma (MM) cell lines and patient cells. Significantly, 2ME2 also inhibits growth and induces apoptosis in MM cells resistant to conventional therapies including melphalan (LR-5), doxorubicin (Dox-40 and Dox-6), and dexamethasone (MM.1R). In contrast to its effects on MM cells, 2ME2 does not reduce the survival of normal peripheral blood lymphocytes. Moreover, 2ME2 enhances Dex-induced apoptosis, and its effect is not blocked by interleukin-6 (IL-6). We next examined the effect of 2ME2 on MM cells in the bone marrow (BM) milieu. 2ME2 decreases survival of BM stromal cells (BMSCs), as well as secretion of vascular endothelial growth factor (VEGF), and IL-6 triggered by the adhesion of MM cells to BMSCs. We show that apoptosis induced by 2ME2 is mediated by the release of mitochondrial cytochrome-c (cyto-c) and Smac, followed by the activation of caspases-8, -9, and -3. Finally, 2ME2 inhibits MM cell growth, prolongs survival, and decreases angiogenesis in a murine model. These studies, therefore, demonstrate that 2ME2 mediates anti-MM activity directly on MM cells and in the BM microenvironment. They provide a framework for the use of 2ME2, either alone or in combination with Dex, to overcome drug resistance and to improve outcome in MM.
X-linked thrombocytopenia with thalassemia from a mutation in the amino finger of GATA-1 affecting DNA binding rather than FOG-1 interaction
Authors: Yu Channing , Niakan Kathy K. , Matsushita Mark , Stamatoyannopoulos George , Orkin Stuart H. , Raskind Wendy H.
Published: August 2002

Transcription factor GATA-1 is essential for the development of erythroid cells and megakaryocytes. Each of its 2 zinc fingers is critical for normal function. The C-terminal finger is necessary for DNA binding. The N finger mediates interaction with FOG-1, a cofactor for GATA-1, and also modulates DNA-binding affinity, notably at complex or palindromic GATA sites. Residues of the N finger–mediating interaction with FOG-1 lie on the surface of the N finger facing away from DNA. Strong sequence conservation of residues facing DNA suggests that this other surface may also have an important role. We report here that a syndrome of X-linked thrombocytopenia with thalassemia in humans is caused by a missense mutation (Arg216Gln) in the GATA-1 N finger. To investigate the functional consequences of this substitution, we used site-directed mutagenesis to alter the corresponding residue in GATA-1. Compared with wild-type GATA-1, Arg216Gln GATA-1 shows comparable affinity to single GATA sites but decreased affinity to palindromic sites. Arg216Gln GATA-1 interacts with FOG-1 similarly with wild-type GATA-1. Arg216Gln GATA-1 supports erythroid maturation of GATA-1 erythroid cells, albeit at reduced efficiency compared with wild-type GATA-1. Together, these findings suggest that residues of the N finger of GATA-1–facing DNA contribute to GATA-1 function apart from interaction with the cofactor FOG-1. This is also the first example of β-thalassemia in humans caused by a mutation in an erythroid transcription factor.
Racial and ethnic differences in survival of children with acute lymphoblastic leukemia
Authors: Bhatia Smita , Sather Harland N. , Heerema Nyla A. , Trigg Michael E. , Gaynon Paul S. , Robison Leslie L.
Published: August 2002

Black children with acute lymphoblastic leukemia (ALL) have poor outcomes, but limited information is available for children from other racial and ethnic backgrounds, such as Hispanic and Asian. We undertook a retrospective cohort study of children with ALL treated on Children's Cancer Group therapeutic protocols to determine outcomes by racial and ethnic backgrounds of patients treated with contemporary risk-based therapy. In total, 8447 children (white, n = 6703; Hispanic, n = 1071; black, n = 506; and Asian, n = 167) with newly diagnosed ALL between 1983 and 1995 were observed for a median of 6.5 years. Analysis of disease outcome was measured as overall survival (OS) and event-free survival (EFS) and was adjusted for known predictors of outcome including clinical features, disease biology, socioeconomic status, and treatment era (1983-1989 vs 1989-1995). There was a statistically significant difference in survival by ethnicity (P < .001). Five-year EFS rates were: Asian, 75.1% ± 3.5%; white, 72.8% ± 0.6%; Hispanic, 65.9% ± 1.5%; and black, 61.5% ± 2.2%. Multivariate analysis revealed that when compared with white children, black and Hispanic children had worse outcomes and Asian children had better outcomes after adjusting for known risk factors. The poorer outcomes among black children were most apparent among patients with standard-risk features (relative risk [RR], 2.0; 95% confidence interval [CI], 1.6-2.5), whereas poorer outcomes in Hispanic children (RR, 1.4; 95% CI, 1.2-1.6) were most evident among patients with high-risk features. Asian children had better outcomes than all racial and ethnic groups among high-risk patients, particularly in the recent era (5-year EFS, 90.9% ± 6.1%). Racial and ethnic differences in OS and EFS persist among children with ALL who receive contemporary risk-based therapy. Future studies should focus on reasons—perhaps compliance or pharmacogenetics—for those differences.
Evidence for a graft-versus-leukemia effect after allogeneic peripheral blood stem cell transplantation with reduced-intensity conditioning in acute myelogenous leukemia and myelodysplastic syndromes
Authors: Martino Rodrigo , Caballero Marı́a Dolores , Pérez Simón José Antonio , Canals Carmen , Solano Carlos , Urbano-Ispı́zua Alvaro , Bargay Joan , Léon Angel , Sarrá Josep , Sanz Guillermo F. , Moraleda José Marı́a , Brunet Salut , San Miguel Jesús , Sierra Jorge
Published: August 2002

We report the results of a prospective study of a reduced-intensity conditioning (RIC) regimen followed by allogeneic peripheral blood stem cell transplantation (PBSCT) from an HLA-identical sibling in 37 patients with acute myeloid leukemia (AML; n = 17) or myelodysplastic syndrome (MDS; n = 20). The median age was 57 years, and 22 (59%) were beyond the early phase of their disease. The incidence of grade II to IV acute graft-versus-host disease (GVHD) was 19% (5% grade III-IV), and the 1-year incidence of chronic extensive GVHD was 46%. With a median follow-up of 297 days (355 days in 24 survivors), the 1-year probability of transplant-related mortality was 5%, and the 1-year progression-free survival was 66%. The 1-year incidence of disease progression in patients with and without GVHD was 13% (95% CI, 4%-34%) and 58% (95% CI, 36%-96%), respectively (P = .008). These results suggest that a graft-versus-leukemia effect plays a crucial role in reducing the risk of relapse after a RIC allograft in AML and MDS.
Disruption of the long-range GPIIIa Cys5-Cys435 disulfide bond results in the production of constitutively active GPIIb-IIIa (αIIbβ3) integrin complexes
Authors: Sun Qi-Hong , Liu Chao-Yan , Wang Ronggang , Paddock Cathy , Newman Peter J.
Published: August 2002

The major platelet integrin αIIbβ3, also known as the platelet glycoprotein (GP) IIb-IIIa complex, mediates platelet aggregation by serving as the receptor for fibrinogen and von Willebrand factor. In addition to its physiologic role, GPIIb-IIIa also bears a number of clinically important alloantigenic determinants. Previous studies have shown that disruption of the long-range Cys5-Cys435 disulfide bond of the β3 subunit results in the production of isoforms that bind some, but not all, anti-PlA1 alloantibodies, suggesting that mutations in this so-called long-range disulfide bond can alter the conformation of GPIIIa. The purpose of this study was to examine the effects of either the Cys5Ala or Cys435Ala substitution of GPIIIa on the adhesive properties of the GPIIb-IIIa complex. We found that both Ala5GPIIIa and Ala435GPIIIa were capable of associating with GPIIb and were expressed normally on the cell surface when cotransfected into Chinese hamster ovary (CHO) cells. CHO cells expressing GPIIb-Ala5GPIIIa or GPIIb-Ala435IIIa bound well-characterized, conformationally sensitive ligand-induced binding site (LIBS) antibodies, and were capable of constitutively binding the fibrinogen-mimetic monoclonal antibodies Pl-55 and PAC-1, as well as soluble fibrinogen. Both GPIIb-Ala5IIIa– and GPIIb-Ala435IIIa–transfected CHO cells also bound more avidly to immobilized fibrinogen and were capable of mediating the tyrosine phosphorylation of pp125FAK on cell adhesion. These data are consistent with the notion that these regions of GPIIIa participate in the conformational change associated with receptor activation. Additionally, these studies may provide a molecular explanation for the previously reported ability of mild reducing agents to activate the GPIIb-IIIa complex and promote platelet aggregation.
Identification of tumor-associated antigens in chronic lymphocytic leukemia by SEREX
Authors: Krackhardt Angela M. , Witzens Mathias , Harig Sabine , Hodi F. Stephen , Zauls A. Jason , Chessia Morgan , Barrett Patrick , Gribben John G.
Published: August 2002

Chronic lymphocytic leukemia (CLL) is associated with a variety of immunologic disturbances. Hypogammaglobulinemia and autoimmune phenomena are both often present in this disease. In contrast, humoral or cellular antitumor responses are rarely observed. It has been previously shown that antigens detected in patients with malignant diseases can provide information regarding intracellular molecules engaged in the transformation process and can identify tumor antigens that may be useful for development of immunotherapeutic strategies. Serologic identification by recombinant expression cloning (SEREX) has been demonstrated to be a useful method to detect tumor and tumor-associated antigens in a variety of malignancies. Although this approach is complicated in CLL, we used a modified SEREX approach and identified 14 antigens (KW-1 to KW-14) using this methodology. Several clones showed a restricted expression pattern in normal tissues. Moreover, distinctive expression of splice variants and aberrant gene expression in malignant tissue were detected. In this study, 6 antigens were detected exclusively in patients with CLL. Eight antigens were detected also in lymphoma patients. Healthy donors showed antibody responses against only 3 of the identified antigens. T cells with specific cytotoxicity against peptides derived from the 2 antigens tested could be generated from healthy donors. These findings demonstrate that humoral and cellular immune responses against CLL-associated antigens can be detected. Ongoing experiments investigate their potential for the development of immunotherapeutic strategies.
Anemia and iron overload due to compound heterozygosity for novel ceruloplasmin mutations
Authors: Bosio Sandra , De Gobbi Marco , Roetto Antonella , Zecchina Gabriella , Leonardo Eugenio , Rizzetto Mario , Lucetti Claudio , Petrozzi Lucia , Bonuccelli Ubaldo , Camaschella Clara
Published: August 2002

Aceruloplasminemia is a recessive disorder characterized by anemia, iron overload, and neurodegeneration, caused by the absence of ceruloplasmin (Cp), a multicopper oxidase important for iron export. Few patients homozygous for loss of function mutations of the Cp gene have been reported. We describe a 62-year-old white woman with heavy liver iron overload, diabetes, anemia, and neurologic symptoms. She was compound heterozygote for 2 novel mutations that result in the absence of hepatocyte Cp: an adenine insertion at nucleotide 2917 causing a truncated protein and a C-G transversion causing a glutamine→glutamic acid substitution at position 146. Although rare in whites, aceruloplasminemia should be considered in the differential diagnosis of unexplained anemia associated with iron overload, because these features anticipate progressive neurologic symptoms. We propose that anemia, secondary to the impaired macrophage iron release, plays a major role in hepatic iron overload through increased absorption mediated by the erythroid regulator.
Prolongation of cardiac repolarization by arsenic trioxide
Authors: Chiang Chern-En , Luk Hsiang-Ning , Wang Tsui-Ming , Ding Philip Yu-An
Published: August 2002

Arsenic trioxide (As2O3; ATO) has recently been found to be very effective for relapsed acute promyelocytic leukemia. Several articles reported prolongation of QT interval or ventricular arrhythmias in patients receiving ATO. However, the QT-prolonging effect has not been confirmed and the direct membrane effect of ATO has never been studied. In the present investigation, using conventional action potential recording technique, we found that ATO dose dependently prolonged action potential duration (APD) in guinea pig papillary muscle with a slow pacing frequency. Parenteral administration of ATO prolonged QT interval and APD in guinea pig hearts. Intravenous infusion of clinically relevant doses of ATO prolonged QT interval and APD dose dependently. These studies suggest that ATO has a direct effect on cardiac repolarization. Patients who are receiving ATO should avoid concomitant administration of other QT-prolonging agents or conditions in favor of delaying cardiac repolarization.
Second malignancy after Hodgkin disease treated with radiation therapy with or without chemotherapy: long-term risks and risk factors
Authors: Ng Andrea K. , Bernardo M. V. Patricia , Weller Edie , Backstrand Kendall , Silver Barbara , Marcus Karen C. , Tarbell Nancy J. , Stevenson Mary Ann , Friedberg Jonathan W. , Mauch Peter M.
Published: August 2002

The excess risk of second malignancy after Hodgkin disease is an increasing problem. In light of the long-term data, guidelines for follow-up of survivors of Hodgkin disease need to be redefined. In this study we attempt to analyze the long-term risks and temporal trends, identify patient- and treatment-related risk factors, and determine the prognosis of patients who develop a second malignancy after radiation treatment with or without chemotherapy for Hodgkin disease. Among 1319 patients with clinical stage I-IV Hodgkin disease, 181 second malignancies and 18 third malignancies were observed. With a median follow-up of 12 years, the relative risk (RR) and absolute excess risk of second malignancy were 4.6 and 89.3/10 000 person-years. The RR was significantly higher with combined chemotherapy and radiation therapy (6.1) than with radiation therapy alone (4.0, P = .015). The risk increased with increasing radiation field size (P = .03) in patients who received combined modality therapy, and with time after Hodgkin disease. After 15 and 20 years, there was a 2.3% and 4.0% excess risk of second malignancy per person per year. The 5-year survival after development of a second malignancy was 38.1%, with the worst prognosis seen after acute leukemia and lung cancer. The excess risk of second malignancy after Hodgkin disease continues to be increased after 15 to 20 years, and there does not appear to be a plateau. Our analysis suggests that the risk may be reduced with smaller radiation fields, as are used in current trials of abbreviated chemotherapy and limited-field radiation therapy.
Effect of hyperhomocysteinemia on protein C activation and activity
Authors: Lentz Steven R. , Piegors Donald J. , Fernández José A. , Erger Rochelle A. , Arning Erland , Malinow M. René , Griffin John H. , Bottiglieri Teodoro , Haynes William G. , Heistad Donald D.
Published: August 2002

Hyperhomocysteinemia has been proposed to inhibit the protein C anticoagulant system through 2 mechanisms: decreased generation of activated protein C (APC) by thrombin, and resistance to APC caused by decreased inactivation of factor Va (FVa). We tested the hypotheses that generation of APC by thrombin is impaired in hyperhomocysteinemia in monkeys and that hyperhomocysteinemia produces resistance to APC in monkeys, mice, and humans. In a randomized crossover study, cynomolgus monkeys were fed either a control diet or a hyperhomocysteinemic diet for 4 weeks. Plasma total homocysteine (tHcy) was approximately 2-fold higher when monkeys were on the hyperhomocysteinemic diet than when they were on the control diet (9.8 ± 2.0 μM versus 5.6 ± 1.0 μM; P < .05). After infusion of human thrombin (25 μg/kg of body weight), the peak level of plasma APC was 136 ± 16 U/mL in monkeys fed the control diet and 127 ± 13 U/mL in monkeys fed the hyperhomocysteinemic diet (P > .05). The activated partial thromboplastin time was prolonged to a similar extent by infusion of thrombin in monkeys fed the control diet and in those fed the hyperhomocysteinemic diet. The sensitivity of plasma FV to human APC was identical in monkeys on control diet and those on hyperhomocysteinemic diet. We also did not detect resistance of plasma FV to APC in hyperhomocysteinemic mice deficient in cystathionine β-synthase (plasma tHcy, 93 ± 16 μM) or in human volunteers with acute hyperhomocysteinemia (plasma tHcy, 45 ± 6 μM). Our findings indicate that activation of protein C by thrombin and inactivation of plasma FVa by APC are not impaired during moderate hyperhomocysteinemia in vivo.
DNB: a partial D with anti-D frequent in Central Europe
Authors: Wagner Franz F. , Eicher Nicole I. , Jørgensen Jan R. , Lonicer Cornelie B. , Flegel Willy A.
Published: August 2002

To improve routine D typing and define transfusion strategy, it is important to establish the frequency of partial D alleles and their susceptibility to anti-D alloimmunization due to transfusion or pregnancy. We identified the partial D DNB that was caused by anRHD(G355S) allele associated with a CDehaplotype and whose phenotype presented a normal D in routine typing. The antigen density was about 6000 D antigens per red blood cell, and the Rhesus index was 0.02. Five anti-D immunization events with allo–anti-D titers up to 128 were observed. Twelve carriers of DNB were whites of Central Europe; the only Danish proband had Austrian ancestry. DNB was the most frequent partial D recognized so far in whites, occurring with frequencies of up to 1:292 in Switzerland. DNB was the underlying partial D phenotype in a relevant fraction of anti-D immunizations occurring in whites.
The caspase inhibitor z-VAD is more effective than CD18 adhesion blockade in reducing muscle ischemia-reperfusion injury: implication for clinical trials
Authors: Iwata Akiko , Harlan John M. , Vedder Nicholas B. , Winn Robert K.
Published: August 2002

Ischemia-reperfusion (I/R) leads to organ injury and organ dysfunction in a variety of clinical disorders. Preclinical investigations examining leukocyte adhesion molecules in I/R provided overwhelming evidence that blocking the function of leukocyte adhesion molecules would be highly effective in improving outcome in clinically relevant diseases. Unfortunately, all 9 of the recently completed phase 2 and 3 clinical trials examining antiadhesion therapy have failed. In this report, we show that a modest increase in ischemic time results in conversion from a CD18-dependent to a CD18-independent injury. This fundamental change in the mechanism of injury can be reduced by inhibition of caspases leading to blockade of apoptosis. Muscle injury resulting from aortic clamping was measured by release of creatine kinase. I/R injury following ischemia of 60 minutes or less and 3 hours of reperfusion was significantly reduced by pretreatment with anti-CD18 monoclonal antibody. However, 90 minutes of ischemia resulted in a marked increase in injury that was not reduced by CD18 blockade. Importantly, the injury resulting from 90 or 120 minutes of ischemia was reduced by the pancaspase inhibitor z-VAD. We propose that the length of ischemia can result in a fundamental change in the mechanism of injury and that all preclinical investigations of I/R must be evaluated with increasing ischemia if they are to model the clinical disease. The result showing CD18-independent I/R injury is not unique; likewise, protection by caspase inhibitors is not unique. However, we show for the first time that caspase inhibitors are effective when CD18 blockade is not.
A genetic and genomic analysis identifies a cluster of genes associated with hematopoietic cell turnover
Authors: de Haan Gerald , Bystrykh Leonid V. , Weersing Ellen , Dontje Bert , Geiger Hartmut , Ivanova Natalia , Lemischka Ihor R. , Vellenga Edo , Van Zant Gary
Published: August 2002

Hematopoietic stem cells from different strains of mice vary widely with respect to their cell cycle activity. In the present study we used complementary genetic and genomic approaches to identify molecular pathways affecting this complex trait. We identified a major quantitative trait locus (QTL) associated with variation in cell proliferation in C57BL/6 and DBA/2 mice to a 10 centimorgan (cM) region on chromosome 11. A congenic mouse model confirmed that a genomic interval on chromosome 11 in isolation confers the proliferation phenotype. To detect candidate genes we performed subtractive hybridizations and gene arrays using cDNA from highly enriched stem cells from parental strains. Intriguingly, a disproportionate number of differentially expressed genes mapped to chromosome 11 and, more specifically, these transcripts occurred in 3 distinct clusters. The largest cluster colocalized exactly with the cell cycling QTL. Such clustering suggested the involvement of genetic variation that affects higher-order chromosomal organization. This hypothesis was reinforced by the fact that differentially expressed genes mapped to recombination “coldspots,” as a consequence of which clustered genes are collectively inherited. These findings suggest the functional interdependence of these closely linked genes. Our data are consistent with the hypothesis that this isolated cell cycle QTL does not result from a mutation in a single gene but rather is a consequence of variable expression of a collection of highly linked genes.
Amelioration of the macrothrombocytopenia associated with the murine Bernard-Soulier syndrome
Authors: Kanaji Taisuke , Russell Susan , Ware Jerry
Published: August 2002

An absent platelet glycoprotein (GP) Ib-IX receptor results in the Bernard-Soulier syndrome and is characterized by severe bleeding and the laboratory presentation of macrothrombocytopenia. Although the macrothrombocytopenic phenotype is directly linked to an absent GP Ib-IX complex, the disrupted molecular mechanisms that produce the macrothrombocytopenia are unknown. We have utilized a mouse model of the Bernard-Soulier syndrome to engineer platelets expressing an α-subunit of GP Ib (GP Ibα) in which most of the extracytoplasmic sequence has been replaced by an isolated domain of the α-subunit of the human interleukin-4 receptor (IL-4Rα). The IL-4Rα/GP Ibα fusion is membrane expressed in Chinese hamster ovary (CHO) cells, and its expression is facilitated by the presence of human GP IX and the β-subunit of GP Ib. Transgenic animals expressing a chimeric receptor were generated and bred into the murine Bernard-Soulier syndrome–producing animals devoid of mouse GP Ibα but expressing the IL-4Rα/GP Ibα fusion sequence. The characterization of these mice revealed a 2-fold increase in circulating platelet count and a 50% reduction in platelet size when compared with platelets from the mouse model of the Bernard-Soulier syndrome. Immunoprecipitation confirmed that the IL-4Rα/GP Ibα subunit interacts with filamin-1 and 14-3-3ζ, known binding proteins to the GP Ibα cytoplasmic tail. Mice expressing the chimeric receptor retain a severe bleeding phenotype, confirming a critical role for the GP Ibα extracytoplasmic domain in hemostasis. These results provide in vivo insights into the structural elements of the GP Ibα subunit that contribute to normal megakaryocyte maturation and thrombopoiesis.
Hydrocortisone reduced in vivo, inflammation-induced slow rolling of leukocytes and their extravasation into human conjunctiva
Authors: Kirveskari Juha , Helintö Maaret , Moilanen Jukka A. O. , Paavonen Timo , Tervo Timo M. T. , Renkonen Risto
Published: August 2002

Hydrocortisone reduces the number of inflammatory leukocytes within tissues, but thus far the site of action on the multistep adhesion cascade leading to leukocyte extravasation has not been identified. We have recently developed a noninvasive in vivo reflected-light confocal microscopy technique to study this at sites of inflammation in human patients. In the present study, we evaluated the effect of preoperative intravenous hydrocortisone treatment on leukocyte trafficking after conjunctival inflammation induced by cataract surgery in human subjects in vivo. The surgery generated leukocyte rolling along the endothelial lining of conjunctival vessels. While preoperative hydrocortisone did not reduce the number of rolling cells, it significantly raised the velocity of individual rolling leukocytes and concomitantly reduced leukocyte emigration into conjunctival tissue. Immunohistology of conjunctival biopsies excised from the individuals studied provided circumstantial evidence that endothelial P-selectin might play a role in the surgery-induced up-regulation of the leukocyte rolling. Furthermore, hydrocortisone reduced surgery-induced P-selectin induction, suggesting a role for this selectin in the regulation of local leukocyte traffic into sites of inflammation in human conjunctiva. Taken together, these results suggest that control of the rolling velocity might be an effective way to adjust leukocyte traffic in vivo in human subjects.
Is psychiatry losing touch with the rest of medicine?
Authors: Kennedy Peter
Published: August 2002

What is happening in and around psychiatric services that raises fears of a harmful separation between psychiatrists and the rest of the medical fraternity? The last National Health Service (NHS) reforms at the beginning of the 1990s split some mental health services into separate organisations from their local acute services. Paragraph 5.14 in the Government's White Paper The New NHS (NHS Executive, 1997a) seems to favour specialist mental health trusts. Combined whole district trusts are under threat. Shotguns are loaded to ‘encourage’ the marriage with social services. All this may seem to conspire against psychiatrists maintaining close professional relationships with physicians and surgeons in general hospitals. Instead it puts the emphasis on aligning psychiatric work much more closely with social services and primary care. Much restructuring is already afoot. It is a time for sober reflection on what is in the best interests of patients now and in the future (see Box 1).
Aspects of basic management of offenders with mental disorders
Authors: Humphreys Martin
Published: August 2002

Mental disorder and mental illness are common. Delinquency, offending and offending behaviour are widespread. The two things therefore occur frequently together, but are not always necessarily causally linked (Humphreys et al, 1994). Access to appropriate psychiatric services and care does not depend solely upon the presence of disturbed behaviour or offending that is obviously directly related to symptoms or signs.
Code of Practice 1999: navigating before reform of the Mental Health Act 1983
Authors: Hughes Julian C. , Lawson Tony
Published: August 2002

Mental health legislation must steer a course between Scylla and Charybdis. Scylla represents the notion of individual liberties; Charybdis represents the notion of safety and, in particular, public safety. At the time of writing a Green Paper is expected in 1999, so reform of the Mental Health Act 1983 is on its way. Meanwhile, does the new Code of Practice (Department of Health & Welsh Office, 1999), in force since 1 April 1999, give us any indication as to the course we might be steering?
Commentary
Authors: Exworthy Tim
Published: August 2002

Dr Humphreys is right to emphasise that the majority of offenders with mental disorder have not committed serious offences (conversely, a number of general psychiatric patients are admitted to hospital after incidents in the community which, in different circumstances, might have attracted official attention) and that the principles of treatment and management are the same as for patients in general psychiatry. However, psychiatric assessment of more serious offending does call for careful consideration of additional issues, such as ‘psychiatric defences’ to criminal charges (including fitness to plead, insanity, automatisms, and in cases of charges of murder, diminished responsibility) and the role of security in their management.
Managing amphetamine dependence
Authors: Bruce Malcolm
Published: August 2002

“Appropriately controlled and structured clinical trials have not yet revealed any pharmacological approach for rehabilitation of stimulant dependent individuals for which the risks have been shown to outweigh the assets.” (Schuckit, 1994)
Commentary
Authors: Cantwell Roch
Published: August 2002

In the sometimes sensational world of illicit drug reportage, there is one unsung villain. While heroin misuse remains the bête noir of tabloid journalism, ecstasy the demon of the dance floors and cocaine caricatured as the choice of the rich and famous, amphetamine misuse has lurked the shadows. Its use defies such simple categorisation and spans several groups in society. Bruce has provided a timely reminder of this neglected area in substance misuse literature and, in the process, has highlighted the relevance of basic information gathering as the most important tool in the armamentorium of drug misuse workers. The lack of prominence given to what they describe as a “hidden epidemic” is striking. Could this be because amphetamine misuse is a less prevalent problem than that of other illicit drugs? Evidence suggests otherwise. Amphetamine is the second most common illicit drug seized in the UK (after cannabis). It is easily produced and used in a variety of modes, and recent research confirms a high prevalence of misuse in this country reflecting that found in North American and Australian literature.
Working with elderly carers of people with learning disabilities and planning for the future
Authors: Hubert Jane , Hollins Sheila
Published: August 2002

The majority of people with learning disabilities in the UK live at home with their families, usually with their parents (Mental Health Foundation, 1996) or – more commonly in later life – with one parent, usually their mother. Nowadays, people with learning disabilities live much longer than they did in the past, with the result that there is also an expanding population of elderly parents who are continuing to care for a son or daughter well into old age.
Diagnosis of depression in elderly patients
Authors: Evans Mavis , Mottram Pat
Published: August 2002

Depression in old age is a pathological process, not a normal reaction to growing older. The majority of people cope with ageing, and many feel happy and fulfilled. However, there is a bias among health professionals and the community in general to accept lower functioning and more symptoms in older people (Alexopoulos, 1992). Depression tends to be denied by the current generation of elderly people, many of whom were raised in an atmosphere where showing feelings was discouraged, and this adds to diagnostic difficulties. Comorbid medical conditions, the tendency of patients to somatise, cognitive deterioration, and multiple life events, often of loss (e.g. bereavement, retirement, moving to smaller housing), all further complicate the diagnostic process.
Safer services: conclusions from the report of the National Confidential Inquiry
Authors: Appleby Louis
Published: August 2002

In May 1999, the National Confidential Inquiry into Suicide and Homicide by People with Mental Illness published its first major report since it was reestablished at the University of Manchester (Appleby et al, 1999a). Entitled Safer Services, it presented findings from a two-year sample of suicides and an 18-month sample of homicides occurring in England and Wales, and on the basis of these put forward 31 recommendations for mental health services (Appleby, 1999b; Shaw et al, 1999). This article, modified from a chapter in Safer Services, discusses how services might deal with the risk of suicide and homicide or serious violence.
Transference and countertransference in communication between doctor and patient
Authors: Hughes Patricia , Kerr Ian
Published: August 2002

“The reasonable man adapts himself to the world; the unreasonable one persists in trying to adapt the world to himself” George Bernard Shaw, Maxims for Revolutionists.
Stigma of mental illness and ways of diminishing it
Authors: Byrne Peter
Published: August 2002

Stigma is defined as a sign of disgrace or discredit, which sets a person apart from others. The stigma of mental illness, although more often related to context than to a person's appearance, remains a powerful negative attribute in all social relations. Sociological interest in psychiatric stigma was given added vigour with the publication ofStigma – Notes on the Management of Spoiled Identity(Goffman, 1963). More recently, psychiatrists have begun to re-examine the consequences of stigma for their patients. In 1989, the American Psychiatric Association's annual meeting's theme ‘overcoming stigma’ was subsequently published as a collection of articles (Fink & Tasman, 1992), and last year saw the launch of the Royal College of Psychiatrists' five-yearChanging Mindsanti-stigma campaign.
Commissioning mental health services: role of the consultant psychiatrist
Authors: Simpson Chris
Published: August 2002

The current National Health Service (NHS) approach to commissioning health services is in flux. The purchasing of care from providers by general practitioner fundholders (GPFHs) and health authorities has changed with the new White Papers. GPFHs no longer exist and the commissioning role is being handed over from health authorities to primary care groups (PCGs). An understanding of the reasons for change and current arrangements will aid the consultant psychiatrist in influencing this process.
Suicide in the elderly
Authors: Cattell Howard
Published: August 2002

Despite the fact that suicide and its prevention continues to be a priority area for health care in the UK, suicide in the elderly remains a neglected subject receiving little interest and research attention. The Green Paper Our Healthier Nation (Secretary of State for Health, 1998) maintained the concept of setting targets for suicide reduction originally proposed in The Health of the Nation strategy. The new target proposes that by the year 2010 the death rate from suicide and undetermined injury will be reduced by at least a further sixth (17%) from the baseline of 1996. The setting of such targets has always been a contentious issue among many psychiatrists, who have concerns that they may be used as a quality measure of psychiatric services, especially as some consider that social influences predominate over health care issues. The subsequent debate has focused on targeting specific at-risk groups, notably severely mentally ill young men, Asian women and those who deliberately harm themselves.
Neuroimaging in dementia and depression
Authors: O'Brien John , Barber Bob
Published: August 2002

Neuroimaging is traditionally divided into structural and functional imaging. Structural imaging looks at brain structure or anatomy and includes computed tomography (CT) and magnetic resonance imaging (MRI). Functional techniques seek to examine the physiological functioning of the brain, either at rest or during activation, and include single photon emission computed tomography (SPECT), positron emission tomography (PET), MRI spectroscopy, functional MRI (fMRI) and encephalographic brain mapping. Although fMRI, MRI spectroscopy and brain mapping are likely to have clinical applications in the near future, the main imaging modalities of current clinical relevance to psychiatrists are CT, MRI and SPECT, which will be the focus of this article.
Community in-patient units and halfway hospitals
Authors: Boardman Anthony , Hodgson Richard
Published: August 2002

There is a current gap in the nomenclature of psychiatric in-patient services. There are few descriptions of types of in-patient care and over recent years the literature has abounded with debates concerning alternatives. However, it may be argued that these debates have been based on the creation of the ‘straw man’ of the psychiatric admission, which is only fit for knocking down. Although a post-war consensus has emerged concerning the need to abandon the Victorian asylums, this has often been misrepresented as the need to avoid in-patient admission. The poorly articulated and emotional concept of community care and its lack of clear and consistent definition in public policy and key legislation have contributed to this (Bulmer, 1987). Recent changes in our view of community care have led to a refining of the concept and a shift from its comforting appellations (Titmus, 1968) to a pragmatic approach that matches it to empirical experiences and new resources. This approach sees psychiatric services for adults as being based locally and provided by a spectrum of services – in-patient, residential and ambulatory (Department of Health, 1996) – based on best available evidence. This article has been written with these issues in mind. We will address the current problems of in-patient care and the current literature on alternatives and supplements to traditional in-patient units.