bluuebunny/crossref_metadata_2025 huggingface dataset:
Proinflammatory Effects in Experimental Mesangial Proliferative Glomerulonephritis of the Immunosuppressive Agent SDZ RAD, a Rapamycin Derivative
Authors: Daniel Catherine , Ziswiler Ruth , Frey Brigitte , Pfister Marc , Marti Hans-Peter
Published: April 2003

Background/Aim: The new immunosuppressant SDZ RAD, a rapamycin derivative, inhibits growth factor driven cell proliferation. SDZ RAD designed for transplantation may also be a candidate agent to treat inflammatory kidney diseases. Therefore, we investigated the effects of SDZ RAD in two different animal models of glomerulonephritis, in anti- Thy1.1 nephritis and in acute puromycin aminonucleoside (PAN) nephrosis. Methods: Eighty-seven male Wistar rats were investigated. Anti-Thy1.1 nephritis: healthy rats (n = 9), SDZ RAD-treated healthy rats (n = 6), nephritic rats (n = 9), SDZ RAD placebo treated nephritic rats (n = 6), SDZ RAD-pretreated nephritic rats (n = 9), and early (n = 6) as well as delayed (n = 6) SDZ RAD-posttreated nephritic rats. PAN nephrosis: healthy rats (n = 6), SDZ RAD-treated healthy rats (n = 6), nephritic rats (n = 12), and SDZ RAD-pretreated nephritic rats (n = 12). In a separate study, 12 male Sprague-Dawley rats were analyzed in anti-Thy1.1 nephritis: healthy rats (n = 3), nephritic rats (n = 3) and pretreated nephritic rats (n = 6). SDZ RAD and SDZ RAD placebo were given at single doses of 2.5 mg/kg body weight per day by gavage. The experiments lasted until days +2 and +9 after induction of anti-Thy1.1 nephritis and until day +13 in the case of PAN nephrosis. Results: In anti-Thy1.1 nephritis, SDZ RAD demonstrated marked proinflammatory effects in a time-dependent manner, as reflected by severe focal damage to glomerular histology including inhibition of mesangial cell proliferation, reduction of creatinine clearance, and increase in plasma creatinine levels as well as proteinuria. Almost identical results were obtained in both rat strains. In contrary, SDZ RAD ameliorated significantly the development of PAN nephrosis. Animals pretreated by this agent showed a significant reduction of proteinuria and of glomerular invasion of monocytes/macrophages. Conclusion: Some caution is warranted for the use of SDZ RAD in inflammatory glomerular diseases, since it accentuated glomerular damage induced by anti-Thy1.1 antibodies.
Acoustic Correlates of Perceived versus Actual Sexual Orientation in Men’s Speech
Authors: Linville Sue Ellen
Published: April 2003

The purpose of this study was to explore possible differences in the speech patterns of gay and straight men, both as a function of perceived and actual sexual orientation. Tape recordings of monologue readings from 5 openly gay men and 4 straight men were played to 25 listeners for judgments of perceived sexual orientation. Monologues were analyzed in terms of /s/ duration, /s/ peak frequency, modal speaking fundamental frequency, speech rate, and long-term average speech spectra. Listeners correctly identified the sexual orientation ofthese speakers at a rate of 79.6%. Multiple regression analysis revealed that gay judgments were significantly associated with higher peak /s/ frequency values and longer /s/ duration values. Acoustic cues associated with perceived sexual orientation generally agreed with acoustic findings as a function of actual sexual orientation. While these results must be interpreted cautiously, findings suggest that members of the community of openly gay men demonstrate certain speech characteristics that are discernible to listeners.
The Influence of Sexual Swelling on the Behaviour of Captive Female Chimpanzees (Pan troglodytes)
Authors: Palagi Elisabetta , Lunardini Antonella
Published: April 2003

Zuckerman [1] suggested that during their fertile phase hamadryas females can influence the social organization of the whole group. Carpenter [2] later suggested that females of Macaca mulatta can achieve a higher social rank during the fertile phase. Recent studies confirm that this particular status can induce various behavioural changes, although these are relatively subtle. For example, female plains baboons do not achieve a higher social rank during the fertile phase, but at this time they are more active, spend more time travelling and sit for only periods of a few minutes. Furthermore, proximity between fertile females and males also increases in non-sexual contexts [3].
Anthropoid Origins and the Modern Symphysis
Authors: Ravosa Matthew J.
Published: April 2003

To highlight adaptive transformations in craniomandibular form during anthropoid origins, symphyseal character states and underlying masticatory loading regimes were investigated vis-à-vis shifts in diet and body size. A study of fossil anthropoids is possible because variation in symphyseal fusion is continuous and directly proportional to the amount of symphyseal stress and because such variation can be considered a series of discrete character states each with unique functional underpinnings. Using recent systematic renderings of Eocene and Oligocene taxa as a template with which to assess character evolution, this analysis indicates when, and in which clade(s), specific masticatory features became fixed and thus diagnostic. A general trend throughout early anthropoid evolution is for descendent taxa to be progressively larger than ancestral forms. Coupled with this pattern is the tendency for larger-bodied fossil anthropoids to have ingested tougher diets variably consisting of thick-coated, unripe fruits and/or leaves. Mastication of mechanically tougher foods entails greater repetitive loading of the mandible and requires relatively larger amounts of balancing-side muscle force, thus resulting in correspondingly greater symphyseal fusion due to elevated dorsoventral shear. With a single exception, these adaptive transformations characterize the evolutionary pathway leading both to parapithecines and a catarrhine:platyrrhine clade (crown anthropoids). While the ancestor of crown anthropoids would have possessed a body size, diet and masticatory adaptations similar to parapithecines, such a common suite of features evolved independently. Moreover, the evolution of an early-fusing symphysis and associated wishboning loading regime of catarrhines and platyrrhines is unique among all anthropoids. Lastly, the apparent lack of reversals in symphyseal fusion indicates the improbability of phylogenetic hypotheses in which a relationship is proposed between ‘ancestral’ taxa with a greater degree of symphyseal fusion and ‘descendent’ anthropoids with a lesser degree of ossification.
A Prisoner’s Dilemma Model of the Evolution of Paternal Care
Authors: Key Catherine , Aiello Leslie C.
Published: April 2003

The heavy energetic demands of gestation, lactation and rearing of offspring mean that studies of paternal care in primates usually focus on female reproductive effort. Here it is shown that both male and female reproductive effort must be considered in order to understand how paternal care evolved. This is done using the Prisoner’s Dilemma, best known as a model of reciprocal altruism. It is found that the relative cost of reproduction for males and females is crucially important in determining co-operative and competitive strategies. In particular, when male reproductive costs are less than female reproductive costs, males co-operate with females even when females do not reciprocate. This surprising behaviour, termed non-reciprocal altruism, is comparable with male investment in a female and her offspring.
Sicherheit und Akzeptanz der Selbstanwendung von Würfel- und Urethraringpessaren
Authors: Kölle D. , Kunczicky V. , Uhl-Steidl M. , Pontasch H.
Published: April 2003

Fragestellung: Mit Zugfäden ausgestattete flexible Würfel- und Ringpessare mit suburethraler Auftreibung wurden zur Therapie von Deszensus bzw. Stressinkontinenz vorgeschlagen. Die Studie untersuchte die Sicherheit der Selbstanwendung und die Akzeptanz der Therapie. Methoden: Achtzehn Frauen (Alter 36–85 Jahre), davon 9 voroperiert, versuchten die Pessartherapie (Würfel n = 6; Ring n = 12). Die Pessargrösse wurde klinisch bestimmt, das Pessar von den Frauen nachts selbst entfernt. Kontrollen erfolgten nach einer Woche und dann monatlich. Ergebnisse: Alle Frauen kamen zur Erstkontrolle. Zwölf von 18 waren sehr zufrieden, 5/18 Patientinnen wollten innerhalb von 2 Monaten die Operation, 3/18 lehnten das Pessar später ab, 1 Frau erschien nicht mehr zur Kontrolle und 9/18 verwendeten die Pessare weiter (mittlere Dauer 11 Monate). In den 107 Behandlungsmonaten wurden lediglich 2 milde Komplikationen ohne Interventionsbedarf beobachtet (milde Erosion, Kontaktblutung). Schlussfolgerungen: Würfel- und Urethraringpessare sind sicher, die Drop-out-Rate liegt jedoch bei etwa 50%. Es handelt sich um eine effiziente Therapiealternative für manche Frauen.
Konnatale Schädelimpression
Authors: Rieger-Fackeldey E. , Kloos S. , Winkler P.A. , Korell M. , Genzel-Boroviczény O. , Schulze A.
Published: April 2003

Ein reifes Neugeborenes einer III. Gravida, II. Para wurde nach unauffälliger Schwangerschaft bei pathologischem Kardiotokogramm, Polysystolie und mekoniumhaltigem Fruchtwasser per Sectio caesarea entwickelt. Der pH des Nabelarterienblutes war 7,03, als Apgar-Werte wurden 1/4/7 nach 1, 5 und 10 min vergeben. Das 3250 g schwere Kind bot eine 5 × 7 cm grosse, 1,5 cm tiefe, feste Impression links temporoparietal, ohne Ödeme oder Hämatome der Weichteile in diesem Bereich bei unauffälligem neurologischem Status. Im kranialen CT fand sich an dieser Stelle keine Frakturlinie, der darunterliegende Kortex wirkte etwas verdichtet. Über ein Bohrloch wurde der eingedrückte Teil der Parietalschuppe im Alter von 11 Tagen gehoben. Das postoperative CT zeigte keine Kontinuitätsunterbrechung der Kalotte. Das Kind konnte 8 Tage nach Operation klinisch unauffällig entlassen werden. Bei der Nachuntersuchung im Alter von 6 Monaten sahen wir ein altersgemäss entwickeltes Kind in gutem Allgemeinzustand. Wir diskutieren anhand dieses Falles die Ätiologie, Diagnostik und Therapie der konnatalen Impression des Schädels.
Experimentelle Forschung und klinische Anwendung der photodynamischen Therapie an der Krankenanstalt Rudolfstiftung
Authors: Wierrani Franz
Published: April 2003

Wir berichten über experimentelle Forschung und klinische Anwendung der photodynamischen Therapie an der Krankenanstalt Rudolfstiftung. An Staphylococcus aureus (Wild type) konnten wir eine hohe Dunkeltoxizität von Mesotetrahydroxyphenylchlorin nachweisen. Hämatoporphyrinderivat zeigte lediglich Toxizität bei Beleuchtung; die photodynamische Aktivität des Farbstoffgemisches erbrachte eine additive Wirkung. Die Entwicklung einer Antibiotikaresistenz von S. aureus nach der Behandlung mit Hämatoporphyrinderivat war lediglich gegenüber Erythromycin zu beobachten. Eine zusätzliche intraoperative photodynamische Therapie bei der chirurgischen Behandlung von Rezidiven gynäkologischer Karzinome scheint eine gute adjuvante Behandlungsmöglichkeit zu sein. Sie wirkt gewebsspezifisch und stellt eine sehr einfache Intervention dar. Die psychische Belastung von Tumorpatienten wird nicht verstärkt. In einer weiteren klinischen Studie haben wir oberflächliche zervikale Dysplasien der Grade I und II und Papillomavirusinfektionen mit 5-Aminolävulinsäure oder mit Mesotetrahydroxyphenylchlorin behandelt. Bei 33 von 38 (86,8%) Patientinnen konnte die Dysplasie zur Abheilung gebracht werden; die Eradikation von Papillomavirusinfektionen konnte in 80% der Fälle erfolgreich durchgeführt werden.
No Correlation between A(–1438)G Polymorphism in 5-HT2A Receptor Gene Promoter and the Density of Frontal Cortical 5-HT2A Receptors in Schizophrenia
Authors: Kouzmenko Alexander P. , Scaffidi Antonietta , Pereira AvrilM. , Hayes Wendy L. , Copolov David L. , Dean Brian
Published: April 2003

The A(–1438)G promoter polymorphism of the 5-hydroxytryptamine 2a receptor (5-HT2AR) gene and its influence on the cortical density of 5-HT2AR was studied using brain tissue donated at autopsy from 58 schizophrenic and 64 non-schizophrenic subjects. A linkage between genotypes for the A(–1438)G and a T102C polymorphic site identified in a previous study was observed. Our data suggest no association of the A(–1438)G polymorphism with schizophrenia and no effect of the promoter genotype upon 5-HT2AR densities in either the schizophrenic or non-schizophrenic groups.
Allelic Frequencies of Six (CA)n Microsatellite Markers of the Dystrophin Gene in the Korean Population
Authors: Kim Un Kyung , Cho Min Soon , Chae Jae Jin , Kim Sung Han , Hong Sung Soo , Lee Sook Hwan , Namkoong Yong , Lee Chung Choo
Published: April 2003

Using the polymerase chain reaction, we examined the allele frequencies and heterozygosities of six (CA)n markers of the dystrophin gene in Koreans. Allele frequencies of these markers were different from those reported for Caucasians. The heterozygosity values for these markers range from 29 to 86%. With the exception of the STR50 marker, these values were lower than those of Caucasians. However, all markers except for the 3′CA marker showed PIC values over 0.5, suggesting a high degree of polymorphism. Therefore, this study will be useful in linkage analysis for Duchenne and Becker muscular dystrophy families in the Korean population.
Haplotype Evolution and Linkage Disequilibrium: A Simulation Study
Authors: Calafell Francesc , Grigorenko Elena L. , Chikanian Alexei A. , Kidd Kenneth K.
Published: April 2003

We simulated the evolution of a three-site haplotype system, two restriction fragment length polymorphisms flanking one short tandem repeat polymorphism, under five different demographic scenarios, three with constant population size and two with population growth. The simulation was designed to observe the effects of population history, recombination fraction, and mutation rate on allele and haplotype frequencies, haplotype diversity, frequency of ancestral alleles, and linkage disequilibrium. The known ancestral haplotypes were often found at low frequencies and even became extinct after 5,000 generations, especially with small effective population sizes. The original linkage disequilibrium was eroded and even reversed.
Genetic Linkage Analysis of Prostate Cancer Families to Xq27–28
Authors: Peters Mette A. , Jarvik Gail P. , Janer Marta , Chakrabarti Lisa , Kolb Suzanne , Goode Ellen L. , Gibbs Mark , DuBois Charles C. , Schuster Eugene F. , Hood Leroy , Ostrander Elaine A. , Stanford Janet L.
Published: April 2003

Objectives: A recent linkage analysis of 360 families at high risk for prostate cancer identified the q27–28 region on chromosome X as the potential location of a gene involved in prostate cancer susceptibility. Here we report on linkage analysis at this putative HPCX locus in an independent set of 186 prostate cancer families participating in the Prostate Cancer Genetic Research Study (PROGRESS). Methods: DNA samples from these families were genotyped at 8 polymorphic markers spanning 14.3 cM of the HPCX region. Results: Two-point parametric analysis of the total data set resulted in positive lod scores at only two markers, DXS984 and DXS1193, with scores of 0.628 at a recombination fraction (θ) of 0.36 and 0.012 at θ = 0.48, respectively. The stratification of pedigrees according to the assumed mode of transmission increased the evidence of linkage at DXS984 in 81 families with no evidence of male-to-male transmission (lod = 1.062 at θ = 0.28). Conclusions: Although this analysis did not show statistically significant evidence for the linkage of prostate cancer susceptibility to Xq27–28, the results are consistent with a small percentage of families being linked to this region. The analysis further highlights difficulties in replicating linkage results in an etiologically heterogeneous, complexly inherited disease.
Ligand Blot Analysis of Insulin-Like Growth Factor-Binding Proteins using Biotinylated Insulin-Like Growth Factor-I
Authors: Grulich-Henn J. , Grulich-Henn J. , Spiess S. , Spiess S. , Heinrich U. , Heinrich U. , Schönberg D. , Schönberg D. , Bettendorf M. , Bettendorf M.
Published: April 2003

A nonradioactive method for the detection of insulin-like growth factor-binding proteins (IGFBPs) was developed utilizing human recombinant insulin-like growth factor-I (IGF-I) biotinylated with N-hydroxysuccinimido-biotin. Human plasma samples were separated by 15% SDS polyacrylamide gel electrophoresis, and proteins were transferred to nitrocellulose by Western blotting. The nitrocellulose sheets were incubated overnight with IGF-I-biotin at 4°C. The next day streptavidin-peroxidase was added for 1 h, and IGFBPs were visualized by an enhanced chemiluminescence system. Five characteristic bands with molecular weights of 41 and 39 (IGFBP-3), 34 (IGFBP-2), 30 (IGFBP-1) and 24 kD (IGFBP-4) were detected. Binding was specific for IGFs, since unlabeled IGF-I inhibited IGF-I-biotin binding. IGFBP ligand blots with biotinylated IGF-I and 125I-IGF-I yielded comparable results. The suitability of the new assay for clinical purposes was demonstrated by several clinical examples. In summary, a rapid, reliable, nonradioactive assay for qualitative analysis of IGFBPs has been developed.
Plasma Leptin Concentrations in Phenylketonuric Patients
Authors: Schulpis Kleopatra H. , Papakonstantinou E.D. , Tzamouranis J.
Published: April 2003

High levels of phenylalanine (Phe) in blood have been shown to reduce dopamine (DA) and noradrenaline (NA) production. Leptin levels rise with increasing adiposity in rodents and humans acting as a negative feedback adipostatic signal to brain centers. The aim of this study was to evaluate leptin plasma levels in phenylketonuria (PKU) patients adhering to their special diet and in those on a ‘loose diet’. Forty-nine patients with classical PKU were divided into two groups. Those in group A (n = 21) adhered very strictly to their diet (Phe: 0.15 ± 0.04 mmol/l) and those in group B (n = 28) were on a ‘loose diet’ (Phe: 0.8 ± 0.04 mmol/l). Thirty healthy children of comparable age served as controls. Both patients and controls were in pubertal stage 0 (Tanner). BMI (kg/m2) was evaluated in all the members of the groups. Their daily nutrients were calculated with a 7-day dietary protocol. Leptin was evaluated by RIA, and Phe and Tyrosine with an amino acid autoanalyser. Adrenaline (A), NA and DA were measured by an HPLC method. Plasma leptin in group B patients (28.4 ± 2.0 ng/ml) was significantly increased as compared to group A patients (16.8 ± 2.6 ng/ml) and controls (17.8 ± 3.0 ng/ml; p < 0.001). Plasma DA, A, and NA in group B was lower than in group A and controls. Additionally, leptin negatively correlated with A and DA, whereas Phe positively correlated with the hormone in all groups. Leptin, also, correlated with BMI only in group A and controls. Additionally, the hormone negatively correlated with the total energy intake only in group A (r = –0.43, p < 0.01) and in controls (r = –0.040, p < 0.01). It is suggested that the disregulation of the neuroendocrine system as well as the high Phe blood levels might play an important role in the increased leptin concentrations in PKU patients on a ‘loose diet’.
Experience with Growth Hormone Therapy in Turner Syndrome in a Single Centre: Low Total Height Gain, No Further Gains after Puberty Onset and Unchanged Body Proportions
Authors: Schweizer R. , Ranke M.B. , Binder G. , Herdach F. , Zapadlo M. , Grauer M.L. , Schwarze C.P. , Wollmann H.A.
Published: April 2003

The experience gained since 1987, through observation of 85 girls with Turner syndrome under growth hormone (GH) treatment, has enabled the analysis of one of the largest cohorts. Our results show that age, karyotype and height reflect the heterogeneity of the patients examined at our growth centre. In 47 girls, followed over 4 years on GH (median dose 0.72 IU/kg/week), the median age was 9.4 years and mean height SDS was –3.55 (Prader) and –0.14 (Turner-specific), while height and other anthropometrical parameters [weight, body mass index, sitting height (SH), leg length (LL) SH/LL, head circumference, arm span] were documented and compared to normative data as well as to Turner-specific references established on the basis of a larger (n = 165) untreated cohort from Tübingen. The latter data are also documented in this article. Although there was a trend towards normalization of these parameters during the observation period, no inherent alterations in the Turner-specific anthropometric pattern occurred. In 42 girls who started GH treatment at a median age of 11.8 years, final height (bone age >15 years) was achieved at 16.7 years. The overall gain in height SDS (Turner) from start to end of GH therapy was 0.7 (± 0.8) SD, but 0.9 (± 0.6) SD from GH start to onset of puberty (spontaneous 12.2 years, induced 13.9 years) and –0.2 (± 0.8) from onset of puberty to end of growth. Height gain did not occur in 12 patients (29%) and a gain of > 5 cm was only observed in 16 patients (38%). Height gain correlated positively with age at puberty onset, duration, and dose of GH, and negatively with height and bone age at the time GH treatment started. Final height correlated positively with height SDS at GH start and negatively with the ratio of SH/LL (SDS). We conclude that, in the future, GH should be given at higher doses, but oestrogen substitution should be done cautiously, owing to its potentially harmful effect on growth. LL appears to determine height variation in Turner syndrome and the potential to treat short stature successfully with GH.
Involvement of the N-Terminus of Der p 2 in IgE and Monoclonal Antibody Binding
Authors: Hakkaart Gerrit A.  J. , Aalberse Rob C. , van Ree Ronald
Published: April 2003

The major house dust mite allergen Der p 2 was expressed as a recombinant fusion protein in Escherichia coli either with glutathione-S-transferase as fusion partner or with a poly-histidine tag. Both recombinant fusion proteins failed to react with 3/14 Der p 2-specific monoclonal antibodies (mAbs). When Der p 2 was expressed in yeast with one alanine linked N-terminally to the allergen, no reactivity was observed. When expressed without any fusion partner, all 14 mAbs showed reactivity. The addition of a single N-terminal alanine also disrupted an important epitope for IgE. In RAST inhibitions, an average decrease in inhibitory potency of 72±32% was observed (n = 16) with a maximum decrease of 91%. These observations suggest that the N-terminus of Der p 2 is involved in an important epitope for IgE that is disrupted by the addition of one single aminoacid. Recombinant Der p 2 molecules should therefore preferably lack any fusion peptide.
Receptor-Mediated Maternofetal Transfer of ImmunoglobulinsInhibition of Transport of Anti-HIV-1 Immunoglobulin by Generic Immunoglobulins in the in vitro Perfused Placenta
Authors: Landor Michael , Rubinstein Arye , Kim Ana , Calvelli Theresa , Mizrachi Yaffa
Published: April 2003

Objectives: The passage of immunoglobulin G (IgG) across the placenta is thought to involve Fc’ receptors on the syncytiotrophoblast. To confirm the receptor dependency of this process we have studied the changes in the tissue content and transfer kinetics of immunoglobulins and hyperimmune serum to HIV (HIVIG) during in vitro dual placental perfusion. Methods: Isolated lobules of term placentae from normal pregnancies were perfused in a model of maternal and fetal circulation. The perfused tissue was compared to fresh tissue samples from the same placenta for the content of IgG, IgG subclasses, IgM, cytokeratin, human placental lactogen and SP1 antigen by immunohistochemistry and by protein elution. Results: The immunoglobulin staining faded by an average of 40% during the 1st hour of perfusion. In contrast, staining for cytokeratin, human placental lactogen and SP1 remained unchanged. During a 4-hour recycling of endogenous immunoglobulins in the maternal circulation, IgG and HIVIG crossed to the fetal side in a steady rate. The transport of HIVIG could be inhibited by preperfusion with an intravenous gammaglobulin preparation (IVIG). Discussion: The transfer of IgG across the placenta occurs in a steady state rate consistent with a receptor-mediated mechanism. Furthermore, inhibition of HIVIG maternofetal transfer by IVIG further establishes the receptor-mediated transfer of immunoglobulins through the placenta.
Role of Antigen-Induced Cytokine Release in Atopic Pruritus
Authors: Lippert U. , Hoer A. , Möller A. , Ramboer I. , Cremer B. , Henz B.  M.
Published: April 2003

In order to further evaluate the role of cytokines in the induction of atopic pruritus, leukocytes from 10 atopic eczema patients or 10 nonallergic controls were stimulated in vitro with mite or birch pollen antigen for 1 and 4 days. Subjects were prick-tested with the supernatants, and whealing and itching were evaluated 20 and 60 min later. The supernatants were also examined for the contents of GM-CSF, IL-2, IL-6 and IL-8 by ELISA and TNFα. Two hours prior to testing, the antihistamine cetirizine (20 mg) or a placebo tablet were given to the patients according to a randomized, double-blind study protocol. After pricking with antigen-stimulated leukocyte supernatants, 6 of 10 patients but no controls reacted mostly at 20 min with whealing and/or pruritus. In the cetirizine-treated group, no decrease in these skin reactions was seen compared to placebo. Analysis for cytokines showed increased levels of IL-8 in allergen-stimulated samples, with no correlation to the induction of itching or whealing by these supernatants. IL-6 levels were low and variable, and GM-CSF, IL-2 and TNFα levels were always below standard values. These data show that leukocytes selectively release IL-8 in response to in vitro antigen stimulation. They furthermore provide additional support for the concept that as yet to be identified products play a role in atopic pruritus.
Effects of Intracellular Cyclic AMP Modulators on Human Eosinophil Survival, Degranulation and CD11b Expression
Authors: Momose T. , Okubo Y. , Horie S. , Suzuki J. , Isobe M. , Sekiguchi M.
Published: April 2003

Background: Brochial asthma is characterized by infiltration of inflammatory cells such as lymphocytes and eosinophils. Theophylline is one of the most widely used drugs in the therapy of bronchial asthma, and phosphodiesterase (PDE) inhibition is thought to be an important mechanism of its anti–inflammatory actions. However, the detailed effects of PDE inhibition on eosinophils still remain unclear. Methods: Eosinophils in peripheral blood obtained from normal subjects and patients with mild off–season allergic rhinitis were purified using CD16 negative selection. The following effects of theophylline (nonselective PDE inhibitor), KF19514 (selective PDE IV inhibitor), mirlinone (selective PDE III inhibitor), procaterol (β2–adrenoceptor agonist) and N6, 2′–O–dibutyryladenosine 3′5′–cyclic monophosphate (dB–cAMP; AMP analogue) on eosinophils were examined: (1) survival in the presence of interleukin–5, (2) degranulation by granulocyte/macrophage colony–stimulating factor (GM–CSF) or platelet–activating factor (PAF), (3) CD11b expression under GM–CSF or PAF stimulation and (4) intracellular cAMP level. Results: Eosinophil survival was inhibited by theophilline, KF19514 or procaterol. GM–CSF– or PAF–induced degranulation was inhibited by theophylline, KF19514, procaterol or dB–cAMP. CD11b up–regulation by PAF was inhibited by theophylline, KF19514 or dB–cAMP, while GM–CSF–stimulated CD11b up–regulation was not significantly inhibited by any of the drugs tested. The levels of intracellular cAMP were increased by theophylline, KF19514 and procaterol. Conclusions: Intracellular cAMP is an important factor in the regulation of eosinophil biological functions. PDE IV inhibitors and β2–agonists are suggested to be useful for the treatment of bronchial asthma through inhibition of eosinophil effector function.
Two Common Polymorphisms in the Coding Part of the CD43 Gene Are Not Associated with Atopy
Authors: Kruse Susanne , Kuehr Joachim , Forster Johannes , Deichmann Klaus A.
Published: April 2003

Background: Recently linkage and association of the chromosomal region 16p12–11 with enhanced IgE responsiveness have been shown. The gene coding for CD43 (sialophorin) has been localized to this region. Sialophorin represents a major sialoglycoprotein on the surface of human lymphocytes, monocytes and granulocytes. It is supposed to play an important role in human mast cell, T– and B–cell regulation and activation and has been described in connection with immunodeficiency diseases such as the Wiskott–Aldrich syndrome. Therefore, it can be designated as a candidate gene for atopy. Methods: Using SSCP analysis and direct genomic sequencing, polymorphisms in the CD43 gene have been looked for and their association with atopy has been tested in a population of 260 largely atopic children and young adults. Results: Three common polymorphisms in the coding part of the CD43 gene were found. Two of them are leading to amino acid exchanges, one from argine to cysteine at amino acid position 337 of the mature gene product and one from leucine to phenylalanine at amino acid position 341. Subsequent association studies revealed no obvious influence of R337C or L341F on IgE regulation (p = 0.47 and 0.43), neither in a cognate nor in an uncognate fashion. Conclusion: We conclude that CD43 polymorphisms are unlikely to account for the observed linkage effect at 16p12–11. Whether the polymorphisms R337C and L341F adjacent to phosphorylation sites in the intracellular region of the protein alter the normal functioning of CD43 remains to be elucidated.
Regulation of Eosinophil Apoptosis: Transduction of Survival and Death Signals
Authors: Simon Hans-Uwe , Alam Rafeul
Published: April 2003

Since eosinophils are prominent in allergic inflammation, investigators became interested in how these cells accumulate in tissues and their role within the inflammatory cascade. There is increasing evidence from several laboratories that eosinophil numbers are regulated in vivo, not only by eosinophil production in the bone marrow, but also by the amount of eosinophil apoptosis. Moreover, it has been directly demonstrated that eosinophil apoptosis is delayed in allergic inflammatory sites, and that this mechanism contributes to the expansion of these cells in tissue. In this article, we review recent studies that shed light on the intracellular pathways that control eosinophil apoptosis.
Effects of Interferon–γ on Mobilization and Release of Eosinophil–Derived RANTES
Authors: Velazquez J.R. , Lacy P. , Mahmudi-Azer S. , Moqbel R.
Published: April 2003

Eosinophils synthesize and release a number of cytokines and chemokines, including RANTES, a potent chemoattractant particularly for memory T cells and eosinophils. Long–term (>12 h) incubation with interferon–γ (IFN–γ) has been shown to activate eosinophils and induce expression of membrane receptors. We hypothesized that IFN–γ mobilizes intracellular RANTES in eosinophils in advance of mediator release. Highly purified peripheral blood eosinophils were obtained from asthmatics and stimulated with IFN–γ at 500 U/ml for time course analysis up to 2h. By specific ELISA, RANTES was detected in supernatants (80±15 pg per 2×106 cells) following 120min of stimulation. Immunoreactive RANTES in resting cells (5×107 eosinophils) was detected in two intracellular compartments in studies of subcellular fractionation by density gradient centrifugation. After 10 min IFN–γ stimulation, RANTES immunoreactivity was confined to crystalloid granules. RANTES was redistributed from secretory granules to light–membrane fractions after 60 min of IFN–γ incubation. Our data suggest that rapid mobilization and release of RANTES occurs from stimulated eosinophils. These findings may have important implications for the role of IFN–γ in activating human eosinophils, particularly in severe chronic asthma or viral exacerbation of asthmatic inflammation, where this cytokine may play a role.
Dual Effects of TNF on Synthesis of Complement Components by a Gastric Cancer–Derived Cell Line, KATO–III
Authors: Kitano Etsuko , Kitamura Hajime
Published: April 2003

Background: Complement components are synthesized extrahepatically, although hepatocytes are the major source of plasma complement. It is now clear that local production of complement is important in homeostasis and immune defense in tissue. Methods: The secretion of complement components was studied in vitro with a gastric cancer–derived cell line, KATO–III (signet–ring cell carcinoma). Complement components C2 and C3 were estimated by functional assay and/or ELISA in culture medium obtained after incubation of KATO–III cells for 3 days in protein–free culture medium, with or without addition of tumor necrosis factor (TNF), in a humidified atmosphere of 5% CO2/95% air at 37°C. Results:  (1) While a higher amount of C3 was detected in medium when KATO–III cells were cultured in the presence of TNF than in medium lacking TNF, higher C2 activity was detected when cultured in medium lacking TNF than in TNF–supplemented medium. (2) TNF suppressed C2 secretion and enhanced C3 secretion in a dose–dependent fashion. (3) C3 secretion remained less than 20 ng/106 cells/24 h but increased from the first day of TNF (10U/ml) addition (concentrations approached 108.6– 115.6 ng/106 cells/24 h on the 3rd day) and decreased on the 1st day without TNF. In contrast, C2 activity, detected when cultured in the absence of TNF, was decreased on the 2nd day of TNF addition and increased again on the 1st day without TNF. The daily secretion of C2 in the absence of TNF was 3.75–6.30×107 effective molecules/106 cells. (4) Reversible inhibition of C2 and C3 secretion was observed when the cell line was cultured in the presence of cycloheximide, indicating that both components were synthesized de novo. Conclusion: It appears that TNF enhances C3 secretion and suppresses C2 secretion by KATO–III.
Molecular Characterization of the Group 4 House Dust Mite Allergen from Dermatophagoides pteronyssinus and Its Amylase Homologue from Euroglyphus maynei
Authors: Mills K.L. , Hart B.J. , Lynch N.R. , Thomas W.R. , Smith W.
Published: April 2003

Background: Of the ten recognised groups of Dermatophagoides pteronyssinus allergens, the group 4 is the only group that has not been characterised at the molecular level. Methods: Primers were designed to PCR amplify Der p 4 (D. pteronyssinus) and Eur m 4 (Euroglyphus maynei) cDNA. These fragments were used to screen the corresponding cDNA libraries and the cDNA clones obtained were subsequently sequenced. The coding regions of Der p 4 and Eur m 4 were cloned into the pET expression vector and recombinant histidine–tagged proteins expressed in Escherichia coli. Results: cDNA clones which included the mature protein coding sequence for Der p 4 and Eur m 4 were sequenced. The Der p 4 and Eur m 4 genes were found to code for 496 amino acid mature proteins with residues important for the function of α–amylase highly conserved. Der p 4 and Eur m 4 were calculated to be 90% identical and a BLAST search of the GenBank database found these sequences to be approximately 50% identical to insect and mammalian α–amylases. The calculated molecular weights of Der p 4 and Eur m 4 were approximately 57,000, although recombinant Der p 4 and Eur m 4 migrate on SDS–PAGE at about 60,000. Der p 4 recombinant protein was found to bind specific IgE in 3 of the 10 house dust mite allergic patients tested. Conclusions: This paper describes the first cDNA sequence of Der p 4 and Eur m 4 confirming that this allergen is house dust mite α–amylase.
Important Animal Allergens Are Lipocalin Proteins: Why Are They Allergenic?
Authors: Virtanen Tuomas , Zeiler Thomas , Mäntyjärvi Rauno
Published: April 2003

Major respiratory allergens of dogs, mice, rats, horses and cows belong to the lipocalin group of proteins. The sequence identity of lipocalins is often less than 20%, but they contain between one and three structurally conserved regions and their three–dimensional structures are similar. Lipocalins share common biological functions, predominantly related to the transport of small hydrophobic molecules, such as vitamins and pheromones. Immune reactivity to lipocalin allergens is not well known. In Bos d 5, the IgE–binding epitopes are spread along the molecule, whereas in Bos d 2, the C terminus appears to contain the human B cell epitopes. Bos d 5 contains several murine T cell epitopes. No information is available on human T cell epitopes. The maximal number of epitopes an allergic patient’s T cells could recognize in Bos d 2 was five. Three of the epitopes were colocalized in the structurally conserved regions of lipocalins. Interestingly, one of the epitopes was recognized by the T cells of all patients and the computer predictions suggested that there would be an epitope in the corresponding parts of human endogenous lipocalins. The proliferative responses of peripheral blood mononuclear cells of Bos d 2–allergic subjects to Bos d 2 were weak. The T cell response was Th2–dominated. To explain these observations, we have proposed that the allergenicity of lipocalins may be a consequence of molecular mimicry between lipocalin allergens and endogenous lipocalins at the T cell level.
Characterization of Tissue Outgrowth Developed in vitro in Patients with Rheumatoid Arthritis: Involvement of T Cells in the Development of Tissue Outgrowth
Authors: Wakisaka Sueshige , Suzuki Noboru , Nagafuchi Hiroko , Takeba Yuko , Kaneko Atsushi , Asai Tomiaki , Sakane Tsuyoshi
Published: April 2003

Background: The aim of this study was to analyze cellular and cytokine interactions governing the development of synovial tissue outgrowth in patients with rheumatoid arthritis (RA). Methods: A single–cell suspension of dissociated synovial tissues of RA patients was cultured for a long period to develop tissue outgrowth. The resulting tissue outgrowth was characterized by immunohistochemical staining and ELISA. Results: The tissue outgrowth developed in vitro included various cell types, such as macrophage–like synovial cells, fibroblast–like synovial cells and lymphocytes. Even after prolonged cultivation, synovial cells devoid of infiltrating T lymphocytes did not form tissue outgrowth. The outgrowth contained CD3+ cells, LeuM3 (CD14)+ cells and HLA–DR+ cells. The T cells expressed lymphocyte function–associated antigen (LFA)–1 and CD2, and the synovial cells expressed intracellular adhesion molecule (ICAM)–1 and LFA–3, suggesting possible interactions via LFA–1/ICAM–1 and CD2/LFA–3. Production of T–cell derived IFN–γ and IL–17 and synovial–cell–derived fibroblast growth factor (FGF)–1 and IL–15 was confirmed in the tissue outgrowth as well as in RA synovial tissue. These cell types stimulate each other by secreting cytokines, leading to the secretion of proinflammatory cytokines and matrix metalloproteinase (MMP)–1 by the tissue outgrowth and proliferation of both lymphocytes and synovial cells. Conclusion: This study emphasizes the importance of cellular interactions between T cells and synovial cells, via adhesion molecules and the secretion of cytokines with stimulatory activity towards other cell types, for the hyperactivity of RA synovial cells.
Inhibitory Effect of Wheat Germ Agglutinin on Mouse Mast Cell Adhesion to Fibronectin
Authors: Wyczółkowska Janina , Weyer Anne , Dastych Jarosław
Published: April 2003

Background: Mast cells play a critical role in allergic and inflammatory responses. The interactions between these cells and extracellular matrix components influence the distribution of mast cells in tissues and their biological responsiveness. It has been reported that the lectin wheat germ agglutinin (WGA) inhibits mast cell mediator release. We decided to investigate whether adhesion to fibronectin (FN), another mast cell function, which is upregulated following FcεRI cross-linking, is also inhibited by WGA. Methods: Mouse bone-marrow-derived mast cell line MCP5/L was used. For FcεRI-dependent mast cell activation, MCP5/L cells were sensitized with mouse IgE antibodies. WGA was added to cell suspensions simultaneously with a challenging agent and, after an appropriate incubation period, β-hexosaminidase release and adhesion to FN were determined. Results: Both FcεRI cross-linking-dependent mast cell adhesion to FN and mediator release were dose-dependently inhibited by WGA; however, the lectin concentrations required to induce maximum inhibition of adhesion were significantly lower. Furthermore, WGA inhibited phorbol-myristate-acetate- and A-23187-mediated mast cell adhesion to FN, i.e. processes that do not engage FcεRI. The effect of WGA on FcεRI-mediated secretion was reversed by GlcNAc. In contrast, combination of GlcNAc and NeuNAc or N,N′-diacetylchitobiose was required to reverse the inhibitory effect of WGA on mast cell adhesion. Conclusion: The characteristics of WGA-mediated inhibition of MCP5/L mast cell adhesion to FN suggest that mast cell integrins are targets of the inhibitory action of WGA and the sugar moieties on these receptors might be important for receptor function.
Human Cytomegalovirus Stimulates Cellular Dihydrofolate Reductase Activity in Quiescent Cells
Authors: Lembo David , Gribaudo Giorgio , Cavallo Rossana , Riera Ludovica , Angeretti Alessandra , Hertel Laura , Landolfo Santo
Published: April 2003

Human cytomegalovirus (HCMV) productively infects quiescent fibroblasts in which the levels of deoxynucleotide triphosphates (dNTPs) and cell functions involved in DNA metabolism are very low. Since sufficient dNTPs levels are essential for human HCMV replication, host cell enzymes involved in the biosynthesis of dNTPs might be expected to be stimulated by viral infection in quiescent cells. We report that HCMV infection of quiescent fibroblasts stimulates the activity of cellular dihydrofolate reductase (DHFR), a key enzyme in DNA precursor synthesis. We also demonstrate that suppression of DHFR activity by the specific inhibitor methotrexate prevents HCMV replication and DNA synthesis. These observations indicate that induction of DHFR activity by HCMV is required for efficient viral replication in quiescent fibroblasts.
Hepatitis B Virus Replication: Novel Roles for Virus-Host Interactions
Authors: Nassal Michael
Published: April 2003

Chronic hepatitis B continues to be one of the most widespread and serious viral infections in humans worldwide. Several fundamental aspects of the molecular biology of its causative agent, hepatitis B virus, are meanwhile understood in some detail. However, recent research has emphasized that the dependence of the viral infectious cycle on cellular factors is far greater than previously anticipated. More and more intracellular interactions between viral and cellular components are discovered, and probably each individual step of genome replication will turn out to involve several host factors. Prominent examples are the activation of the viral reverse transcriptase, P protein, by chaperones, and the nucleocytoplasmic trafficking of viral nucleic acids by as yet unidentified components of the host machinery. Some of these new developments will be described here but many more can be expected to follow. Identifying these host factors and characterizing their interactions with the viral components will certainly reveal novel targets for specific antiviral strategies.
Existence of TT Virus DNA in Extracellular Body Fluids from Normal Healthy Japanese Subjects
Authors: Matsubara Hiroshi , Michitaka Kojiro , Horiike Norio , Yano Mariko , Akbar S.M. Fazle , Torisu Masamoto , Onji Morikazu
Published: April 2003

Although recent studies indicate a high prevalence (12–92%) of TT virus (TTV) DNA in sera of healthy Japanese individuals, there is a paucity of information regarding the route of transmission of this virus. Analyzing the nucleotide sequences of the existing polymerase chain reaction (PCR) primers of TTV DNA, we developed a set of noble primers (HM-1) and looked for the prevalence of TTV DNA in sera from 39 normal healthy Japanese individuals using PCR. The existence of TTV DNA was also checked in saliva, urine, sweat, stool, and tears from 11 and in semen from 10 serum TTV-positive normal subjects. TTV DNA was detected in sera from 23 of 39 (59.0%) normal subjects. TTV DNA was also detected in saliva, stool, semen and tears from all cases with TTV-DNA-positive serum, but not in body fluids from subjects with TTV-DNA-negative serum. TTV DNA remained undetected in urine and sweat from all cases. Data from these experiments showing the existence of TTV DNA in different body fluids suggest that the high rates of prevalence of TTV among normal healthy subjects might be due to a possible fecal-oral, droplet, or sexual route of transmission of TTV.
Angiotensin II Type 1 Receptor Gene Polymorphism Is Associated with an Increased Vascular Reactivity in the Human Mammary Artery in vitro
Authors: Henrion Daniel , Amant C. , Benessiano Joëlle , Philip I. , Plantefève G. , Chatel D. , Hwas Ulrich , Desmont J.M. , Durand Geneviève , Amouyel P. , Lévy Bernard I.
Published: April 2003

A gene polymorphism of the angiotensin II (AII) type 1 receptor has been described previously (A to C transversion at position 1166). Besides the epidemiological studies needed to determine a possible relationship between the polymorphism and some cardiovascular diseases, no study has been conducted to determine the impact of the polymorphism on vascular functions. At subthreshold concentrations, within the physiological range, AII potentiates α-adrenergic-dependent vascular tone. We investigated phenylephrine-induced tone and its amplification by AII (10 pmol/l) in human internal mammary artery rings mounted in organ baths. We performed concentration-response curves to phenylephrine (0.1–100 µmol/l) before and after pretreatment with AII (10 pmol/l). Patients had the genotype AA (n = 20) or the A to C transversion (AC/CC, n = 30). Contractions to phenylephrine (0.1–100 µmol/l) were significantly higher in rings from AC/CC than from AA patients (maximum response: 1.47±0.07 vs. 1.22±0.06 mN/mg, p < 0.001). AII (10 pmol/l) induced a significant potentiation of phenylephrine-induced contraction (e.g. 58.9% increase in tone with 1 µmol/l phenylephrine, p < 0.001) which was significantly lower in the AC/CC than in the AA group (46±9 vs. 66±7% with 1 µmol/l phenylephrine, p < 0.01). Contractions to AII (1 or 100 nmol/l) were not significantly affected by the genotype. Although the study was performed in arteries from patients with a coronary artery disease, these changes in vascular reactivity might be of interest in the understanding of the relationship between a possible higher probability of cardiovascular disorder and the genetic polymorphism of the AII type 1 receptor.