bluuebunny/crossref_metadata_2025 huggingface dataset:
Impact of antigenemia on the bioactivity of infused anti-Tac antibody: Implications for dose selection in antibody immunotherapies
Authors: Junghans R. P. , Carrasquillo J. A. , Waldmann T. A.
Published: July 2002

In patients with malignancies and immune disorders expressing Tac (α chain of the interleukin 2 receptor; CD25), physiologic shedding of this receptor may lead to high blood levels of the soluble form (sTac). This system was used to model the interaction of soluble antigen with antibody in therapeutic settings and to develop rational principles to optimize the delivery of antibody to tumor target cells. First, we confirmed that sTacin vivocan block anti-Tac binding sites and diminish antibody binding to Tac+ cells. Second, the bioactivity of antibodyin vivocorrelated directly with the amount of antibody infused and inversely with the sTac concentration. Third, bindability of antibody declined in the hours and days after anti-Tac infusion in patients. Finally, tumor targeting was achieved even in the presence of excess sTac, demonstrating a partition of antibody between soluble and cell-bound antigen. A role is proposed for the Brambell receptor (FcRB) to delay saturation of human or chimeric antibodies via differential catabolism of antigen–antibody complexes. Principles are developed for predicting activity of administered antibody in the presence of soluble antigen to assist in dose selection in passive, radioimmuno and immunotoxin therapies.
SR proteins are sufficient for exon bridging across an intron
Authors: Stark Jeremy M. , Bazett-Jones David P. , Herfort Manfred , Roth Mark B.
Published: July 2002

We have developed a defined system to characterize the role of SR proteins and exonic enhancers in directly promoting splice-site interactions across an intron. Using RNA affinity chromatography, we find that SR proteins alone are sufficient to promote the specific association of the enhancer-containing exon 5 with the adjoining exon 6 from avian cardiac troponin-T. Direct visualization of this exon/exon association by electron spectroscopic imaging shows it to be highly specific. Furthermore, using
in vivo
characterized mutants of exon 5, we also show that this exon/exon association depends on the splicing enhancer within exon 5. These results suggest a model by which SR proteins may function through exonic enhancers to directly promote exon bridging.
Human T cell lymphotropic virus type I Tax protein trans-activates interleukin 15 gene transcription through an NF-κB site
Authors: Azimi Nazli , Brown Keith , Bamford Richard N. , Tagaya Yutaka , Siebenlist Ulrich , Waldmann Thomas A.
Published: July 2002

Interleukin 15 (IL-15) mRNA is expressed in a wide variety of tissue types. However, with the exception of some T cell lines, IL-15 transcript expression has not been described in T cells. Herein we demonstrate that IL-15 mRNA can be detected in freshly isolated normal T cells and T cell lines. Furthermore, its expression is 3- to 4-fold higher in human T cell lymphotropic virus type I (HTLV-I)-infected T cells. By using reporter constructs bearing the 5′ regulatory region of the IL-15 gene, we observed a positive correlation between HTLV-I Tax protein expression and IL-15 promoter activity in HTLV-I-infected T cells. Additionally, by using a Jurkat T cell transfectant that expresses Tax under an inducible promoter, we demonstrated that the expression of IL-15 mRNA increased 3-fold as Tax was expressed, suggesting that the Tax protein activates IL-15 transcription. An NF-κB consensus sequence is located at the −75 and −65 region of the IL-15 5′ regulatory region. Mutations in the NF-κB motif or deletion of this sequence abrogated the promoter activity in both HTLV-I-positive and Jurkat Tax-transfectant cells. These data represent evidence for trans-activation of the IL-15 gene by the HTLV-I Tax protein through an NF-κB motif and suggest a potential role for IL-15 in HTLV-I-associated diseases such as adult T cell leukemia and HTLV-I-associated myopathy/tropical spastic paraparesis.
Increased platelet sensitivity to ADP in mice lacking platelet-type 12-lipoxygenase
Authors: Johnson Eric N. , Brass Lawrence F. , Funk Colin D.
Published: July 2002

Arachidonic acid metabolism is one of several mechanisms culminating in the production of an agonist for platelet activation and recruitment. Although the proaggregatory role of thromboxane A
2
, a product of the aspirin-inhibitable cyclooxygenase, is well established, relatively little is known regarding the biological importance of arachidonic acid metabolism via the 12-lipoxygenase (P-12LO) pathway to 12-hydro(pero)xyeicosatetraenoic acid. We observed that platelets obtained from mice in which the P-12LO gene has been disrupted by gene targeting (P-12LO
−/−
) exhibit a selective hypersensitivity to ADP, manifested as a marked increase in slope and percent aggregation in
ex vivo
assays and increased mortality in an ADP-induced mouse model of thromboembolism. The hyperresponsiveness to ADP is independent of dense granule release, cyclooxygenase-derived eicosanoid synthesis, and protein kinase C activity. The addition of 12-hydroxyeicosatetraenoic acid to P-12LO
−/−
platelet-rich plasma rescues the hyperresponsive phenotype resulting in a diminished ADP-induced aggregation profile. The enhanced ADP sensitivity of P-12LO
−/−
mice appears to reveal a mechanism by which a product of the P-12LO pathway suppresses platelet activation by ADP.
The trihelix DNA-binding motif in higher plants is not restricted to the transcription factors GT-1 and GT-2
Authors: Smalle Jan , Kurepa Jasmina , Haegman Mira , Gielen Jan , Van Montagu Marc , Straeten Dominique Van Der
Published: July 2002

GT-2 is a plant transcriptional activator that contains two separate, but similar, trihelix DNA-binding domains. GT-1 is similar to GT-2, although it contains only one of such domains. cDNAs that encode GT-2 were isolated from rice (
OS-GT2
) and
Arabidopsis
(
AT-GT2
). Evidence is presented for the existence of an
Arabidopsis
gene family that is structurally related to
AT-GT2
. Two members of this
GT2
-like family,
AT-GTL1
and
AT-GTL2
, have been isolated and characterized. Their sequences suggest that they evolved by a recent gene duplication event. Both
AT-GT2
and
AT-GTL
genes contain an intron in the amino-terminal trihelix motif, indicating that this DNA-binding domain resulted from exon shuffling. RNA gel blot analysis using
AT-GTL1
as a probe revealed four transcripts in the aerial part of the plant. All mRNA levels were significantly higher in siliques, suggesting that this gene family may function in fruit and/or seed development. To date, DNA-binding proteins characterized by the trihelix motif have been described only in plants, and may therefore be involved in plant-specific processes. Our results show that in
Arabidopsis thaliana
, the trihelix motif is not restricted to the GT-1 and GT-2 DNA-binding proteins.
Escherichia coli
rho factor induces release of yeast RNA polymerase II but not polymerase I or III
Authors: Lang Walter H. , Platt Terry , Reeder Ronald H.
Published: July 2002

Purified RNA polymerase II (pol II) from the yeast
Saccharomyces cerevisiae
pauses without releasing at many locations during
in vitro
transcription. Pausing can be induced by intrinsic DNA sequence as well as by specific DNA bound proteins such as the RNA pol I termination factor, Reb1p, or
lac
repressor. Addition of rho termination factor from
E. coli
induces RNA pol II to release at all of these pause sites. Rho-induced release of pol II requires both a rho binding site in the transcript upstream of the pause sites as well as hydrolysis of ATP. In contrast, rho factor has no effect on either pausing or release by RNA pol I or III. When combined with previous observations, these results suggest that RNA pol II may terminate by a mechanism closely related to the rho-dependent mechanism of prokaryotes. In contrast, pol I and III appear to utilize a mechanism more related to the rho-independent terminators of prokaryotes.
Formation of heteromeric gap junction channels by connexins 40 and 43 in vascular smooth muscle cells
Authors: He Ding Sheng , Jiang Jean X. , Taffet Steven M. , Burt Janis M.
Published: July 2002

Connexin (Cx) 43 and Cx40 are coexpressed in several tissues, including cardiac atrial and ventricular myocytes and vascular smooth muscle. It has been shown that these Cxs form homomeric/homotypic channels with distinct permeability and gating properties but do not form functional homomeric/heterotypic channels. If these Cxs were to form heteromeric channels, they could display functional properties not well predicted by the homomeric forms. We assessed this possibility by using A7r5 cells, an embryonic rat aortic smooth muscle cell line that coexpresses Cxs 43 and 40. Connexons (hemichannels), which were isolated from these cells by density centrifugation and immunoprecipitated with antibody against Cx43, contained Cx40. Similarly, antibody against Cx40 coimmunoprecipitated Cx43 from the same connexon fraction but only Cx40 from Cx (monomer) fractions. These results indicate that heteromeric connexons are formed by these Cxs in the A7r5 cells. The gap junction channels formed in the A7r5 cells display many unitary conductances distinct from homomeric/homotypic Cx43 or Cx40 channels. Voltage-dependent gating parameters in the A7r5 cells are also quite variable compared with cells that express only Cx40 or Cx43. These data indicate that Cxs 43 and 40 form functional heteromeric channels with unique gating and conductance properties.
Tumor necrosis factor α is a key component in the obesity-linked elevation of plasminogen activator inhibitor 1
Authors: Samad Fahumiya , Uysal K. Teoman , Wiesbrock Sarah M. , Pandey Manjula , Hotamisligil Gökhan S. , Loskutoff David J.
Published: July 2002

Obesity is associated with a cluster of abnormalities, including hypertension, insulin resistance, hyperinsulinemia, and elevated levels of both plasminogen activator inhibitor 1 (PAI-1) and transforming growth factor β (TGF-β). Although these changes may increase the risk for accelerated atherosclerosis and fatal myocardial infarction, the underlying molecular mechanisms remain to be defined. Although tumor necrosis factor α (TNF-α) has been implicated in the insulin resistance associated with obesity, its role in other disorders of obesity is largely unknown. In this report, we show that in obese (ob/ob) mice, neutralization of TNF-α or deletion of both TNF receptors (TNFRs) results in significantly reduced levels of plasma PAI-1 antigen, plasma insulin, and adipose tissue PAI-1 and TGF-β mRNAs. Studies in which exogenous TNF-α was infused into lean mice lacking individual TNFRs indicate that TNF-α signaling of PAI-1 in adipose tissue can be mediated by either the p55 or the p75 TNFR. However, TNF-α signaling of TGF-β mRNA expression in adipose tissue is mediated exclusively via the p55 TNFR. Our results suggest that TNF-α is a common link between the insulin resistance and elevated PAI-1 and TGF-β in obesity. The chronic elevation of TNF-α in obesity thus may directly promote the development of the complex cardiovascular risk profile associated with this condition.
VanX, a bacterial
d
-alanyl-
d
-alanine dipeptidase: Resistance, immunity, or survival function?
Authors: Lessard Ivan A. D. , Walsh Christopher T.
Published: July 2002

The zinc-containing
d
-alanyl-
d
-alanine
(
d
-Ala-
d
-Ala) dipeptidase VanX has been
detected in both Gram-positive and Gram-negative bacteria, where it
appears to have adapted to at least three distinct physiological roles.
In pathogenic vancomycin-resistant enterococci,
vanX
is
part of a five-gene cluster that is switched on to reprogram cell-wall
biosynthesis to produce peptidoglycan chain precursors terminating in
d
-alanyl-
d
-lactate
(
d
-Ala-
d
-lactate) rather than
d
-Ala-
d
-Ala. The modified peptidoglycan
exhibits a 1,000-fold decrease in affinity for vancomycin, accounting
for the observed phenotypic resistance. In the glycopeptide antibiotic
producers
Streptomyces toyocaensis
and
Amylocatopsis orientalis
, a
vanHAX
operon
may have coevolved with antibiotic biosynthesis genes to provide
immunity by reprogramming cell-wall termini to
d
-Ala-
d
-lactate as antibiotic biosynthesis is
initiated. In the Gram-negative bacterium
Escherichia
coli
, which is never challenged by the glycopeptide antibiotics
because they cannot penetrate the outer membrane permeability barrier,
the
vanX
homologue (
ddpX
) is
cotranscribed with a putative dipeptide transport system
(
ddpABCDF
) in stationary phase by the transcription
factor RpoS (σ
s
). The combined action of DdpX and the
permease would permit hydrolysis of
d
-Ala-
d
-Ala
transported back into the cytoplasm from the periplasm as cell-wall
crosslinks are refashioned. The
d
-Ala product could then be
oxidized as an energy source for cell survival under starvation
conditions.
CD48-deficient mice have a pronounced defect in CD4+T cell activation
Authors: González-Cabrero Jesús , Wise Catherine J. , Latchman Yvette , Freeman Gordon J. , Sharpe Arlene H. , Reiser Hans
Published: July 2002

We have generated mice deficient in the expression of the lymphocyte cell surface antigen CD48 (Blast-1, BCM1, sgp-60) by gene targeting in embryonic stem cells. Mice homozygous for the CD48 mutation (CD48−/−mice) are severely impaired in CD4+T cell activation. Proliferative responses to mitogens, anti-CD3 mAb, and alloantigen are all reduced. Experiments in which T cells and antigen-presenting cells from either wild-type or CD48−/−mice were cocultured reveal that CD48 is important on both T cells and antigen-presenting cells. The most dramatic impairment was observed in experiments in which highly purified T cells were stimulated through the T cell receptor in the presence of the phorbol ester, phorbol 12-myristate 13-acetate. The results of these experiments raise the possibility that CD48 plays a role in signaling through the T cell receptor.
Thyroid hormone receptor-associated proteins and general positive cofactors mediate thyroid hormone receptor function in the absence of the TATA box-binding protein-associated factors of TFIID
Authors: Fondell Joseph D. , Guermah Mohamed , Malik Sohail , Roeder Robert G.
Published: July 2002

Coactivators previously implicated in ligand-dependent activation functions by thyroid hormone receptor (TR) include p300 and CREB-binding protein (CBP), the steroid receptor coactivator-1 (SRC-1)-related family of proteins, and the multicomponent TR-associated protein (TRAP) complex. Here we show that two positive cofactors (PC2 and PC4) derived from the upstream stimulatory activity (USA) cofactor fraction act synergistically to mediate thyroid hormone (T
3
)-dependent activation either by TR or by a TR-TRAP complex in an
in vitro
system reconstituted with purified factors and DNA templates. Significantly, the TRAP-mediated enhancement of activation by TR does not require the TATA box-binding protein-associated factors of TFIID. Furthermore, neither the pleiotropic coactivators CBP and p300 nor members of the SRC-1 family were detected in either the TR-TRAP complex or the other components of the
in vitro
assay system. These results show that activation by TR at the level of naked DNA templates is enhanced by cooperative functions of the TRAP coactivators and the general coactivators PC2 and PC4, and they further indicate a potential functional redundancy between TRAPs and TATA box-binding protein-associated factors in TFIID. In conjunction with earlier studies on other nuclear receptor-interacting cofactors, the present study also suggests a multistep pathway, involving distinct sets of cofactors, for activation of hormone responsive genes.
Rational design of a scytalone dehydratase-like enzyme using a structurally homologous protein scaffold
Authors: Nixon Andrew E. , Firestine Steven M. , Salinas Frank G. , Benkovic Stephen J.
Published: July 2002

The generation of enzymes to catalyze specific reactions is one of the more challenging problems facing protein engineers. Structural similarities between the enzyme scytalone dehydratase with nuclear transport factor 2 (NTF2) suggested the potential for NTF2 to be re-engineered into a scytalone dehydratase-like enzyme. We introduced four key catalytic residues into NTF2 to create a scytalone dehydratase-like active site. A C-terminal helix found in scytalone dehydratase but absent in NTF2 also was added. Mutant NTF2 proteins were tested for catalytic activity by using a spectroscopic assay. One of the engineered enzymes exhibited catalytic activity with minimal
k
cat
and
K
m
values of 0.125 min
−1
and 800 μM, respectively. This level of catalytic activity represents minimally a 150-fold improvement in activity over the background rate for substrate dehydration and a dramatic step forward from the catalytically inert parent NTF2. This work represents one of the few examples of converting a protein scaffold into an enzyme, outside those arising from the induction of catalytic activity into antibodies.
Disruption of estrogen signaling does not prevent progesterone action in the estrogen receptor α knockout mouse uterus
Authors: Curtis Sylvia W. , Clark James , Myers Page , Korach Kenneth S.
Published: July 2002

Estrogen is known to increase progesterone receptor (PR) levels in the wild-type mouse uterus, and this estrogen induction was thought to be important for progesterone action through the PR. The estrogen receptor α knockout (ERKO) mouse uterus was observed to express PR mRNA that cannot be induced by estrogen. Progesterone action was characterized to determine whether it was diminished in ERKO mice. The PR protein is present in the ERKO uterus at 60% of the level measured in a wild-type uterus. The PR-A and PR-B isoforms are both detected on Western blot, and the ratio of isoforms is the same in both genotypes. Although the level of PR is reduced in the ERKO uterus, the receptor level is sufficient to induce genomic responses, since both calcitonin and amphiregulin mRNAs were increased after progesterone treatment. Finally, the ERKO uterus can be induced to undergo a progesterone-dependent decidual response. Surprisingly, the decidual response is estrogen independent in the ERKO, although it remains estrogen dependent in a wild type. These results indicate that estrogen receptor α modulation of PR levels is not necessary for expression of the PR or genomic and physiologic responses to progesterone in the ERKO uterus.
Calcium influx is required for endocytotic membrane retrieval
Authors: Vogel Steven S. , Smith Robert M. , Baibakov Boris , Ikebuchi Yoshihide , Lambert Nevin A.
Published: July 2002

Cells use endocytotic membrane retrieval to compensate for excess surface membrane after exocytosis. Retrieval is thought to be calcium-dependent, but the source of this calcium is not known. We found that, in sea urchin eggs, endocytotic membrane retrieval required extracellular calcium. Inhibitors of P-type calcium channels—cadmium, ω-conotoxin MVIIC, ω-agatoxin IVA, and ω-agatoxin TK—blocked membrane retrieval; selective inhibitors of N-type and L-type channels did not. Treatment with calcium ionophores overcame agatoxin inhibition in a calcium-dependent manner. Cadmium blocked membrane retrieval when applied during the first 5 minutes after fertilization, the period when the membrane potential is depolarized. We conclude that calcium influx through ω-agatoxin-sensitive channels plays a key role in signaling for endocytotic membrane retrieval.
Asynchronous oscillations of two zebrafish CLOCK partners reveal differential clock control and function
Authors: Cermakian Nicolas , Whitmore David , Foulkes Nicholas S. , Sassone-Corsi Paolo
Published: July 2002

Most clock genes encode transcription factors that interact to elicit cooperative control of clock function. Using a two-hybrid system approach, we have isolated two different partners of zebrafish (zf) CLOCK, which are similar to the mammalian BMAL1 (brain and muscle arylhydrocarbon receptor nuclear translocator-like protein 1). The two homologs, zfBMAL1 and zfBMAL2, contain conserved basic helix–loop–helix-PAS (Period-Arylhydrocarbon receptor-Singleminded) domains but diverge in the carboxyl termini, thus bearing different transcriptional activation potential. As for
zfClock
, the expression of both
zfBmal
s oscillates in most tissues in the animal. However, in many tissues, the peak, levels, and kinetics of expression are different between the two genes and for the same gene from tissue to tissue. These results support the existence of independent peripheral oscillators and suggest that zfBMAL1 and zfBMAL2 may exert distinct circadian functions, interacting differentially with zfCLOCK at various times in different tissues. Our findings also indicate that multiple controls may be exerted by the central clock and/or that peripheral oscillators can differentially interpret central clock signals.
Prolonging the half-life of human interferon-α2 in circulation: Design, preparation, and analysis of (2-sulfo-9-fluorenylmethoxycarbonyl)7- interferon-α2
Authors: Shechter Yoram , Preciado-Patt Liana , Schreiber Gideon , Fridkin Mati
Published: July 2002

Polypeptide drugs are generally short-lived species in circulation. In this study, we have covalently linked seven moieties of 2-sulfo-9-fluorenylmethoxycarbonyl (FMS) to the amino groups of human interferon-α2. The derivative thus obtained (FMS7–IFN-α2) has ≈4% the biological potency and 33 ± 4% the receptor binding capacity of the native cytokine. Upon incubation, FMS7–IFN-α2 undergoes time-dependent spontaneous hydrolysis, generating active interferon witht1/2values of 24 ± 2 h at pH 8.5 and 98 ± 10 h at pH 7.4. When native IFN-α2 is intravenously administered to mice, circulating antiviral activity is maintained for a short duration and then declines witht1/2= 4 ± 0.5 h, reaching undetectable values at ≈18 h after administration. With intravenously administered FMS7–IFN-α2, there is a lag period of 2 h, followed by a progressive elevation in circulating antiviral-active protein, which peaked at 20 h and declined witht1/2= 35 ± 4 h. FMS7–IFN-α2 is resistant to α-chymotrypsin digest and to proteolytic inactivation by human serum proteasesin vitro. We have thus introduced here an inactive IFN-α2 derivative, which is resistant toin situinactivation and has the capability of slowly reverting to the native active protein at physiological conditionsin vivoandin vitro. Having these attributes, FMS7–IFN-α2 maintains prolonged circulating antiviral activity in mice, exceeding 7–8 times the activity of intravenously administered native cytokine.
Bothrops asper snake venom and its metalloproteinase BaP–1 activate the complement system. Role in leucocyte recruitment
Authors: Farsky Sandra H. P. , Gonçalves Luís Roberto C. , Gutiérrez José M. , Correa Adriana P. , Rucavado Alexandra , Gasque Philippe , Tambourgi Denise V.
Published: July 2002

The venom of the snake Bothrops asper, the most important poisonous snake in Central America, evokes an inflammatory response, the mechanisms of which are not well characterized. The objectives of this study were to investigate whether B. asper venom and its purified toxins – phospholipases and
metalloproteinase – activate the complement system and the contribution of the effect on leucocyte recruitment.
In vitro chemotaxis assays were performed using Boyden′s chamber model to investigate the ability of serum incubated with venom and its purified toxins to induce neutrophil migration. The complement consumption by the venom was evaluated using an in vitro haemolytic assay. The importance of complement activation by the venom on neutrophil migration was investigated in vivo by injecting the venom into the peritoneal cavity of C5‐deficient mice. Data obtained demonstrated that serum incubated with crude venom and its purified metalloproteinase BaP–1 are able to induce rat neutrophil chemotaxis, probably mediated by agent(s) derived from the complement system. This hypothesis was corroborated by the capacity of the venom to activate this system in vitro. The involvement of C5a in neutrophil chemotaxis induced by venom‐activated serum was demonstrated by abolishing migration when neutrophils were pre‐incubated with antirat C5a receptor antibody. The relevance of the complement system in in vivo leucocyte mobilization was further demonstrated by the drastic decrease of this response in C5‐deficient mice. Pre‐incubation of serum with the soluble human recombinant complement receptor type 1 (sCR 1) did not prevent the response induced by the venom, but abolished the migration evoked by metalloproteinase‐activated serum. These data show the role of the complement system in bothropic envenomation and the participation of metalloproteinase in the effect. Also, they suggest that the venom may contain other component(s) which can cause direct activation of C5a.
Augmentation of NKT and NK cell‐mediated cytotoxicity by peptidoglycan monomer linked with zinc
Authors: Mrakovčić-Šutić Ines , Radošević-Stašić Biserka , Šimin Marija , Muhvić Damir , Rukavina Daniel
Published: July 2002

Background: Peptidoglycan monomer (PGM), which was originally prepared by biosynthesis from culture fluids of penicillin‐treated Brevibacterium divaricatum, is an immunostimulator, the activities of which might be improved by addition of zinc (Zn) to the basic molecule.

Methods: To test the possible cytotoxic effects of this new analogue, we analyzed the ability of PGM‐Zn and PGM to change the phenotypic profile of hepatic and splenic mononuclear lymphatic cells and to affect the growth of malignant T‐cell line YAC‐1 and syngeneic thymocytes.

Results: Pretreatment of C57BL/6 mice primarily with PGM‐Zn over 6 days (10/mg/kg intraperitoneally) significantly enhanced the proportions of NK1.1high+, CD4-CD8-, CD69+, and CD3intermediate/NK1.1
+/IL2R‐β+ (NKT) cells in the liver, and major histocompatibility complex class II+, CD69+, and CD8+ cells in the spleen. Both types of cells were highly cytotoxic against YAC‐1 and syngeneic thymocytes, increasing the destruction of YAC‐1 by 70% on addition of hepatic cells and by 30% on addition of splenic cells. Destruction of thymocytes increased by 10 and 50%,
respectively.

Conclusion: The results point to PGM‐Zn as a potent cytotoxicity‐inducing agent, which also generates autoreactive NKT cells.
(1→3)‐β‐D‐glucan in patients with pulmonary aspergilloma
Authors: Yuasa Kazumi , Goto Hajime
Published: July 2002

To elucidate the role of (1→3)‐β‐D‐glucan in pulmonary aspergilloma, serum concentrations of (1→3)‐β‐D‐glucan were measured repeatedly for as long as 10 months in eight patients. In four patients with inactive disease, concentrations of (1→3)‐β‐D‐glucan were in the normal range.The concentrations of (1→3)‐β‐D‐glucan increased in two patients, although the disease was inactive. This increase might show the earliest stage of the invasive process of the disease. In two other patients with active disease, (1→3)‐β‐D‐glucan increased. Other parameters, such as galactomannan, immunodiffusion and a radio‐allergosorbent test, as well as inflammatory m arkers such as C‐reactive protein and the leukocyte count, did not show any consistent tendency in regard to the activity of the disease. Thus, a (1→3)‐β‐D‐glucan assay may add valuable data for evaluating the disease activity and understanding the disease process of pulmonary aspergilloma.
Pentoxifylline inhibits the fibrogenic activity of pleural effusions and transforming growth factor‐β
Authors: Entzian P. , Schlaak M. , Seitzer U. , Acil Y. , Ernst M. , Zabel P.
Published: July 2002

Physiopathology of organ fibrosis is far from being completely understood, and the efficacy of the available therapeutic strategies is disappointing. We chose pleural disease for further studies and addressed the questions of which cytokines are relevant in pleural fibrosis and which drugs might interrupt its development. We screened pleural effusions for mediators thought to interfere with fibrogenesis (transforming growth factor‐β (TGF‐β), tumour necrosis factor α (TNFα), soluble TNF‐receptor p55 (sTNF‐R)) and correlated the results with patient clinical outcome in terms of extent of pleural thickenings. We found pleural thickenings correlated with TGF‐β (p < 0.005)
whereas no correlations could be observed with TNFα and sTNF‐R. Further, we were interested in finding out how TGF‐β effects on fibroblast growth could be modulated. We found that pentoxifylline is able to inhibit both fibroblast proliferation and collagen synthesis independently of the stimulus. We conclude that, judging from in vitro studies, pentoxifylline might offer a new approach in the therapy of pleural as well as pulmonary fibrosis.
Interleukin‐6 and interleukin‐6 receptor secretion by chronic lymphatic leukaemia and normal B lymphocytes: effect of PMA and PWM
Authors: Drucker I. , Klajman A. , Revel M. , Manor Y. , Ben-Efraim S. , Novick D.
Published: July 2002

Interleukin‐6 (IL‐6) and soluble interleukin‐6 receptor (sIL‐6R) were detected in supernatants of cultures of B chronic lymphatic leukaemia (CLL) lymphocytes. Phorbol‐12‐myristate 13 acetate (PMA) caused a decrease in the levels of IL‐6 in 14 out of 16 cultures and an increase in levels of sIL6R in all 15 cases. The effect of pokeweed mitogen (PWM) was variable and not significant. The levels of IL‐6 were below the detection limit (60 pg/ml) in sera of 13 CLL patients whereas sIL‐6R was detected (13 ng/ml to 97 ng/ml) in the 13 sera. IL6 was not detected in cultures of unstimulated or stimulated with PMA or PWM normal human B cells. Levels of sIL‐6R were minimal in cultures of normal B lymphocytes and were increased in PMA stimulated cultures. The results are consistent with the view that B‐CLL cells produce spontaneously IL‐6 which could act in an autocrine fashion to cause shedding of surface IL‐6R and account for the correlation found between serum levels of sIL‐6R and B‐CLL lymphocyte numbers. The fall in levels of IL‐6 in PMA stimulated CLL cultures might express masking or degradation of IL‐6 after combination with the receptor.
Nitric oxide decreases intestinal haemorrhagic lesions in rat anaphylaxis independently of mast cell activation
Authors: Tavares J. Carvalho , Moreno A. , Crespo M. Sánchez
Published: July 2002

The purpose of this study is to assess the role of nitric oxide (NO) in the intestinal lesions of passive anaphylaxis, since this experimental model resembles necrotizing enterocolitis. Sprague‐Dawley rats were sensitized with IgE anti‐dinitrophenol monoclonal antibody. Extravasation of protein‐rich plasma and haemorrhagia were measured in the small intestine. Plasma histamine was measured to assess mast cell activation. The effect of exogenous NO on the lesions was assessed by using two structurally unrelated NO‐donors: sodium nitroprusside and S‐nitroso‐Nacetyl‐penicillamine (SNAP). An increased basal production of NO was observed in cells taken after anaphylaxis, associated with a reduced response to platelet‐activating factor, interleukin 1beta, and IgE/DNP‐bovine serum albumin complexes. The response to bacterial lipopolysaccharide and dibutyryl cyclic adenosine monophosphate (AMP) was enhanced 24 h after challenge, but at earlier times was not significantly different from that observed in controls. Treatment with either sodium nitroprusside or SNAP produced a significant reduction of the haemorrhagic lesions, which are a hallmark of rat anaphylaxis. The extravasation of protein‐rich plasma was not influenced by NO‐donors. The increase of plasma histamine elicited by the anaphylactic challenge was not influenced by SNAP treatment. NO‐donors protect intestinal haemorrhagic lesions of rat anaphylaxis by a mechanism apparently independent of mast cell histamine release.
Functional roles of Phe12 of deacetyl‐thymosin β4 in the impaired blastogenic response of uraemic T‐lymphocytes
Authors: Abiko T. , Sekino H.
Published: July 2002

Phe12 of deacetyl‐thymosin β4 is one of the structural essentials for restorative effect on the impaired blastogenic response of uraemic T‐lymphocytes. In order to evaluate the functional roles of this phenyl group in the restorative effect on impaired T‐lymphocytes, two analogues, [1‐ Nal12]deacetyl‐thymosin β4 and [Cha12]deacetyl4 thymosin β4, were synthesized by a solid‐phase method and evaluated for restorative effect on the impaired blastogenic response of uraemic T‐lymphocytes. The results indicated that
[1‐Nal12]deacetyl‐thymosin β4 which had a bulky naphthyl ring showed a stronger restorative effect than that of deacetyl‐thymosin β4, but it was slightly weaker than that of [Phe(4F)12]deacetyl‐thymosin β4. However, [Cha12]deacetyl‐thymosin β4 showed no restorative effect on the impaired blastogenic response of uraemic T‐lymphocytes.
Immunomodulating activities of a natural α1 ‐acid glycoprotein and its carbohydrate chains attached to the Protein‐Free Polymer
Authors: Pukhalsky A. L. , Shyian S. D. , Kalashnikova E. A. , Shmarina G. V. , Pukhalskaya D. A. , Bovin N. V.
Published: July 2002

Immunomodulating effects of a neoglycoconjugate created on the basis of α1 ‐acid glycoprotein (AGP) carbohydrate chains and synthetic protein‐free carrier have been investigated. It was demonstrated that this pseudo‐AGP suppressed PHA‐ or anti‐CD3 antibody‐induced lymphocyte proliferation in a dosedependent manner. Pseudo‐AGP revealed a similar antiproliferative effect as the natural AGP samples. Stimulation of the LPS‐induced proinflammatory cytokine production by m ononuclear cells treated with both natural and pseudo‐AGP has been also demonstrated. These data show that carbohydrate chains of AGP play a crucial role in the studied biological effect realization.
Inflammation and CFTR: Might Neutrophils be the Key in Cystic Fibrosis?
Authors: Witko-Sarsat V. , Sermet-Gaudelus I. , Lenoir G. , Descamps-Latscha B.
Published: July 2002

The aim of this hypothesis is to provide new insights into the still unclear mechanisms governing airway inflammation in cystic fibrosis. Although the genetic basis of cystic fibrosis as well as the molecular structure of cystic fibrosis transmembrane regulator (CFTR), the mutated protein which causes the disease, have been well defined, a clear relationship between the genetic defect and the pulmonary pathophysiology, especially chronic infections and neutrophildominated airway inflammation has not been established. Cystic fibrosis is thus a unique pathological situation in that neutrophils can be depicted as both an antiinfectious and a proinflammatory cell. In cystic fibrosis there is an emerging picture of an imbalance between these two roles with both a reduction in the antiinfectious efficacy and an augmentation of the proinflammatory functions. Better knowledge of fundamental defects in neutrophil function in cystic fibrosis as well as a novel cellular function of CFTR, which will be reviewed, will allow identification of potentially new clinical targets and aid selective therapeutic action aimed at counteracting the lethal neutrophil‐induced airway inflammation. The rationale for colchicine therapy is a significant example of a drug which might act both at the molecular levels on CFTR expression in epithelial cells and on neutrophils to mediate antiinflammatory effects. Preliminary results are presented in this issue (Med Inflamm 1999; 8: 13‐15).
Differential Modulation of Annexin I Binding Sites on Monocytes and Neutrophils
Authors: Euzger H. S. , Flower R. J. , Goulding N. J. , Perretti M.
Published: July 2002

Specific binding sites for the anti‐inflammatory protein annexin I have been detected on the surface of human monocytes and polymorphonuclear leukocytes (PMN). These binding sites are proteinaceous in nature and are sensitive to cleavage by the proteolytic enzymes trypsin, collagenase, elastase and cathepsin G. When monocytes and PMN were isolated independently from peripheral blood, only the monocytes exhibited constitutive annexin I binding. However PMN acquired the capacity to bind annexin I following co‐culture with monocytes. PMN incubation with sodium azide, but not protease inhibitors, partially blocked this process. A similar increase in annexin I binding capacity was also detected in PMN following adhesion to endothelial monolayers. We propose that a juxtacrine activation rather than a cleavage‐mediated transfer is involved in this process. Removal of annexin I binding sites from monocytes with elastase rendered monocytes functionally insensitive to full length annexin I or to the annexin I‐derived pharmacophore, peptide Ac2‐26, assessed as suppression of the respiratory burst. These data indicate that the annexin I binding site on phagocytic cells may have an important function in the feedback control of the inflammatory response and their loss through cleavage could potentiate such responses.
An Overview of Controlled Studies of Adolescent Substance Abuse Treatment
Authors: Deas Deborah , Thomas Suzanne E.
Published: July 2002

Although several treatments for adolescents with substance use disorders are available, there are few well‐controlled studies in the extant literature that compare these treatments for efficacy. This paper provides an overview of controlled treatment studies for adolescents with substance use disorders. It focuses specifically on five main treatment modalities: family‐based and multi‐systemic interventions, behavioral therapy, cognitive behavioral therapy, pharmacotherapy, and twelve step approaches. Examples of adolescent‐specific standardized assessment instruments are also provided, the inclusion of which may improve future treatment comparison studies. While the results look especially promising for cognitive behavioral therapy and family‐based/multi‐systemic therapies for adolescents with SUDs, most of the relevant studies fail to utilize validated outcome measures, making it difficult to conclude that one treatment approach is more effective than another.
Impaired sleep during the post‐alcohol withdrawal period in alcoholic patients
Authors: Watanabe Kei‐ICHIRO , Ogihara‐Hashizume Aki , Kobayashi Yaweko , Mitsushio Hiroshi , Komiyama Tokutaro
Published: July 2002

Twenty‐seven patients who suffered from alcohol dependence syndrome and who were admitted to the Department of Psychiatry at NCNP were investigated for impaired sleep. The incidence of “insomnia night”, when a patient has trouble getting sufficient sleep even after taking additional drugs for insomnia, was used as the index for impaired sleep. “Insomnia night” was observed among 51.9% (14 of 27) of the patients during administration; 18.5% (nine of 27) experienced “insomnia night” only during the withdrawal period. The mean incidence was 3.5 nights of 10 nights for 0–9th day after abstinence and 2.0 nights of 10 nights for 10–19th day after abstinence; 33.3% (nine of 27) claimed “insomnia night” even after the withdrawal period and the highest incidence was the 160–169th day after abstinence, average frequency being 4.4 nights of 10 nights. Impaired sleep emerging after the withdrawal period neared the clinical course of Komiyama's protracted withdrawal symptoms, thus suggesting that it might be one of the protracted withdrawal symptoms.
Local Excision Without Radiation for High‐Grade Soft‐Tissue Sarcoma ofthe Extremity and Superficial Trunk
Authors: Weir Lorna M. , Vanbergeyk Anthony B. , Masri Bassam A. , Grafton Clive A. , Duncan Clive P. , Goddard Karen J. , Joe Howard A.
Published: July 2002

Purpose. Limb‐sparing surgery combined with radiation treatment
has become the accepted treatment for patients with high‐grade soft‐tissue sarcoma.
Adjuvant radiation was not routinely used at this institution for patients with clear margins
after surgery.This retrospective review analyses the outcome of this group of patients.

Patients and methods. Patients studied were referred from 1984 to 1995,
were over 16 years of age, were diagnosed with primary high‐grade soft‐tissue sarcoma
of the extremity or superficial trunk, had clear margins after excision and did not receive
radiation as a part of their initial treatment. A total of 46 patients were identified.

Results. At 5 years, the local control rate was 87%, disease‐specific
survival was 75% and overall survival was 68%. Of the 6 local recurrences, 3 were located
in the buttock (from a total of 7 patients with primary tumours of the buttock), 3 had a
primary size of ≥ 10 cm (from a total of 8 primary tumours of that size) and all were deep
tumours.

Discussion. Our data, and those from other reports, suggest that in
carefully selected patients appropriate surgery alone results in acceptable local control
and survival, and that the morbidity of radiation can be avoided.
Leiomyosarcoma as a Second Metachronous Malignant Neoplasm Following Colon Adenocarcinoma. A Case Report and Review of the Literature
Authors: Mavrodontidis A. , Zalavras Ch. , Skopelitou A. , Karavasilis V. , Briasoulis E.
Published: July 2002

Long‐term cancer survivors are at increased risk for the development of second primary malignancies. This is usually associated
with common genetic and etiologic factors and the treatment modality used for the primary cancer. In this paper we
describe the case of a patient who developed a leiomyosarcoma in his left arm 5 years after he had a colon adenocarcinoma
resected. Both primary tumours were treated successfully with surgical resection alone. The literature regarding second
primary neoplasms, specifically focused on sarcomas, is briefly reviewed.
EORTC Group Phase II Study of Oral Etoposide for Pretreated Soft Tissue Sarcoma
Authors: Keizer H. Jan , Crowther Derek , Nielsen Ole Steen , Oosterom Allan T. Van , Muguiro Javier Hornedo , Pottelberghe Christine Van , Somers Renier , Tursz Thomas
Published: July 2002

Purpose. This study investigates the efficacy and toxicity of daily oral etoposide in
chemotherapy for non‐heavily pretreated advanced and metastatic soft tissue sarcoma (STS).

Subjects. Twenty‐seven patients with progressive and measurable disease were treated. Median age was 53 years
(range 20–71 years) and performance status WHO 0 or 1. Histologies included mainly leiomyosarcoma (8),
malignant fibrous histiocytoma (4), rhabdomyosarcoma (4), liposarcoma (2) and synovial sarcoma (2). Fifteen
patients had received prior radiotherapy, of whom three included sites with haematopoiesis. All patients had received
prior chemotherapy, including adjuvant therapy (7) and mostly consisted of one two‐drug schedule (ifosfamide and
doxorubicin) or two single‐drug regimens.

Methods. Chemotherapy consisted of etoposide (VP16‐213), 50 mg m−2
day−1 × 21 q 4 weeks. Blood cell counts
were done weekly. Dose reductions and a maximum delay of 2 weeks was allowed depending on cell counts during
treatment and at the start of a new 4‐week treatment cycle.

Results. No objective response was observed. Progressive disease was observed after two treatment cycles in 17/27
patients (68%) and after three cycles in 22/27 patients (81%). The other patients received three to five cycles.
Twenty‐four patients went off study due to progressive disease. Grade 3 and 4 neutropenia was observed in eight
and one patients, respectively. Thrombocytopenia grade 3 was seen in two patients. Non‐haematological toxicity
grade 3 (nausea, diarrhoea or alopecia) was observed in three patients, and grade 4 (dyspnea, hypotension or
haemorrhage) in three patients.

Discussion. No objective response was obtained. Oral etoposide at a dose of 50 mg m−2
day−1 × 21 q 4 weeks is
inactive in chemotherapy of pretreated STS. Disease progression occurred within three cycles in the majority (81%)
of patients. Toxicity of this regimen in non‐heavily pretreated patients is low.
Clinical and Histopathological Prognostic Factors in Chondrosarcomas
Authors: Daugaard Søren , Myhre-Jensen Olaf , Schiødt Torben , Jurik Anne G. , Keller Johnny , Mouridsen Henning T. , Lund Bjarne
Published: July 2002

Purpose. In an attempt to identify clinical and histopathological factors of prognostic
importance in chondrosarcomas, 115 cases of malignant and borderline chondromatous tumours were reviewed.

Patients/methods. Histopathological features tested for prognostic information as well
as reproducibility included cellularity, nuclear pleomorphism, multinucleated cells, mitotic activity and grade.
Eleven patients had a biopsy only, and a short survival (median 2.0 years); these were excluded from further analysis.
The remaining 104 patients who had received intended curative treatment had a median survival of 14.7 years.

Results. In univariate analysis, tumour size, extra‐compartmental growth, surgical margin and sex
were significantly correlated to recurrence‐free survival (RFS); sex was marginally significant while age, site and pathological
parameters were not significant. Overall survival (OAS) was likewise found to be independent of pathological features as well as site,
size and surgical margin; but age, sex and extra‐compartmental growth were statistically significant. However, when the
same parameters were entered into a stepwise Cox (multivariate) analysis, only surgical margin, cellularity and pleomorphism
were significantly related to RFS; margin, grade, pleomorphism and age to OAS. Overall inter‐observer agreement
on grade was relatively low: 0.54, with a Kappa value of 0.32. It was not better for the other histological parameters, with
the exception of the mitotic count. However, acceptable values were achieved when the material was divided into
low‐grade (grade I and below) vs high‐grade (grade II and III) lesions: overall agreement 0.79, Kappa 0.56.

Discussion. Although the grading of chondrosarcomas is in need of improvement, its replacement by
semiquantitative evaluation of individual histopathological parameters as performed in this study offers no advantage.
Among the clinical parameters, only the adequacy of the surgical treatment and the patient′s age appear to be important.
Fine‐Needle Aspiration Cytology of Epithelioid Leiomyoblastoma
Authors: Thompson Karen Sue , Jensen Joanne , Reyes Cesar V.
Published: July 2002

Purpose. Epithelioid leiomyoblastomas comprise the majority of gastric sarcomas
and are uncommon in other parts of the gastrointestinal tract. Diagnosis of this lesion
by fine‐needle aspiration cytology has been occasionally described in the literature.
Two additional cases are herein reported.

Subjects . A 66‐year old male with an omental mass and a 47‐year old male with a
perihepatic tumor.

Results and Discussion. Cytologic materials in both cases showed predominantly
round or epithelioid cells, along with polygonal to spindle cells, occuring singly and in
clusters, with oval to spindle‐shaped nuclei.The nuclei were monotonous, usually banal,
and centrally‐located with only focal suggestion of pleomorphism and rare mitosis.
Eosinophilic cytoplasm was noted in most of the cells, some demonstrating vacuolation.
Electron microscopy supported a primitive smooth cell derivation of the neoplastic cells.

Conclusions. The cytomorphology of the tumors of the two cases reported here is not
adequately known. More cases need to be collected and studied.