bluuebunny/crossref_metadata_2025 huggingface dataset:
Characteristics of Umbilical Cord Blood Units Collected from Preterm Deliveries
Authors: Solves Pilar , López Mara , Mirabet Vicente , Blanquer Amando , Roig Roberto , Perales Alfredo
Published: August 2009

Background: Umbilical cord blood (UCB) banking is a well-established activity supporting the increasing number of UCB transplantations in haematological diseases. Our aim was to analyse the UCB characteristics of UCB units from preterm deliveries and compare them to full-term deliveries. Material and Methods: A prospective study in 194 preterm deliveries occurring at the La Fe University Hospital in Valencia was performed. Patients between 25 and 37 weeks of gestation were included. Those cases were compared to a full-term deliveries control group. Results: The cases were grouped according to the gestational age: between 25 and 33 weeks (group 1), between 34 and 37 weeks (group 2) and between 38 and 42 weeks (group 3). Among obstetric variables, only arterial pH and maternal age variables were similar for all the groups. Higher CD34+ cell counts were observed in the group 2, while the clonogenic efficiency was higher for the most preterm deliveries. Discussion: UCB from deliveries of at least 34 weeks of gestation contain sufficient hematopoietic stem cell content for unrelated banking and transplantation, even containing higher CD34+ cell content than UCB units from full-term deliveries. However, UCB from deliveries of less than 33 weeks’ gestation contain only sufficient progenitors for children under 20 kg.
JNK MAPK Signaling Contributes in vivo to Injury-Induced Corneal Epithelial Migration
Authors: Okada Yuka , Saika Shizuya , Shirai Kumi , Yamanaka Osamu , Kitano Ai , Wang Zheng , Yang Hua , Reinach Peter
Published: August 2009

Purpose: Injury-mediated corneal epithelial wound healing in vivo is mediated through different cell signaling pathways depending on whether or not the basement membrane is removed. Given this dependence, we ascertained if c-jun N-terminal kinase (JNK/SAPK) mitogen-activated protein kinase (MAPK) cell signaling mediates this response in vivo and in vitro, irrespective of the presence or absence of the basement membrane. Furthermore, in vitro the relative contribution was determined by the JNK/SAPK pathway to that of its p38 and ERK MAPK counterparts in mediating injury-induced increases in cell migration. Methods: Corneal epithelial debridement was performed in C57BL/6 mice and their organ-cultured eyes without removal of the basement membrane. In rabbits, following basement membrane removal by keratectomy, fluorescein-staining monitored reepithelialization was performed as in the mice. Immunohistochemistry evaluated changes in JNK phosphorylation status and localization. JNK inhibitor I and its inactive analogue determined if JNK signaling activation contributes to wound healing. BrdU staining assessed cell proliferation. A scratch wound assay of healing rates in SV40-immortalized human corneal epithelial cell line (HCEC) evaluated the relative contributions by p38 and ERK and JNK MAPK signaling activation to wound healing. A TUNEL assay probed for apoptosis after wound closure of HCEC. MTT assay evaluated corneal epithelial viability. Results: Two hours following mice corneal epithelial debridement, phospho-JNK was transiently upregulated in the nucleus, whereas total JNK was constitutively expressed. JNK inhibitor I suppressed epithelial spreading in organ-cultured mouse eyes and rabbit corneal blocks, irrespective of the presence or absence of basement membrane. No proliferation was detected at the wound edges. In HCEC, a p38 (SB203580) and a JNK pathway inhibitor (JNK inhibitor I) inhibited migration rates more than U0126-induced ERK, whereas the JNK inhibitor I inactive analogue had no effect. JNK pathway inhibition wound closure in this region was not associated with either any TUNEL or BrdU-positive cells. Cell viability was unaffected by any of these MAPK inhibitors. Conclusion: JNK/SAPK pathway activation stimulates wound healing in vitro and in vivo, irrespective of the presence or absence of the basement membrane. Therefore, studies on how wound closure is elicited in HCEC are relevant for identifying how MAPK signaling mediates this response in vivo and in organ-cultured eyes. This realization suggests that the JNK signaling system has a role in vivo that is intermediate to those of ERK and p38 in mediating increases in cell migration.
Epidemiology of Pediatric Eye Injury in Split-Dalmatia County
Authors: Karaman Ksenija , Znaor Ljubo , Lakos Venera , Olujic Ivana
Published: August 2009

Aims: The aim of this study was to assess the etiological factors and circumstances associated with the occurrence of eye injuries in children in Split-Dalmatia County, and to assess the visual outcomes of such injuries. Methods: All 106 cases of pediatric eye trauma presenting to the Clinical Hospital Split, during a 4-year period, were identified by a medical record search. Results: The annual calculated incidence of serious eye injuries in pediatric patients of Split-Dalmatia County that required hospitalization was 22.5 per 100,000 in the underage population. The most common causes of ocular injury were blunt objects and missiles, which mostly caused contusions and lamellar lacerations, and sharp objects, which mostly caused penetrating eye injury. In 99 (90.83%) eyes, the injuries were mechanical. Open globe injuries were significantly severer than closed globe injuries. Final visual acuity was better than upon admission in 78.10% of cases. The annual incidence of pediatric monocular blindness in Split-Dalmatia County was 0.63 per 100,000 in the underage population. Conclusions: Eye trauma in children occurs most frequently outdoors and generally results from the use of blunt objects. Prevention of ocular injury requires greater education of both children and their parents and adult supervision of children, especially while at play.
Noninvasive Determination of Cardiac Output by the Inert-Gas-Rebreathing Method – Comparison with Cardiovascular Magnetic Resonance Imaging
Authors: Saur Joachim , Fluechter Stephan , Trinkmann Frederik , Papavassiliu Theano , Schoenberg Stefan , Weissmann Joerg , Haghi Dariusch , Borggrefe Martin , Kaden Jens J.
Published: August 2009

Background: An easy, noninvasive and accurate technique for measuring cardiac output (CO) would be desirable for the diagnosis and therapy of cardiac diseases. Innocor, a novel inert-gas-rebreathing (IGR) system, has shown promising results in smaller studies. An extensive evaluation in a larger, less homogeneous patient collective is lacking. Methods: We prospectively assessed the accuracy and reproducibility of CO measurements obtained by IGR in 305 consecutive patients as compared to the noninvasive gold standard, cardiovascular magnetic resonance (CMR) imaging. Results: Bland-Altman analysis showed a good correspondence of the two methods for CO measurement with an average deviation of 0.2 ± 1.0 liters/min (mean ± SD) and a good reproducibility with a mean bias of 0.2 ± 0.5 liters/min. The accuracy of the present measurements at rest was significantly better in the physiological range than in higher or lower CO ranges. The error levels set forth by current recommendations were exceeded. Conclusion: The data show that IGR measurements are easy to perform and show good agreement with CMR; however, the technique appears to be less accurate in extreme CO ranges at rest. The clinical importance of the IGR method remains to be proven by further studies.
Analysis of the Antibody Responses Induced by Subcutaneous Injection Immunotherapy with Birch and Fagales Pollen Extracts Adsorbed onto Aluminum Hydroxide
Authors: Gadermaier E. , Flicker S. , Aberer W. , Egger C. , Reider N. , Focke M. , Vrtala S. , Kundi M. , Valenta R.
Published: August 2009

Background: Allergen-specific subcutaneous immunotherapy (SCIT) is an antigen-specific therapy of IgE-mediated allergies. In the present study, we analyze the epitope specificities of antibody responses induced by SCIT with allergen extracts from pollen of trees belonging to the order Fagales (birch, alder, hazel) adsorbed onto aluminum hydroxide. Methods: The IgE, IgG1–4 and IgA responses to defined recombinant allergens (birch pollen: Bet v 1; alder pollen: Aln g 1; hazel pollen: Cor a 1; apple: Mal d 1) as well as to Bet v 1-derived recombinant fragments and synthetic peptides were analyzed in sera from patients who had undergone SCIT for different periods of time. Results: Long-term SCIT (>1 year; cumulative dose >1,000,000 SQ units) induced more pronounced IgG1, IgG2 and IgG4 responses to Bet v 1 and Bet v 1-related allergens according to the degree of sequence homology (Bet v 1>Aln g 1>Cor a 1>Mal d 1) than short-term SCIT (<1 year; cumulative dose <1,000,000 SQ units). In contrast to patients treated for <1 year, patients treated for >1 year mounted distinct IgG1, IgG2 and IgG4 responses against sequential Bet v 1 epitopes. No relevant allergen-specific IgA or IgG3 responses were induced by short- or long-term SCIT. Using a competitive ELISA assay, it could be shown that serum IgG from patients undergoing long-term SCIT inhibited IgE reactivity to Bet v 1 better than IgG from patients undergoing short-term SCIT. Conclusion: SCIT with allergen extracts adsorbed onto aluminum hydroxide induces IgG responses against new epitopes that block IgE binding and cross-react with structurally related allergens depending, among other factors, on duration of treatment and cumulative injected dose.
Locus 1q21 Gene Expression Changes in Atopic Dermatitis Skin Lesions: Deregulation of Small Proline-Rich Region 1A
Authors: Jarzab Jerzy , Filipowska Barbara , Zebracka Jadwiga , Kowalska Małgorzata , Bozek Andrzej , Rachowska Regina , Gubala Elzbieta , Grzanka Alicja , Hadas Ewa , Jarzab Barbara
Published: August 2009

Background: Discovery of the significant impact of filaggrin (FLG) mutations on the genetic predisposition to atopic dermatitis (AD) focused attention on the 1q21 locus, where not only FLG but also other epidermal genes are located. In the present study, we compared 1q21 gene expression in lesional versus nonlesional AD skin. Methods: A real-time quantitative PCR analysis of 10 1q21 genes, selected on the basis of a previous microarray study, was performed in skin biopsies from 33 individuals with AD. Three alternative pathway keratins were also evaluated. Results: In chronic AD skin lesions, we observed an increase in RNA encoding involucrin, S100 calcium-binding proteins A2 and A7–A9 and small proline-rich region (SPRR) proteins 1A and 2C, with fold changes ranging from 2.0 for S100A2 to 15.4 for S100A8 (p < 0.001, Bonferroni corrected), in parallel to the overexpression of the alternative pathway keratins 6A, 6B and 16. The loricrin (LOR) expression level was significantly decreased in lesional AD skin (fold change 0.5; p < 0.01). The expression of the majority of 1q21 genes and alternative keratins was closely correlated; however, for SPRR1A (and SPRR2C) in lesional skin, the correlation with other genes was lost. Conclusions: We hypothesize that the deregulated increase in SPRR1A expression in chronic atopic skin lesions reflects an insufficient rise in SPRR transcripts, unable to compensate for the lack of LOR and thus contributing to the persistence of chronic AD skin lesions. Turning off the stress response in the skin may be regarded as a goal in the treatment of AD skin lesions, and SPRR genes might be targets for such an approach.
Rupatadine Inhibits Proinflammatory Mediator Secretion from Human Mast Cells Triggered by Different Stimuli
Authors: Vasiadi Magdalini , Kalogeromitros Dimitris , Kempuraj Duraisamy , Clemons Anthony , Zhang Bodi , Chliva Caterina , Makris Michael , Wolfberg Adam , House Michael , Theoharides Theoharis C.
Published: August 2009

Background: Mast cells are involved in allergy and inflammation by secreting multiple mediators including histamine, cytokines and platelet-activating factor. Certain histamine 1 receptor antagonists have been reported to inhibit histamine secretion, but the effect on cytokine release from human mast cells triggered by allergic and other stimuli is not well known. We investigated the ability of rupatadine, a potent histamine 1 receptor antagonist that also blocks platelet-activating factor actions, to also inhibit mast cell mediator release. Methods: Rupatadine (1–50 μM) was used before stimulation by: (1) interleukin (IL)-1 to induce IL-6 from human leukemic mast cells (HMC-1 cells), (2) substance P for histamine, IL-8 and vascular endothelial growth factor release from LAD2 cells, and (3) IgE/anti-IgE for cytokine release from human cord blood-derived cultured mast cells. Mediators were measured in the supernatant fluid by ELISA or by Milliplex microbead arrays. Results: Rupatadine (10–50 μM) inhibited IL-6 release (80% at 50 μM) from HMC-1 cells, whether added 10 min or 24 h prior to stimulation. Rupatadine (10–50 μM for 10 min) inhibited IL-8 (80%), vascular endothelial growth factor (73%) and histamine (88%) release from LAD2 cells, as well as IL-6, IL-8, IL-10, IL-13 and tumor necrosis factor release from human cord blood-derived cultured mast cells. Conclusion: Rupatadine can inhibit histamine and cytokine secretion from human mast cells in response to allergic, immune and neuropeptide triggers. These actions endow rupatadine with unique properties in treating allergic inflammation, especially perennial rhinitis and idiopathic urticaria.
Impact of Urbanization on the Proteome of Birch Pollen and Its Chemotactic Activity on Human Granulocytes
Authors: Bryce M. , Drews O. , Schenk M.F. , Menzel A. , Estrella N. , Weichenmeier I. , Smulders M.J.M. , Buters J. , Ring J. , Görg A. , Behrendt H. , Traidl-Hoffmann C.
Published: August 2009

Background: Epidemiologic studies reveal a dramatic increase in allergies in the last decades. Air pollution is considered to be one of the factors responsible for this augmentation. The aim of this study was to analyze the impact of urbanization on birch pollen. The birch pollen proteome was investigated in order to identify differences in protein abundance between pollen from rural and urban areas. The allergenicity of birch pollen from both areas was evaluated by assessing its chemotactic potency as well as its protein and allergen contents. Methods: Difference gel electrophoresis (DIGE) was used to analyze the pollen proteome. The chemotactic activity of aqueous pollen extracts was determined by migration assays of human neutrophils. Results: DIGE revealed 26 differences in protein spot intensity between pollen from urban and rural areas. One of these proteins was identified by de novo sequencing as the 14-3-3 protein, which resembles a stress-induced factor in other plant species. Furthermore, extracts from pollen collected in urban areas had higher chemotactic activity on human neutrophils compared to pollen from rural sites. Conclusions: The present study points to an impact of air pollution on allergen carrier proteome and release of chemotactic substances. The increment in proinflammatory substances such as pollen-associated lipid mediators might contribute to the described urban-rural gradient of allergy prevalence. Furthermore, our study suggests that allergenicity is determined by more than the sole allergen content.
Asthma Severity According to Global Initiative for Asthma and Its Determinants: An International Study
Authors: Cazzoletti Lucia , Marcon Alessandro , Corsico Angelo , Janson Christer , Jarvis Deborah , Pin Isabelle , Accordini Simone , Bugiani Massimiliano , Cerveri Isa , Gislason David , Gulsvik Amund , de Marco Roberto
Published: August 2009

Background: The identification of the factors associated with severe asthma may shed some light on its etiology and on the mechanisms of its development. We aimed to describe asthma severity using the Global Initiative for Asthma (GINA) classification and to investigate its determinants in a cross-sectional, population-based sample in Europe. Methods: In the European Community Respiratory Health Survey II (1999–2002), 1,241 adults with asthma were identified. Severity was assessed using the 2002 GINA classification (intermittent, mild persistent, moderate persistent, severe persistent) and it was related to potential determinants by a multinomial logistic model, using the intermittent group as the reference category for relative risk ratios. Results: About 30% of asthmatic subjects were affected by moderate-to-severe asthma. Sensitization to Cladosporium was associated with a more than 5-fold greater risk of having (mild, moderate or severe) persistent asthma than intermittent asthma. Persistent asthma was positively associated with sensitization to house dust mite, nonseasonal asthma, an older age at asthma onset, and chronic cough and phlegm. Sensitization to cat increased the risk of severe asthma only. Smoking was more strongly associated with asthma severity in men, while rhinitis was more strongly associated with asthma severity in women. Conclusions: One third of the asthmatic population have moderate-to-severe asthma. Sensitization to perennial indoor allergens, particularly Cladosporium, is strongly associated with asthma severity. The role of smoking and rhinitis in determining asthma severity may differ between the sexes, and it should be further investigated.
Pathophysiological Role of Skin Mast Cells in Wound Healing after Scald Injury: Study with Mast Cell-Deficient W/WV Mice
Authors: Shiota Naotaka , Nishikori Yoriko , Kakizoe Eiichi , Shimoura Keiko , Niibayashi Tomomi , Shimbori Chiko , Tanaka Tetsuya , Okunishi Hideki
Published: August 2009

Background: The major role of mast cells in wound healing process has not been identified. In this study, we used mast cell-deficient W/WV mice and their congenic control (+/+) mice to examine the role of mast cells in scald wound healing. Methods: The size of the scald wound, thickness of the dermis, collagen deposition, vascularization, number of mast cells and chymase activity were measured before and at 3, 7, 14 and 21 days after inducing scald injury. Results: Although the process of wound closure and re-epithelialization was not markedly different between W/WV mice and +/+ mice, the degree of fibrous proliferation at the wound edge and wound vascularization in the proliferative phase was significantly lower in W/WV mice than in +/+ mice, and no vascular regression in the late remodeling phase was observed in W/WV mice. Mast cells producing chymase, FGF2, TGF-β1 and VEGF were highly accumulated at the edge of scald wound in +/+ mice during the proliferative and remodeling phases at days 14 and 21. Chymase activity in the injured tissues of +/+ mice decreased in the acute phase, but recovered to no-injury level at days 14 and 21. The number of mast cells and chymase activity were very low in the injured tissues of W/WV mice throughout the experiment. Conclusions: Wound healing after skin scald injury was partially impaired in mast cell-deficient mice. Mast cells may contribute to the wound healing process, especially in the proliferative and remodeling phases after scald injury.
Fusiform Aneurysm Model in Rabbit Carotid Artery
Authors: Reinald Nicoleta , Fournier Benjamin , Naveau Adrien , Couty Ludovic , Lemitre Mathilde , Seguier Sylvie , Coulomb Bernard , Gogly Bruno , Lafont Antoine , Durand Eric
Published: August 2009

Aims: To develop a reproducible and accessible model of elastase-induced fusiform aneurysm in carotid rabbit arteries. Methods: Elastase, at a concentration of 1–30 U, was incubated into the lumen of carotid rabbit arteries. Four weeks later, angiography, histomorphometry, immunohistochemistry and zymography were performed. Results: The optimal concentration of elastase in this model was 3 U according to the balance between mortality and thrombosis rates. Indeed, at 3 U, external carotid diameter increased from 1.9 ± 0.1 to 3.1 ± 0.4 mm (p < 0.0001) associated with degradation of elastic fibers, matrix metalloproteinase-9 secretion, apoptosis and macrophage infiltration. Conclusions: Our study underlines that abdominal aortic aneurysm can be reliably duplicated in an elastase-induced aneurysm in carotid artery, a much more accessible vessel.
Management of Esophageal Perforation: Experience from a Tertiary Center in India
Authors: Amudhan Anbalagan , Rajendran Shanmugasundaram , Vimal Raj Vellayudham , Rajarathinam Govindhasamy , Jyotibasu D. , Ravichandran Palanisamy , Jeswanth Satyanesan , Balachandar Tirupporur Govindaswamy , Kannan Devy Gounder , Surendran Rajagopal
Published: August 2009

Background/Aim: The management of esophageal perforations remains controversial in large part due to variability in etiology, time of presentation, location, and underlying esophageal disease. We reviewed our experience in treating patients with esophageal perforations and evaluated the etiology, management and outcome of intervention in a tertiary center. Methods: Between 1999 and 2007, 48 patients with esophageal perforation were treated in a tertiary referral center in southern India. Postdilatation corrosive stricture perforations constituted the major etiology. Results: Thirty (62.5%) patients were diagnosed early (<24 h) and the remaining 18 (37.5%) were late (>24 h). The 30-day mortality was 6.2%, and mean hospital stay was 13 ± 9.3 days. Comparing outcomes between early and late groups, statistically significant difference was observed, with increased mortality (p = 0.02) and hospital stay (p = 0.04) following late diagnosis. Conclusion: This report suggests that early diagnosis decreases mortality and hospital stay in esophageal perforation. Preservation of esophagus may be attempted while treating esophageal perforations.
A Prospective Bicenter Study Investigating the Diagnostic Value of Procalcitonin in Patients with Acute Appendicitis
Authors: Sand M. , Trullen X.V. , Bechara F.G. , Pala X.F. , Sand D. , Landgrafe G. , Mann B.
Published: August 2009

Background: Procalcitonin (PCT) is an established laboratory marker for disease severity in patients with infection and sepsis. In addition, PCT has been shown to be an effective marker for a limited number of localized infections. However, whether or not PCT has any diagnostic value for acute appendicitis, still remains unclear. The purpose of this prospective bicenter study was, therefore, to determine whether or not the PCT levels in the serum of patients with acute appendicitis have any diagnostic value. Methods: This prospective study included 103 patients who received an appendectomy, based on the clinical diagnosis of acute appendicitis, in a surgical department of an academic teaching hospital in Germany or in a county hospital in Spain. White blood cell count (WBC), C-reactive protein (CRP) and procalcitonin (PCT) values were determined preoperatively. All appendectomy specimens were sent for routine histopathological evaluation. Based on this information, the patients were assigned to 1 of 5 groups that reflected the severity of the appendicitis. Results: Of the 103 patients who were included in the study, 98 had appendicitis. Fourteen (14.3%) showed an increase in PCT values. Of those 14, 4 had a serum PCT >0.5 ng/ml, 9 had a PCT value >2–10 ng/ml and 1 had a PCT value >10 ng/ml. The sensitivity of PCT was calculated to be 0.14. The mean WBC value was 13.0/nl (± 5.2, 3.4–31), and for CRP it was 8.8 mg/dl (± 13, 0–60.2). The values of CRP, WBC and PCT increased with the severity of the appendicitis. Conclusions: PCT is potentially increased in rare cases of severe inflammation and, in particular, after appendiceal perforation or gangrenous appendicitis. However, its remarkably low sensitivity prohibits its routine use for the diagnosis of appendicitis.
Human Parvovirus B19 (B19V) Infection in Systemic Sclerosis Patients
Authors: Zakrzewska Krystyna , Corcioli Fabiana , Carlsen Karen M. , Giuggioli Dilia , Fanci Rosa , Rinieri Alessio , Ferri Clodoveo , Azzi Alberta
Published: August 2009

Background: Our previous reports suggested a possible association between parvovirus B19 (B19V) infection and systemic sclerosis (SSc), based on higher prevalence of B19V DNA in SSc patients in respect to controls. Methods: In the present study, to further evaluate the differences in the pattern of B19 infection in SSc, skin biopsies and bone marrow samples from patients and controls were analysed for B19V DNA detection, genotyping and viral expression. Results: B19V DNA was detected in skin biopsies from 39/49 SSc patients and from 20/28 controls. Bone marrow showed positive in 17/29 SSc patients, 5/21 haematological patients and 0/10 healthy controls. Genotype 1 was more frequent in skin and bone marrow from patients than from controls. Simultaneous persistence of 2 genotypes was detected in SSc skin and bone marrow samples, never in controls. Viral mRNA for capsid protein was detected in the skin of genotype 1-positive patients and not in control skins. Conclusion: The results outline some differences in the rate of persistence of B19V DNA, in the simultaneous persistence of 2 genotypes and in the pattern of viral expression among SSc patients and controls.
Complement Components, C1 Activation and Disease Activity in SLE
Authors: Sturfelt G. , Sjöholm A.G. , Svensson B.
Published: August 2009

Laboratory parameters were studied in 8 systemic lupus erythematosus patients during periods of high and low disease activity, mainly as defined by clinical criteria. Renal manifestations were present in 6 patients 5 of which showed antibodies to native DNA. C-reactive protein was raised in 3 patients. Only 1 of these showed a superimposed bacterial infection. Markedly high concentrations of CĪr-CĪs-CĪ inactivator complexes (CĪr-CĪs-CĪ IA) in the sera provided direct evidence of C1 activation independent of disease activity. During active disease, CĪr-CĪs-CĪ IA were correlated with C1q binding immune complexes as measured by solid phase, but not by fluid phase assay. Immunochemical concentrations of C1q, C4 and C3 and functional C2 were decreased in active SLE, consistent with sequential activation of the classical pathway. Discrepancies were noted between functional and immunochemical assays for C2 but not for factor B. Although essentially within the normal range, the levels of C1s, C4 binding protein, C5 and properdin were lower during active than during inactive disease. The concentrations of the factors B, I and H did not suggest involvement of the alternative pathway. 1 exceptional patient showed low factor B, a relative decrease of factor I and the presence of Bb fragments in plasma during active SLE. Markedly high factor D values were found. This could partly be explained by reduced renal function.
Effect of Agents Which Block Steroid Receptor-DNA Interaction on Anti-DNA Antibody Titers in Sera of Patients with Systemic Lupus Erythematosus
Authors: Kelly R.H. , Ostergren K.A. , Rosenbaum L.
Published: August 2009

We studied whether agents capable of blocking the interaction between steroid receptors and DNA could inhibit the serologic reactivity of anti-nuclear, and other, autoantibodies found in sera of humans with autoimmune disease. Conventional immunofluorescence procedures were used for both detection and titration of these autoantibodies. Parallel studies were conducted with sera diluted in buffers containing the inhibitory agents; KC1, Na2MoO4 or 1,10-phenanthroline, at optimal or suboptimal levels for blocking the steroid receptor-DNA interaction. In most cases these agents had no significant effect on the titer of autoantibodies present in human sera. However, there was one notable exception, that of autoantibodies to native DNA found in sera of patients’ with systemic lupus erythematosus and detected by indirect immunofluorescence using Crithidia luciliae kinetoplast DNA as substrate. Here these agents caused either total inhibition of serologic reactivity at optimal blocking concentrations, or a significant reduction in titer, in all twelve specimens studied. These results give rise to at least two possible interpretations: (1) antibodies to native DNA are unusual with respect to the ease with which their binding to antigen may be inhibited by these three chemically unrelated compounds, and (2) steroid receptors are somehow involved in the binding of these antibodies to native DNA.
A New Method of Testing Anti-Allergic Drugs
Authors: Koltai M.
Published: August 2009

Rats were sensitized with a single intraperitoneal dose of bovine serum albumin in alhydrogel plus Bordetella pertussis vaccine, and local anaphylaxis was elicited in the paw by soluble antigen 2 weeks later. The response was mainly due to IgE-type antibodies and proved to be highly sensitive to beta-adrenoceptor agonists. Dexamethasone inhibited the response after a lag phase. Methysergide, disodium chromoglycate, diethylcarbamazine, BW 775/c, nordihydroguajaretic acid, and FPL 55712 were also suppressive, while mepyramine was without effect. A synergism between methysergide and FPL 55712 was shown. Active local paw anaphylaxis appears to be adequate and easily applicable for large-scale screening of anti-allergic drugs.
LDH Isoenzyme Distribution in Human Eosinophils
Authors: De Simone C. , Ferrari M. , Sorice F.
Published: August 2009

The lactate dehydrogenase (LDH) isoenzyme pattern has been determined in human eosinophils isolated from the peripheral blood of healthy donors and patients with parasitic diseases. Almost equal LDH1 and LDH5 values appear to be a characteristic of the eosinophils obtained from healthy subjects. Eosinophilic granulocytes which have been isolated from the peripheral blood of patients with eosinophilia due to parasitic infestations have reduced values of LDH5 accompanied by higher LDH1 as compared with eosinophils from healthy donors. Centrifugation of cells through multiple discontinuous gradients of polyvinylpyrrolidone-coated silica particles made it possible to isolate distinct eosinophil subpopulations of differing densities which showed differences in cell surface markers, K-cell activity and also LDH patterns. Analysis of the lactate dehydrogenase isoenzyme patterns showed that EAG+ and EAC+ eosinophilic leukocytes are characterized by very low values of LDH5.
Studies in Immunodermatology
Authors: Dabski Krzysztof , Beutner Ernst H. , Jablonska Stefania
Published: August 2009

The presence of allospecificities of stratum corneum (SC) antibodies was suggested by the observation that 5 normal human sera tested by indirect immunofluorescence (IF) on 5 unrelated normal human skins yielded variable titers on some skin specimens. Titers of SC antibodies of normal sera absorbed with ground callus collected from one donor in IF tests on 5 unrelated normal human skin specimens provided further evidence that allospecific SC antibodies are present in human sera and that SC antigens in different human skins are not identical. Since SC antibodies in human sera do not give significantly different titers on autologous skin substrates as compared to homologous ones, they behave both like auto- and alloantibodies.
The Antigens of Pigeon Breeder’s Disease
Authors: Krause-Hooyman Laurel , Fredricks Walter W.
Published: August 2009

Two esterase activities were detected in pigeon dropping extracts (PDE). The first exhibited the ability to cleave p-tosyl-L-arginine methyl ester (TAMe), and the second utilized α-naphthyl acetate as its substrate. Both enzymes were stable at a temperature of 60°C. Although the enzymes could be separated from each other through gel filtration chromatography, both were associated with PDE1 (a major antigen found in pigeon droppings) throughout the fractionation procedure. In addition, it was shown that the TAMe esterase exhibited protease activity by virtue of its ability to hydrolyze α-casein.
Cross-Reactivity of Cat and Dog Allergen Extracts
Authors: Viander M. , Valovirta E. , Vanto T. , Koivikko A.
Published: August 2009

The commercial cat and dog allergen extracts are traditionally prepared from pelt, fur or dander. However, there is increased evidence of the allergenicity of saliva and urine of the animals. We have investigated 25 asthmatic children with a positive cat and/or dog RAST result. All 20 subjects with a positive cat RAST gave a positive skin prick test result to cat saliva, cat urine and cat hair. Analogously, all 20 subjects with a positive dog RAST had a positive skin reaction to dog saliva, urine and dander. In RAST inhibition experiments with dog and cat allergen discs, dog saliva appeared to be at least as potent as a commercial dog dander and hair extract, while cat saliva was less potent than the respective commercial extract. Both dog and cat salivas were clearly more potent than the respective urine. Significant cross-reactivity was observed between cat hair and dog dander in the RAST inhibition, whereas saliva and urine were shown to be more species-specific. An experimental dog dander preparation had about the same specificity as, and even higher allergenic activity than, that of dog saliva or urine. Our results suggest that saliva actually may be the best source of cat and dog allergen preparations. The importance of urine warrants further investigation.
Effect of Diet on Adjuvant-Induced Disease and Mitogenic Responses of Fisher Rats
Authors: Yiagou M. , Hadjipetrou-Kourounakis L.
Published: August 2009

Fisher rats from a inbred colony, when fed on a salt-free high-protein diet, developed only a mild arthritis after adjuvant injection. Their spleen cells failed to respond in vitro to concanavalin A (a T-cell mitogen), although they possessed a B-cell function of plaque formation to sheep red blood cells. When a full salt supplement was included in the diet, or magnesium or copper or zinc was included in the drinking water, adjuvant-induced arthritis was severe and the response to the T-cell mitogen was restored. The above results suggest that these trace elements may stabilize or activate certain cell populations needed for some immune responses in rats.
Experimental Asthma in Guinea Pigs Revisited
Authors: Kallós Paul , Kallós Liselotte
Published: August 2009

Experimental asthma in guinea pigs, as defined in 1937 by Kallós and Pagel, and the pertinent literature are reviewed. The relation between guinea pig asthma and human asthma bronchiale is established. The underlying mechanism in both processes is the misdirected (aberrant) formation of anaphylactic or reaginic (IgE) antibody against innocuous antigens, leading at renewed exposure to the antigen to release of mast cell mediators (such as histamine, SRS-A, eosinotactic peptides, PAF), bronchoconstriction, vasodilation and eosinophilic inflammation at the sites of antigen-antibody reactions in the tissues of the respiratory organs.
Experimental Erythrocyte Autoimmunity
Authors: Cox K.O. , Samcewicz Basia , Howles Anne
Published: August 2009

Mice injected with rat erythrocytes (RBC) produce RBC autoantibodies and antibodies against rat RBC. Transfer of spleen cells from autoantibody-producing mice to syngeneic recipients before the series of rat RBC injections causes a significant delay in autoantibody production although the response against rat RBC is elevated. Here it is shown that antibodies against rat RBC are markedly increased in recipients exposed to 350 rad of whole-body irradiation before transfer of spleen cells and the injections of rat RBC. In contrast, autoantibody production remained significantly suppressed. This shows that the anti-rat RBC response is regulated, at least in part, by cells in normal mice that are abrogated by 350 rad of whole-body irradiation.
The Effects of Cyclophosphamide on Delayed Hypersensitivity in Rats
Authors: de Brito F.B. , Hanahoe T.H.P.
Published: August 2009

Pretreatment of Tuck Wistar rats with cyclophosphamide (100 mg kg––1) intraperitoneally prevents the development of adjuvant-induced arthritis, experimental allergic encephalomyelitis and the delayed skin reactions after ovalbumin and Freund’s incomplete adjuvant. The resistance of these rats to these stimuli is similar to that found in untreated NR Wistar rats which are resistant to dextran. However, cyclophosphamide suppresses the responses of Tuck Wistar rats to collagen-induced arthritis and sheep red blood cell agglutination, stimuli to which untreated NR Wistar rats respond. Nevertheless, the reactions of Tuck animals to the purified protein derivative of M. tuberculosis, the skin sensitisation to oxazolone, and the development of the delayed reaction after ovalbumin and Freund’s complete adjuvant remain unchanged after cyclophosphamide pretreatment. The results indicate the possible existence of two separate types of delayed hypersensitivity reactions.
Mast Cell Secretion
Authors: Berlin Gösta , Enerbäck Lennart
Published: August 2009

Mast cell secretion is closely associated with extrusion of granules from the cell, and results in rapid release of both amines and heparin. Amines may also be released from intracellular granules communicating with the extracellular environment through channels formed by membrane fusions. We have recently reported the use of the fluorescent, cationic dye berberine as a vital stain marking amine-releasing granules located within the domains of the cell. The present report concerns early membrane events and the fate of such granules after termination of the secretory process. Purified mast cells were stimulated to a moderate secretory response by a low concentration of polymyxin B. The cells were then kept in a medium without the secretagogue. The fluorescence intensity of individual cells after vital berberine staining (reflecting the number of intracellular granules in contact with the extracellular environment) decreased rapidly after terminated mast cell secretion. The heparin content (reflecting the total number of intracellular granules) and the histamine content of the cells, both reduced by the secretory process, remained constant over a period of 1 h after terminated mast cell secretion. These findings suggest that mast cell membranes are rapidly restored after secretion, permitting selective amine release without accompanying release of heparin or other matrix components.
Staphylococcus aureus Inhibits Contact Sensitivity to Oxazolone by Activating Suppressor B Cells in Mice
Authors: Benedettini G. , De Libero G. , Mori L. , Campa M.
Published: August 2009

Killed Staphylococcus aureus strain Cowan I cells inhibit contact sensitivity to oxazolone in mice, when given intravenously 24–72 h before sensitization. With transfer experiments it was found that the cells responsible for the suppression are antigen-specific, nylon-adherent, resistant to antitheta serum + C, and sensitive to anti-mouse Ig serum + C. These suppressor B cells bear anti-oxazolone immunoglobulins and appear to exert their suppressive activity by preventing the contact sensitizer from reaching the specific reactive T cells.
Isolation, Cloning and Characterization of a Tyrosinase with Improved Activity in Organic Solvents from Bacillus megaterium
Authors: Shuster Vered , Fishman Ayelet
Published: August 2009

A tyrosinase-expressing bacterium was isolated from soil, and extracellular enzymatic activity was induced by the presence of tyrosine and CuSO4. Amplification of the 16S rDNA genes revealed a high similarity with Bacillus megaterium. The enzyme was over-expressed in Escherichia coli BL21 and purified using an affinity column. The tyrosinase was composed of 297 amino acids and was determined to be a monomer with a relative molecular mass of 31 kDa according to gel filtration. The Km values for 3,4-dihydroxy-L-phenylalanine (L-DOPA) and L-tyrosine were 0.35 and 0.075 mM, respectively, and the kcat/Km values were 28.9·103 and 32.9·103 (s–1·M–1). The maximum activity for both monophenolase and diphenolase was observed at 50°C and pH 7.0. Enzymatic activity was enhanced in the presence of 10–50% water-miscible organic solvents, which included ethanol, methanol, 2-propanol and dimethyl sulfoxide (DMSO). The activity in 30% DMSO was 170% of the activity in water and the enantioselectivity towards L-DOPA decreased by 40%. The residual activity following an incubation period of 17 h in 0–70% methanol was constant. This newly isolated and characterized tyrosinase may have potential applications in organic synthesis due to its high activity and stability at typically denaturing conditions.
Recruitment of Thymocytes from G1 into S Phase by a Thymic Factor
Authors: Söder O. , Ernström U.
Published: August 2009

A thymic factor previously shown to stimulate the DNA synthesis of immature thymocytes was found to recruit G1 cells synchronously into S phase within 1.5–2 h. The S + G2 + M duration of the cultured thymocytes was not affected. In the absence of the thymic factor, the responding thymocytes seemed to be blocked in G1 at a putative restriction point 1.5 h prior to S phase. When the addition of the thymic factor to cultured cells was delayed, the responsiveness rapidly declined, indicating that target cells were transferred into an unresponsive state. A stimulating effect of the DNA synthesis of cultured thymocytes was obtained also after a transient exposure to the thymic factor for 2–3 h. The stimulating activity was completely absorbed from the medium by incubation with thymocytes at both 4 and 37 °C, but not by lymph node lymphocytes which are unresponsive to the thymic factor. Our interpretation of the results is that the thymic factor acts on thymocytes in G1, triggering the responsive cells via membrane receptors to proceed from G1 into S phase. The thymic factor is proposed to be a progression factor for the rapidly cycling immature thymocytes and to be involved in the regulation of the intense thymic growth in vivo.
Protection against Experimental Cholera in the Rat
Authors: Lange Stefan , Lönnroth Ivar , Nygren Håkan
Published: August 2009

The importance of antibodies to cholera toxin (CT) versus desensitization of intestinal adenylate cyclase for protection against experimental cholera in the rat was investigated. Animals were immunized five times with CT either perorally or intravenously; antitoxic antibodies were measured in both serum and bile, and intestinal anti-CT-containing plasma cells (ACC) as well as eosinophilic leucocytes were counted. Both peroral and intravenous immunizations induced high levels of serum antibodies, while antibodies in bile appeared only after peroral immunization. The number of eosinophilic leucocytes in the intestinal mucosa increased in response to peroral immunization. CT-induced stimulation of intestinal adenylate cyclase was significantly suppressed, i.e., desensitized, after peroral immunization, but was equally stimulated in intravenous immunized and in control animals. A constant and long-lasting protection against CT-induced secretion was obtained after peroral immunization. This protection correlated neither to the concentrations of antitoxic antibodies in serum, nor to those in bile, nor to the number of ACC, but did correlate to the desensitization of adenylate cyclase.
In vitro Antigen-Induced Increase in Motility and Degranulation of Basophilic Granulocytes from Atopic Asthmatics, Studied by Microscopic Motion Pictures
Authors: Kimura Ikuro , Tanizaki Yoshiro , Sasaki Yoshihide
Published: August 2009

Increase in motility and degranulation of basophilic granulocytes by stimulation with antigen were studied by phase-contrast microscopic motion pictures. There was no difference in random movement between sensitized (atopic asthmatics) and nonsensitized basophilic granulocytes (healthy subjects). The incidence of basophilic granulocytes with oriented movement before antigen stimulation was significantly lower in atopic asthmatics compared to that in healthy subjects, but was significantly increased at 3–6 min and reached the peak at 12–15 min after addition of antigen. Degranulation of sensitized basophilic granulocytes began to occur at 3–6 min with the maximum at 12–15 min after antigen stimulation.
Analytic Study of the Differential Anticomplementary Effects of Dextran Sulphate and Heparin in the Assay for the Mouse Alternative Pathway
Authors: Klerx Jac P.A.M. , Van Dijk Hans , Van der Maaden Walter J. , Willers Jan M.N.
Published: August 2009

In a recent paper, a linkage between immunological adjuvant activity in mice and in vitro anticomplementary (alternative pathway assay) effects was described for different polyanions. This connection was found only if mouse serum was used as a complement (C) source. In order to investigate the possible role of C in adjuvant activity, the differential effects of polyanions on mouse C were studied in detail. For this study, substances with different activity were selected, namely dextran sulphate with strong C-regulatory and immunoadjuvant activities, and heparin, which was weakly anticomplementary and devoid of adjuvant effect. In general, studies of mouse C are complicated by the inavailability of isolation procedures for the C-components involved. This difficulty was circumvented by making use of C5-deficient serum and of the haemolytic activity of mouse membrane attack complexes formed in the fluid phase. With yeast cells as alternative pathway activators it was shown that the effect of heparin on this pathway was restricted to activation of the terminal route. In contrast, dextran sulphate also caused a functional decay of a yeast-bound alternative pathway C5-convertase and interfered with the haemolytic activity of fluid-phase membrane attack complexes as well. Further studies will be needed to decide whether these specific effects of dextran sulphate are related to the immunological adjuvant activity of the substance in mice.
Appearance of Mast Cells in Bone Marrow, Peripheral Blood and Spleen of Immunized Rats
Authors: Ahlstedt S. , Olaisson E. , Thellin J. , Björkstén B.
Published: August 2009

The appearance of mast cells and their progenitors was studied in rats. The animals were immunized according to a protocol utilizing 2-week periods of daily administration of small doses of ovalbumin either as aerosol or as subcutaneous injection without adjuvant. These procedures are known to induce allergic responses of the IgE-mediated type. The immunization procedures resulted in increased numbers of mature mast cells in bone marrow. There were also increased numbers of progenitors, as indicated by increased maturation of mast cells in cultures of bone marrow cells from immunized animals compared with nonimmunized controls. Bone marrow cells in cultures from immunized and nonimmunized animals showed similarly increased maturation of mast cell progenitors when stimulated with conditioned medium from mixed lymphocyte reaction cultures. The number of progenitors was considerably lower in peripheral blood and spleen than in bone marrow. However, cultures of cells from peripheral blood and spleen of aerosol-immunized rats contained cells which matured into mast cells after addition of medium from mixed lymphocyte reaction cultures. The study shows that in immune responses similar to those seen in allergic individuals mast cells are normal constituents.
Induced Regional Differences in Cutaneous Cell-Mediated Immunity in the Mouse
Authors: Green Jon A. , Spruance Spotswood L.
Published: August 2009

Single applications of 1-fluoro-2,4-dinitrobenzene (DNFB) or oxazolone to one ear of a mouse resulted in brisk local cell-mediated immunity (CMI) upon antigenic rechallenge of the same ear. However, only a minimal CMI reaction occurred when the other ear was treated with antigen. The relative unresponsiveness of the contralateral ear persisted for periods as long as 14 weeks. The depressed CMI of the contralateral ear was eliminated when cyclophosphamide was given prior to initial antigenic exposure, and therefore, appeared to be mediated by suppressor cells. These observations indicate that regional variations in the intensity of cutaneous CMI can be induced in mice. The significance of these observations is discussed.