bluuebunny/crossref_metadata_2025 huggingface dataset:
Interaction of the Ras-related protein associated with diabetes Rad and the putative tumor metastasis suppressor NM23 provides a novel mechanism of GTPase regulation
Authors: Zhu Jianhua , Tseng Yu-Hua , Kantor Jason D. , Rhodes Christopher J. , Zetter Bruce R. , Moyers Julie S. , Kahn C. Ronald
Published: July 2002

Rad is the prototypic member of a new class of Ras-related GTPases. Purification of the GTPase-activating protein (GAP) for Rad revealed nm23, a putative tumor metastasis suppressor and a development gene in
Drosophila
. Antibodies against nm23 depleted Rad-GAP activity from human skeletal muscle cytosol, and bacterially expressed nm23 reconstituted the activity. The GAP activity of nm23 was specific for Rad, was absent with the S105N putative dominant negative mutant of Rad, and was reduced with mutations of nm23. In the presence of ATP, GDP⋅Rad was also reconverted to GTP⋅Rad by the nucleoside diphosphate (NDP) kinase activity of nm23. Simultaneously, Rad regulated nm23 by enhancing its NDP kinase activity and decreasing its autophosphorylation. Melanoma cells transfected with wild-type Rad, but not the S105N-Rad, showed enhanced DNA synthesis in response to serum; this effect was lost with coexpression of nm23. Thus, the interaction of nm23 and Rad provides a potential novel mechanism for bidirectional, bimolecular regulation in which nm23 stimulates both GTP hydrolysis and GTP loading of Rad whereas Rad regulates activity of nm23. This interaction may play important roles in the effects of Rad on glucose metabolism and the effects of nm23 on tumor metastasis and developmental regulation.
The stretch-activation response may be critical to the proper functioning of the mammalian heart
Authors: Vemuri Ramesh , Lankford Edward B. , Poetter Karl , Hassanzadeh Shahin , Takeda Kazuyo , Yu Zu-Xi , Ferrans Victor J. , Epstein Neal D.
Published: July 2002

The “stretch-activation” response is essential to the generation of the oscillatory power required for the beating of insect wings. It has been conjectured but not previously shown that a stretch-activation response contributes to the performance of a beating heart. Here, we generated transgenic mice that express a human mutant myosin essential light chain derived from a family with an inherited cardiac hypertrophy. These mice faithfully replicate the cardiac disease of the patients with this mutant allele. They provide the opportunity to study the stretch-activation response before the hearts are distorted by the hypertrophic process. Studies disclose a mismatch between the physiologic heart rate and resonant frequency of the cardiac papillary muscles expressing the mutant essential light chain. This discordance reduces oscillatory power at frequencies that correspond to physiologic heart-rates and is followed by subsequent hypertrophy. It appears, therefore, that the stretch-activation response, first described in insect flight muscle, may play a role in the mammalian heart, and its further study may suggest a new way to modulate human cardiac function.
Lethal mutagenesis of HIV with mutagenic nucleoside analogs
Authors: Loeb Lawrence A. , Essigmann John M. , Kazazi Farhad , Zhang Jue , Rose Karl D. , Mullins James I.
Published: July 2002

The human immunodeficiency virus (HIV) replicates its genome and
mutates at exceptionally high rates. As a result, the virus is able to
evade immunological and chemical antiviral agents. We tested the
hypothesis that a further increase in the mutation rate by promutagenic
nucleoside analogs would abolish viral replication. We evaluated
deoxynucleoside analogs for lack of toxicity to human cells,
incorporation by HIV reverse transcriptase, resistance to repair when
incorporated into the DNA strand of an RNA⋅DNA hybrid, and
mispairing at high frequency. Among the candidates tested,
5-hydroxydeoxycytidine (5-OH-dC) fulfilled these criteria. In seven of
nine experiments, the presence of this analog resulted in the loss of
viral replicative potential after 9–24 sequential passages of HIV in
human CEM cells. In contrast, loss of viral replication was not
observed in 28 control cultures passaged in the absence of the
nucleoside analog, nor with other analogs tested. Sequence analysis of
a portion of the HIV reverse transcriptase gene demonstrated a
disproportionate increase in G → A substitutions, mutations predicted
to result from misincorporation of 5-OH-dC into the cDNA during reverse
transcription. Thus, “lethal mutagenesis” driven by the class of
deoxynucleoside analogs represented by 5-OH-dC could provide a new
approach to treating HIV infections and, potentially, other viral
infections.
Synergy between an antiangiogenic integrin αvantagonist and an antibody–cytokine fusion protein eradicates spontaneous tumor metastases
Authors: Lode Holger N. , Moehler Thomas , Xiang Rong , Jonczyk Alfred , Gillies Stephen D. , Cheresh David A. , Reisfeld Ralph A.
Published: July 2002

The suppression and eradication of primary tumors and distant metastases is a major goal of alternative treatment strategies for cancer, such as inhibition of angiogenesis and targeted immunotherapy. We report here a synergy between two novel monotherapies directed against vascular and tumor compartments, respectively, a tumor vasculature-specific antiangiogenic integrin αvantagonist and tumor-specific antibody–interleukin 2 (IL-2) fusion proteins. Simultaneous and sequential combination of these monotherapies effectively eradicated spontaneous liver metastases in a poorly immunogenic syngeneic model of neuroblastoma. This was in contrast to controls subjected to monotherapies with either an antiangiogenic integrin αvantagonist or antibody–IL-2 fusion proteins, which were only partially effective at the dose levels applied. Furthermore, simultaneous treatments with the integrin αvantagonist and tumor-specific antibody–IL-2 fusion proteins induced dramatic primary tumor regressions in three syngeneic murine tumor models, i.e., melanoma, colon carcinoma, and neuroblastoma. However, each agent used as monotherapy induced only a delay in tumor growth. A mechanism for this synergism was suggested because the antitumor response was accompanied by a simultaneous 50% reduction in tumor vessel density and a 5-fold increase in inflammatory cells in the tumor microenvironment. Subsequently, tumor necrosis was demonstrated only in animals receiving the combination therapy, but not when each agent was applied as monotherapy. The results suggest that these synergistic treatment modalities may provide a novel and effective tool for future therapies of metastatic cancer.
Structure and function in rhodopsin: Kinetic studies of retinal binding to purified opsin mutants in defined phospholipid–detergent mixtures serve as probes of the retinal binding pocket
Authors: Reeves Philip J. , Hwa John , Khorana H. Gobind
Published: July 2002

In the current standard procedure for preparation of mammalian rhodopsin mutants, transfected COS-1 cells expressing the mutant opsin genes are treated with 5 μM 11-
cis-
retinal before detergent solubilization for purification. We found that binding of 11-
cis-
retinal to opsin mutants with single amino acid changes at Trp-265 (W265F,Y,A) and a retinitis pigmentosa mutant (A164V) was far from complete and required much higher concentrations of 11-
cis-
retinal. By isolation of the expressed opsins in a stable form, kinetic studies of retinal binding to the opsins
in vitro
have been carried out by using defined phospholipid–detergent mixtures. The results show wide variation in the rates of 11-
cis-
retinal binding. Thus, the
in vitro
reconstitution procedure serves as a probe of the retinal-binding pocket in the opsins. Further, a method is described for purification and characterization of the rhodopsin mutants after retinal binding to the opsins
in vitro
.
Selective killing of transformed cells by cyclin/cyclin-dependent kinase 2 antagonists
Authors: Chen Ying-Nan P. , Sharma Sushil K. , Ramsey Timothy M. , Jiang Li , Martin Mary S. , Baker Kayla , Adams Peter D. , Bair Kenneth W. , Kaelin William G.
Published: July 2002

Recent studies identified a short peptide motif that serves as a docking site for cyclin/cyclin-dependent kinase (cdk) 2 complexes
.
Peptides containing this motif block the phosphorylation of substrates by cyclin A/cdk2 or cyclin E/cdk2. Here we report that cell membrane-permeable forms of such peptides preferentially induced transformed cells to undergo apoptosis relative to nontransformed cells. Deregulation of E2F family transcription factors is a common event during transformation and was sufficient to sensitize cells to the cyclin/cdk2 inhibitory peptides. These results suggest that deregulation of E2F and inhibition of cdk2 are synthetically lethal and provide a rationale for the development of cdk2 antagonists as antineoplastic agents.
The first-nucleotide binding domain of the cystic-fibrosis transmembrane conductance regulator is important for inhibition of the epithelial Na
+
channel
Authors: Schreiber Rainer , Hopf Anna , Mall Marcus , Greger Rainer , Kunzelmann Karl
Published: July 2002

The cystic-fibrosis transmembrane conductance regulator (CFTR) functions as a cAMP-regulated Cl
−
channel and as a regulator of other membrane conductances. cAMP-dependent activation of CFTR inhibits epithelial Na
+
channels (ENaC). The specificity of interaction between CFTR and ENaC was examined by coexpression of ENaC and ATP-binding cassette (ABC) proteins other than CFTR. In addition, we identified domains within CFTR that are of particular importance for the inhibition of ENaC. To that end, two-electrode voltage-clamp experiments were performed on
Xenopus
oocytes coexpressing ENaC together with CFTR, the multidrug resistance protein MDR1, the sulfonyl urea receptor SUR1, or the cadmium permease YCF1. Except for CFTR, none of the other ABC proteins were able to inhibit ENaC. Several truncated versions of CFTR were examined for their inhibitory effects on ENaC. In fact, it is shown that C-terminal truncated CFTR is able to inhibit ENaC on activation by intracellular cAMP. Moreover, the data also show that an intact first-nucleotide binding domain (NBF-1) is important for inhibition of ENaC. We conclude that NBF-1 of CFTR contains a CFTR-specific regulatory site that down-regulates ENaC. It is speculated that this regulatory site also is needed for CFTR-mediated interactions with other membrane proteins and that it is not present in NBF-1 of other ABC proteins.
MEK kinase 1 is critically required for c-Jun N-terminal kinase activation by proinflammatory stimuli and growth factor-induced cell migration
Authors: Xia Ying , Makris Constantin , Su Bing , Li Erguang , Yang Jianhua , Nemerow Glen R. , Karin Michael
Published: July 2002

Exposure of eukaryotic cells to extracellular stimuli results in activation of mitogen-activated protein kinase (MAPK) cascades composed of MAPKs, MAPK kinases (MAP2Ks), and MAPK kinase kinases (MAP3Ks). Mammals possess a large number of MAP3Ks, many of which can activate the c-Jun N-terminal kinase (JNK) MAPK cascade when overexpressed, but whose biological function is poorly understood. We examined the function of the MAP3K MEK kinase 1 (MEKK1) in proinflammatory signaling. Using MEKK1-deficient embryonic stem cells prepared by gene targeting, we find that, in addition to its function in JNK activation by growth factors, MEKK1 is required for JNK activation by diverse proinflammatory stimuli, including tumor necrosis factor α, IL-1, double-stranded RNA, and lipopolysaccharide. MEKK1 is also essential for induction of embryonic stem cell migration by serum factors, but is not required for activation of other MAPKs or the IκB kinase signaling cascade.
The vav exchange factor is an essential regulator in actin-dependent receptor translocation to the lymphocyte–antigen-presenting cell interface
Authors: Wülfing Christoph , Bauch Angela , Crabtree Gerald R. , Davis Mark M.
Published: July 2002

During the interaction of a T cell with an antigen-presenting cell (APC), several receptor ligand pairs, including the T cell receptor (TCR)/major histocompatibility complex (MHC), accumulate at the T cell/APC interface in defined geometrical patterns. This accumulation depends on a movement of the T cell cortical actin cytoskeleton toward the interface. Here we study the involvement of the guanine nucleotide exchange factor vav in this process. We crossed 129 vav
−/−
mice with B10/BR 5C.C7 TCR transgenic mice and used peptide-loaded APCs to stimulate T cells from the offspring. We found that the accumulation of TCR/MHC at the T cell/APC interface and the T cell actin cytoskeleton rearrangement were clearly defective in these vav
+/−
mice. A comparable defect in superantigen-mediated T cell activation of T cells from non-TCR transgenic 129 mice was also observed, although in this case it was more apparent in vav
−/−
mice. These data indicate that vav is an essential regulator of cytoskeletal rearrangements during T cell activation.
Direct evidence for local oscillatory current sources and intracortical phase gradients in turtle visual cortex
Authors: Prechtl James C. , Bullock Theodore H. , Kleinfeld David
Published: July 2002

Visual stimuli induce oscillations in the membrane potential of neurons in cortices of several species. In turtle, these oscillations take the form of linear and circular traveling waves. Such waves may be a consequence of a pacemaker that emits periodic pulses of excitation that propagate across a network of excitable neuronal tissue or may result from continuous and possibly reconfigurable phase shifts along a network with multiple weakly coupled neuronal oscillators. As a means to resolve the origin of wave propagation in turtle visual cortex, we performed simultaneous measurements of the local field potential at a series of depths throughout this cortex. Measurements along a single radial penetration revealed the presence of broadband current sources, with a center frequency near 20 Hz (γ band), that were activated by visual stimulation. The spectral coherence between sources at two well-separated loci along a rostral–caudal axis revealed the presence of systematic timing differences between localized cortical oscillators. These multiple oscillating current sources and their timing differences in a tangential plane are interpreted as the neuronal activity that underlies the wave motion revealed in previous imaging studies. The present data provide direct evidence for the inference from imaging of bidirectional wave motion that the stimulus-induced electrical waves in turtle visual cortex correspond to phase shifts in a network of coupled neuronal oscillators.
Heat shock protein 60 sequence comparisons: Duplications, lateral transfer, and mitochondrial evolution
Authors: Karlin Samuel , Brocchieri Luciano
Published: July 2002

Heat shock proteins 60 (GroEL) are highly expressed essential
proteins in eubacterial genomes and in eukaryotic organelles. These
chaperone proteins have been advanced as propitious marker sequences
for tracing the evolution of mitochondrial (Mt) genomes. Similarities
among HSP60 sequences based on
significant segment pair
alignment
calculations are used to deduce associations of
sequences taking into account GroEL functional/structural domain
differences and to relate HSP60 duplications pervasive in
α-proteobacterial lineages to the dynamics of lateral transfer and
plasmid integration. Multiple alignments with consensuses are
determined for 10 natural groups. The group consensuses sharpen the
similarity contrasts among individual sequences. In particular, the Mt
group matches best with the classical α-proteobacteria and closely
with
Rickettsia
but significantly worse with the
rickettsial groups
Ehrlichia
and
Orientia
. However, across broad protein sequence
comparisons, there appears to be no consistent prokaryote whose protein
sequences align best with animal Mt genomes. There are plausible
scenarios indicating that the nuclear-encoded HSP60 (and HSP70)
sequences functioning in Mt are results of lateral transfer and are
probably derived from an α-proteobacterium. This hypothesis relates
to the plethora of duplicated HSP60 sequences among the classical
α-proteobacteria contrasted with no duplications of HSP60 among other
clades of proteobacterial genomes. Evolutionary relations are
confounded by differential selection pressures, convergence, variable
mutational rates, site variability, and lateral gene transfer.
Actin coating of secretory granules during regulated exocytosis correlates with the release of rab3D
Authors: Valentijn Jack A. , Valentijn Karine , Pastore Lisa M. , Jamieson James D.
Published: July 2002

The present study describes a novel phenomenon in pancreatic acinar cells undergoing regulated exocytosis. When acinar cell preparations were challenged with the secretagogue carbamylcholine, a subpopulation of zymogen granules became coated with filamentous actin. These zymogen granules were always in proximity of the acinar cell apical membrane (the site of exocytosis) but did not appear to have fused yet. They were distinct from regular zymogen granules not only because of their association with filamentous actin, but also because the majority of them lacked the zymogen granule marker rab3D, a small GTPase implicated in regulated exocytosis. The apparent loss of rab3D, presumed to result from the release of rab3D from the granule membranes, could be prevented by agents that modulate the actomyosin system as well as by GTP[γS]. These data suggest that zymogen granules engaging in exocytosis become coated with actin before fusion and that this actin coating is tightly coupled to the release of rab3D. We propose that rab3D is involved in the regulation of actin polymerization around secretory granules and that actin coating might facilitate the movement of granules across the subapical actin network and toward their fusion site.
Inhibition of PC cell-derived growth factor (PCDGF, epithelin/granulin precursor) expression by antisense PCDGF cDNA transfection inhibits tumorigenicity of the human breast carcinoma cell line MDA-MB-468
Authors: Lu Runqing , Serrero Ginette
Published: July 2002

PC-cell derived growth factor (PCDGF) is an 88-kDa growth factor originally purified from the highly tumorigenic teratoma PC cell line and corresponds to the epithelin/granulin precursor. In teratoma cells, PCDGF expression was shown to be essential for tumorigenicity. We have reported that PCDGF was expressed in estrogen receptor-positive (ER
+
) human mammary epithelial cells in an estrogen-dependent fashion. In this study, we have investigated PCDGF expression in human mammary epithelial cell lines ranging from immortalized nontumorigenic cells to ER
+
and ER
−
breast carcinoma cells. Northern and Western blot analyses indicated that PCDGF mRNA and protein expression was low in nontumorigenic cells and increased in human breast carcinomas cell lines in a positive correlation with their tumorigenicity. Treatment of the ER
−
MDA-MB-468 cells with anti-PCDGF neutralizing antibody resulted in a dose-dependent inhibition of their proliferation, suggesting that secreted PCDGF acted as an autocrine growth factor for breast carcinoma cells. We then examined the
in vitro
and
in vivo
growth properties of MDA-MB-468 cells, where PCDGF expression had been inhibited by antisense PCDGF cDNA transfection. Inhibition of PCDGF expression resulted in a reduced proliferation rate
in vitro
and a 60–80% reduction in colony formation. Tumor formation
in vivo
was dramatically inhibited in antisense cells with a 90% inhibition of tumor incidence and tumor weight. These results demonstrate the importance of PCDGF overexpression for the proliferation and tumorigenicity of ER
−
breast carcinomas and suggest that PCDGF overexpression may play an important role in human breast cancer.
Does clutch size evolve in response to parasites and immunocompetence?
Authors: Martin Thomas E. , Møller A. P. , Merino S. , Clobert J.
Published: July 2002

Parasites have been argued to influence clutch size evolution, but past work and theory has largely focused on within-species optimization solutions rather than clearly addressing among-species variation. The effects of parasites on clutch size variation among species can be complex, however, because different parasites can induce age-specific differences in mortality that can cause clutch size to evolve in different directions. We provide a conceptual argument that differences in immunocompetence among species should integrate differences in overall levels of parasite-induced mortality to which a species is exposed. We test this assumption and show that mortality caused by parasites is positively correlated with immunocompetence measured by cell-mediated measures. Under life history theory, clutch size should increase with increased adult mortality and decrease with increased juvenile mortality. Using immunocompetence as a general assay of parasite-induced mortality, we tested these predictions by using data for 25 species. We found that clutch size increased strongly with adult immunocompetence. In contrast, clutch size decreased weakly with increased juvenile immunocompetence. But, immunocompetence of juveniles may be constrained by selection on adults, and, when we controlled for adult immunocompetence, clutch size decreased with juvenile immunocompetence. Thus, immunocompetence seems to reflect evolutionary differences in parasite virulence experienced by species, and differences in age-specific parasite virulence appears to exert opposite selection on clutch size evolution.
History of modern psychology in Germany1 in 19th‐ and 20th‐century thought and society
Authors: Sprung Lothar , Sprung Helga
Published: July 2002

This entry surveys the characteristics, stages, and lines of development of modern psychology as an empirical psychology in Germany during the 19th and 20th centuries. These developments occurred in the context of very different political periods in German history, during a time span extending from the period of the Restoration after the Vienna Congress of 1814/15 to the present‐day “Berlin Republic”. At the centre lies the question of continuities and discontinuities in the evolution of psychology during the profound sociocultural changes that marked the politically heterogeneous developmental phases of German history. The stages are indicated by headings treating the development of psychology during the period of Restoration after 1814/15, in the German Empire (1871–1918), in the Weimar Republic (1919–1933), during the time of National Socialism (1933–45), in the Federal Republic of Germany (FRG, 1949–1990), in the German Democratic Republic (GDR, 1949–1990), and in united Germany after 1990. The result will be to demonstrate how psychology in 19th‐ and 20th‐century Germany developed from modest beginnings into a richly elaborated scientific field with diverse institutions and a differentiated professional structure. The fate of individual German psychologists during this time can be only tangentially considered.
How new is the New Deal? A qualitative study of the New Deal for Young People on minority ethnic groups in Oldham
Authors: Fieldhouse Dr. Edward A. , Kalra Dr. Virinder S. , Alam Saima
Published: July 2002

Despite a clear commitment to equality of opportunity under the New Deal, some have questioned whether the New Deal for Young People has served ethnic minority communities as well as the white population. This research is based primarily on a qualitative longitudinal study of ethnic minority New Deal participants (and non-participants) in Oldham in Greater Manchester, and also on analysis of the New Deal Evaluation Database. We fin d that objective outcomes, particularly employment, can be problematic, but that there is strong evidence of positive impacts on employability, especially through work experience. In these terms the NDYP clearly represents a departure from the YTS and has had a generally positive reception from participants. However, our research emphasizes the fact that participants are a self-selected section of young unemployed people, and that there are major differences between participants and ‘avoiders’. Most notably, non-participants were much more disaffected with the labour market and more negative about the New Deal.
Permeability characteristics of human endothelial monolayers seeded on different extracellular matrix proteins
Authors: Nooteboom Arjan , Hendriks Thijs , Ottehöller Irene , Van Der Linden Cees. J.
Published: July 2002

Objective: To investigate whether endothelial monolayer permeability changes induced by inflammatory mediators are affected by the extracellular matrix protein used for cell seeding.

Methods: Human umbilical venular endothelial cells (HUVEC) were grown to confluent monolayers on membranes coated with either collagen, fibronectin or gelatin. The permeability to albumin and dextran was then assessed, both under normal conditions and after treatment with tumor necrosis factor‐alpha (TNF‐α) and bacterial
lipopolysaccharide (LPS).

Results: With any of the three protein coatings, tight junctions were formed all over the monolayers. The permeability of the coated membranes to albumin and dextran was reduced strongly by confluent monolayers; the relative reduction was similar for the three matrix proteins used. Pre‐incubation of the monolayers with either TNF‐α or LPS increased permeability dose dependently. However, the relative increase due to either treatment was independent of the protein used for membrane coating.

Conclusion: The extracellular matrix protein used for initial seeding of endothelial cultures plays a minor role in determining the permeability changes induced in HUVEC monolayers by inflammatory mediators.
Analysis of local and systemic inflammatory responses induced by polymicrobial peritonitis in mice
Authors: Fröde Tânia Silvia , Ferreira Silvia Ines Alejandra Cordoba Pires , Medeiros Yara Santos
Published: July 2002

Background: Abdominal sepsis induces a local production of proinflammatory mediators that may trigger both septic shock and organ‐system dysfunction.

Aims: The present study analyzed exudation, cell migration, and CD11a and CD18 subset cells of both local and systemic responses induced by fecal peritonitis in mice.

Methods: Animals were anesthetized and, after performing a midline incision in the abdomen, the cecum was ligated and punctured twice with a needle. Sham‐operated animals were included. Some groups were previously treated with Evans blue dye (intravenously) to further evaluate the amount of tissue and abdominal cavity leakages.

Results: Fecal peritonitis triggered a local inflammatory reaction with an increased number of leukocytes and exudation between 6 and 48 h (p < 0.01). Although CD11a/CD18‐positive cells in the abdomen peaked after 24 h, a significant decrease of them was detected after 48 h (p < 0.05). At the studied period of time (6‐48 h), different degrees of exudation in several organs occurred, whereas a significant late recruitment (24 h) of CD11a/CD18 cells into the lungs was observed.

Conclusions: In this model, cell migration and exudation at the site of injury occurred in parallel. However, in the lungs, the recruitment of leukocytes that express CD11a/CD18 adhesion molecules constitutes a non‐dependent event in relation to fluid leakage accumulation at this site.
T cells expressing the γδ receptor are essential for Th2‐mediated inflammation in patients with acute exacerbation of asthma
Authors: Hamzaoui Agnès , Kahan Alice , Ayed Khaled , Hamzaoui Kamel
Published: July 2002

Objective: T lymphocytes have a central regulatory role in the pathogenesis of asthma. The objective of this study was to characterize immunologically the activation stage of asthma and the functional profile of lymphocytes from induced sputum, with particular emphasis on γδ T cells.

Methods: Induced sputum was collected from 10 patients with acute exacerbation of asthma, and from healthy controls. The expression of activation markers on freshly isolated induced sputum lymphocytes and T‐cell subsets was analyzed by double immunofluorescent staining and flow cytometry. Fas ligand (FasL) was determined by reverse transcriptase‐polymerase chain reaction analysis. The phenotype of γδ T‐cell subpopulations was tested by A13 and BB3 monoclonal antibodies. In this context, the functional profile of γδ T cells was tested in a chromium releasing test.

Results: A significantly decreased proportion of αβ T cells and an increased proportion of γδ T cells,
CD56+ cells and CD8+ γδ T cells were found in asthma patients compared with healthy controls. In asthmatic patients, there is a significantly increased proportion of T cells expressing CD69 and CD25 antigen. After stimulation of γδ T cells, an increased expression of intracellular tumour necrosis factor‐α, interleukin (IL)‐4 and IL10 cytokines were found at higher levels than controls. Interferon‐γ was observed at similar levels in asthma patients and healthy controls. Freshly isolated T‐cell receptor
(TCR) γδ+ cells exhibited an increased percentage of FasL in our patient group. FasL mRNA was detected in (TCR) γδ+ cells before and after IL2 stimulation. (TCR) γδ+ cells were cytotoxic against the K562 cell line. This natural killer activity was mediated by the A13‐positive subpopulation.

Conclusion: The presence of cytokines producing γδ cells in induced sputum of asthmatic patients is consistent with regulatory activities. These cells display also cytotoxic function.
Susceptibility of lung epithelium to neutrophil elastase: protection by native inhibitors
Authors: Bingle L. , Richards R. J. , Fox B. , Masek L. , Guz A. , Tetley T. D.
Published: July 2002

The development of emphysema is thought to be due to an imbalance of proteases (especially neutrophil elastase [NE]) and antiproteases with loosening of the respiratory epithelium as an early event. We investigated the effect of NE on respiratory epithelial cell adherence in vitro , in the presence of varying concentrations and combinations of native inhibitors, α‐1‐proteinase inhibitor (PI) and secretory leukoprotease inhibitor (SLPI). SLPI was two to 12 times more effective than PI at preventing the effects of NE, especially when enzyme:inhibitor ratios were almost equivalent. Even when the concentration of SLPI was only 10% of the total (as in normal peripheral lung secretions), it gave greater protection than PI alone. This suggests that SLPI plays an important role in controlling neutrophil elastaseinduced inflammation and tissue damage.
Carrageenan‐induced acute inflammation in the mouse air pouch synovial model. Role of tumour necrosis factor
Authors: Romano M. , Faggioni R. , Sironi M. , Sacco S. , Echtenacher B. , Di Santo E. , Salmona M. , Ghezzi P.
Published: July 2002

We used the mouse air pouch model of inflammation to study the interaction between cytokines,
prostaglandin E2 (PGE2) and cell migration during the various phases of acute local inflammation induced by carrageenan. In serum, the levels of interleukin 1 (IL‐1), interleukin 6 (IL‐6), tumour necrosis factor (TNF), serum amiloid‐A (SAA) and Fe++ were never different from controls, indicating that no systemic inflammatory changes were induced. Locally the exudate volume and the number of leukocytes recruited into the pouch increased progressively until 7 days after carrageenan. The same was true for PGE2 production. We could not measure IL‐1 but the production of IL‐6 and TNF reached a maximum after 5‐24 h then quickly decreased. Anti‐TNF antibodies inhibited cell migration by 50% 24 h after treatment. Pretreatment with interleukin 10 (IL‐10) inhibited TNF production almost completely and cell migration by 60%. Carrageenan‐induced inflammation was modulated by anti‐inflammatory drugs. Pretreatment with dexamethasone (DEX) or indomethacin (INDO) inhibited cell migration and reduced the concentration of TNF in the exudate. Production of
PGE2 or vascular permeability did not correlate with the number of cells in the pouch. Local TNF seems to play an important role in this model, particularly for leukocyte migration in the first phase of the inflammatory process. In conclusion, the air pouch seems to be a good model for studying the regulation of the early events of local inflammation, particularly the role of cytokines and cell migration.
Eicosanoid and cytokine levels in plasma of patients during mesenteric infarction
Authors: Nathan N. , Denizot Y. , Feiss P.
Published: July 2002

Multible organ failure (MOF) induced by mesenteric infarction is associated with a high mortality rate. This study reports eicosanoid and cytokine levels in the blood of three atherosclerotic patients who ultimately died from MOF induced by mesenteric infarction. High plasma levels of 6‐ keto‐prostaglandin (PG) F1α (the stable metabolite of
PGI2), interleukin (IL)‐6 and IL‐8 are observed whereas plasma tumour necrosis factor alpha (TNFα),
TxB2 (the stable metabolite of TxA2), PGE2, leukotrienes (LT)B4 and LTC4, and whole blood platelet‐activating factor levels are not different from values obtained in similarly severe atherosclerotic patients. This short report questioned the clinical involvement of TNFα during such a pathology where a persistent translocation of endotoxin has been observed through the gut endothelial barrier. Activation of phospholipase A2 is suggested by the increase in the stable metabolite of PGI2 and might be by itself or through lipidic metabolites, a major systemic stimulus of IL‐6 and IL‐8 production.
Protective activity of propofol, Diprivan® and intralipid against active oxygen species
Authors: Mathy-Hartert M. , Deby-Dupont G. , Hans P. , Deby C. , Lamy M.
Published: July 2002

We separately studied the antioxidant properties of propofol (PPF), Diprivan® (the commercial form of PPF) and intralipid (IL) (the vehicle solution of PPF in Diprivan® ) on active oxygen species produced by phorbol myristate acetate (10‐6M)‐stimulated human polymorphonuclear leukocytes (PMN: 5 × 10 5 cells/ assay), human endothelial cells (5 × 105 cells/assay) or cell‐free systems (NaOCl or H2O2/peroxidase systems), using luminol (10-4M)‐enhanced chemiluminescence (CL). We also studied the protective effects of Diprivan® on endothelial cells submitted to an oxidant stress induced by H2O2/MPO system: cytotoxicity was assessed by the release of preincorporated 51Cr. Propofol inhibited the CL produced by stimulated PMN in a dose dependent m anner (until 5 × 10-5 M, a clinically relevant concentration), while Diprivan® and IL were not dose‐dependent inhibitors. The CL produced by endothelial cells was dosedependently inhibited by Diprivan® and PPF, and weakly by IL (not dose‐dependent). In cell free systems, dose‐dependent inhibitions were obtained for the three products with a lower effect for IL. Diprivan® efficaciously protected endothelial cells submitted to an oxidant stress, while IL was ineffective. By HPLC, we demonstrated that PPF was not incorporated into the cells. The drug thus acted by scavenging the active oxygen species released in the extracellular medium. IL acted in the same manner, but was a less powerful antioxidant.
Drug therapy: dose‐response relationship of oral mesalazine in inflammatory bowel disease
Authors: Mulder C. J. J. , Van Den Hazel S. J.
Published: July 2002

Mesalazine is widely used in the treatment of inflammatory bowel disease. Little is known about the doseresponse relationship and about possible dose related side effects. In ulcerative colitis higher dosages of mesalazine (3 g) are more effective in maintaining a remission than lower dosages (1.5 g). In mild to moderately active ulcerative colitis, studies also indicate that higher dosages might be more effective in inducing remission. Dose‐comparing studies in Crohn′s disease are even more sparse, but the available results indicate higher efficacy at higher dose levels.

None of the known side effects of mesalazine are clearly dose‐related. A pH‐dependent release system, however, can cause a sudden release of high doses of mesalazine. Consequent peak levels in serum have been implicated in mesalazine induced nephrotoxicity. In conclusion, despite the current practice of using increasing dosages of mesalazine in inflammatory bowel disease, both efficacy and safety have been established tentatively.
Release of prostaglandin D2 by murine mast cells: importance of metabolite formation for antiproliferative activity
Authors: Haberl C. , Hültner L. , Flügel A. , Falk M. , Geuenich S. , Wilmanns W. , Denzlinger C.
Published: July 2002

Prostaglandin (PG) D2 , PGJ2 and Δ12 ‐PGJ2 are antiproliferative eicosanoids. We investigated the production of PGD2 by murine bone marrow‐derived mast cells (BMMC) taking into consideration metabolism of PGD2 to PGD2 and Δ12‐PGJ2. PG‐metabolites were quantified by high performance liquid chromatography (HPLC) combined with radioimmunoassay (RIA). Stimulated with calcium ionophore A23187 BMMC released eight‐fold more PGJ2 and Δ12‐PGJ2 than PGD2. Conversion of endogenously produced PGD2 to PGJ2 and Δ12‐PGJ2 proceeded rapidly in contrast to metabolism of exogenously added PGD2. The antiproliferative potency of these prostaglandins is demonstrated in vitro. We conclude that determination of PGD2 production by mast cells must take into consideration rapid conversion to active derivatives, which may play a significant role in growth regulation.
Interest of Colchicine for the Treatment of Cystic Fibrosis Patients. Preliminary Report
Authors: Sermet-Gaudelus I. , Stoven V. , Annereau J. P. , Witko-Sarsat V. , Reinert P. , Guyot M. , Descamps-Latscha B. , Lallemand J. Y. , Lenoir G.
Published: July 2002

Cystic fibrosis (CF) lung disease is characterized by persistent inflammation. Antiinflammatory drugs, such as corticosteroids and ibuprofene, have proved to slow the decline of pulmonary function although their use is limited because of frequent adverse events. We hypothesized that colchicine could be an alternative treatment because of its antiinflammatory properties and upregulatory effect on cystic fibrosis transmembrane regulator (CFTR) closely related proteins. We herein present results obtained in an open study of eight CF children treated with colchicine for at least 6 months. Clinical status was better in all patients and respiratory function tests significantly improved in five. Median duration of antibiotherapy decreased significantly. These preliminary results support our hypothesis of a beneficial effect of colchicine in CF patients and stress the need for a controlled therapeutic trial.
Prevention of Antigen‐Induced Bronchial Hyperreactivity and Airway Inflammation in Sensitized Guinea‐Pigs by Tacrolimus
Authors: e Silva J. R. Lapa , Ruffié C. , Lefort J. , Nahori M-A. , Vargaftig B. B. , Pretolani M.
Published: July 2002

We examined the effect of the immunosuppressive agent, tacrolimus (FK506), on antigen‐induced bronchial hyperreactivity to acetylcholine and leukocyte infiltration into the airways of ovalbumin‐challenged guinea‐pigs. Subcutaneous injection of 0.5 mg/kg of FK506, 1 h before and 5 h after intra‐nasal antigen challenge prevented bronchial hyperreactivity to aerosolized acetylcholine, eosinophilia in bronchoalveolar lavage (BAL) fluid and bronchial tissue and the invasion of the bronchial wall by CD4+ T‐lymphocytes. FK506 also suppressed ovalbumininduced increase in the number of leukocytes adhering to the pulmonary vascular endothelium and expressing α4‐integrins. Inhibition by FK506 of antigen‐induced bronchial hyperreactivity in sensitized guinea‐pigs may thus relate to its ability to prevent the emergence of important inflammatory components of airway inflammation, such as eosinophil accumulation, as well as CD4+ T‐lymphocyte infiltration into the bronchial tissue.
Effects of Sulfur Mustard on Cytokines Released from Cultured Human Epidermal Keratinocytes
Authors: Kurt Elien M. , Schafer Robert J. , Arroyo Carmen M.
Published: July 2002

The release of the cytokines interleukin (IL)-1α, IL-1β, IL-6, IL-8, and tumor necrosis factor alpha (TNF-α was measured from epiderm alkeratinocytes in an attempt to characterize the immunologic response in keratinocytes following exposure to bis (2-chloroethyl)sulfide (sulfur mustard, HD). Enzyme-linked immunosorbentassay (ELISA) was used to measure cytokine levels in adult and neonatal culture human epidermal keratinocytes (HEK) 3 h after exposure to 0.50 and 1.0 m M HD. A two-way analysis of variance was carried out for cell type and HD concentration. That analysis showed significant differences between cell types for IL-1α and IL-1β(p =.001 and p =.015, respectively). In both of these cytokines, release in neonatal HEK decreased less than in adult HEK. A significant effectof HD concentration was shown only for IL-1β (P <.001), with cytokine release decreasing with increasing HD dose. In addition, a significant cell donor type by HD concentration interaction effect was found for IL-1β under the experimental conditions described in materials and methods. With increasing HD concentration, the relative decrease in cytokine release was much greater for adult than for neonatal HEK.
Subchronic Toxicity and Developmental Toxicity Studies in Rats With Aquateric® Aqueous Enteric Coating
Authors: Kotkoskie Lois A. , Freeman Christine , Palmieri Mark A.
Published: July 2002

Studies were conducted to evaluate the subchronic and developmental toxicity of Aquateric® Aqueous Enteric Coating. Homogeneity and stability studies were conducted over a range of 5,000 to 50,000 ppm Aquateric in the diet. In the subchronic study, groups of Sprague-Dawley rats (20/sex/group) received 0 (control), 5,000, 25,000, or 50,000 ppm Aquateric in the diet for 90 consecutive days. No mortality, clinical signs of toxicity or adverse toxicological effects on hematology or serum chemistry parameters, body weights, feed consumption, ophthalmological examinations, or histological evaluation of tissues were noted in any treatment group. In the developmental toxicity study, groups of pregnant Sprague-Dawley rats (25/group) received 0 (control), 5,000, 25,000 or 50,000 ppm Aquateric in the diet on gestational days 6-15. No evidence of maternal toxicity or fetotoxicity or embryotoxicity was noted. The no observed adverse effect level (NOAEL) exceeds 50,000 ppm in the diet, which represents a dose range of approximately 3600 to 4100 mg/kg/day. The results of these studies demonstrate the low toxicity of Aquateric. The estimated human intake is approximately 4 mg/kg/day. Based on the NOAEL from the subchronic study of 3604 mg/kg/day, the margin of safety is 900.
The ‘Anglo-American’ model, privatization and the transformation of private capitalism in Italy
Authors: McCann Dermot
Published: July 2002

Italian capitalism has long been distinguished by the large size of the public enterprise sector and the power of large family firms, underpinned by powerful networks of inter-firm alliance orchestrated by the merchant bank, Mediobanca. This article seeks to analyse the extent to which the current programme of privatization is serving not only to shrink the size of, but to transform the structure of, power in the private sector. While some reformers have seen in privatization an instrument to encourage the adoption of Anglo-Saxon forms of corporate governance in Italy, it is argued here that the evidence of recent changes indicates the emergence of a more differentiated pattern of corporate governance rather than the triumph of any single model.
Cerebral Metastases from Malignant Fibrous Histiocytoma of Bone
Authors: Rogers Susanne J. , Whelan Jeremy S.
Published: July 2002

Four patients with malignant fibrous histiocytoma of bone (MFH‐B) metastasizing to brain are reported. In two cases, signs of cerebral involvement developed between 4 and 28 months after diagnosis. Both patients had known pulmonary or bony metastases. As a consequence of this experience, two further patients were subsequently identified, one with a definite cerebral metastasis and one who had an asymptomatic supratentorial lesion, possibly metastatic. It is suggested that patients with MFH‐B and widespread metastatic disease at presentation or developing within a short interval should undergo cerebral imaging.
Conformal Radiotherapy Facilitates the Delivery of ConcurrentChemotherapy and Radiotherapy: A Case of Primitive NeuroectodermalTumour of the Chest Wall
Authors: Coles C. E. , Twyman N. , Earl H. M. , Burnet N. G.
Published: July 2002

We illustrate the principle of conformal radiotherapy by discussing the case of a patient with a primitive neuroectodermal
tumour of the chest wall. Recent advances in radiotherapy planning enable precise localization of the planning target volume
(PTV) and normal organs at risk of irradiation. Customized blocks are subsequently designed to produce a treatment field
that ‘conforms’ to the PTV. The use of conformal radiotherapy (CRT) in this case facilitated the delivery of concurrent
chemotherapy and radiotherapy by significantly reducing the volume of red marrow irradiated.The lack of acute and late
toxicities was attributed to optimal exclusion of normal tissues from the treatment field, made possible by CRT.
Leiomyosarcoma of the Cephalic Vein
Authors: Jefferson Kieran P. , Dixon John H.
Published: July 2002

A 78‐year‐old man presented with a mass on his right forearm. A 5 x 4 x 3 cm3 mass was excised en bloc with extensions along
the course of the cephalic vein and its tributaries. Histological analysis revealed the mass to be a high‐grade leiomyosarcoma
arising within the cephalic vein. The tumour was controlled locally and distally until the patient died 10 months later, from
an unrelated illness. This is the first reported case of a venous leiomyosarcoma of the cephalic vein.
A Phase I/II Study of a 72‐h Continuous Infusion of Etoposide in Advanced Soft Tissue Sarcoma
Authors: Crawley Charles R. , Judson Ian R. , Verrill Mark , Hill Catherine , Raynaud Florence I.
Published: July 2002

Purpose. The study was performed to assess the antitumour activity and toxicity of a 72‐h continuous infusion of
single‐agent etoposide as second‐line treatment for patients with locally advanced or metastatic soft tissue sarcoma (STS),
following reports of substantial activity using this schedule of etoposide administration as first‐line treatment in
combination with ifosfamide.

Patients/method. This was an open phase I/II trial performed at a single institution in patients with metastatic or locally
advanced STS who had failed first‐line treatment with doxorubicin + ifosfamide combination chemotherapy or, less
commonly, single‐agent treatment with doxorubicin or ifosfamide. Etoposide was given as a continuous intravenous
infusion over 72 h. The starting dose level was
200 mgm−2
day−1 × 3 escalating in 10% steps in cohorts of three patients
until dose‐limiting toxicity was encountered.

Results. Seventeen patients were treated, median age 47 years (range 26–71 years). No responses were seen in 16
assessable patients despite etoposide levels in the cotoxic range. The steady‐state plasma concentration exceeded
8 μg ml−1 in all patients and in patients treated at ≥ 600 mg m−2 the mean steady‐state level was 14.4 μg ml−1. The
median event‐free survival was 6 weeks (95% confidence interval (CI) 3.31–8.69) and the overall survival 16 weeks (95%
CI 9.28–22.72). The maximum tolerated dose in this pretreated patient group was 200 mg m−2
day−1 × 3. The dose‐limiting toxicity was myelosuppression.

Discussion. Etoposide given by 72‐h infusion is inactive as second‐line chemotherapy in STS.
It is associated with significant toxicity when given in these doses, in this patient group.
Multi‐Focal, Multi‐Centric Angiosarcoma of Bone
Authors: Kakouri Eleni , Whelan Jeremy S. , Coltart Stewart , Smith Mark E. F. , Souhami Robert L.
Published: July 2002

A multi‐focal multi‐centric, malignant tumour of vascular origin arising in bone in a 55‐year‐old man is described. The presenting symptoms were pain and weight loss. Radiologically, multiple lytic lesions were demonstrated in the long bones of both legs and throughout the pelvis. Histological examination demonstrated an angiosarcoma which was predominantly low grade in nature but with focal areas of intermediate grade. Turnout cells expressed the endothelial markers CD31, CD34 and von Willebrand′s factor. There was rapid radiological progression of disease with no response to radiotherapy. Pain abated within a few days of institution of doxorubicin, 75 mg m−2, but the patient died of massive pulmonary thromboembolism 14 days later, 11 months after the first symptoms.