bluuebunny/crossref_metadata_2025 huggingface dataset:
Prolonged Antigen Exposure Ameliorates Airway Inflammation But Not Remodeling in a Mouse Model of Bronchial Asthma
Authors: Sakai Kimiko , Yokoyama Akihito , Kohno Nobuoki , Hamada Hironobu , Hiwada Kunio
Published: April 2003

Background: In naive rodents, repeated exposure to aerosolized antigen induces suppression of the Th2 response to the antigen. We hypothesized that more prolonged exposure of established asthma model to antigen aerosols may downregulate asthmatic phenotype. Methods: After establishing an ovalbumin (OVA)-induced asthma model, mice were further exposed to OVA (prolonged exposure group) or phosphate-buffered saline (positive controls) 3 days per week for 6 weeks. During week 7, the mice of both groups were finally challenged with OVA. Results: Prolonged OVA exposure resulted in marked suppression of serum OVA-specific immunoglobulin E (IgE) antibody levels, eosinophilia of the airway, and airway hyperresponsiveness (AHR). However, airway remodeling characterized by goblet cell hyperplasia and airway fibrosis was observed to the same degree in both groups. These effects were accompanied by diminished production of Th2 cytokines such as interleukin-4 (IL-4), IL-5 and IL-13 in bronchoalveolar lavage fluid (BALF) and cultured supernatant of splenocytes. Furthermore, prolonged exposure markedly increased IL-12 levels in BALF. Conclusions: Prolonged antigen exposure has inhibitory effects on eosinophilic inflammation, AHR and IgE response to antigen, but not on airway remodeling, presumably via inhibition of Th2 cytokines and increased IL-12 production in the lungs.
Characterization of Human IgE and Mouse IgG1 Responses to Allergens in Three Mosquito Species by Immunoblotting and ELISA
Authors: Jeon Soung-Hoo , Park Jung Won , Lee Bong Hee
Published: April 2003

Background: IgE-mediated allergic reactions caused by mosquito bites are a common problem all over the world. This study was undertaken to determine IgE levels in subjects, to elucidate human IgE and mouse IgG1 binding patterns and to investigate the cross-reactivity of salivary gland antigens with three mosquitoes. Methods: Mosquito larvae of Aedes togoi, Culex tritaeniorhynchus and Culex pipiens pallens were collected and maintained in the laboratory. Salivary gland extracts (SGE) and whole-body extracts (WBE) were prepared from female mosquitoes of each species. Mosquito-specific IgE levels in 17 subjects were measured by ELISA. Polypeptide patterns were analyzed by SDS-PAGE. Immunoblotting was performed with sensitized human and immune mouse sera, and elucidated human IgE and mouse IgG1 binding patterns to SGE. For the determination of cross-reactivity to the three types of mosquitoes, ELISA inhibition tests were performed using sera from mice sensitized by biting of A. togoi. Results: The 9 sera out of 12 with positive skin reactions to SGE of A. togoi by skin prick test showed significantly higher anti-mosquito SGE IgE levels than in those without skin reactions. Protein band patterns of the SGE and WBE of the three species were different from one another. Specific human IgE reacted to the protein in SGE of 30.5, 33, 37 and 57.5 kD from A. togoi, of 38, 43 an 68 kD from C. tritaeniorhynchus, and of 23, 33, 34, 43, 44, 60, 74 and 93 kD from C. pipiens pallens. There were specific mouse IgG1 reactions to the bands of 30.5, 33, 37 and 57.5 kD in the SGE of A. togoi. The ELISA inhibition studies disclosed almost no cross-reactivities between A. togoi, C. tritaeniorhynchus and C. pipiens pallens.Conclusions: Immunoblot analysis disclosed that allergenic proteins in the SGE of mosquitoes and their patterns were remarkably similar between human and mouse sera to the SGE of A. togoi. Species-shared allergens may not exist among the three mosquito species prevalent in Korea.
Extraction of IgE-Binding Components of Helicobacter pylori by Immunoblotting Analysis in Chronic Urticaria Patients
Authors: Liutu M. , Kalimo K. , Uksila J. , Savolainen J.
Published: April 2003

Background: An association between Helicobacter pylori and chronic urticaria has been suspected previously. An IgE-mediated pathway might be a possible link between H. pylori infection and chronic urticaria, and therefore we wanted to prepare an optimal H. pylori antigen to detect H. pylori-specific IgE antibodies in chronic urticaria patients. Methods:H. pylori antigen extracts were prepared in different ways to find the optimal antigen extract to be used in the assays. Immunoblotting was used to detect IgE-binding bands. The results were applied in an H. pylori RAST assay for specific H. pylori IgE antibodies in patient sera. Results: In immunoblotting, the largest number of IgE-stained bands were visualized in the washing fluids and sonicated extracts, while strong heating and denaturing treatments destroyed the epitopes for IgE binding, suggesting that they belonged to the flagellar structures of H. pylori. However, in H. pylori-specific RAST analysis, specific IgE was found only in 1 of 25 H. pylori-infected patients. Conclusions: Our findings suggest that although IgE-binding epitopes were found in H. pylori, H. pylori-specific IgE antibodies are not common in chronic urticaria, and the clinical significance of the IgE response is unclear.
Identification and Characterization of the Allergens in the Tomato Fruit by Immunoblotting
Authors: Kondo Yasuto , Urisu Atsuo , Tokuda Reiko
Published: April 2003

Background: Although the tomato fruit (Lycopersicon esculentum) has been widely investigated for breeding purposes, there have been few studies on tomato allergenicity. We attempted to identify the tomato fruit allergens and to compare the concentrations of IgE-binding proteins among the different growth stages with sodium dodecylsulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and immunoblotting. Methods: An immunoblot experiment on tomato fruit extracts was performed using sera from 11 patients with oral allergy syndrome (OAS) to tomatoes. Bands reacting with IgE from more than half of the OAS patients’ sera were excised and subjected to determination of N-terminal amino acid sequences using the automated Edman degradation method. Moreover, we compared the concentrations of these proteins at each growth stage of the tomato fruit with SDS-PAGE and immunoblotting. Results: Four proteins binding with IgE from more than half of the OAS patients’ sera were determined to be polygalacturonase 2A (PG2A), β-fructofuranosidase, superoxide dismutase (SOD) and pectinesterase (PE). The concentrations of PG2A, β-fructofuranosidase and PE were highest in the red ripening stage with both SDS-PAGE and immunoblotting. Conclusion: The concentrations of 3 of 4 tomato allergens increased during ripening.
TU-572, a Potent and Selective CD45 Inhibitor, Suppresses IgE-Mediated Anaphylaxis and Murine Contact Hypersensitivity Reactions
Authors: Hamaguchi Takuya , Takahashi Akiko , Manaka Akira , Sato Masakazu , Osada Hiroyuki
Published: April 2003

Background: CD45, receptor-type protein tyrosine phosphatases (PTPases) are essential components of signaling through both the T cell receptor and the B cell antigen receptor. However, the functional significance of CD45 in the signaling pathway through the high-affinity immunoglobulin (Ig) E receptor has not yet been established. In this study, we demonstrate that the potent CD45 inhibitor negatively regulates IgE-dependent anaphylaxis and contact hypersensitivity reactions. Method: We have previously found that TU-572, 2-[(4-methylthiopyridin-2-yl)methylsulfinyl]-5-isopropoxybenzimidazole, had a potent and selective inhibitory effect against PTPase activity of CD45. Using a CD45 inhibitor, we examined in vitro and in vivo IgE-mediated responses. Results: TU-572 potently inhibited histamine release from rat peritoneal mast cells and mouse systemic anaphylaxis reaction using monoclonal anti-dinitrophenyl (DNP) IgE and DNP-BSA. TU-572 also suppressed the immediate-type hypersensitivity response induced by repeated epicutaneous application of trinitrochlorobenzene in BALB/c mice. Conclusion: These findings revealed that the PTPase activity of CD45 played a critical role in signal transduction of IgE-mediated anaphylaxis in vitro and in vivo. PTPase inhibitors such as TU-572 are useful in the treatment of allergic diseases.
Hymenoptera-Venom-Induced Upregulation of the Basophil Activation Marker Ecto-Nucleotide Pyrophosphatase/Phosphodiesterase 3 in Sensitized Individuals
Authors: Platz Imogen J. , Binder Marc , Marxer Anke , Lischka Gerd , Valent Peter , Bühring Hans-Jörg
Published: April 2003

Background: Bee and wasp venoms are potent allergens capable of inducing severe clinical reactions. To detect immediate-type hypersensitivity to these allergens, a rapid in vitro test was developed that relies on the upregulation of ecto-nucleotide pyrophosphatase/phosphodiesterase 3 (E-NPP3) on activated basophils. Methods: Blood basophils of 13 healthy donors and 22 patients allergic to bee or wasp venom were analyzed for E-NPP3 (CD203c) expression using monoclonal antibody 97A6. Basophils were analyzed by flow cytometry after activation with anti-IgE antibody or allergen. Venom-induced E-NPP3 upregulation on basophils was compared with diagnostic parameters, including skin tests and assessment of specific IgE. In selected samples, the increase in E-NPP3 expression on activated basophils was compared with histamine release and CD63 upregulation. Results: In 20/22 patients sensitized to wasp or bee venom, E-NPP3 expression on basophils was upregulated in response to activation by allergen or anti-IgE. The maximum increase in E-NPP3 expression (above ten times of baseline) was achieved after 15 min of stimulation with 1 µg/ml of allergen or anti-IgE antibody. Sensitized individuals who failed to upregulate E-NPP3 in response to IgE receptor cross-linking also failed to induce histamine release and CD63 upregulation. Conclusions: Flow cytometric determination of hymenoptera-venom-induced upregulation of E-NPP3 is a novel in vitro test to identify sensitized individuals.
Cecal Ligation and Puncture as a Model of Sepsis in the Rat: Influence of the Puncture Size on Mortality, Bacteremia, Endotoxemia and Tumor Necrosis Factor Alpha Levels
Authors: Otero-Antón E. , González-Quintela A. , López-Soto A. , López-Ben S. , Llovo J. , Pérez L.F.
Published: April 2003

Background: Cecal ligation and puncture is a widely used experimental model of sepsis. Aim of the Study: The present study was aimed to evaluate the influence of the size of the cecal puncture on mortality, bacteremia, endotoxemia and plasma TNF-α levels. Materials and Methods: Female Sprague-Dawley rats underwent cecal ligation and puncture, divided into the following groups, defined by the diameter of the cecal puncture: 0.5-cm blade incision (n = 25), 13-gauge (n = 25), 16-gauge (n = 25), 18-gauge puncture (n = 25) and 4 punctures with a 22-gauge needle (n = 25). A sham operation was performed in another 25 rats. Three animals of each group were sacrificed 5 h after the procedure for blood cultures as well as determination of plasma endotoxin and TNF-α. The remaining animals were followed up for a week after cecal ligation and puncture for evaluation of mortality. Results: Five hours after cecal ligation and puncture, bacteremia was present in all animals, independently of the puncture size. Endotoxemia and plasma TNF levels tended to increase along with the diameter of the cecal puncture. Mortality gradually increased with the puncture size, from 27% with a 22-gauge needle to 95% with the blade incision. Conclusions: The severity of sepsis obtained with cecal ligation and puncture in rats can be easily modulated varying the size of the puncture.
Hepatic Indocyanine Green Uptake and Excretion in a Rabbit Model of Steatosis
Authors: Seifalian A.M. , El-Desoky A. , Davidson B.R.
Published: April 2003

Transplantation of a fatty liver is associated with a higher incidence of primary non-function of the graft. Indocyanine green (ICG) has been used for assessing hepatic dysfunction but not for quantifying liver steatosis. New Zealand white rabbits were fed a normal diet (group A) or a high-cholesterol (2%) diet for 4, 8, and 12 weeks in groups B, C, and D, respectively. Laparotomy was performed for liver exposure. Hepatic artery, portal vein, and total blood flow, hepatic microcirculation, portal pressure, liver function parameters, and blood cholesterol levels were measured. The hepatic ICG concentration was measured using near-infrared spectroscopy, and its uptake and excretion rates were calculated. The severity of steatosis was assessed from liver biopsy specimens by a semiquantitative grading system. Cholesterol feeding resulted in mild steatosis after 4 weeks and in moderate steatosis after 8 and 12 weeks. Mild steatosis was associated with insignificant changes in haemodynamic parameters, liver function, and ICG handling as compared with controls. Moderate steatosis caused a significant reduction in portal and total hepatic blood flow and microcirculation with a significant increase in hepatic artery flow and portal pressure. These haemodynamic changes were associated with a significant alteration in liver function tests. With moderate steatosis, ICG uptake and excretion were significantly reduced. The ICG uptake rate significantly correlated with total blood flow and microcirculation. The ICG excretion rate significantly correlated with the changes in bilirubin, liver enzymes, and albumin. Direct ICG quantification by near-infrared spectroscopy could be used to assess the severity of hepatic steatosis by reflecting impaired parenchymal perfusion and liver dysfunction.
Evaluation of Splanchnic Perfusion and Oxygenation during Positive End-Expiratory Pressure Ventilation in Relation to Subcutaneous Tissue Gases and pH
Authors: Jedlińska Barbara , Mellström Å. , Månsson P. , Hartmann M. , Jönsson K.
Published: April 2003

Evaluation of splanchnic perfusion and oxygenation was performed by measurements of serosal tissue oxygen tension (PserO2) and intramucosal pH (pHi) in relation to subcutaneous oxygen tension (PscO2), subcutaneous carbon dioxide tension (PscCO2) and subcutaneous pH (pHsc) in pigs subjected to oleic acid-induced lung injury during ventilation with increasing levels of positive end-expiratory pressure (PEEP). Lung injury resulted in a general hypoxia and redistribution of perfusion away from the subcutaneous and splanchnic tissues, illustrated by a decrease in PaO2 from 93 to 37 mm Hg (p < 0.01), PscO2 from 45 to 17 mm Hg (p < 0.01), PserO2 from 80 to 30 mm Hg (p < 0.01) and pHi from 6.84 to 6.74 (p < 0.05) and a decrease of porta flow from 0.77 to 0.57 l/min. Application of PEEP up to 10–15 cm H2O resulted in an increase of portal vein oxygen tension (PportaO2) from 21 to 34 mm Hg (p < 0.01), PscO2 from 17 to 26 mm Hg (p < 0.05) and PserO2 from 30 to 55 mm Hg (p < 0.05). At PEEP 20 cm H2O PserO2 decreased to 47 mm Hg (p < 0.05). Porta flow decreased continuously with increasing levels of PEEP. PserO2 correlated with PportaO2 (r = 0.7, p < 0.001). pHi correlated poorly with PportaO2 (r = 0.2) and porta flow (r = 0.4). PscO2 and PserO2 correlated well (r = 0.8, p < 0.001). In summary, splanchnic perfusion and oxygenation was better reflected by serosal oxygen tension than pHi in the colon. Changes in serosal oxygenation of the colon paralleled changes in subcutaneous tissue oxygenation.
Involvement of Soluble Adhesion Molecules in Acute Pancreatitis
Authors: Nakae H. , Endo S. , Sato N. , Wakabayashi G. , Inada K. , Sato S.
Published: April 2003

The relationship between inflammatory cytokines and adhesion molecules in cases of acute pancreatitis was studied. Twenty-five patients were divided into three groups based on the severity of their disease as rated by the Ranson score. They were also divided into two groups according to the presence or absence of the complications of the multiorgan dysfunction syndrome (MODS) and the prognosis. As the severity of acute pancreatitis increased, the plasma levels of TNF-α, slCAM-1, sE-selectin, and sVCAM-1 also increased. The levels of all of these soluble adhesion molecules were significantly higher in the MODS-complicated group than in the non-MODS-complicated group. The levels of TNF-α, slCAM-1, and sVCAM-1 were significantly higher in patients who eventually died than in patients who survived. Significant correlations were observed between the levels of TNF-α and slCAM-1, between those of TNF-α and sVCAM-1, and between the levels of slCAM-1 and sVCAM-1. These results suggest that inflammatory cytokines stimulate the expression of adhesion molecules on the vascular endothelium and that mediators, including these adhesion molecules, are closely involved in the pathogenesis of acute pancreatitis. The plasma levels of these soluble adhesion molecules reliably reflected the severity of the disease.
A New Mesangial Triumvirate: Sulfonylureas, Their Receptors and Endosulfines
Authors: Yee Jerry , Szamosfalvi Balázs
Published: April 2003

Although sulfonylureas have long been therapeutically utilized for their hypoglycemic properties in type 2 diabetic patients, there is a paucity of clinical or experimental data that suggests that this pharmacotherapeutic class confers a benefit on the course of diabetic renal disease. Because the mesangial compartment is central to the fibrogenic response that evolves during the course of diabetic nephropathy, determining the metabolic influence of sulfonylureas on mesangial cells is important. In this article, the current knowledge regarding the metabolic and functional consequences of a mesangial triumvirate of sulfonylureas, their sulfonylurea receptors and sulfonylurea-like ligands termed endosulfines will be reviewed.
Increased Osteopontin Expression following Renal Ablation Is Attenuated by Angiotensin Type 1 Receptor Antagonism
Authors: Cao Zemin , Cox Alison , Bonnet Fabrice
Published: April 2003

Osteopontin is an extracellular matrix protein that is upregulated in renal injury. The aim of this study was to explore the renal expression of osteopontin in a model of progressive renal injury following subtotal nephrectomy (STNx) in rats and the effects of angiotensin type1 (AT1) receptor antagonist irbesartan on osteopontin expression. STNx or a sham operation was performed in 8-week-old Sprague-Dawley rats. STNx rats were given either irbesartan (15 mg/g) or no treatment for 12 weeks. Upregulation of osteopontin mRNA expression was observed in injured renal tubules as assessed by in situ hybridization (42 ± 8 dpm/mm2 v.s. control 7.7 ± 0.6 dpm/mm2, p < 0.01). Increased osteopontin expression was closely related to infiltration of monocytes/macrophages and increased cellular proliferation. Double immunohistochemical staining demonstrated co-existence of proliferating cell nuclear antigen and osteopontin positive staining in individual cells in kidney sections from STNx rats. The increase in osteopontin expression was inhibited by the AT1 receptor antagonist irbesartan (6.9 ± 1.2 dpm/mm2), associated with attenuation of impaired renal function and pathology as well as decreased monocyte/macrophage infiltration and cellular proliferation. These findings suggest that osteopontin is upregulated in STNx rats and is reduced by AT1 receptor antagonism.
Glucagon Acutely Inhibits but Chronically Activates Na+/H+ Antiporter 3 Activity in OKP Cells
Authors: Amemiya Morimasa , Kusano Eiji , Muto Shigeaki , Tabei Kaoru , Ando Yasuhiro , Alpern Robert J. , Asano Yasushi
Published: April 2003

Background: We previously found that the Na+/H+ exchanger 3 (NHE3) is localized in the apical membrane of the rat renal proximal tubule and thick ascending limb of Henle. In the present study, we examined the direct effect of glucagon on the opossum kidney P (OKP) cell Na+/H+ antiporter, encoded by NHE3. Methods: Na+/H+ antiporter activity was measured as the rate of cell pH recovery from an acid load using 2′,7′-bis(carboxyethyl)-5(6)-carboxyfluorescein. Northern blot and Western blot analyses were performed using OKP NHE3 cDNA and anti-OKP-NHE3 antibodies. Results: Glucagon (1 ng/ml) acutely (1 h) inhibited, but chronically (24 h) activated NHE3 activity in OKP cells. These effects were blocked by either KT5720 or RpcAMP [protein kinase A (PKA) inhibitors], and mimicked by 10–4M dibutyryl-cAMP. Both NHE3 mRNA and protein abundance increased with the 24-hour incubation in glucagon or dibutyryl-cAMP. Cycloheximide did not prevent a significant increase in NHE3 activity at 24 h. We therefore examined NHE3 protein abundance in the surface membrane by the biotinylation mehtod. cAMP or glucagon significantly increased NHE3 protein abundance in the surface membrane when incubated with cycloheximide for 24 h. Conclusions: Glucagon acutely inhibits but chronically activates NHE3 activity in OKP cells via a PKA-dependent pathway. Both protein-synthesis-dependent and -independent mechanisms play important roles in the chronic activation of NHE3.
Gene Expression Profiling in silico: Relative Expression of Candidate Angiogenesis Associated Genes in Renal Cell Carcinomas
Authors: Gerritsen Mary E. , Peale Jr. Franklin V. , Wu Thomas
Published: April 2003

Recent advances in gene expression profiling have led to the development of comprehensive databases which can be queried in various manners. In the present report, we have taken a list of genes previously associated with angiogenesis, either in in vivo or in in vitro models, and queried a commercial database established by GeneLogic® to determine the relative expression of these candidate genes in normal kidneys and in renal cell carcinomas (RCC). We identified a number of genes, including CXCR4, matrix metalloproteinase 9, thrombospondin 2, and vascular endothelial growth factor, that were highly expressed in RCC versus normal tissue. One gene, hevin, appears to be selectively upregulated in RCC in contrast to downregulation of this gene in lung and colon tumors. This approach provides a powerful means to identify potential markers of tumor vascularization.
Microarray Studies of Gene Expression in Circulating Leukocytes in Kidney Diseases
Authors: Alcorta David , Preston Gloria , Munger William , Sullivan Pamela , Yang Jia Jin , Waga Iwao , Jennette J. Charles , Falk Ronald
Published: April 2003

The molecular characterization of changes in mRNA expression in renal tissue during disease is hampered by the acquisition of sufficient mRNA to do genomewide expression profiling. In many renal diseases, such as systemic lupus erythematosus, IgA nephropathy, antineutrophil cytoplasmic autoantibody (ANCA) associated glomerulonephritis, and small-vessel vasculitis (ANCA disease), circulating leukocytes play a role in onset, progression, and severity of the condition. Circulating leukocytes are readily isolated and supply sufficient mRNA for analysis, allowing molecular investigation into their involvement in the disease process. Our laboratory has undertaken a systematic study of the genomewide expression profiles of the circulating leukocytes from patients with a variety of renal diseases (ANCA disease, IgA nephropathy, lupus nephritis, focal segmental glomerulosclerosis), using the Affymetrix high-density gene chip array technology. Analysis of the data showed clustering of expressed genes unique for each individual disease group. These results imply that significant gene expression changes occur in leukocytes that are circulating in patients with renal diseases. In addition, gene expression has been studied in leukocytes activated in vitro by mechanisms that mimic pathogenic events in vivo. The expression levels of genes identified in in vitro studies were compared with the patient leukocyte gene expression to determine whether similar pathological events were occurring in vivo.
Genes Involved in Atherosclerosis
Authors: Laukkanen Johanna , Ylä-Herttuala Seppo
Published: April 2003

Atherosclerosis is a multifactorial disease that involves several genes and proteins. The purpose of this article is to focus on the arterial wall and to review lipoprotein receptors, growth factors, cytokines, chemokines, matrix metalloproteinases, adhesion molecules, and apoptosis genes and their involvement in atherogenesis.
Chimpanzee Band 3 Protein Polymorphism and the Origin of the Band 3 Memphis in Humans
Authors: Kimura Masako , Ishida Takafumi
Published: April 2003

//8RBand 3 protein, a major erythrocyte membrane protein, serves as an anion exchanger and as anchorage sites for erythrocyte skeletal proteins and enzymes [1]. Among band 3 protein variants, the most common is the band 3 Memphis [2]. The band 3 Memphis was documented as an asymptomatic variant with a retarded electrophoretic mobility on SDS-PAGE and its molecular basis is an amino acid substitution (Lys56Glu: AAG → GAG) [3]. The Memphis variant is observed in a polymorphic status among human populations ubiquitously, however, it remains as a minor allele [2]. There was a report suggesting the possible presence of a Memphis-like band 3 protein variant among chimpanzees [4]. This implied an ancient origin of the band 3 Memphis polymorphism that could be traced back to a common ancestor of man and chimpanzee. To elucidate the origin of the human Memphis polymorphism, as well as to confirm the presence of the Memphis variant in the chimpanzee, we studied African great ape band 3 protein and genes.
Hormonal Therapy for the Subfertility of Cryptorchidism
Authors: Huff Dale S. , Snyder III Howard M. , Rusnack Susan L. , Zderic Stephen A. , Carr Michael C. , Canning Douglas A.
Published: April 2003

Background: The subfertility of cryptorchidism correlates with severely reduced total germ cell counts in prepubertal testicular biopsies of undescended testes. Reduced total germ cell counts are associated with defects in the two prepubertal steps in maturation and proliferation in germ cells: first, the transformation of the fetal stem cell pool (gonocytes) into the adult stem cell pool (adult dark spermatogonia) at two to three months of age and, second, the transformation of adult dark spermatogonia into primary spermatocytes (meiosis) at 4–5 years. The defects in maturation are associated with blunting of the normal surges in gonadotropins and testosterone. Prepubertal treatment with gonadotropin-releasing hormones would theoretically trigger normal germ cell maturation and proliferation and thereby improve total germ cell counts and improve fertility. Prepubertal treatment of cryptorchidism with the GnRH analogue Buserelin has resulted in improved total germ cell counts and improved spermiograms. The purpose of this report is to describe the results of treatment of cryptorchidism with the GnRH analogue Naferelin. Patients: Twelve boys with cryptorchidism, 6 unilateral and 6 bilateral, and severely reduced germ cell counts in testicular biopsies were treated with Naferelin following successful orchidopexy and bilateral testicular biopsies. Response of the total germ cell counts was assessed in follow-up bilateral biopsies within 5 months of completing the hormonal therapy. Results: Eight of the 12 boys (5 of the 6 with unilateral and 3 of the 6 with bilateral cryptorchidism) showed improvement in the total germ cell counts in one or both testes. All 8 had a poor prognosis for fertility pretreatment and a good prognosis for fertility posttreatment. Of the 5 with unilateral cryptorchidism who improved, 2 showed improvement in both testes; and 3, only in the contralateral descended testes. All 3 of the boys with bilateral cryptorchidism who improved showed improvement in both testes. Testes with absence of germ cells and older patients tended to show no improvement. Of the 6 contralateral descended, 5 (83%) improved, and of the 18 undescended testes, 8 (44%) improved. Conclusions: In this preliminary study, Naferelin therapy appears to induce improvement in the total germ cell counts and the prognosis for future fertility in 75% of patients.
Radiological Signs of Leri-Weill Dyschondrosteosis in Turner Syndrome
Authors: Binder Gerhard , Fritsch Heidi , Schweizer Roland , Ranke Michael B.
Published: April 2003

Background/Aims: Leri-Weill dyschondrosteosis (LWD), a mesomelic short stature syndrome with Madelung deformity, was recently reported to be caused by SHOX (short stature homeobox-containing gene) haploinsufficiency. The loss of SHOX on Xp22.32, also called PHOG (pseudoautosomal homeobox-containing osteogenic gene), through structural aberrations of the X chromosome was also implicated in the short stature phenotype and some additional stigmata of Turner syndrome. The aim of this study was to systematically examine left-hand radiographs from Turner girls for the presence of signs of LWD. Methods: We retrospectively studied 168 left-hand radiographs from 54 patients with Turner syndrome (bone age >10.5 years) who were treated with rhGH and seen during the last 10 years in our clinic. For comparison, we analyzed 7 radiographs from 5 patients with LWD and 52 radiographs from 20 patients with GH deficiency. The shape of the distal radial epiphysis (triangularisation index = TI) and the carpal angle were quantitatively measured. In addition, we screened for the presence of a premature cleft fusion or an ulnar deviation of the articular surface of the distal radial epiphysis and for fourth metacarpal shortening. One of 54 Turner girls (2%) was affected with LWD and presented with Madelung deformity. Results: No milder forms of Madelung deformity were detected. However, there was a significant trend to a triangular shape of the distal radial epiphysis in Turner syndrome: the median TI was 2.7 in normal controls (range 1.8–3.7), 3.1 in Turner girls (range 2.0–6.3) (p < 0.001 against controls), and 6.0 in patients with LWD (range 3.5–11.0) (p < 0.001 against controls). Conclusions: The triangularisation index did not correlate with the carpal angle (median 122.5
Frasier Syndrome with Childhood-Onset Renal Failure
Authors: Buzi F. , Mella P. , Pilotta A. , Felappi B. , Camerino G. , Notarangelo L.D.
Published: April 2003

Background: The Wilms’ tumour 1 (WT1) gene encodes a protein which is believed to exert transcriptional and tumour-suppressor activities. Mutations of this gene have occasionally been associated with Wilms’ tumour (<15% of cases) and, more consistently, with three syndromes characterized by urogenital abnormalities (WAGR, Denys-Drash and Frasier syndrome). Subject/Method: A 25-year-old phenotypic female with a 46,XY karyotype presented with amenorrhoea. An ultrasound scan showed streak gonads and a rudimentary uterus. The patient had a history of post-streptococcal glomerulonephrosis, when aged 4 years, which had rapidly progressed to kidney failure, requiring transplantation at age 8. Result: Frasier syndrome was suspected and confirmed by genetic analysis. In fact, direct sequencing of the PCR product of the intron 9 donor splice site revealed a substitution of guanine for adenine in position +5. Conclusion: Besides being one of the few Frasier syndrome cases to be genetically characterized, this case is interesting because of the unusually early-onset renal failure.
Potential Consequences of Maternal Hypothyroidism on the Offspring: Evidence and Implications
Authors: Glinoer Daniel
Published: April 2003

The adequate functioning of both the maternal and fetal thyroid glands plays important roles to ensure that the fetal neuropsychointellectual development progresses normally. Three sets of clinical disorders ought to be envisaged, potentially leading to impaired brain development: defective glandular ontogenesis (leading to congenital hypothyroidism), maternal hypothyroidism (usually related to chronic autoimmune thyroiditis), and finally iodine deficiency (affecting both the maternal and fetal thyroid functions). The present review will be focused mainly on maternal hypothyroidism, where both the severity and temporal occurrence of maternal thyroid underfunction drive the resulting repercussions for an impaired fetal neuronal development: such clinical situations may take place during early gestation (in women with known but untreated hypothyroidism) or appear only during later gestational stages (in women with thyroid antibodies, who remain euthyroid during the first half of gestation). Recent available evidence and its implications are discussed, as well as our present concepts relating to the complexities of the fetomaternal thyroid relationships, and the potential impact of maternal thyroid function abnormalities on the ideal offspring’s development.
Juvenile Hypertension, the Role of Genetically Altered Steroid Metabolism
Authors: Ferrari Paolo , Bianchetti Mario , Frey Felix J.
Published: April 2003

The importance of hypertension in the pediatric population is not as well appreciated as in adults. This might be related in part to the lower prevalence of high blood pressure in this age group. As with height and weight, blood pressure increases with age during childhood. The underlying causes of significant hypertension in children differ considerably from those in adults: while the prevalence of hypertension in pediatrics is lower than in adults, clinically identifiable causes of hypertension are common. Abnormalities in steroid biosynthesis have been known for years to cause hypertension in some cases of congenital adrenal hyperplasia. In these patients, hypertension usually accompanies a characteristic phenotype with abnormal sexual differentiation. Recently, the molecular basis of four forms of severe hypertension transmitted on an autosomal basis has been elucidated: (a) the glucocorticoid-remediable aldosteronism (GRA), (b) the syndrome of apparent mineralocorticoid excess (AME), (c) activating mutation of the mineralocorticoid receptor and (d) Liddle’s syndrome. All these conditions are characterized primarily by low or low-normal plasma renin, normal or low serum potassium and salt-sensitive hypertension, indicating an increased mineralocorticoid effect. These forms of juvenile hypertension are a consequence of abnormal biosynthesis, metabolism or action of steroid hormones: (a) GRA is due to expression of a chimeric gene produced by fusion of 11β-hydroxylase aldosterone-synthase genes. Expression of the chimeric enzyme occurs in the zona fasciculata of the adrenal cortex under the control of ACTH and can be suppressed by administration of glucocorticoids. (b) AME is caused by mutations of the 11β-hydroxysteroid dehydrogenase type 2 enzyme, an enzyme that metabolizes cortisol into its receptor inactive keto-form cortisone, thus protecting the mineralocorticoid receptor (MR) from occupation by glucocorticoids. (c) The activating mutation of the MR results in constitutive MR activity and alters receptor specificity, with progesterone and other steroids lacking 21-hydroxyl groups becoming potent agonists. (d) Liddle’s syndrome is due to mutations in the β or γ chain of the epithelial sodium channel in distal renal tubule cells. The hyperactivity of this channel caused by the mutations results in increased sodium reabsorption. With the advent of molecular biology in clinical practice it has become evident that some genetic defect may present with a more discrete phenotype, with only moderate hypertension with or without hypokalemia as presenting feature. Considering that hypertension in children and adolescents is often ‘nonessential’, a search for disorders should be integral part of the diagnostic work-up in young patients with hypertension.
Hormonal Treatment for Unilateral Inguinal Testis: Comparison of Four Different Treatments
Authors: Bertelloni Silvano , Baroncelli Giampiero I. , Ghirri Paolo , Spinelli Claudio , Saggese Giuseppe
Published: April 2003

Background: Hormonal treatment of cryptorchidism has been used since the 30s, but controversies persist on its efficacy. It is also unclear whether there are differences with the use of different hormonal trials. Aims: To evaluate the efficacy of four hormonal treatments on testicular descent in a homogeneous group of cryptorchid boys. Patients: 155 patients (age 10–48 months) with unilateral inguinal palpable testis were studied. Methods: The patients were subdivided into four groups according to hormonal treatment: group 1 = hCG [500 IU/week (if the chronological age was <2 years) or 1,000 IU/week (if the chronological age was >2 years) for 6 weeks]; group 2 = hCG + hMG (hCG as in group 1 + hMG 75 IU/week for 6 weeks); group 3 = GnRH (1,200 µg/daily for 28 days); group 4 = GnRH + hCG (1,200 µg/daily for 28 days + 1,500 IU/week for 3 weeks, respectively). The results were evaluated at the end of the treatment period and 6 months later to exclude temporarily positive results. Results: At the end of the hormonal therapy, scrotal testicular descent was present in 30 of 155 boys (success rate 19.3%). Seven testes relapsed during follow-up (23.3%). The long-term success rate was 14.8% (23/155 testes). No significant differences were observed in success rates as well as in relapse rates among the four groups. Conclusions: Hormonal therapy induced permanent testicular descent in a minority of young cryptorchid boys with inguinal palpable testis. Similar results were obtained with four different trials.
Effects of Alcohol on Blood Pressure and Production of Vascular Aldosterone and Corticosterone
Authors: Zhang Yongsheng , Sun Tiejun , Jiang Feng , Chu Liyun , Yu Wengone , Wei Li , Pang Jien , Liu Yong , Su Guangzhi
Published: April 2003

Aims: The aims of this study were to search for the role of alcohol in blood pressure regulation in rats and to investigate the effects of alcohol on the production of vascular aldosterone and corticosterone. Methods: Male Wistar rats received alcohol 0.7 g · kg–1 · day–1 (alcohol-treated group 1) or 1.4 g · kg–1 · day–1 (alcohol-treated group 2) or 2.1 g · kg–1 · day–1 (alcohol-treated group 3), orally, for 3 months, and blood pressure was monitored by a pressure transducer. Systolic blood pressure increased in Wistar rats treated with alcohol compared to control rats. Mesenteric artery perfusion ex vivo was performed and pressor responses to norepinephrine were determined. The pressor responses to norepinephrine in mesenteric arteries treated with alcohol were significantly increased. The perfusate from the mesenteric arteries was collected and applied to a Sep-Pak C 18 cartridge column for reverse-phase high-performance liquid chromatography and aldosterone and corticosterone levels were determined by radioimmunoassay, aldosterone was decreased but corticosterone was increased in the perfusate from arteries treated with alcohol. Results: Reverse transcriptase polymerase chain reaction showed that alcohol inhibited the expression of 11β-HSD2 and CYP11B2 mRNA in mesenteric arteries. Conclusions: These results reveal that alcohol is able to induce hypertension and provide evidence that alcohol inhibits the transcriptions of both 11β-HSD2 and CYP11B2 in the vasculature, leading to lower aldosterone and higher corticosterone production in vessels, and increased vasoconstrictor responses to norepinephrine.
Fertility after Unilateral Cryptorchidism
Authors: Miller Karyn D. , Coughlin Michael T. , Lee Peter A.
Published: April 2003

Objective: To further evaluate whether fertility is decreased among a cohort of men with previous unilateral cryptorchidism as compared with a control group of men. Subjects and Methods: Formerly unilateral cryptorchid men who had undergone orchiopexy between the years of 1955 and 1975 at the Children’s Hospital of Pittsburgh and a control group of men who were matched for age of an unrelated surgery at the same institution were evaluated by review of medical records and by completion of a questionnaire. 359 previously cryptorchid men were identified as having attempted paternity. Of these men, 320 had information concerning preoperative testicular location and 163 for preoperative testicular size. 106 of these men had levels of testosterone, inhibin B, FSH, and LH measured, while 95 of the men had semen analyses. Results: Among men who had attempted paternity, there was no statistical difference in success of paternity between the previously unilateral group (89.7%) and the control group (93.7%). There was no difference in the mean time to conception (7.1 ± 0.7 months for the unilateral group vs. 6.9 ± 2.3 for the control group). Within the unilateral group in regard to success at paternity, no difference was found compared with the age of orchiopexy, preoperative testicular location, or preoperative testicular size. Inhibin B levels were lower among the unilateral group. FSH, LH, testosterone, sperm density, motility and morphology were not different, but considerable variation was noted within the cryptorchid group. Conclusions: In this continued evaluation of a cohort of previously cryptorchid men who had undergone unilateral orchiopexy, paternity does not appear to be significantly compromised after unilateral cryptorchidism. Unilateral cryptorchidism appears to be one of several factors contributing to infertility, similar to those found in the general population. No correlation was found between success at paternity and the age of orchiopexy, preoperative testicular size or preoperative testicular location. Inhibin B levels were lower while FSH, LH, T and sperm parameters did not differ.
Should the Definition of Micropenis Vary According to Ethnicity?
Authors: Cheng Pik Shun , Chanoine Jean-Pierre
Published: April 2003

Objective: We determined whether the existing reference values for the diagnosis of micropenis are appropriate for optimal care of neonates in a multiethnic environment like Vancouver. Methods: The stretched penile length and width were measured in 105 full-term newborn males of Caucasian (n = 40), Chinese (n = 40) and East-Indian origin (n = 25). Results: Mean length –2.5 SD was used for the definition of micropenis and was 2.6, 2.5 and 2.3 cm for Caucasian, East-Indian and Chinese babies, respectively (p < 0.05). This is close to the widely accepted recommendation that a penile length of 2.4– 2.5 cm be considered as the lowest limit for the definition of micropenis. Conclusion: Mean penile length and diameter are slightly but significantly smaller in newborns of Chinese origin compared to newborns of Caucasian and East-Indian origins.
Decreased Tubular Uptake of L-3,4-Dihydroxyphenylalanine in Streptozotocin-Induced Diabetic Rats
Authors: Carranza Andrea , Karabatas Liliana , Barontini Marta , Armando Ines
Published: April 2003

Background/Aims: This study determined alterations in renal dopamine production in streptozotocin-treated rats and explored the mechanisms underlying this alteration. Methods: Streptozotocin (65 mg/kg) or vehicle was administered to 3-month-old male Wistar rats. Treated animals had hyperglycemia, glycosuria and increased diuresis, natriuresis and excretion of L-dopa. Urinary dopamine and dihydroxyphenylacetic acid were similar to those in control animals. The production of dopamine by renal cortex slices from treated rats was significantly less than that from control animals. The addition of glucose (8.4–18.4 mM) to the incubation medium decreased about 40% the uptake of L-dopa by isolated proximal tubular cells. Scatchard analysis of the saturation curves obtained in this condition showed a decrease in the Vmax without changes in the Km. Results: Our results confirm previous studies suggesting a renal dopaminergic deficiency in insulin-dependent diabetes and provide evidence strongly suggesting that a decrease in the number of tubular L-dopa transport sites is the underlying defect of this deficiency. Conclusion: These results highlight the role of the uptake of dopa as an important modulator of renal dopamine synthesis.
Therapeutic Vaccines for Human Papillomaviruses
Authors: Gissmann Lutz , Osen Wolfram , Müller Martin , Jochmus Ingrid
Published: April 2003

Although papillomavirus infections are not very immunogenic there is evidence that the immune system controls the spread of virus and the development of diseases associated with such infections. Certain types of human papillomaviruses (HPV) are the major cause of premalignant and malignant diseases of the anogenital tract, most notably cancer of the uterine cervix, a major health care problem worldwide. Since the viral oncoproteins E6 and E7 are constitutively expressed within the tumor cells, they are considered as suitable targets for attack by T lymphocytes. Several approaches to specifically trigger a cell-mediated immune response have been successful in experimental animals, leading to suppression of HPV-induced tumors. First clinical trials have been completed which raise hopes that a similar effect can also be achieved by therapeutic vaccination of humans.
Inhibition of Mayaro Virus Replication by Prostaglandin A1 in Vero Cells
Authors: Burlandy F.M. , Rebello M.A.
Published: April 2003

Prostaglandins exhibit antiviral activity against a wide variety of RNA and DNA viruses. In the present report, we describe the effect of cyclopentenone prostaglandin A1 (PGA1) on Mayaro virus replication in Vero cells. Virus yield was significantly reduced at nontoxic concentrations which did not suppress DNA, RNA or protein synthesis in uninfected or infected cells. Antiviral action decreased if PGA1 was added at later times after infection. In Mayaro virus-infected cells, PGA1 inhibited the synthesis of virus proteins. This effect is accompanied by the induction of heat shock proteins (HSPs). Actinomycin D treatment not only inhibited the induction of HSPs but also partially prevented PGA1 antiviral activity.
Viral Dynamics and Pharmacokinetics in Combined Interferon Alfa-2b and Ribavirin Therapy for Patients Infected with Hepatitis C Virus of Genotype 1b and High Pretreatment Viral Load
Authors: Tsubota Akihito , Akuta Norio , Suzuki Fumitaka , Suzuki Yoshiyuki , Someya Takashi , Kobayashi Masahiro , Arase Yasuji , Saitoh Satoshi , Ikeda Kenji , Kumada Hiromitsu
Published: April 2003

Objective: The aim of this study was to clarify the viral dynamics and single- and multiple-dose pharmacokinetics of ribavirin and interferon (IFN) alfa-2b in the virologic response to combination therapy with both compounds in patients infected with hepatitis C virus (HCV) genotype 1b and high viral load. Methods: Fourteen patients received high-dose daily induction therapy followed by intermittent maintenance therapy with IFN alfa-2b and daily oral ribavirin for 24 weeks, and followed up for 24 weeks after treatment. Single- and multiple-dose pharmacokinetic studies and viral dynamics were assessed by serial measurements of serum concentrations of both compounds and HCV RNA, respectively, at weeks 1 and 24. Results: During treatment, all 14 patients showed biochemical response (i.e., normalization of serum alanine transaminase activity), while 11 showed virologic response (i.e., undetectable serum HCV RNA level by qualitative polymerase chain reaction assay). Sustained biochemical and virologic response after cessation of treatment was noted in 8 and 2 patients, respectively. Serum ribavirin concentrations asymptoted by 4 weeks of treatment. Serum ribavirin concentrations in steady state, and maximum concentration and accumulation rate of ribavirin after multiple dosing were significantly higher in the presence of sustained virologic response. HCV-related parameters were not significantly associated with sustained virologic response. Conclusions: Continuous exposure and tissue accumulation of ribavirin may be necessary for sustained virologic response to combination therapy in chronic hepatitis C with genotype 1b and high viral load. Pharmacokinetic analysis of ribavirin provides information on its mechanism of action and for developing more rational treatment for IFN-resistant HCV.
Splenic Rupture following ESWL for a Pancreatic Duct Calculus
Authors: Leifsson Björn Geir , Borgström Anders , Ahlgren Göran
Published: April 2003

Extracorporeal shock wave lithotripsy (ESWL) is an established and extensively used treatment alternative for urinary calculi. It is also an established method of dealing with pancreatic duct calculi complementing endoscopic techniques in selected cases. Three reports of splenic injury following and probably caused by ESWL for urinary calculi have previously been published. We report a case of splenic rupture presenting with life-threatening hemorrhage 6 days after a single ESWL therapy session for pancreatic duct calculi.