bluuebunny/crossref_metadata_2025 huggingface dataset:
Patch Testing to Exclude Allergic Contact Dermatitis Caused by Povidone-Iodine
Authors: Kozuka Takehito
Published: April 2003

Povidone-iodine (polyvinylpyrrolidone-iodine, PVP-I) solution is an effective and safe disinfectant agent used throughout the world; however, prolonged exposure to PVP-I solution may cause irritation or rarely severe skin reactions. When PVP-I as is in the aqueous state is used for patch testing, a false-positive result is likely to arise due to irritation. Patch testing to exclude allergic contact dermatitis caused by PVP-I should be done with dried 10% PVP-I solution, 10% PVP-I gel, 8% sodium thiosulfuric acid and a mixture of 10% PVP-I solution and 8% sodium thiosulfuric acid.
Recombinant Marker Allergens: Diagnostic Gatekeepers for the Treatment of Allergy
Authors: Kazemi-Shirazi Lili , Niederberger Verena , Linhart Birgit , Lidholm Jonas , Kraft Dietrich , Valenta Rudolf
Published: April 2003

During the past decade an increasing number of recombinant allergens have become available, representing a significant proportion of the epitope complexity of natural allergen extracts. Component-resolved diagnosis with recombinant allergens reveals the antibody reactivity profile of allergic patients and identifies the disease-eliciting allergen molecules. This article exemplifies how recombinant allergen molecules with high cross-reactive potential can be used as marker allergens to identify allergic patients who are cross-sensitized to a variety of allergen sources. It further demonstrates how the use of allergens with a restricted distribution in a certain group of allergen sources may allow the identification of patients who have been genuinely sensitized by a particular allergen molecule. Drawing from those examples, it is suggested how diagnostic tests based on such recombinant marker allergens may be used to improve the choice and monitoring of currently available forms of specific immunotherapy.
Induction of a Proliferative Response of T Cells by a High-Molecular Antigen in Dermatophagoides farinae Feces
Authors: Tategaki Airo , Kawamoto Seiji , Hashimoto Kanako , Okuda Takahide , Aki Tsunehiro , Suzuki Osamu , Ono Kazuhisa , Shigeta Seiko
Published: April 2003

Background: We have previously demonstrated that high-molecular mite antigen (HM1) from Dermatophagoides farinae feces is an allergen which binds to mite-allergic patients IgE. HM1 also induced a proliferative response in lymph node cells from mite-immunized mice as well as nonimmunized mice. In the present study, we demonstrated that HM1 induced T cell proliferation and investigated the HM1-stimulated T cell proliferative pathways using nonallergic human peripheral blood mononuclear cells (PMBC). Methods: Blood samples were obtained from 10 healthy donors. Using primary culture, T cell response stimulated with HM1 was performed on purified T cells, CD19+ cell-depleted PBMC and CD11b+ cell-depleted PBMC. In addition, interleukin (IL)-5 and interferon (IFN)-γ produced by mite-allergic and healthy donors stimulated with HM1 were estimated by enzyme immunoassay. Results: T cell proliferation was detected only in CD19+ cell-depleted PBMC. When T cells were cocultured with CD11b+ cells they recovered their proliferative response to HM1. In addition, the pathway of HM1-stimulated T cell proliferation did not involve HLA class II restriction. Both activated CD11b+ cells and their conditioned media were needed to induce HM1-stimulated T cell proliferation. Furthermore, HM1 induced IFN-γ production in both healthy and allergic donors. Conclusion: The high-molecular mite antigen, HM1, induced a proliferative response of T cells in healthy as well as allergic donors, without HLA class II restriction. Our results suggest that further investigation of HM1 could constitute a valuable avenue of research into complex allergic diseases.
Elevated Nasal Mucosal G Protein Levels and Histamine Receptor Affinity in a Guinea Pig Model of Nasal Hyperresponsiveness
Authors: Chiba Yoshihiko , Saitoh Nobuyuki , Matsuo Kensuke , Misawa Miwa
Published: April 2003

Background: Nasal hyperresponsiveness is a common feature of allergic rhinitis, but the underlying mechanisms have yet to be elucidated. The effects of repeated antigen inhalation on the characteristics of histamine H1 receptors and expression levels of heterotrimeric guanosine 5′-triphosphate-binding proteins in nasal mucosa were investigated to understand the mechanisms of the pathogenesis of nasal hyperresponsiveness in allergic rhinitis. Methods: Male Hartley guinea pigs were sensitized by the inhalation of dinitrophenylated ovalbumin antigen (10 mg of protein/ml) and repeatedly challenged by inhaling aerosolized dinitrophenylated ovalbumin antigen for 3 weeks. Twenty-four hours after the last antigen inhalation, in vivo nasal responsiveness to histamine was measured. [3H]Mepyramine binding assays and immunoblotting for α subunits of the Gq protein were also performed using membrane preparations of isolated nasal mucosae. Results: The histamine-induced increase in intranasal pressure was significantly augmented after repeated antigen challenge, indicating that nasal hyperresponsiveness was achieved. In saturation binding studies, no significant change was observed in the density and antagonist affinity of H1 receptors in the hyperresponsive animals. On the other hand, the affinity of histamine for high-affinity agonist binding sites in the hyperresponsive group, measured by histamine competition binding studies, was much greater than that in control animals, and these results were affected by guanosine 5′-O-(3-thiotriphosphate) in both groups. Moreover, Gαq levels in nasal mucosal homogenates were significantly increased after repeated antigen challenge. Conclusions: Elevated G protein levels in nasal mucosa might induce an increased binding affinity of histamine to its receptors, resulting in an augmented nasal response to histamine, that is, nasal hyperresponsiveness, in guinea pigs.
Protection of Monkeys against Shiga Toxin Induced by Shiga Toxin-Liposome Conjugates
Authors: Suzaki Yuriko , Ami Yasushi , Nagata Noriyo , Naito Seishiro , Kato Hiroshi , Taneichi Maiko , Takahashi Motohide , Komiya Takako , Satoh Sachihiro , Gondaira Fumio , Sugiyama Junichi , Nakano Yoshio , Mori Masahito , Komuro Katsutoshi , Uchida Tetsuya
Published: April 2003

Background: We previously reported that the purified Shiga toxins (Stx) Stx1 and Stx2, when coupled with liposomes, induced substantial production of anti-Stx1 and anti-Stx2 IgG antibody, respectively, in mice. The levels of anti-Stx antibody in the sera of mice immune to Stx-liposome correlated well with the protection against subsequent challenge with Stx. Furthermore, mice immunized with a mixture of Stx1-liposome and Stx2-liposome were successfully protected against oral infection with cytotoxin-producing Escherichia coli O157:H7. Methods: In this study, the induction of protection against Stx2 by Stx2-liposomes was evaluated in monkeys. Results: Stx2-liposomes induced a substantial amount of anti-Stx2 IgG antibodies as well as Stx2 neutralizing antibodies in monkeys. Test monkeys were successfully protected against challenge with lethal doses of Stx2. Moreover, these monkeys showed no apparent symptoms, while nonimmunized control monkeys died within 4 days with hemorrhagic gastroenteritis and renal disorder. In addition, as shown by other cases involving antigen-liposome conjugates, Stx2-liposome did not induce anti-Stx2 IgE antibody production, though it stimulated the production of a substantial amount of anti-Stx2 IgG antibodies. Conclusion: These results suggest that Stx-liposome conjugates may serve as candidate vaccines to induce protection against death caused by cytotoxin-producing E. coli infection.
Substance P Induced Preprotachykinin-A mRNA, Neutral Endopeptidase mRNA and Substance P in Cultured Normal Fibroblasts
Authors: Bae Sang-Jae , Matsunaga Yoshitaka , Takenaka Motoi , Tanaka Yoichi , Hamazaki Yoichiro , Shimizu Kazuhiro , Katayama Ichiro
Published: April 2003

In certain skin diseases, stress can modulate the induction and/or progression of cutaneous manifestations. However, little is known about the circuit in neuroendocrine and in the immune systems of the skin. To address this question, we have analyzed the regulatory mechanisms of autocrine induction of substance P (SP) by cultured normal human fibroblasts that compose the major population of the skin and might augment stress-induced skin inflammatory responses. In nonstimulated conditions, normal fibroblasts express a moderate amount of preprotachykinin-A (PPT-A), a precursor of SP mRNA, and exogenous SP significantly upregulated PPT-A mRNA expression. Maximum response of SP peptide and SP mRNA in fibroblasts was observed 1–3 h after stimulation with SP. In contrast, the expression of neutral endopeptidase (NEP), a cell surface peptide with hydrolyzing activity of SP, was increased in fibroblasts stimulated with SP after 24 h. The administration of NEP inhibitor (phosphoramidon) to the fibroblasts induced higher SP production. In addition, the neurokinin (NK) receptor antagonists (spantide, FK224 and FK888) and protein synthesis inhibitor (cycloheximide) inhibited SP production by 30–40% of control response. In immunostaining study, specific cytoplasmic staining of SP was observed in fibroblasts stimulated with SP. Finally, we confirmed that the nucleotide sequence of the PPT-A expressed in fibroblasts perfectly corresponded to the gene bank human PPT-A cDNA. This is the first report that SP mRNA, NEP mRNA and SP peptide can be induced by normal human skin fibroblasts in response to exogenous SP, and that fibroblast-derived SP might play an important role in the induction and acceleration of certain cutaneous diseases.
Oral Cancer: Prevention and Detection
Authors: Joseph Bobby K.
Published: April 2003

Researchers in oral cancer agree that the early diagnosis of oral carcinoma greatly increases the probability of cure with minimum impairment and deformity. Primary prevention which involves reducing the exposure to tobacco, alcohol and betel quid has been shown to be effective in reducing the incidence of oral cancer. Secondary prevention involves screening for the early detection of oral cancer. Oral cancer screening can take many forms. Clinical examination and biopsy allow the early detection of premalignant and early oral cancers. Screening can be made more efficient by inspecting high-risk sites – the floor of the mouth, the ventrolateral surface of the tongue and the soft palate. Due to the cost of population screening, it is advisable to initially target high-risk groups, those over 40 years of age, including smokers and heavy drinkers. It is recommended that dentists perform an annual visual oral cancer examination on all their patients and obtain a specialist opinion for suspicious oral lesions. OraTest with toluidine blue may be used as an adjunct to soft tissue examination to highlight any invisible, asymptomatic lesions. Exfoliative cytology can detect early oral cancer and can be performed by dentally untrained personnel. It is rapid and relatively non-invasive and therefore may be useful in population-based oral cancer screening programmes. Recently, based on various studies, the oral CDx brush biopsy technique has been proposed as a highly accurate method of detecting oral precancerous and cancerous lesions. More frequent oral cancer examinations are recommended for treated oral cancer patients to monitor the development of secondary tumours. Family members of patients with oral cancer are also at high risk and therefore should be examined more frequently. Whatever screening method is used, a positive screening result must be confirmed by biopsy. A public awareness programme that stresses the importance of at least one annual dental examination, identification of warning signs of oral cancer and recognition of the hazards of tobacco and alcohol use is necessary to reverse the high morbidity and mortality rates associated with this disease. In the future, the identification of oncogene and tumour suppressor gene mutations in biopsy specimens may give a clearer indication of the likely behaviour of suspicious oral lesions.
Oral Health in Kuwait before the Gulf War
Authors: Behbehani Jawad M. , Shah Nasra M.
Published: April 2003

Objective: The aim of this oral health survey was to determine the oral health status and oral health behavior in the whole population in Kuwait. This survey was part of the Kuwait Health Survey, which was conducted by the Ministry of Public Health. The original data were destroyed during the Gulf War (1990/1991), but the aim of this report is to publish the main findings concerning oral health and related factors. Methodology: The data were collected between April 1984 and April 1985. The sample consisted of 3,358 households and 26,530 individuals. The survey consisted of interviews and clinical examinations (>12-year-olds). WHO (1977) criteria were used for examinations. Results: Females were more often brushing their teeth at least once a day than males and non-Kuwaitis slightly more often than Kuwaitis. Over one third (39%) had visited a dentist during the previous 12 months. The proportion of subjects with soft deposit was 66%, calculus 45%, intensive gingivitis 46% and advanced periodontitis 18%. The caries experience was prevalent (52%) in primary dentition (<8-year-olds) and in permanent dentition, highest (95%) among the 60- to 64-year-olds. Conclusion: Preventive programs for periodontal diseases and for dental caries are urgently needed. The coverage of curative care of both diseases also needs to be improved. Health behavior improvement should be targeted by oral health promotion activities.
A Case against Spiral Ligament Atrophy as a Cause of Ménière’s Disease
Authors: Harris Jeffrey P. , Keithley Elizabeth M.
Published: April 2003

The etiology of Ménière’s disease and endolymphatic hydrops has been very difficult to determine. In order to develop rational therapies for patients with Ménière’s disease the etiology needs to be established. In the current study an examination was made of the spiral ligament in temporal bones from individuals with Ménière’s and endolymphatic hydrops and normal control temporal bones. No evidence of atrophy in either the length or width or the fibrocyte cell density was seen in the temporal bones with Ménière’s disease. The role of the spiral ligament in the development of endolymphatic hydrops is still unclear.
A Case of Cutaneous Delayed-Type Allergy to Oral Dexamethasone and to Betamethasone
Authors: Nucera Eleonora , Buonomo Alessandro , Pollastrini Emanuela , De Pasquale Tiziana , Del Ninno Massimiliano , Roncallo Chiara , Schiavino Domenico , Patriarca Giampiero
Published: April 2003

Corticosteroids are drugs that may cause allergic contact dermatitis, but systemic allergic reactions to these drugs are rare. A 29-year-old man developed a maculopapular rash during an oral therapy with betamethasone. Patch tests demonstrated a delayed-type allergy to dexamethasone, betamethasone and fluocortolone. Oral, intramuscular or topical provocation tests with other corticosteroids – deflazacort, hydrocortisone, methylprednisolone, fluticasone dipropionate, triamcinolone and prednisone – were all negative. This demonstrates that a patient with a systemic allergy to a group of corticosteroids can tolerate those of other groups.
Suppression of Puberty in Girls with Short-Acting Intranasal versus Subcutaneous Depot GnRH Agonist
Authors: Tuvemo Torsten , Gustafsson Jan , Proos Lemm A.
Published: April 2003

Background/Aim: Central precocious puberty (CPP) is more common in females than in males. During the last few decades a new group of patients with CPP has been seen frequently in Northern Europe, namely children adopted from developing countries. GnRH analogue preparations, administered either as intranasal spray or as depot preparations, are the drugs of choice for inhibiting the release of gonadotropins. The aim of this study was to compare the effect of buserelin given by intranasal spray with that of the same compound given as a subcutaneous depot preparation. Methods: The study group comprised 46 pubertal girls below the age of 9.5 years, adopted from a developing country. During the first 2 years, the treatment used was buserelin acetate 300 µg 6 times daily as a nasal spray. During the third year the treatment was changed to Suprefact Depot, 6.3 mg, given as a subcutaneous implant every 8 weeks. Half of the girls were randomized to growth hormone treatment in addition to the pubertal inhibition. Results: GnRH provocation tests after 6 weeks, 1 year and 2 years of treatment with intranasal GnRH analogue showed suppression of gonadotropin secretion except in 1 case of noncompliance. During the third year, when the long-acting depot preparation was used, suppression was more pronounced. The peak LH response, especially, was considerably lower than during treatment with the nasal spray preparation. In all cases the clinical inhibition of puberty was adequate both during the first two years and during the third year. Conclusion: Even though the clinical suppression of puberty was adequate with both modes of administration, the effect of the depot preparation, in this study Suprefact Depot, was more pronounced in terms of gonadotropin suppression and less dependent on patient compliance.
Pulsatile Gonadotropin-Releasing Hormone Treatment of Men with Idiopathic Hypogonadotropic Hypogonadism
Authors: Christiansen Peter , Skakkebæk Niels E.
Published: April 2003

Objectives/Methods: To induce testicular growth and spermatogenesis, 11 patients with idiopathic hypogonadotropic hypogonadism were treated with long-term subcutaneous pulsatile gonadotropin-releasing hormone (GnRH) administration. Three patients had a history of undescended testes. Patients who did not respond to therapy with a sufficient increase in serum testosterone or spermatogenesis were offered additional injections with hCG or, after discontinuation of GnRH, either combined therapy with hCG and hMG or recombinant FSH. Results: During treatment testicular volume and serum levels of FSH, LH and testosterone increased. Semen analysis revealed the presence of spermatogenesis in 9 of the 11 patients (8 on GnRH alone and in 1 when hCG/hMG was subsequently instituted), and 7 pregnancies have resulted thus far. Conclusion: Pulsatile GnRH therapy is a well-tolerated and effective therapy for the induction of spermatogenesis in some men with idiopathic hypogonadotropic hypogonadism. It appears that a significant fraction of them should be treated for a minimum of 1–2 years to maximize testicular growth and achieve spermatogenesis. Cryptorchidism was a negative prognostic factor.
Induction of Puberty in the Hypogonadal Girl – Practices and Attitudes of Pediatric Endocrinologists in Europe
Authors: Kiess W. , Conway G. , Ritzen M. , Rosenfield R. , Bernasconi S. , Juul A. , van Pareren Y. , de Muinck Keizer-Schrama S.M.P.F. , Bourguignon J.-P.
Published: April 2003

The management of children and adolescents with hypogonadism and in particular the induction of puberty in the hypogonadal girl is subject to controversy. Therefore, under the auspices and through organization of the Drugs and Therapeutics Committee of the European Society of Paediatric Endocrinology (ESPE), an interactive voting session and workshop was held at the 39th ESPE Annual Meeting in Brussels to discuss these topics. Common practice in Europe and attitudes of pediatric endocrinologists in Europe were questioned and recorded in the 1.5-hour program. We now report on some of the results of the questionnaires and discussions of that session to further the discussion on and knowledge of current concepts of induction of puberty in the hypogonadal girl in Europe. It became clear from the data accumulated here that the start of treatment, the aims of therapy and the modalities of how to treat the hypogonadal girl vary amongst pediatric endocrinologists in Europe. For example, a chronological age ≧11 years was considered appropriate for the start of estrogen therapy by 40.4% (out of 188 answers), while 47.8 and 7.5% felt that a chronological age ≧13 and ≧15 years respectively was appropriate. In respect to the form and route of estrogen administration, the audience was asked for their common estrogen replacement practice: 31.9% used oral 17β-estradiol treatment, while 10% would prescribe 17β-estradiol transdermal patches. Another 12.2% would recommend conjugated estrogens (e.g. Premarin®) orally, 4.8% use oral estradiol valerate and 39.3% ethinylestradiol orally. Only 1.8% out of 229 physicians answering were undecided. In addition, counseling of patients and their families is quite variable and perceptions for example regarding potential pregnancies in affected women are also not uniform. In this report the authors do not want to provide their own personal views but rather reflect current practice in Europe. It is hoped that a more uniform picture will emerge once European and international guidelines on how to treat the girl with hypogonadism will be available and even more discussions amongst doctors from different countries have been led.
Effect of Subclinical Hypothyroidism and Obesity on Whole-Body and Regional Bone Mineral Content
Authors: Bertoli A. , Fusco A. , Andreoli A. , Magnani A. , Tulli A. , Lauro D. , de Lorenzo A.
Published: April 2003

Objective: The present investigation was aimed to evaluate the effect of subclinical hypothyroidism and obesity on bone mineral content (BMC) in different body segments. Methods: Thirty-two premenopausal women (age: 37 ± 9.9 years), with a wide range in body mass index (BMI), were studied. Subclinical hypothyroidism was defined by a basal TSH ≧4 µU/l and/or a TRH-stimulated peak ≧30 µU/l. For each subject, weight, height, BMI (weight/height2) and the waist/hip ratio were measured. Total BMC, total bone mineral density (BMD), leg BMC, leg BMD, trunk BMC, trunk BMD, arm BMC and arm BMD were determined using dual-energy X-ray absorptiometry. Thyroid function (basal and TRH-stimulated TSH, free T3 and free T4) were determined from fasting blood samples for all subjects. Results: Anova was conducted within all the groups to observe the effect of thyroid status and/or obesity on BMC and BMD. There was no statistical difference for age. Total BMC was affected by obesity (p < 0.05) but not by thyroid status, BMD of the legs was significantly influenced both by thyroid function and obesity (p < 0.01); total BMD was affected by hypothyroid status (p < 0.05). A direct relationship between leg BMD and TSH was demonstrated. Conclusion: Subclinical thyroid hypofunction and obesity seem to affect BMD differently in the body segments. An influence of gravitational force seems necessary in order to make evident the effect of subclinical hypothyroidism on bone. A condition of subclinical hypothyroidism should be considered when evaluating subjects for osteoporosis, since a BMD measured at the femoral neck may induce underestimation of initial osteoporosis.
Functional IL-18 Is Produced by Primary Pancreatic Mouse Islets and NIT-1 Beta Cells and Participates in the Progression towards Destructive Insulitis
Authors: Frigerio Simona , Holländer Georg A. , Zumsteg Urs
Published: April 2003

Background: Preclinical stages of type 1 diabetes are characterized by infiltrating T cells and by the peri- and intra-islet accumulation of pro-inflammatory mediators, such as IFNγ. Methods/Results: Using quantitative PCR we demonstrated that mRNA specific for the IFNγ-inducing cytokine IL-18 is upregulated in NIT-1 beta cells and intact mouse islets upon exposure to IL-1β, IFNγ and TNFα. The biological activity of IL-18 was blocked using caspase inhibitors and anti-IL-18 antibodies. Increased IL-18 expression was also detected in islets during advanced stages of insulitis and correlated with elevated transcripts for IFNγ and for the IL-18 receptor. Conclusion: Thus, beta cells produce bioactive IL-18 in the course of insulitis and actively contribute to the exacerbation of inflammation leading to their own demise.
Long-Term GH Therapy: Epidemiology and Auxologic Outcome
Authors: Audí L. , Gilabert A. , Lloveras G. , Martí-Henneberg C. , Rodríguez-Hierro F. , Vilardell E. , Webb S. , Canela J. , Sedano E. , Vicens-Calvet E.
Published: April 2003

Objectives: Epidemiologic and auxologic characteristics of patients treated with GH during childhood and adolescence and entered in a national registry in Catalonia were studied between 1988 and 1997. At the end of 1997, prevalence was 53.2 treatments/100,000 inhabitants aged 0–14 years. Maximum annual incidence rates were observed in 1990 and 1991 (34.0–35.6 cases/100,000 inhabitants aged 0–14 years). Study Design: Analysis of treatments terminated in 1993 (n = 548) revealed, for the three principal reasons for cessation of treatment (‘near-final height’, ‘adequate height but further growth potential’, and ‘poor growth response’), that males began and ended treatment at older ages with a better auxologic situation in SDS than girls at the beginning and end of therapy in the first two subgroups, with a similar duration of therapy. Severe GH deficiency (GHD) [both multiple pituitary hormone deficiency (MPHD) and the most severe isolated GHD (IGHD-A)] was more frequent in the group ending treatment at ‘near-final height’, whereas cessation of therapy because of ‘poor growth response’ was more frequent in the group with ‘other causes of short stature’ and no demonstrable GHD by routine tests. In the near-final height group, after excluding Turner’s syndrome, MPHD and GHD cases secondary to brain tumors and GH deficiencies associated with malformative syndromes, positive linear correlations were observed between HSDS at the end of treatment and HSDS at the beginning, predicted adult height SDS (PAHSDS) and target height SDS (THSDS). Multiple regression analysis showed that in this group of patients, 41.4% of the variability in HSDS increment can be explained by the equation: HSDS increment = –0.33 + 0.29 THSDS – 0.68 HSDS at the beginning of treatment. Results: The outcome showed a reasonable use of GH, since good-response cases generally continued treatment until final height whereas therapy was suspended in doubtful cases.
Testing Association of Statistically Inferred Haplotypes with Discrete and Continuous Traits in Samples of Unrelated Individuals
Authors: Zaykin Dmitri V. , Westfall Peter H. , Young S. Stanley , Karnoub Maha A. , Wagner Michael J. , Ehm Margaret G.
Published: April 2003

There have been increasing efforts to relate drug efficacy and disease predisposition with genetic polymorphisms. We present statistical tests for association of haplotype frequencies with discrete and continuous traits in samples of unrelated individuals. Haplotype frequencies are estimated through the expectation-maximization algorithm, and each individual in the sample is expanded into all possible haplotype configurations with corresponding probabilities, conditional on their genotype. A regression-based approach is then used to relate inferred haplotype probabilities to the response. The relationship of this technique to commonly used approaches developed for case-control data is discussed. We confirm the proper size of the test under H₀ and find an increase in power under the alternative by comparing test results using inferred haplotypes with single-marker tests using simulated data. More importantly, analysis of real data comprised of a dense map of single nucleotide polymorphisms spaced along a 12-cM chromosomal region allows us to confirm the utility of the haplotype approach as well as the validity and usefulness of the proposed statistical technique. The method appears to be successful in relating data from multiple, correlated markers to response.
Detecting Gene-Gene Interactions Using Affected Sib Pair Analysis with Covariates
Authors: Holmans Peter
Published: April 2003

Interest has recently focussed on allowing for interactions between loci as a way to increase power to detect linkage. In this paper, a simplified logistic regression method was used to perform affected sib pair analyses allowing for the inclusion of data from other loci. A systematic search of two-locus disease models was carried out to determine the situations in which this was advantageous. If IBD information is available (e.g. from a genome scan), it is unlikely that allowing for interactions will give a large lod score in the absence of linkage evidence from sinlge-locus analysis. Furthermore, allowing for interactions rarely gave a significant increase in power to detect linkage over a single-locus analysis, except for heterogeneity models with low KP. Conversely, the availability of disease-associated genotypes may greatly increase the power both to detect linkage to a second locus and interaction between the loci. These results indicate that when only IBD information is available, two-locus analysis of genome scan data should be restricted to regions giving peaks under single-locus analysis. If disease-associated genotypes are available, it may be worth re-analysing the whole genome.
Epicutaneous Application of Protein Antigens Incorporated into Cosmetic Cream Induces Antigen-Nonspecific Unresponsiveness in Mice and Affects the Cell-Mediated Immune Response
Authors: Ptak Włodzimierz , Szczepanik Marian , Bryniarski Krzysztof , Tutaj Monika , Ptak Maria
Published: April 2003

Background: Protein antigens applied epicutaneously by the patch method induce allergic dermatitis mediated by IgE antibodies in mice and simultaneously significant suppression of Th1-mediated delayed-type hypersensitivity (DTH) reactions. Methods: We developed a method in which protein antigens (calf collagen, elastin, keratin, TNP-substituted mouse Ig) were homogenized with neutral cream. Animals treated epicutaneously with such preparations were tested for contact sensitivity to TNP hapten or DTH hypersensitivity to hemocyanin. Antigen specificity of induced unresponsiveness was tested in vivo in ‘transfer-in’ and ‘transfer-out’ experiments. The influence of skin-induced regulatory cells on in vitro [3H]thymidine uptake by immune cells as well as the possible mode of their action using anticytokine antibodies were tested. Results: Our procedure in which different protein antigens are applied on the skin in the form of cream induces a state of antigen-nonspecific unresponsiveness affecting cell-mediated immune responses in mice. Cream alone has no such effect. Suppression is transferable in vivo by TCR-αβ lymphocytes, while Tγδ cells show no activity. In vitro, lymphoid cells of skin-tolerized mice suppress [3H]-thymidine incorporation by immune cells and this effect can be abolished by adding anti-TGF-β but neither anti-IL-4 nor anti-IL-10 antibodies. Conclusions: Lack of antigen-specifity of unresponsiveness induced by epicutaneous deposition of cream containing protein antigens resembles ‘determinant spreading’ in oral tolerance induced by antigen feeding. This may suggest that similar immunoregulatory mechanisms operate on these two bodily surfaces.
Modification of Peanut Allergen Ara h 3: Effects on IgE Binding and T Cell Stimulation
Authors: Rabjohn Pat , West C. Michael , Connaughton Cathie , Sampson Hugh A. , Helm Ricki M. , Burks A. Wesley , Bannon Gary A.
Published: April 2003

Background: Peanut allergy is a major health concern due to the increased prevalence, potential severity, and chronicity of the reaction. The cDNA encoding a third peanut allergen, Ara h 3, has been previously cloned and characterized. Mutational analysis of the Ara h 3 IgE-binding epitopes with synthetic peptides revealed that single amino acid changes at critical residues could diminish IgE binding. Methods: Specific oligonucleotides were used in polymerase chain reactions to modify the cDNA encoding Ara h 3 at critical IgE binding sites. Four point mutations were introduced into the Ara h 3 cDNA at codons encoding critical amino acids in epitopes 1, 2, 3 and 4. Recombinant modified proteins were used in SDS-PAGE/Western IgE immunoblot, SDS-PAGE/Western IgE immunoblot inhibition and T cell proliferation assays to determine the effects of these changes on in vitro clinical indicators of peanut hypersensitivity. Results: Higher amounts of modified Ara h 3 were required to compete with the wild-type allergen for peanut-specific serum IgE. Immunoblot analysis with individual serum IgE from Ara-h-3-allergic patients showed that IgE binding to the modified protein decreased ∼35–85% in comparison to IgE binding to wild-type Ara h 3. Also, the modified Ara h 3 retained the ability to stimulate T cell activation in PBMCs donated by Ara-h-3-allergic patients. Conclusions: The engineered hypoallergenic Ara h 3 variant displays two characteristics essential for recombinant allergen immunotherapy; it has a reduced binding capacity for serum IgE from peanut-hypersensitive patients and it can stimulate T-cell proliferation and activation.
Rat Monoclonal Anti-Murine IgE Antibody Removes IgE Molecules Already Bound to Mast Cells or Basophilic Leukemia Cells, Resulting in the Inhibition of Systemic Anaphylaxis and Passive Cutaneous Anaphylaxis
Authors: Miyajima Hiroaki , Watanabe Naohiro , Ovary Zoltan , Okumura Ko , Hirano Takao
Published: April 2003

IgE plays a central role in allergic reactions. Some anti-IgE antibodies (HMK-12, 6HD5) inhibit the binding of IgE to the FcΕRI of mast cells/basophilic leukemia cells (PT-18, RBL/2H3), but less inhibition is seen with the anti-allotypic JKS-6 and the anti-idiotypic Eb-1. Anti-IgE HMK-12 can detach bound IgE molecules from the FcΕRI. When mast cells or basophils were incubated with monoclonal anti-DNP-IgE SPE-7, washed and treated with anti-IgE HMK-12, anti-IgE/IgE complexes were found in the supernatant. Similar results were obtained with the Fab fragment of HMK-12. Mice injected with anti-DNP-IgE SPE-7 and later with DNP-BSA had the typical systemic anaphylactic shock. However, if they were injected with the anti-IgE antibody (HMK-12) before the challenge, they did not get an anaphylactic shock. In the sera of mice injected with monoclonal IgE SPE-7 and anti-IgE antibody (HMK-12), IgE/anti-IgE complexes were detected. No passive cutaneous anaphylaxis occurred if the rats were injected with anti-IgE antibodies before the challenge. In summary, anti-IgE antibodies can remove IgE antibodies from the FcΕRI; anti-IgE/IgE complexes can be detected in vitro and in vivo, and anti-IgE antibodies can inhibit IgE-mediated systemic or local anaphylactic reactions.
Serum Eosinophil Cationic Protein, Myeloperoxidase, Tryptase, Eotaxin and Th2-Like Cytokines in Dermatitis herpetiformis
Authors: Caproni Marzia , Cardinali Carla , D’Agata Alberino , Selvaggi Walter , Fabbri Paolo
Published: April 2003

Background: Dermatitis herpetiformis (DH) is a subepidermal bullous disease characterized by a neutrophilic and eosinophilic infiltrate, together with activated lymphocytes that show preferential polarization towards the Th2-like phenotype. To date, the role of neither soluble mediators, such as cytokines, nor serum levels of myeloperoxidase (MPO), eosinophil cationic protein (ECP) and tryptase (TRY) have been studied in the peripheral blood of DH. In this study we investigated 40 DH subjects for possible alterations in MPO, ECP, TRY, eotaxin, interleukin (IL) 4 and IL-5 concentrations to evaluate the role of neutrophils, eosinophils and mast cells in the induction of skin lesions. Sera from 20 healthy subjects were tested as controls. Methods: Eotaxin, IL-4 and IL-5 serum contents were determined by a solid-phase enzyme-linked immunosorbent assay. A double antibody radioimmunoassay was used to measure serum levels of MPO, ECP and TRY. Results: Normal levels of eotaxin and IL-4 were found in all DH subjects; increased IL-5 levels were found only in 1 subject. The MPO levels were significantly higher in DH subjects than in healthy controls (p < 0.01). There was also a significant difference between the serum ECP levels of DH and healthy controls (p < 0.05). TRY was not revealed in the serum of DH subjects. Conclusions: The determination of Th2-like cytokines and TRY serum levels is not important in quantifying the involvement of activated lymphocytes and mast cells in DH; however, the finding of high MPO and ECP serum levels seems to confirm that these products are good markers of the presence of activated neutrophils and eosinophils.
Endotoxin-Free Purification of Monocytes for Dendritic Cell Generation via Discontinuous Density Gradient Centrifugation Based on Diluted Ficoll-Paque Plus®
Authors: Lehner M. , Holter W.
Published: April 2003

Background: For the investigation of the biology of human monocyte-derived dendritic cells (DCs), and also for further clinical use of these cells, it is important to separate large numbers of monocytes without isolation-induced activation. Methods: Monocytes were purified from mononuclear cells (MNCs) by density gradient centrifugation using Ficoll-Paque Plus® adjusted to a density of 1,068 g/ml by dilution with phosphate-buffered saline (PBS) and compared to Percoll® isolated monocytes. DCs were generated by culturing monocytes with interleukin (IL)-4 plus granulocyte-macrophage colony-stimulating factor (GM-CSF), and they were functionally tested for IL-12 p70 production by stimulation with lipopolysaccharide (LPS) plus interferon-γ (IFN-γ). Results: Density gradient centrifugation using diluted Ficoll-Paque Plus® resulted in a monocyte purity of more than 75% and a recovery of 70%. DCs derived from such monocytes produced high amounts of IL-12 p70 in response to LPS plus IFN-γ, thus demonstrating an important functional capacity. Conclusion: Since Ficoll-Paque Plus® is commercially available in an endotoxin-free form, this modified Ficoll gradient offers a simple modality for isolating large amounts of monocytes under endotoxin-free conditions without introducing further potential monocyte-activating agents or procedures.
Prevalence of Primary Helicobacter pylori Resistance to Several Antimicrobials in a Saudi Teaching Hospital
Authors: Eltahawy A.T.
Published: April 2003

Objective: To determine the resistance rate of the most commonly used antimicrobial agents amongst Helicobacter pylori isolates. Methods: The agar disk diffusion method (Kirby Bauer) was utilized to determine the susceptibility of 223 H. pylori strains isolated before treatment. Isolates were tested against metronidazole (5 µg), clarithromycin (15 µg), amoxycillin (10 µg), and tetracycline (30 µg). Results: The resistance rate was 80% for metronidazole and 4% for clarithromycin. Tetracycline and amoxycillin showed very low degree of resistance with 1 (0.4%) and 3 (1.3%) of the strains resistant to these antibiotics, respectively. Age, sex and ethnicity had a remarkable effect on the resistance rate. Conclusion: The results indicate that metronidazole and clarithromycin should not be used as the only antimicrobial agents in the treatment of H. pylori infection. Susceptibility testing using the disk diffusion method is cost-effective in the screening of antimicrobial resistance against H. pylori.
Nasal and Bronchial Histamine Responsiveness in Pollen-Exposed Patients with Seasonal Rhinitis
Authors: Ehnhage Anders , Kölbeck Karl-Gustav , Mossberg Björn , Juto Jan-Erik
Published: April 2003

In a previous study, we found an increased nasal responsiveness as measured by rhinostereometry and histamine challenge out of season in a sample of 12 patients suffering mainly from hay fever. The aim of the present study was to investigate whether airway responsiveness in these patients was further increased after direct pollen exposure, after a single nasal pollen provocation as well as by repeated exposure during the pollen season. In spite of increased allergic symptoms, the basal degree of nasal mucosal swelling was unchanged before histamine challenge under these circumstances. After histamine challenge, nasal mucosal swelling was increased in the same way over the seasons. Also bronchial responsiveness was unchanged during the pollen season. It correlated to frequent sneezing following nasal histamine challenge during the season (p = 0.0071, r = –0.74). We interpret the results as an indication of a continuos airway inflammation regardless of season in these patients with pollen allergy, with acute symptoms added on direct exposure to the allergen.
Polymorphisms of Arylamine N-Acetyltransferase (NAT1 and NAT2) and Larynx Cancer Susceptibility
Authors: Varzim Graça , Monteiro Eurico , Silva Regina , Pinheiro Carlos , Lopes Carlos
Published: April 2003

Our aim was to examine the role of NAT1 and NAT2 polymorphisms in human larynx cancer susceptibility. Genotype tests for NAT1 alleles *4, *10 and *11, and NAT2 alleles *4, *5, *6A and *7A, using PCR-RFLP analysis, were performed in 172 healthy Portuguese individuals and 88 patients with squamous cell carcinoma of the larynx. NAT1 and NAT2 genotype frequencies were correlated between patients and control groups, using the chi-square test. Odds ratios and 95% confidence intervals were calculated from 2 × 2 tables with the Fisher’s exact model. The statistical analysis of NAT1 and NAT2 genotype frequencies revealed a significant difference of NAT1*10/*11 (p = 0.038) and NAT2*5/*7 (p = 0.003) genotype distribution between cases and controls. We also observed differences concerning tumor location, since NAT1*10/*11 genotype frequency was significantly different when comparing normal control individuals with the glottic subgroup of patients. The present data suggest that NAT1 and NAT2 polymorphisms may be correlated with an increased risk of larynx cancer.
Exclusive Surgery versus Postoperative Radiotherapy for Supraglottic Cancer
Authors: Esposito Erik , Motta Sergio , Motta Gaetano
Published: April 2003

Aim of the Study: To verify the effectiveness of prophylactic postoperative radiotherapy for supraglottic cancer. Patients and Methods: 97 patients underwent supraglottic horizontal laryngectomy with bilateral neck dissection: 35 patients (group A) received postoperative radiotherapy (60–70 Gy, 2 Gy fractions daily); 62 patients (group B) received only surgery. Results: Overall 5-year actuarial survival and corrected actuarial survival rates were 74 and 90% in group A and 61 and 80% in group B (p = 0.2 and 0.4, respectively). As for tumor extent, no significant differences were observed between the two groups. In N0 patients overall actuarial survival rate was significantly higher in group A as compared to group B (p = 0.01); most likely this difference was due to errors in clinical staging for the presence of reactive lymphadenitis and micrometastases. Conclusions: The present study did not document the effectiveness of postoperative radiotherapy. Radiation therapy should be avoided in those patients in whom surgery was proven to be curative while it could be considered in combination with surgery when the resection margins are dubious and/or inadequate.
Histopathological Examination of Ruptured Carotid Artery after Irradiation
Authors: Okamura Hiro-oki , Kamiyama Ryosuke , Takiguchi Yoshitaka , Kimizuka Kouki , Ishikawa Norihiko , Kishimoto Seiji
Published: April 2003

We report 2 cases of carotid artery rupture after irradiation that was performed 1 year and 17 years before the ruptures. When irradiation-induced arterial rupture occurs, it usually does so within a few months following irradiation. However, the histopathological sections obtained in the present cases revealed carotid artery necrosis that was presumably induced by irradiation. Carotid artery rupture is sudden, massive hemorrhage that ranks among the most dreaded complications in the head and neck. However, several patients have been saved by hospital personnel who discovered the rupture in time to take appropriate measures such as cleaning of the wound and protection with myocutaneous or myofascial flaps. Therefore, it is important to be aware of the possibility of rupture or perforation of major vessels after irradiation, even when the radiation therapy was performed a long time ago.
Chronic Caffeine Treatment Causes Changes in Cardiac Adenosine Receptor Function in Rats
Authors: White P.J. , Nguyen T.T.
Published: April 2003

Caffeine exerts a number of cardiovascular effects via antagonism of adenosine A1 and A2a receptors, and chronic caffeine intake has been suggested to be a preventable risk factor for cardiovascular disease. We attempted, therefore, to determine the effects of chronic caffeine treatment on adenosine receptor function and the heart rate and blood pressure of anaesthetized rats. Rats were given two weeks of drinking water containing either caffeine (0.2%) or tap water alone. Rats were then anaesthetized and blood pressure and heart rate was assessed, as well as cardiovascular responses to adenosine receptor agonists and antagonists. Following 2 weeks caffeine treatment, both the resting blood pressure and heart rate of the treated rats were significantly increased compared to the non-caffeine treated (147 ± 5 compared to 161 ± 3 mm Hg). Negative chronotropic responses to A1 receptor agonist N6 cyclopentyladenosine occurred at lower concentrations in caffeine-treated rats than control (2.5-fold decrease in the ED50 of bradycardic responses to N6 cyclopentyladenosine), whilst vasodilator and reflex tachycardic responses to A2a receptor agonist CGS 21680 were unchanged. A shorter duration of caffeine- treatment resulted in moderate increases in heart rate and bradycardic responses to N6 cyclopentyladenosine. In summary, we found changes in resting heart rate and blood pressure in caffeine-treated rats which were concurrent with changes in adenosine receptor function.
Effect of Hindlimb Suspension and Clenbuterol Treatment on Polyamine Levels in Skeletal Muscle
Authors: Abukhalaf Imad K. , von Deutsch Daniel A. , Wineski Lawrence E. , Silvestrov Natalia A. , Abera Saare A. , Sahlu Sinafikish W. , Potter David E.
Published: April 2003

Polyamines are unbiquitous, naturally occurring small aliphatic, polycationic, endogenous compounds. They are involved in many cellular processes and may serve as secondary or tertiary messengers to hormonal regulation. The relationship of polyamines and skeletal muscle mass of adductor longus, extensor digitorum longus, and gastrocnemius under unloading (hindlimb suspension) conditions was investigated. Unloading significantly affected skeletal muscle polyamine levels in a fiber-type-specific fashion. Under loading conditions, clenbuterol treatment increased all polyamine levels, whereas under unloading conditions, only the spermidine levels were consistently increased. Unloading attenuated the anabolic effects of clenbuterol in predominately slow-twitch muscles (adductor longus), but had little impact on clenbuterol’s action as a countermeasure in fast- twitch muscles such as the extensor digitorum longus. Spermidine appeared to be the primary polyamine involved in skeletal muscle atrophy/hypertrophy.