bluuebunny/crossref_metadata_2025 huggingface dataset:
Anthropomorphism and Anthropomorphic Selection—Beyond the "Cute Response"
Authors: Serpell James
Published: April 2003

This article explores the origin and evolutionary implications of anthropomorphism in the context of our relationships with animal companions. On the human side, anthropomorphic thinking enables animal companions' social behavior to be construed in human terms, thereby allowing these nonhuman animals to function for their human owners or guardians as providers of nonhuman social support. Absence of social support is known to be detrimental to human health and well being. Therefore, anthropomorphism and its corollary, pet keeping, have obvious biological fitness implications. On the animal side, anthropomorphism constitutes a unique evolutionary selection pressure, analogous to sexual selection, which has molded the appearance, anatomy, and behavior of companion animal species so as to adapt them to their unusual ecological niche as social support providers. Although such species undoubtedly have benefited numerically from the effects of this process, the consequences of anthropomorphism are less benign when viewed from the perspective of individual animals. Indeed, anthropomorphic selection probably is responsible for some of the more severe welfare problems currently found in companion animals.
Psychologism and Phenomenological Psychology Revisited, Part II: The Return to Positivity
Authors: Cosgrove Lisa , Davidson Larry
Published: April 2003

The last in a series of examinations, this paper articulates Husserl's mature position on the nature of a phenomenologically informed human science. Falling between the naïve positivity of a naturalistic approach to psychology and the transcendental view of consciousness at the base of phenomenological philosophy, we argue that a human scientific psychology—while not itself transcendental in nature needs to re-arise upon the transcendental ground as an empirical—but no longer transcendentally naïve—discipline through Husserl's notion of the "return to positivity." This notion of the return allows us to avoid "transcendental psychologism," differentiating psychological from transcendental subjectivity but from a transcendental, rather than naïve perspective. In this way, the return to positivity reclaims psychology as a worldly, but no longer naïve, discipline. To facilitate an understanding of the different perspectives in question, and the process of leaving the naturalistic perspective in order to return to it once armed with a transcendental understanding and its associated tools, we continue to develop the illustrative example of anorexia provided in the first part of this series. In conclusion, we discuss the implications of this framework for transcendental reforms both of clinical practice and of psychological research.
Lessons for the Future from the Margins of Psychology
Authors: Giorgi Amedeo
Published: April 2003

Having spent 40 years as a psychologist in academia with a minority perspective at odds with the culture of his profession, the author was requested to reflect upon his experiences in order to offer advice to younger colleagues of the same persuasion. There are indeed prices to be paid when one's values place one outside the established view within the discipline of psychology, but remaining true to oneself is never theless posited as the highest value. The chief drawback of marginality is the lack of the possibility of being placed within a normative university setting where the training of the next generation of scholars with similar values can take place. Basically, one encounters the resistances that sociology of knowledge factors offer rather than intrinsic resistances to the minority theoretical perspective. Five theoretical notions that met political resistance are listed and seven advisory comments are offered for young potentially marginal psychologists.
Phenomenology and the Incest Taboo
Authors: Hadreas Peter
Published: April 2003

It is argued that traditional functional explanations of the incest taboo do not sufficiently supply causal conditions. It is widely acknowledged that the incest taboo, although universal among human societies, is largely a feature of human behavior. Husserl's investigations of intentionality are introduced to supply the particularly human element by which the taboo may be understood. So as to illumine the contrast between the conflicting intentionalities, a classical Aristotelian contrast between eros and parent/ child philia is drawn. Parent/child philia and eros, although both requisite for the survival of the species, are shown to be crosspurposeful in several ways. Husserl's understanding of 'negation,' as it applies to affective and practical intentionalities, is reconsidered. It is thereby explained how parental/child affection and erotic love, are 'incompossible' and not only conflict, but 'nullify' and 'cancel out' each other, generating the 'taboo'.
The Phenomenology of Shame, Guilt and the Body in Body Dysmorphic Disorder and Depression
Authors: Fuchs Thomas
Published: April 2003

From a phenomenological viewpoint, shame and guilt may be regarded as emotions which have incorporated the gaze and the voice of the other, respectively. The spontaneous and unreflected performance of the primordial bodily self has suffered a rupture: In shame or guilt we are rejected, separated from the others, and thrown back on ourselves. This reflective turn of spontaneous experience is connected with an alienation of primordial bodiliness that may be described as a "corporealization": The lived-body is changed into the objective, corporeal body or "body-for-others." The polarity of "bodiliness" and "corporeality" may further a phenomenological understanding of several mental disorders connected with shame and guilt. This is shown by the examples of body dysmorphic disorder and melancholic depression.
Anti-epiphany and the Jungian Manikin: Toward a Theory of Prepsychotic Perceptual Alterations
Authors: Arnold Kyle
Published: April 2003

This paper articulates a psychodynamically informed phenomenological reading of prepsychotic perceptual alterations, which the author calls anti-epiphanies. Several of Carl Jung's experiences of the anti-epiphany, as described in his autobiography Memories, Dreams, Reflections (1961), are taken as exemplar cases. These anti-epiphanies are viewed through a critical psychobiographical lens, in an interpretationwhich tacks back and forth between Jung's childhood, psychological theories, and later prepsychotic experience. It is claimed that Jung's anti-epiphanies are linked to his use of schizoid-narcissistic forms of transitional selfobjects, referred to as Jungian manikins. Such Jungian manikins, it is argued, function to defend the subject against annihilation anxieties related to psychological engulfment, penetration, and finalization. When these anxieties become especially pronounced, the subject's entire perceptual world may be defensively used as a Jungian manikin, creating an anti-epiphany. The author conjectures that similar patterns of experience may be operative in the prepsychotic perceptual alterations had by those with schizoid-narcissistic character pathology.
The Sociology of Law as an Empirical Theory of Validity
Authors: Schluchter Wolfgang
Published: April 2003

Contrary to current tendencies, the founders of sociology as a discipline regarded the sociology of law as an integral part of social theory. Law and its historical variations were treated by them as a constitutive component of social life. This can be demonstrated especially with regard to Émile Durkheim and Max Weber. It also provides an opportunity to highlight the differences between these authors, not only in methodological but also in substantive terms. In the essay it is shown how both authors treated law in comparative and developmental perspectives and varied in assessing the role of penal law and human rights in the course of history. Connections to the current theoretical debate in Germany are also drawn.
From The Rules of Sociological Method to The Polish Peasant
Authors: Guth Suzie , Schrecker Cherry
Published: April 2003

Having translated into French the ‘Methodological Note’ that introduces The Polish Peasant in Europe and in America by William Isaac Thomas and Florian Znaniecki, the present authors compare this theoretical statement with that of Émile Durkheim laid out in The Rules of Sociological Method. In both works sociology is considered to be a nomothetic and teleological science. The major difference between the two works lies in the analysis of causality. The sociological method is based on different types of reasoning (inductive/deductive) and different types of data (objective and general in the first case, specific and individual in the second). Their differential definitions of science lead Durkheim, on the one hand, and Thomas and Znaniecki, on the other, in directions, which, up to a point, are mutually exclusive. This being said, there are similarities between the Durkheimian school (which includes Bouglé and Halbwachs) and Florian Znaniecki’s later (1934) work.
Sociology and Philosophy in the Work of Pierre Bourdieu, 1965-75
Authors: Robbins Derek
Published: April 2003

The paper first offers a brief account of the competition between the Durkheimian sociological tradition and German philosophy in the period in which Bourdieu was a student at the École Normale Supérieure. It indicates the intellectual influences of the early years that Bourdieu subsequently acknowledged and then examines his use of the work of Weber in his first book, Sociologie de l’Algérie (1958). The paper then focuses on the development of Bourdieu’s thought from the mid-1960s to the mid-1970s, a period in which he strategically presented himself as an anti-humanist sociologist whilst also articulating a view of science that was in tune with phenomenological and ontological philosophy. Bourdieu’s ‘Sociology and Philosophy in France since 1945’ (1967a) receives particular attention since his analysis of sociology and philosophy in France in the post-war period was a key element in his own position-taking in respect of the two disciplines. The paper then examines Bourdieu’s critiques of Weber at this time and suggests that his dissatisfaction with Weber’s epistemology logically became a dissastisfaction with the claims of sociological explanation as such. There followed an attempt to reconcile a commitment to social science with an allegiance to elements of phenomenological thought. The outcome was a willingness on Bourdieu’s part to see reflexivity as a means to problematiz- ing sociological explanation more than as a means to refining it or making it more sophisticated. The consequence was that commitments to phenomenological ontology and social science co-existed in this period. The balance was to change again subsequently in Bourdieu’s thought, and his responsivenesss to changing conditions exemplifies how we should ourselves rethink the relations between possible future social theories and the classical theories of Western sociology.
Mutually Contested Concepts and Their Standard General Use
Authors: Smith Kenneth
Published: April 2003

In this essay I claim to find within W.B. Gallie’s deservedly well-known discussion of ‘essentially contested concepts’ at least two other closely related concepts, which Gallie himself called ‘mutually contested concepts’ and their widely agreed upon ‘standard general use’. I argue that these two concepts together provide us with the ‘space’ to develop a rival argument to Gallie’s idea of essentially contested concepts. The essence of this rival view is that agreement on the proper use of most concepts within the social sciences is not that difficult to come by once we situate different mutually contested concepts within their agreed-upon standard general use. Viewed in this way, I argue, most concepts within the social sciences are not essentially contested. In order to understand this argument, I argue that it is helpful to view the standard general use of mutually contested concepts as complex multi-dimensional objects.
Impaired germinal center formation and recall T-cell–dependent immune responses in mice lacking the costimulatory ligand B7-H2
Authors: Wong Siew-Cheng , Oh Edwin , Ng Chee-Hoe , Lam Kong-Peng
Published: April 2003

B7-H2, which is expressed constitutively on B cells and binds the inducible costimulator (ICOS) on antigen-activated T cells, is a member of the B7 family of costimulatory ligands. We have inactivated B7-H2 in the mouse. B7-H2–/– mice generate normal populations of B and T cells in their various lymphoid organs but have lower basal levels of heavy chain class–switched antibodies in their sera. These mice are able to mount normal immune responses to both type I and type II T-cell–independent antigens. However, their pattern of responses to a T-cell–dependent antigen is altered, with greatly reduced production of antigen-specific heavy chain class–switched antibodies, the levels of which could not be elevated even with repeated immunizations. This suggests a critical role for B7-H2 in the recall phases of the immune response. Germinal center formation is also impaired in the mutant mice. While B cells from the mutant mice could response normally to anti-IgM, anti-CD40, and lipopolysaccharide stimulation, the production of T-helper–type II cytokines such as interleukin-4 (IL-4) and IL-10 by primed CD4+ T cells from mutant mice were reduced. This indicated that the defects in humoral responses and germinal center formation in B7-H2–deficient mice are due to the lack of T-cell–mediated help to the B cells. Hence, B7-H2 on B cells is important for recruiting T-cell help via its interaction with ICOS and plays a critical role in costimulating humoral immune responses.
Vaccination of patients with cutaneous T-cell lymphoma using intranodal injection of autologous tumor-lysate–pulsed dendritic cells
Authors: Maier Tanja , Tun-Kyi Adrian , Tassis Anatoli , Jungius Karl-Peter , Burg Günter , Dummer Reinhard , Nestle Frank O.
Published: April 2003

Cutaneous T-cell lymphoma (CTCL) is a lymphoproliferative skin disease with limited therapeutic options. Ten CTCL patients were treated with once-weekly intranodal injection of 1 × 106 mature monocyte-derived dendritic cells (DCs) pulsed with 100 μg/mL tumor lysate protein equivalent and keyhole limpet hemocyanin (50 μg/mL). Tumor-specific delayed-type hypersensitivity (DTH) reactions developed in 8 of 8 patients challenged with tumor-lysate-pulsed DCs and in 3 of 8 patients challenged with tumor lysate alone. Three of 5 patients showed significant tumor-lysate-specific increases of in vitro peripheral blood lymphocyte proliferation coinciding with increased interferon-α (IFN-α) production. Five of 10 (50%) patients had objective responses. Four patients had partial responses (PRs). Two are still in PR, and the other 2 patients had a mean PR duration of 10.5 months. One patient had a complete response (CR) for 19 months that is ongoing. The remaining 5 patients had progressive disease. In the 5 responder patients, 6.8 ± 1.4 vaccinations were necessary to induce an objective clinical response. Response was associated with low tumor burden. Continuation of vaccinations with new tumor lysate derived from progressive lesions reinduced treatment responses in 2 patients in PR. Selected patients had massive infiltration of CD8+ and TIA+ cytotoxic T cells at the site of regressing lesions and molecular remission after therapy. Intranodal injection of autologous tumor-lysate-pulsed DCs is well-tolerated and achieves immunologic and objective clinical responses in selected CTCL patients. (Blood. 2003;102:2338-2344)
The coupling of TEL/PDGFβR to distinct functional responses is modulated by the presence of cytokine: involvement of mitogen-activated protein kinases
Authors: Wheadon Helen , Welham Melanie J.
Published: April 2003

The TEL/PDGFβR oncogenic fusion protein is the product of the t(5;12)(q33; p13) translocation recurrently found in patients with chronic myelomonocytic leukemia (CMML). To investigate the coupling of molecular signaling events activated by TEL/PDGFβR to functional responses, we expressed TEL/PDGFβR in interleukin 3 (IL-3)–dependent BaF/3 cells using the tetracycline-regulated expression system. Induction of TEL/PDGFβR expression led to increased cell survival following IL-3 withdrawal and constitutive activation of protein kinase B (PKB), signal transducer and activator of transcription 5 (STAT5), extracellular signal-regulated kinases 1/2 (ERK1/2), Jun N-terminal kinases 1/2 (JNK1/2), and p38 mitogenactivated protein kinase (MAPK) pathways. However, inducible expression of TEL/PDGFβR failed to generate factor-independent cells, whereas constitutive expression of TEL/PDGFβR did, albeit at low frequency, suggesting the duration of TEL/PDGFβR expression is important for transformation. Surprisingly, in cells induced to express TEL/PDGFβR, IL-3–dependent growth was dramatically reduced as a result of increased apoptosis of cells receiving combined IL-3 and TEL/PDGFβR signals. We demonstrate that TEL/PDGFβR expression augmented IL-3–induced activation of PKB, STAT5, ERK1/2, p38, and JNK1/2. Inhibition of neither phosphoinositide-3 kinases nor p38 MAPKs reduced the inhibition of IL-3–driven proliferation observed when TEL/PDGFβR was expressed. However, inhibition of MEKs or JNKs partially reversed the combined inhibitory effects of TEL/PDGFβR and IL-3 on proliferation and survival. These results suggest that the combination of TEL/PDGFβR and IL-3–induced signals activate apoptosis through ERK and JNK MAPK-dependent pathways. Given that in vivo hematopoietic cells are in contact with a variety of cytokines, our results have important implications for cellular responses in the pathogenesis of CMML.
Performance of a new clinical grading system for chronic graft-versus-host disease: a multicenter study
Authors: Akpek Görgün , Lee Stephanie J. , Flowers Mary E. , Pavletic Steven Z. , Arora Mukta , Lee Shing , Piantadosi Steven , Guthrie Katherine A. , Lynch James C. , Takatu Alessandra , Horowitz Mary M. , Antin Joseph H. , Weisdorf Daniel J. , Martin Paul J. , Vogelsang Georgia B.
Published: April 2003

We recently reported 3 risk factors (RFs) at diagnosis of chronic graft-versus-host disease (cGVHD) that were significantly associated with increased nonrelapse mortality. These included extensive skin involvement (ESI), thrombocytopenia (TP), and progressive type of onset (PTO). The hazard ratio (HR) for mortality of the patients with prognostic score (PS) between 0 and 2 (intermediate-risk; 1 RF) compared to those with PS 0 (favorable-risk; 0 RF) was 3.7 (95% CI, 1.4, 9.3); the HR for patients with PS equal to or more than 2 (high-risk; > 1 RF) compared with intermediate-risk group was 6.9 (3.8, 12.4). A rare presentation of TP and PTO without ESI yielded a PS of 1.8 (intermediate-risk). This paper reports the performance of the prognostic model and the individual RFs using data from an additional 1105 patients from University of Nebraska (n = 60), International Bone Marrow Transplantation Registry (n = 708), Fred Hutchinson Cancer Research Center (n = 188), and University of Minnesota (n = 149). The extent of skin involvement was quantified in 3 cohorts using the available data collected in different formats before the analysis. Although the HR for mortality of the patients in the intermediate-risk group versus those in the favorable-risk group ranged from 2.3 to 8.9 across the centers, it was between 1.6 to 6.9 for patients in the high-risk group versus those in the intermediate-risk group. Although TP itself was uniformly associated with increased risk of mortality across all test samples, ESI and PTO showed statistically significant associations with mortality in 1 and 2 cohorts, respectively. In conclusion, the model was predictive of cGVHD-specific survival, but the mortality hazard associated with ESI was lower in each of these test samples compared with the learning sample. Although the new clinical grading based on the model is promising because of its utility across multiple independent data sets, prospective validation is needed.
Hepatitis C virus and B-cell non-Hodgkin lymphomas: an Italian multicenter case-control study
Authors: Mele Alfonso , Pulsoni Alessandro , Bianco Elvira , Musto Pellegrino , Szklo Andrè , Sanpaolo Maria Grazia , Iannitto Emilio , De Renzo Amalia , Martino Bruno , Liso Vincenzo , Andrizzi Cristina , Pusterla Simona , Dore Fausto , Maresca Maddalena , Rapicetta Maria , Marcucci Fabrizio , Mandelli Franco , Franceschi Silvia
Published: April 2003

The existence of an association between infection with hepatitis C virus (HCV) and B-cell non-Hodgkin lymphoma (B-NHL) remains controversial, largely because previous studies were based on prevalent case series or comparisons with less than optimal control groups. This hospital-based case-control study was conducted from January 1998 through February 2001 to evaluate the association between HCV infection and B-NHL of different types. Cases were consecutive patients with a new diagnosis of B-NHL; controls were patients from other departments of the same hospitals. Both groups were interviewed using a standardized questionnaire. The prevalence of HCV infection was calculated by histologic type of B-NHL and clinical behavior (indolent or aggressive). Adjusted odds ratio (OR) and HCV-attributable risk (AR) were estimated. HCV prevalence was 17.5% among the 400 lymphoma patients and 5.6% among the 396 controls. The OR of B-NHL (patients vs controls), adjusted by age, sex, level of education, and place of birth, was 3.1 (95% confidence interval [CI], 1.8-5.2); an OR indicative of positive association was found for indolent and aggressive B-NHL. The estimated AR was 4.6%. This study confirms an association between HCV and B-NHL. In Italy, 1 of 20 instances of B-NHL may be attributable to HCV infection and may, thus, benefit from antiviral treatment.
Interleukin-7 improves T-cell recovery after experimental T-cell–depleted bone marrow transplantation in T-cell–deficient mice by strong expansion of recent thymic emigrants
Authors: Broers Annoek E. C. , Posthumus-van Sluijs Sandra J. , Spits Hergen , van der Holt Bronno , Löwenberg Bob , Braakman Eric , Cornelissen Jan J.
Published: April 2003

Interleukin-7 (IL-7) has been shown to enhance thymic output of newly developed T cells following bone marrow transplantation (BMT) in mice. In addition, IL-7 may affect peripheral expansion of T cells. In order to study the relative contribution of thymopoiesis versus peripheral T-cell expansion in the setting of compromised thymopoiesis, we have applied IL-7 in an experimental stem cell transplantation model using T cell–deficient RAG-1–/– mice. C57BL/6 RAG-1–/– mice received transplants of syngeneic T-cell–depleted (TCD) bone marrow (Ly5.1) with or without supplemented T cells (Ly5.2). IL-7 was administered until day 63 after BMT. Peripheral blood T- and B-cell recovery was quantified by flow cytometry and thymopoiesis was studied by quantification of T-cell receptor rearrangement excision circles (TRECs). In mice receiving a T-cell–replete BMT, IL-7 selectively expanded mature CD45.2+ T cells without affecting the recovery of new bone marrow–derived CD45.1+ T cells. In contrast, IL-7 significantly enhanced the recovery of bone marrow–derived T cells after TCD BMT. Quantification of TRECs in mice receiving a TCD BMT revealed that enhanced T-cell recovery following IL-7 treatment resulted from a strong expansion of newly developed naive T cells. These results suggest that peripheral expansion of recent thymic emigrants or mature T cells may be a preferential mechanism by which IL-7 enhances T-cell recovery after BMT.
Hmgb3: an HMG-box family member expressed in primitive hematopoietic cells that inhibits myeloid and B-cell differentiation
Authors: Nemeth Michael J. , Curtis David J. , Kirby Martha R. , Garrett-Beal Lisa J. , Seidel Nancy E. , Cline Amanda P. , Bodine David M.
Published: April 2003

Hmgb3 is a member of a family of chromatin-binding proteins that can alter DNA structure to facilitate transcription factor binding. We identified the Hmgb3 cDNA in a subtractive hybridization screen for transcripts that are preferentially expressed in hematopoietic stem cells. We inserted an internal ribosomal entry site–green fluorescence protein cassette into the 3′ untranslated region of the X-linked Hmgb3 locus to identify Hmgb3-expressing cells. In adult mice, Hmgb3 mRNA is detected in bone marrow cells, primitive Lin–, c-kit+, Sca-1+, IL-7Rα– cells, and Ter119+ erythroid cells. We observed that long-term repopulating ability is entirely contained in the subpopulation of Lin–, c-kitHI cells that express Hmgb3. Most common lymphoid and myeloid progenitors express Hmgb3. Introduction of a retrovirus containing the Hmgb3 cDNA into mouse bone marrow stem cells demonstrated that enforced expression of Hmgb3 inhibited B-cell and myeloid differentiation. We conclude that down-regulation of Hmgb3 protein levels is an important step for myeloid and B-cell differentiation.
Homeostatic control of T-cell generation in neonates
Authors: Schönland Stefan O. , Zimmer Julia K. , Lopez-Benitez Consuelo M. , Widmann Thomas , Ramin Kirk D. , Goronzy Jörg J. , Weyand Cornelia M.
Published: April 2003

T cells are produced through 2 mechanisms, thymopoiesis and proliferative expansion of postthymic T cells. Thymic output generates diversity of the pool, and proliferation achieves optimal clonal size of each individual T cell. To determine the contribution of these 2 mechanisms to the formation of the initial T-cell repertoire, we examined neonates of 30 to 40 weeks' gestation. Peripheral T cells were in a state of high proliferative turnover. In premature infants, 10% of T cells were dividing; the proliferation rates then declined but were still elevated in mature newborns. Throughout the third trimester, concentrations of T-cell–receptor excision circles (TRECs) were 10 per 100 T cells. Stability of TREC frequencies throughout the period of repertoire generation suggested strict regulation of clonal size to approximately 10 to 20 cells. Neonatal naive CD4+ and CD8+ T cells were explicitly responsive to IL-7; growth-promoting properties of IL-15 were selective for newborn CD8+ T cells. Neonatal T cells expressed telomerase and, in spite of the high turnover, built up a telomeric reserve. Thus, proliferative expansion, facilitated by increased cytokine responsiveness, and thymopoiesis complement each other as mechanisms of T-cell production in neonates. Maintaining optimal clonal size instead of filling the space in a lymphopenic host appears to regulate homeostatic T-cell proliferation during fetal development.
Anti-CD40 monoclonal antibody therapy in combination with irradiation results in a CD8 T-cell–dependent immunity to B-cell lymphoma
Authors: Honeychurch Jamie , Glennie Martin J. , Johnson Peter W. M. , Illidge Timothy M.
Published: April 2003

The mechanisms of interaction between anti-CD40 monoclonal antibody (mAb) therapy and external beam irradiation were investigated in 2 syngeneic B-cell lymphoma models. We have established doses of anti-CD40 mAb and irradiation which, although ineffective when given singly, were capable of providing long-term protection when used in combination. Furthermore, such treatment was not only critically dependent on the dose of mAb and irradiation but also on tumor load, with greater efficacy only occurring at higher tumor burden. Using blocking antibody, the potency of treatment was shown to be totally dependent on CD8+ T cells, with protective levels of CD8+ cells occurring only in mice receiving the combination of anti-CD40 and irradiation. Interestingly, the ratio of T cells (CD8+) to tumor cells in mice receiving combination treatment was between 10 and 15 times that seen in animals given anti-CD40 or irradiation alone. In vivo tracking experiments revealed a 2-phase decrease in tumor burden, the first resulting directly from the external irradiation and the second, occurring 5 days later, concomitant with the rise in tumor-specific CD8+ cells. We suggest that the external irradiation induced an initial kill of lymphoma cells, probably by apoptosis, which releases tumor antigens and slows the progression of the malignancy to allow generation of a curative cytotoxic T lymphocyte (CTL) response promoted by the anti-CD40 mAb. Combining irradiation with immunomodulatory mAb as described here appears to provide a powerful new approach to the management of cancer.
The disintegrin-like metalloproteinase ADAM10 is involved in constitutive cleavage of CX3CL1 (fractalkine) and regulates CX3CL1-mediated cell-cell adhesion
Authors: Hundhausen Christian , Misztela Dominika , Berkhout Theo A. , Broadway Neil , Saftig Paul , Reiss Karina , Hartmann Dieter , Fahrenholz Falk , Postina Rolf , Matthews Vance , Kallen Karl-Josef , Rose-John Stefan , Ludwig Andreas
Published: April 2003

The CX3C chemokine fractalkine (CX3CL1) exists as a membrane-expressed protein promoting cell-cell adhesion and as a soluble molecule inducing chemotaxis. Transmembrane CX3CL1 is converted into its soluble form by defined proteolytic cleavage (shedding), which can be enhanced by stimulation with phorbol-12-myristate-13-acetate (PMA). PMA-induced CX3CL1 shedding has been shown to involve the tumor necrosis factor-α–converting enzyme (TACE), whereas the constitutive cleavage in unstimulated cells remains elusive. Here we demonstrate a role of the closely related disintegrin-like metalloproteinase 10 (ADAM10) in the constitutive CX3CL1 cleavage. The hydroxamate GW280264X, capable of blocking TACE as well as ADAM10, proved to be an effective inhibitor of the constitutive and the PMA-inducible CX3CL1 cleavage in CX3CL1-expressing ECV-304 cells (CX3CL1–ECV-304), whereas GI254023X, preferentially blocking ADAM10 but not TACE, reduced the constitutive cleavage only. Overexpression of ADAM10 in COS-7 cells enhanced constitutive cleavage of CX3CL1 and, more importantly, in murine fibroblasts deficient of ADAM10 constitutive CX3CL1 cleavage was markedly reduced. Thus, ADAM10 contributes to the constitutive shedding of CX3CL1 in unstimulated cells. Addressing the functional role of CX3CL1 shedding for the adhesion of monocytic cells via membrane-expressed CX3CL1, we found that THP-1 cells adhere to CX3CL1–ECV-304 cells but detach in the course of vigorous washing. Inhibition of ADAM10-mediated CX3CL1 shedding not only increased adhesive properties of CX3CL1–ECV-304 cells but also prevented de-adhesion of bound THP-1 cells. Our data demonstrate that ADAM10 is involved in the constitutive cleavage of CX3CL1 and thereby may regulate the recruitment of monocytic cells to CX3CL1-expressing cell layers.
Expression of CD52 on plasma cells in plasma cell proliferative disorders
Authors: Kumar Shaji , Kimlinger Teresa K. , Lust John A. , Donovan Kathleen , Witzig Thomas E.
Published: April 2003

Multiple myeloma (MM) and primary systemic amyloidosis (AL) remain incurable disorders, and new treatments targeted to the malignant plasma cells are needed. Alemtuzumab is a humanized monoclonal antibody to CD52 and has activity in chronic lymphocytic leukemia. We examined the CD52 expression on CD45+ and CD45- plasma cell populations to evaluate the potential for using alemtuzumab for these disorders. Bone marrows from 61 patients (29 AL, 23 MM, and 9 MGUS [monoclonal gammopathies of undetermined significance]) were studied using 3-color (CD38/45/52) flow cytometry. Among those with MGUS, MM, and AL, 67%, 52%, and 35%, respectively, were positive for CD52 expression. The CD52 expression was predominantly confined to the clonal CD38+/CD45+ plasma cell fraction with median expression of 68%, 88%, and 82% in MGUS, MM, and AL, respectively, compared with 18%, 6%, and 9% among the CD45- plasma cell population. Clinical trials are warranted in these diseases to learn the therapeutic benefit of anti-CD52 immunotherapy.
The immunoglobulin heavy-chain locus hs3b and hs4 3′ enhancers are dispensable for VDJ assembly and somatic hypermutation
Authors: Morvan Caroline Le , Pinaud Eric , Decourt Catherine , Cuvillier Armelle , Cogné Michel
Published: April 2003

The more distal enhancers of the immunoglobulin heavy-chain 3′ regulatory region, hs3b and hs4, were recently demonstrated as master control elements of germline transcription and class switch recombination to most immunoglobulin constant genes. In addition, they were shown to enhance the accumulation of somatic mutations on linked transgenes. Since somatic hypermutation and class switch recombination are tightly linked processes, their common dependency on the endogenous locus 3′ enhancers could be an attractive hypothesis. VDJ structure and somatic hypermutation were analyzed in B cells from mice carrying either a heterozygous or a homozygous deletion of these enhancers. We find that hs3b and hs4 are dispensable both for VDJ assembly and for the occurrence of mutations at a physiologic frequency in the endogenous locus. In addition, we show that cells functionally expressing the immunoglobulin M (IgM) class B-cell receptor encoded by an hs3b/hs4-deficient locus were fully able to enter germinal centers, undergo affinity maturation, and yield specific antibody responses in homozygous mutant mice, where IgG1 antibodies compensated for the defect in other IgG isotypes. By contrast, analysis of Peyer patches from heterozygous animals showed that peanut agglutinin (PNAhigh) B cells functionally expressing the hs3b/hs4-deficient allele were dramatically outclassed by B cells expressing the wild-type locus and normally switching to IgA. This study thus also highlights the role of germinal centers in the competition between B cells for affinity maturation and suggests that membrane IgA may promote recruitment in an activated B-cell compartment, or proliferation of activated B cells, more efficiently than IgM in Peyer patches.
The vitamin K–dependent γ-glutamyl carboxylase gene contains a TATA-less promoter with a novel upstream regulatory element
Authors: Romero Elizabeth E. , Marvi Umaima , Niman Zachary E. , Roth David A.
Published: April 2003

The expression of the vitamin K–dependent γ-glutamyl carboxylase gene in liver is developmentally regulated. Since the gene product catalyzes an essential post-translational modification of the vitamin K–dependent blood coagulation proteins, the regulation of carboxylase expression is critical for hemostasis. We analyzed the activity of the rat carboxylase gene 5′-regulatory DNA sequences in rat hepatoma cell lines at different states of differentiation. These studies demonstrated that the 2.6-kb 5′-flanking sequence has differentiation-dependent transcriptional activity. Transient gene expression assays, examining the effects of nested deletions and site-directed mutagenesis of putative regulatory sequences, together with electrophoretic mobility shift assays (EMSAs) were used to identify sequences critical for the developmentally regulated transcription of the rat carboxylase gene. We identified a DNA sequence (–76 to –65; GTTCCGGCCTTC) not known to bind to transcription factors, yet which functions as an upstream promoter element. In vivo genomic DNA footprinting confirms the presence of nuclear protein–DNA interactions at this site in the endogenous carboxylase gene in differentiated hepatoma cells. Therefore, this DNA sequence has specific nuclear protein–binding activity and functional properties consistent with a regulatory element that plays a critical role in the developmental expression of the carboxylase gene, and hence the regulation of vitamin K–dependent blood coagulation protein synthesis.
Expression and function of the endothelial protein C receptor in human neutrophils
Authors: Sturn Daniel H. , Kaneider Nicole C. , Feistritzer Clemens , Djanani Angela , Fukudome Kenji , Wiedermann Christian J.
Published: April 2003

Activation of protein C by thrombin bound to thrombomodulin is enhanced by endothelial protein C receptor. This pathway may inhibit inflammation. We investigated effects of protein C and activated protein C on neutrophils as well as whether an endothelial protein C receptor is involved in mediating protein C effects. Neutrophils were from venous blood of healthy donors. Cell migration, respiratory burst, phagocytic activity, and apoptosis were studied by micropore filter assays and fluorometry. Receptor expression was investigated by reverse transcriptase–polymerase chain reaction (PCR) for mRNA, sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS-PAGE) and autoradiography of immunoprecipitated receptor protein, and fluorescence-activated cell-sorter scanner (FACS) analysis using the anti–endothelial protein C receptor antibody RCR-252. Neither protein C nor activated protein C induced migration, yet both of them inhibited neutrophil chemotaxis triggered by interleukin-8, formyl-Met-Leu-Phe, antithrombin, or C5a. A protein C activation–blocking antibody against endothelial protein C receptor diminished inhibitory effects of protein C or activated protein C on migration. No effect of either protein C preparation was seen in neutrophil's respiratory burst, bacterial phagocytosis, or apoptosis assays. Endothelial protein C receptor immunoreactivity was confirmed on neutrophils by FACS. De novo synthesis is suggested by endothelial protein C receptor mRNA expression as demonstrated by reverse transcriptase PCR and immunoprecipitation SDS-PAGE analyses. Data suggest that an endothelial protein C receptor is expressed by human neutrophils whose active site ligation with either protein C or activated protein C arrests directed cell migration. Inhibitory effects of these components of the protein C pathway on neutrophil function may play a role in the protein C–based treatment of severe sepsis.
Factor V New Brunswick: Ala221Val associated with FV deficiency reproduced in vitro and functionally characterized
Authors: Steen Mårten , Miteva Maria , Villoutreix Bruno O. , Yamazaki Tomio , Dahlbäck Björn
Published: April 2003

Factor V (FV) deficiency, also known as parahemophilia, is a rare bleeding disorder. Herein we investigate the first reported missense mutation associated with FV deficiency, Ala221Val, assigned as FV New Brunswick. To elucidate the molecular pathology associated with the Ala221Val substitution, the mutation was recreated in a recombinant system together with 3 FV mutants (Ala221Gly, Glu275Gln, and Cys220Ala/Cys301Ala) designed to help explain the Ala221Val phenotype. The expression pattern was analyzed by pulse-chase experiments and an FV-specific enzyme-linked immunosorbent assay (ELISA), the results suggesting the Ala221Val mutation not to interfere with the synthesis or secretion. The functional properties of the recombinant FV New Brunswick were evaluated in both plasma clotting and purified systems. The Ala221Val mutation did not affect the factor Xa (FXa) cofactor function; nor did it interfere with the activated protein C (APC)–mediated down-regulation of activated FV (FVa) activity. However, FV New Brunswick demonstrated reduced stability at 37°C due to an increased rate of dissociation of light and heavy chains of FVa. In conclusion, this in vitro study of FV New Brunswick suggests the Ala221Val mutation not to impair synthesis and expression of procoagulant activity, indicating overall proper folding of the mutant molecule. Rather, the Ala221Val substitution appears to interfere with the stability of the activated FVa mutant, the reduced stability possibly explaining the deficiency symptoms associated with the mutation.
Mobilization of hematopoietic stem cells during homeostasis and after cytokine exposure
Authors: Abkowitz Janis L. , Robinson Abigail E. , Kale Sujata , Long Michael W. , Chen Jing
Published: April 2003

We created parabiotic mice, joining ROSA26 and PeP3b animals, to study the trafficking of hematopoietic stem cells (HSCs) from marrow to blood and their return to marrow. The transfer of HSCs was assayed by secondary marrow transplantation and was 1.0% to 2.5% after 3, 6, 8, and 12 weeks. Thus, HSC homeostasis is primarily maintained by the retention of stem cells derived from replication events within the marrow, not the homing and engraftment of HSCs from the circulation. Of interest, the phenotypes of marrow progenitors and granulocytes were similar to those for HSCs, implying that the marrow functions as an intact compartment where differentiating cells derive from endogenous HSC. In contrast, 50% of splenic granulocytes and progenitor cells derived from the parabiotic partner, suggesting splenic progenitor cells were in constant equilibrium with progenitors in blood. In additional studies, animals were exposed to granulocyte–colonystimulating factor (G-CSF) and stem cell factor at days 17 to 20 of parabiosis and were studied 3 weeks later; 10.1% of marrow HSCs derived from the parabiotic partner. These data imply that HSCs, mobilized to the blood in response to cytokine exposure, are destined to later return to marrow, an observation that supports the concept that the mobilized peripheral blood stem cells used in clinical transplantation function physiologically.
New insights into the structural basis of integrin activation
Authors: Xiong Jian-Ping , Stehle Thilo , Goodman Simon L. , Arnaout M. Amin
Published: April 2003

Integrins are cell adhesion receptors that communicate biochemical and mechanical signals in a bidirectional manner across the plasma membrane and thus influence most cellular functions. Intracellular signals switch integrins into a ligand-competent state as a result of elicited conformational changes in the integrin ectodomain. Binding of extracellular ligands induces, in turn, structural changes that convey distinct signals to the cell interior. The structural basis of this bidirectional signaling has been the focus of intensive study for the past 3 decades. In this perspective, we develop a new hypothesis for integrin activation based on recent crystallographic, electron microscopic, and biochemical studies.
RANTES (CCL5) uses the proteoglycan CD44 as an auxiliary receptor to mediate cellular activation signals and HIV-1 enhancement
Authors: Roscic-Mrkic Branka , Fischer Marek , Leemann Christine , Manrique Amapola , Gordon Cynthia J. , Moore John P. , Proudfoot Amanda E. I. , Trkola Alexandra
Published: April 2003

The CC-chemokine RANTES (regulated on activation normal T-cell expressed and secreted; CCL5) transduces multiple intracellular signals. Like all chemokines, it stimulates G protein–coupled receptor (GPCR) activity through interaction with its cognate chemokine receptor(s), but in addition also activates a GPCR-independent signaling pathway. Here, we show that the latter pathway is mediated by an interaction between RANTES and glycosaminoglycan chains of CD44. We provide evidence that this association, at both low, physiologically relevant, and higher, probably supraphysiologic concentrations of RANTES, induces the formation of a signaling complex composed of CD44, src kinases, and adapter molecules. This triggers the activation of the p44/42 mitogen-activated protein kinase (MAPK) pathway. By specifically reducing CD44 expression using RNA interference we were able to demonstrate that the p44/p42 MAPK activation by RANTES requires a high level of CD44 expression. As well as potently inhibiting the entry of CCR5 using HIV-1 strains, RANTES can enhance HIV-1 infectivity under certain experimental conditions. This enhancement process depends in part on the activation of p44/p42 MAPK. Here we show that silencing of CD44 in HeLa-CD4 cells prevents the activation of p44/p42 MAPK and leads to a substantial reduction in HIV-1 infectivity enhancement by RANTES.
Association of CIB with GPIIb/IIIa during outside-in signaling is required for platelet spreading on fibrinogen
Authors: Naik Ulhas P. , Naik Meghna U.
Published: April 2003

Platelet spreading on immobilized fibrinogen (Fg) involves progression through a number of morphologic stages that, although distinctive, are not well understood mechanistically. Here we demonstrate that an association between GPIIb/IIIa and calcium- and integrin-binding protein (CIB) is required for the process of platelet spreading. Upon platelet adhesion to immobilized Fg, CIB localizes to the transiently formed filopodia and then redistributes diffusely along the membrane periphery of spread platelets. Immunoprecipitation analyses indicate that CIB and glycoprotein IIb/IIIa (GPIIb/IIIa) interact with each other as platelets adhere to immobilized Fg, and together they associate with the platelet cytoskeleton. Introduction of anti-CIB antibody or GPIIb cytoplasmic peptide into platelets blocks lamellipodia but not filopodia formation. GPIIb peptide–induced inhibition of platelet spreading is recovered by the incorporation of recombinant CIB protein, suggesting that interaction between CIB and GPIIb/IIIa is required for progression from filopodial to spread morphologies. Further, anti-CIB– or GPIIb peptide–induced inhibition of platelet spreading can be overcome by the addition of exogenous adenosine diphosphate (ADP). These data suggest that formation of the CIB-GPIIb/IIIa complex may be necessary for initiation of downstream signaling events, such as ADP secretion, that lead to platelet spreading.
Molecular characterization of Turkish patients with pyrimidine 5′ nucleotidase-I deficiency
Authors: Balta Gunay , Gumruk Fatma , Akarsu Nurten , Gurgey Aytemiz , Altay Cigdem
Published: April 2003

Pyrimidine 5′ nucleotidase-I (P5N-I) deficiency is a rare autosomal recessive disorder associated with hemolytic anemia, marked basophilic stippling, and accumulation of high concentrations of pyrimidine nucleotides within the erythrocyte. Recently, the structure and location of the P5N-I gene have been published. This paper presents the results of a study characterizing the molecular pathologies of P5N-I deficiency in a total of 6 Turkish patients from 4 unrelated families of consanguineous marriages. Mutation analysis in the P5N-I gene led to the identification of 3 novel mutations in these patients. In 4 patients from 2 families, a homozygous insertion of double G at position 743 was detected in exon 9 (743-744insGG), leading to premature termination of translation 23 bp downstream. In one family, a homozygous T to G transition at position 543 (543T>G) in exon 8 resulted in the replacement of tyrosine (Tyr) with a stop codon (Tyr181Stop). In another family, a homozygous insertion of a single A in exon 7 (384-385insA) created a stop signal at the codon nearby. In all families, the parents were heterozygous for the relevant mutations. None of these changes was detected in 200 chromosomes from a healthy Turkish population. These mutations were not correlated with any particular phenotype.
Hemoglobin metabolites mimic benzodiazepines and are possible mediators of hepatic encephalopathy
Authors: Ruscito Brian J. , Harrison Neil L.
Published: April 2003

Liver failure is often accompanied by cognitive impairment and coma, a syndrome known as hepatic encephalopathy (HE). The administration of flumazenil, a benzodiazepine (BZ) antagonist, is effective in reversing the symptoms of HE in many patients. These clinical observations gave rise to notions of an endogenous BZ-like mechanism in HE, but to date no viable candidate compounds have been characterized. We show here that the hemoglobin (Hb) metabolites hemin and protoporphyrin IX (PPIX) interact with the BZ site on the γ-aminobutyric acid (GABAA) receptor and enhance inhibitory synaptic transmission in a manner similar to diazepam and zolpidem. This finding suggests that hemin and PPIX are neuroactive porphyrins capable of acting as endogenous ligands for the central BZ site. The accumulation of these porphyrins under pathophysiologic conditions provides a potentially novel mechanism for the central manifestations of HE.