bluuebunny/crossref_metadata_2025 huggingface dataset:
In vitro Evaluation of Abrasion of Eroded Enamel by Different Manual, Power and Sonic Toothbrushes
Authors: Wiegand A. , Begic M. , Attin T.
Published: May 2008

This study aimed to evaluate the susceptibility of eroded enamel to brushing abrasion performed by manual, power or sonic toothbrushes. Bovine enamel samples were subjected to 5 cycles, each consisting of 5 min demineralisation, 15 min remineralisation and 10 min brushing in a machine. Toothbrushing with the activated electric devices was supplemented with 20 linear strokes/min. Furthermore, enamel specimens were brushed with 20 linear strokes/min or 80 linear strokes/min with the electric toothbrushes without their individual operating action. A manual brush was applied at 100, 20 or 80 linear strokes/min. Specimens of the control group were not brushed after demineralisation. Loss of enamel was determined by profilometry. For all groups, substrate loss for linear brushing treatment applying 20 or 80 strokes/min did not differ significantly from the control (4.97 ± 1.49 µm). Three power toothbrushing treatments significantly increased abrasion compared to linear brushing treatment with 20 or 80 strokes/min in their inactivated condition. The results indicate that brushing treatment with power or sonic toothbrushes may lead to significantly higher loss of demineralised enamel compared to toothbrushing without power or sonic support.
Renal Acid-Base Transport: Old and New Players
Authors: Wagner Carsten A. , Kovacikova Jana , Stehberger Paul A. , Winter Christian , Benabbas Chahira , Mohebbi Nilufar
Published: May 2008

Systemic acid-base homeostasis is the product of complex interactions between metabolism, regulated exhalation of CO2 by the lungs and acid or base excretion by the kidneys. The importance of renal acid-base transport has been highlighted by mutations identified in several proteins involved in this task in patients with inborn forms of renal tubular acidosis. The underlying mechanisms of disease have been further studied in genetically altered mouse models and cell culture. An interesting field of research has focused on the question how changes in metabolism or acid-base homeostasis are sensed and result in altered excretion of acid or bases by the kidney. Several hormonal pathways including aldosterone and endothelin were implicated, a novel subfamily of proton-sensing receptors has been identified, and signaling molecules described that are activated by changes in pH.
Sodium Intake Modifies the Negative Prognostic Value of Renal Damage prior to Treatment with ACE Inhibitors on Proteinuria Induced by Adriamycin
Authors: Kramer Andrea B. , Bos Hendrik , van Goor Harry , Navis Gerjan J.
Published: May 2008

Background: Antiproteinuric treatment by ACE inhibition (ACEi) provides renoprotection. However, resistance to antiproteinuric intervention occurs frequently, resulting in progressive renal damage. The extent of renal damage prior to treatment with ACEi reversely correlates with the antiproteinuric effects of ACEi in established adriamycin nephrosis. Sodium restriction enhances the response to ACEi, but whether it can overcome the negative predictive value of preceding renal damage on the therapeutic response is unknown. We studied the impact of preceding renal damage on the efficacy of ACEi in adriamycin nephrosis on different oral sodium loads. Methods: Male Wistar rats were randomly assigned to a low (LS), normal (NS) or high (HS) sodium diet, initiated 1 week before adriamycin induction. At week 6, proteinuria was stabilized (195 ± 172 mg/24 h), a renal biopsy was performed for analysis of preceding damage and rats were instituted on lisinopril for 6 weeks until sacrifice at week 12. Results: ACEi reduced proteinuria in LS and NS animals. On univariate analysis, the antiproteinuric response was significantly predicted by preceding renal damage (focal glomerulosclerosis, interstitial macrophages and interstitial α-smooth muscle cell actin expression). On multivariate analysis, both sodium intake and preceding renal damage independently predicted residual proteinuria during ACEi (R2 model: 80% and 75% for data after 3 and 6 weeks of therapy, respectively). Conclusion: Our data confirm the predictive value of pretreatment renal damage for the antiproteinuric response to ACEi, despite the fact that the renal damage prior to the ACEi was very mild. The impact of pretreatment damage on the therapeutic response, however, was overcome by low sodium. Thus, the impact of pretreatment damage does not warrant therapeutic nihilism, but rather optimization of therapy response by dietary sodium restriction. Further studies are needed to elucidate whether this also applies to more severe damage, and whether combining ACEi with low sodium diet can improve long-term renal outcome in human.
Dexamethasone Alters Vascular Reactivity by Enhancing COX-Related Vasodilatation in Fetal Ovine Carotids
Authors: Angeles Danilyn M. , Chang Melody , Leong Valerie , Oberg Kerby C. , Pearce William J.
Published: May 2008

Based on preliminary studies that contractile efficacy was altered in vertebral and basilar arteries from neonatal donors treated with postnatal glucocorticoids, we examined the hypothesis that postnatal dexamethasone (DEX), a glucocorticoid used for respiratory disease in neonates, can alter vascular reactivity. Using near-term fetal lamb carotids, we measured 5-hydroxytryptamine (5-HT) dose-response relationship in DEX-treated and untreated arteries. We found that DEX incubation for 1 h had no effect on 5-HT sensitivity and agonist affinity but significantly reduced 5-HT contractile efficacy, a response that became even more pronounced after 4 h of DEX treatment. Coincubation of DEX-treated arteries with INDO for 4 h reversed this DEX-induced attenuation in 5-HT contractile efficacy, although DEX had no significant effects on cyclooxygenase (COX)-1 and COX-2 protein abundance. This data suggests that DEX alters vascular reactivity through a COX-related mechanism, with possible repercussions to neurological injury.
Topological Predictions for Integral Membrane Permeases of the Phosphoenolpyruvate:Sugar Phosphotransferase System
Authors: Nguyen Thai X. , Yen Ming-Ren , Barabote Ravi D. , Saier Jr. Milton H.
Published: May 2008

We report bioinformatic analyses of the largest superfamily of integral membrane permeases of the bacterial phosphotransferase system (PTS), the Enzyme IIC constituents of the Glc superfamily. Phylogenetic analyses reveal that this superfamily consists of five equally distant families, the Glucose (Glc), β-Glucoside (Bgl), Fructose (Fru), Mannitol (Mtl) and Lactose (Lac) families. Average hydropathy, amphipathicity and similarity plots were generated for these five families as well as for the entire superfamily. Charged residue distribution was analyzed, and the most conserved sequence motif, common to all five families, was identified. The results show that the members of all five families exhibit similar average hydropathy plots with regions of average amphipathicity and relative conservation also being similar. Evidence is presented suggesting that the Glucitol (Gut) family of Enzyme IIC constituents is a distant member of the Glc superfamily. Based on our analyses we offer a topological model that resembles, but differs in detail from the two previously proposed models.
Total Nitrogen and Free Amino Acid Losses and Protein Calorie Malnutrition of Hemodialysis Patients: Do They Really Matter?’
Authors: Prado de Negreiros Nogueira Maduro Isolda , Elias Natacha Maria , Nonino Borges Carla Barbosa , Padovan Gilberto João , Cardeal da Costa José Abrão , Marchini Júlio Sérgio
Published: May 2008

Background/Aims: Protein calorie malnutrition (PCM) in patients on hemodialysis (HD) is multifactorial; however, HD per se induces nutrient losses. The aim of the present study was to characterize the losses of total nitrogen (TN) and free amino acids (FAs) through the dialysate and to determine the relationship between this loss and PCM, food ingestion, and the characteristics of the hemodialyzer in patients on HD. Methods: In a prospective study, 21 patients submitted to low-flux HD 3 times a week were evaluated within a period of 6 months regarding nutritional status, dietary calorie and protein intake, and losses through the dialysate of TN, FA, and urea nitrogen (UN). The type, surface area and reuses (up to 12) of the dialysis membrane were determined on each occasion, and the adequacy of dialysis was estimated by Kt/V. Results: 50% of the patients were considered malnourished, although the mean protein and energy intakes were similar for the malnourished and nourished patients. Mean TN losses through the dialysate were 16 g/session (60% UN). FA losses varied from 3.8 to 4.2 g/total volume. TN and FA in the dialysate did not differ significantly between malnourished and nourished patients. There was a positive correlation between membrane (polysulfone) area and TN (p <0.05) and ultrafiltrate volume and TN (p < 0.05), and a nonsignificant correlation between reuse of the dialysis membrane and TN. Conclusions: TN and FA losses through the dialysate were similar for malnourished and non-malnourished patients on chronic HD, thus they do not act as indicators of nutritional status impairment.
DNA Fragmentation in Chronic Glomerulonephritis: An Immunohistological Analysis
Authors: Ott U. , Aschoff A. , Pocock J. , Fünfstück R. , Jirikowski G. , Stein G. , Wolf G.
Published: May 2008

Background: Experimental data suggest that apoptosis plays an important pathophysiological role in glomerulonephritis by restoring tissue structure after proliferation of intrinsic renal cells and infiltration of leukocytes. Relatively little is known of apoptosis in human glomerulonephritis, particularly in predicting renal function during follow-up. Methods: In order to colocalize different markers for cell damage in renal tissue from patients with different forms of glomerulonephritis (GN), a series of semithin sections from 34 kidney biopsies were studied retrospectively. Normal kidney from a nephrectomy specimen with a small renal adenocarcinoma served as a control. DNA fragmentation, expression of tissue transglutaminase II, BAX and BCL-2 were visualized immunohistochemically. In some renal biopsies, immunohistochemical staining for activated caspase 3 was performed. Proinflammatory markers (C-reactive protein, leukocytes), serum creatinine, creatinine clearance, total proteinuria, albuminuria, α1-microglobulin and IgG excretion were determined at the time of biopsy. Serum creatinine and total proteinuria were assessed 6 and 12 months after renal biopsy. Results: Nuclei with different degrees of DNA fragmentation were mainly found in epithelial cells of tubules, but also in glomerular cells, regardless of the form of GN studied. Transglutaminase II expression was found only in cells with a strong staining for DNA fragmentation. DNA fragmentation localized to glomerular cells was more pronounced in proliferative than in non-proliferative forms of GN, being most abundant in patients with rapid progressive GN. Staining for activated caspase 3 in selected biopsies confirmed the presence of apoptosis. BAX and BCL-2 staining was detected within the same cells, but exhibited a different intracellular distribution. In proliferative GN, the extent of DNA damage in tubular epithelial cells significantly corresponds with the concentration of serum creatinine (p < 0.04) and with urinary excretion of α1-microglobulin (p < 0.01) at the time of biopsy. A significant correlation (p < 0.01) was seen between glomerular DNA fragmentation and follow-up total proteinuria 12 months after biopsy for proliferative forms of GN. The damaged glomerular area (e.g. mesangial sclerosis) significantly correlated with DNA fragmentation in proliferative, but not in nonproliferative GN at the time of biopsy. Furthermore, glomerular damaged showed a significant correlation with tubular DNA damage in proliferative GN. Conclusion: In glomerular cells, apoptosis may be important for the clearance of proliferating cells whereas in tubules, cell damage showed dependence on the degree of tubular injury mediated by inflammation and/or proteinuria. Although the degree of apoptosis in tubular cells correlates with serum creatinine in proliferative GN at the time of biopsy, it is of limited use to predict future renal function.
Allergic Sensitization to Common Airborne Allergens among Adults in Estonia
Authors: Raukas-Kivioja Aet , Raukas Evi-Saidi , Meren Mari , Loit Helle-Mai , Rönmark Eva , Lundbäck Bo
Published: May 2008

Background: Allergies and asthma exhibit a growing health problem in the world. Sparse data are available about the prevalence of allergic sensitization in Estonia and Eastern Europe as well. We studied the prevalence of allergic sensitization, the associations between respiratory symptoms and allergic sensitization, and the influence of age, gender, area of residence, number of siblings and other demographic attributes on allergic sensitization in the population aged 17–69 years. Methods: We conducted a cross-sectional study with 1,346 randomly selected participants distributed in 3 geographically and culturally distinct areas of Estonia. Sensitization to 15 aeroallergens was assessed by skin prick tests. The study also included a structured interview. Results: The total prevalence of allergic sensitization was 33.0%. When estimated in patients aged 20–44 years, the prevalence was 38.5%. The most common sensitizer was cockroach followed by storage mites, while sensitization to cat, dog or pollen (common sensitizers in Scandinavian countries) was low. Allergic sensitization was significantly more common in urban and suburban compared with rural areas. Storage mites were the most common sensitizers in rural areas. Living in urban or suburban areas before the age of 5 significantly increased the risk for positive skin prick tests to several allergens. Conclusions: A higher prevalence of allergic sensitization than previously believed was found. Cockroach and storage mite allergens are suggested to be included in the routine investigation panel in Estonia.
Frequency and Predictors of Symptomatic Intracerebral Hemorrhage in Patients with Ischemic Stroke Treated with Recombinant Tissue Plasminogen Activator outside Clinical Trials
Authors: Martí-Fàbregas Joan , Bravo Yolanda , Cocho Dolores , Martí-Vilalta Josep-Lluis , Díaz-Manera Jordi , San Román Luis , Puig Mireia , Blanco Miguel , Castellanos Mar , Millán Mónica , Roquer Jaume , Obach Víctor , Maestre José
Published: May 2008

Background: To determine the frequency and predictors of symptomatic intracerebral hemorrhage (SICH) in patients treated with recombinant tissue plasminogen activator (rt-PA). Methods: We reviewed the databases of 7 tertiary hospitals that treated ischemic stroke patients with intravenous rt-PA. We recorded demographic data, vascular risk factors, time between onset and treatment, dose, the NIHSS score, body temperature, blood pressure, platelet count, blood glucose, antiplatelet treatment, and CT data. We also registered the study protocol used for treatment and deviations from the accepted protocol. A control CT was performed on all patients. SICH was diagnosed if a parenchymal hematoma was detected within the 36 h after rt-PA and was associated with an increase of ≧4 in the NIHSS score. Bivariate analyses were performed followed by a logistic regression analysis. Results: A total of 347 patients were studied, whose mean age was 68 ± 10.9 years; 56% were men. Thirty-two patients (9.2%) exhibited a parenchymal hematoma, and 8 patients (2.3%) suffered a SICH. Patients with SICH had a higher frequency of previous transient ischemic attack (p = 0.04), early signs of ischemia (p = 0.003), hyperdense arterial sign (p = 0.008), and deviations (p = 0.002). Early signs of ischemia (OR 8.5, 95% CI 1.6–45.4, p = 0.01) and deviation from the protocol (OR 11.1, 95% CI 2.4–50, p = 0.002) were independent predictors of SICH. Conclusions: SICH is infrequent in patients with ischemic stroke treated with rt-PA outside of a clinical trial. Its frequency increases in the presence of early signs of ischemia on the noncontrast CT scan and deviations from the recommended protocol.
Multiparametric Iterative Self-Organizing Data Analysis of Ischemic Lesions Using Pre- or Post-Gd T1 MRI
Authors: Soltanian-Zadeh Hamid , Bagher-Ebadian Hassan , Ewing James R. , Mitsias Panayiotis D. , Kapke Alissa , Lu Mei , Jiang Quan , Patel Suresh C. , Chopp Michael
Published: May 2008

Background: The purpose of this work was to evaluate effects of Gd-diethylenetriaminepentacetic acid (DTPA) injection on T1-weighted images of stroke and lesion segmentation and characterization results generated by our multiparametric iterative self-organizing data (ISODATA) method. The post-Gd image incorporates vasculature information into the analysis. Methods: Either a pre-Gd T1-weighted image (T1WI) or a post-Gd T1WI was used along with diffusion-, T2- and proton-density-weighted images in the analysis. ISODATA is a data-driven method that segments and characterizes tissue damage in stroke using multiparametric MRI. Results: Experimental results in both animal and human studies showed that the use of post-Gd T1WI modified the segmentation and characterization results on the periphery of the lesion. The peripheral region that changes with Gd-DTPA has a higher permeability compared to the rest of the lesion. Either of the data sets (including pre- or post-Gd T1WI) was used to estimate the tissue recovery and generated consistent results. Conclusions: This study shows that our multiparametric ISODATA approach consistently identifies and characterizes the core of the ischemic lesion. It also shows that the inclusion of post-Gd T1WI results in the segmentation and characterization of the lesion periphery if it has a higher permeability compared to the rest of the lesion. Finally, it confirms that the multiparametric ISODATA MRI characterizes tissue damage and recovery in stroke.
Improved Survival after Stroke: Is Admission to Hospital the Major Explanation? Trend Analyses of the Auckland Regional Community Stroke Studies
Authors: Carter Kristie N. , Anderson Craig S. , Hackett Maree L. , Barber P. Alan , Bonita Ruth
Published: May 2008

Background: There is uncertainty regarding the impact of changes in stroke care and natural history of stroke in the community. We examined factors responsible for trends in survival after stroke in a series of population-based studies. Methods: We used statistical models to assess temporal trends in 28-day and 1-year case fatality after first-ever stroke cases registered in 3 stroke incidence studies undertaken in Auckland, New Zealand, over uniform 12-month calendar periods in 1981–1982 (n = 1,030), 1991–1992 (1,305) and 2001–2002 (1,423). Cox proportional hazards regression was used to evaluate the significance of pre-defined ‘patient’, ‘disease’ and ‘service/care’ factors on these trends. Results: Overall, there was a 40% decline in 28-day case fatality after stroke over the study periods, from 32% (95% confidence interval, 29–35%) in 1981–1982 to 23% (21–25%) in 1991–1992 and then 19% (17–21%) in 2002–2003. Similar relative declines were seen in 1-year case fatality. In regression models, the trends were still significant after adjusting for patient and disease factors. However, further adjustment for care factors (higher hospital admission and neuroimaging) explained most of the improvement in survival. Conclusions: These data show significant downwards trends in case fatality after stroke in Auckland over 20 years, which can largely be attributed to improved stroke care associated with increases in hospital admission and brain imaging during the acute phase of the illness.
Peroxisome Proliferator-Activated Receptor-γ Agonists Diminish Peritoneal Functional and Morphological Changes Induced by Bioincompatible Peritoneal Dialysis Solution
Authors: Yao Qiang , Pawlaczyk Krzysztof , Ayala Ernesto Rodríguez , Kuzlan Malgorzata , Styszynski Arkadiusz , Breborowicz Andrzej , Heimbürger Olof , Qian Jiaqi , Stenvinkel Peter , Axelsson Jonas , Lindholm Bengt
Published: May 2008

Background: To evaluate if peroxisome proliferator-activated receptor-γ (PPAR-γ) agonists have a potential protective effect on the peritoneum changes induced by bioincompatible peritoneal dialysis (PD) solution in vivo. Methods: Male Wistar rats were dialyzed three times daily for 28 days with 1.36% Dianeal® (two groups: with (D+R) or without (D) rosiglitazone) or 1.36% Physioneal® (two groups: with (P+R) or without (P) rosiglitazone). Peritoneal transport of fluid and small solutes was assessed. Nine rats that did not receive dialysis served as controls. Results: Significant morphological changes were found in the D group compared with controls. Additional use of rosiglitazone in the D+R group resulted in less morphological changes and expression of collagen I as well as an increased drainage volume. The expression of VEGF was inhibited by rosiglitazone while no apparent effect was found regarding TGF/Smad pathway. Conclusions: The addition of rosiglitazone to standard dialysis fluids can maintain the peritoneal morphology and increase ultrafiltration in a PD rat model.
Chromosome ‘speed dating’ during meiosis of polyploid Brassica hybrids and haploids
Authors: Nicolas S.D. , Leflon M. , Liu Z. , Eber F. , Chelysheva L. , Coriton O. , Chèvre A.M. , Jenczewski E.
Published: May 2008

Given their tremendous importance for correct chromosome segregation, the number and distribution of crossovers are tightly controlled during meiosis. In this review, we give an overview of crossover formation in polyploid Brassica hybrids and haploids that illustrates or underscores several aspects of crossover control. We first demonstrate that multiple targets for crossover formation (i.e. different but related chromosomes or duplicated regions) are sorted out during meiosis based on their level of relatedness. In euploid Brassica napus (AACC; 2n = 38), crossovers essentially occur between homologous chromosomes and only a few of them form between homeologues. The situation is different in B. napus haploids in which crossovers preferentially occur between homeologous chromosomes and a few can then form between more divergent duplicated regions. We then provide evidence that the frequency of crossovers between a given pair of chromosomes is influenced by the karyotypic and genetic composition of the plants that undergo meiosis. For instance, genetic evidence indicates that the number of crossovers between exactly the same pairs of homologous A chromosomes gets a boost in Brassica digenomic tetraploid (AACC) and triploid (AAC) hybrids. Increased autosyndesis within B. napus haploids as compared to monoploid B. rapa and B. oleracea is another illustration of this process. All these observations may suggest that polyploidization overall boosts up crossover machinery and/or that the number of crossovers is modulated through inter-bivalents or univalent-bivalent cross-talk effects. The last part of this review gives an up-to-date account of what we know about the genetic control of homologous and homeologous crossover formation among Brassica species.
Chromosome numbers in plant cytotaxonomy: concepts and implications
Authors: Guerra M.
Published: May 2008

Chromosome number is the karyotype feature most commonly used in cytotaxonomical analyses. The chromosome number can be a plesiomorphic characteristic of a large clade or a recurrent trait which arose independently in two or more clades. Some concepts regarding chromosome number variation, such as base number, aneuploidy, paleopolyploidy, and neopolyploidy have been used by different authors in quite different ways. Therefore, its use in cytotaxonomy and karyotype evolution deserves much attention. In this paper, these terms are reappraised and their meaning and implication for plant cytotaxonomy are discussed.
Repetitive DNA and chromosomal rearrangements: speciation-related events in plant genomes
Authors: Raskina O. , Barber J.C. , Nevo E. , Belyayev A.
Published: May 2008

Chromosomal change is one of the more hotly debated potential mechanisms of speciation. It has long been argued over whether – and to what degree – changes in chromosome structure contribute to reproductive isolation and, ultimately, speciation. In this review we do not aim to completely analyze accumulated data about chromosomal speciation but wish to draw attention to several critical points of speciation-related chromosomal change, namely: (a) interrelations between chromosomal rearrangements and repetitive DNA fraction; (b) mobility of ribosomal DNA clusters; and (c) rDNA and transposable elements as perpetual generators of genome instability.
Cytogenetics of Festulolium (Festuca × Lolium hybrids)
Authors: Kopecký D. , Lukaszewski A.J. , Doležel J.
Published: May 2008

Grasses are the most important and widely cultivated crops. Among them, ryegrasses (Lolium spp.) and fescues (Festuca spp.) provide high quality fodder for livestock, are used for turf and amenity purposes, and play a fundamental role in environment protection. Species from the two genera display complementary agronomic characteristics and are often grown in mixtures. Breeding efforts to combine desired features in single entities culminated with the production of Festuca × Lolium hybrids. The so called Festuloliums enjoy a considerable commercial success with numerous cultivars registered all over the world. They are also very intriguing from a strictly cytogenetic point of view as the parental chromosomes recombine freely in hybrids. Until a decade ago this phenomenon was only known in general quantitative terms. The introduction of molecular cytogenetic tools such as FISH and GISH permitted detailed studies of intergeneric chromosome recombination and karyotyping of Festulolium cultivars. These tools were also invaluable in revealing the origin of polyploid fescues, and facilitated the development of chromosome substitution and introgression lines and physical mapping of traits of interest. Further progress in this area will require the development of a larger set of cytogenetic markers and high-resolution cytogenetic maps. It is expected that the Lolium – Festuca complex will continue providing opportunities for breeding superior grass cultivars and the complex will remain an attractive platform for fundamental research of the early steps of hybrid speciation and interaction of parental genomes, as well as the processes of chromosome pairing, elimination and recombination.
The use of cytogenetic tools for studies in the crop-to-wild gene transfer scenario
Authors: Benavente E. , Cifuentes M. , Dusautoir J.C. , David J.
Published: May 2008

Interspecific hybridization in plants is an important evolutionary phenomenon involved in the dynamics of speciation that receives increasing interest in the context of possible gene escapes from transgenic crop varieties. Crops are able to cross-pollinate with a number of wild related species and exchange chromosome segments through homoeologous recombination. In this paper, we review a set of cytogenetic techniques that are appropriate to document the different steps required for the stable introgression of a chromosome segment from a donor species (i.e., the crop) into a recipient species (i.e., the wild). Several examples in hybrids and derivatives are given to illustrate how these approaches may be used to evaluate the potential for gene transfer between crops and wild relatives. Different techniques, from classical chromosome staining methods to recent developments in molecular cytogenetics, can be used to differentiate genomes and identify the chromosome regions eventually involved in genetic exchanges. Some clues are also given for the study of fertility restoration in the interspecific hybrid forms.
Autoimmune Pancreatitis Accompanied by Cholecystitis, Periaortitis and Pseudotumors of the Liver
Authors: Matsubayashi Hiroyuki , Furukawa Hiroyoshi , Uesaka Katsuhiko , Sasaki Keiko , Ono Hiroyuki , Hruban Ralph H.
Published: May 2008

A variety of extrapancreatic lesions have been associated with autoimmune pancreatitis (AIP), and these lesions can be difficult to diagnose. We report a patient referred to Shizuoka Cancer Center with the diagnosis of a possible biliary carcinoma with liver metastasis who was shown to have AIP accompanied by pseudotumors of liver. Clinical imaging revealed diffuse enlargement of the head of the pancreas with irregular narrowing of the main pancreatic duct and inferior common bile duct, multiple liver masses, mediastinal lymphadenopathy, and thickening of the wall of the gallbladder and abdominal aorta. Cytology and biopsy from the pancreaticobiliary tract was negative for malignancy. Serum carcinoembryonic antigen (CEA) and carbohydrate antigen (CA19-9) levels were in the normal range, but soluble interleukin 2 receptor (sIL2R), IgG4 and antinuclear antibody were abnormally high (sIL2R: 2,550 U/ml; IgG4: 764 mg/dl). Corticosteroid therapy was effective and these abnormal findings all improved. This case demonstrates the clinical importance of AIP accompanied by other systemic disorders in the differential diagnosis of patients with a pancreatic mass lesion.
Airway Eosinophilic Inflammation Is Attenuated in Conserved Noncoding Sequence-1-Deficient Mice
Authors: Ohtomo Takayuki , Miyatake Shoichiro , Kajiyama Yuichiro , Umezu-Goto Makiko , Kobayashi Noriaki , Kaminuma Osamu , Mori Akio
Published: May 2008

Background: Conserved noncoding sequence-1 (CNS-1) is an important regulatory element for T helper 2 cytokine expression. IL-4, IL-5 and IL-13 expression as well as serum IgE level were attenuated in CNS-1–/– mice. Method: CNS-1–/– and CNS-1+/+ mice were sensitized with ovalbumin (OVA) followed by antigen challenge. The number of eosinophils and T helper 2 cytokine concentration in the bronchoalveolar lavage fluid, OVA-specific IgE antibody (Ab) in the serum and bronchial responsiveness to methacholine were examined. Results: Bronchoalveolar lavage fluid eosinophilia was significantly attenuated in CNS-1–/– mice compared to CNS-1+/+ mice, which were sensitized with OVA/aluminum once. OVA-specific IgE Ab was also attenuated. When mice were sensitized with OVA/aluminum twice, induction of eosinophilia and OVA-specific IgE Ab was not significantly different between CNS-1–/– and CNS-1+/+ mice. Conclusion: CNS-1 locus regulates eosinophilic inflammation in vivo.
Mechanism for the Decrease in the FIP1L1-PDGFRα Protein Level in EoL-1 Cells by Histone Deacetylase Inhibitors
Authors: Ishihara Kenji , Kaneko Motoko , Kitamura Hajime , Takahashi Aki , Hong Jang Ja , Seyama Toshio , Iida Koji , Wada Hiroshi , Hirasawa Noriyasu , Ohuchi Kazuo
Published: May 2008

Background: Acetylation and deacetylation of proteins occur in cells in response to various stimuli, and are reversibly catalyzed by histone acetyltransferase and histone deacetylase (HDAC), respectively. EoL-1 cells have an FIP1L1-PDGFRA fusion gene that causes transformation of eosinophilic precursor cells into leukemia cells. The HDAC inhibitors apicidin and n-butyrate suppress the proliferation of EoL-1 cells and induce differentiation into eosinophils by a decrease in the protein level of FIP1L1-PDGFRα without affecting the mRNA level for FIP1L1-PDGFRA. In this study, we analyzed the mechanism by which the protein level of FIP1L1-PDGFRα is decreased by apicidin and n-butyrate. Methods: EoL-1 cells were incubated in the presence of the HDAC inhibitors apicidin, trichostatin A or n-butyrate. The protein levels of FIP1L1-PDGFRα and phosphorylated eIF-2α were determined by Western blotting. Actinomycin D and cycloheximide were used to block RNA synthesis and protein synthesis, respectively, in the chasing experiment of the amount of FIP1L1-PDGFRα protein. Results: When apicidin- and n-butyrate-treated EoL-1 cells were incubated in the presence of actinomycin D, the decrease in the protein level of FIP1L1-PDGFRα was significantly enhanced when compared with controls. In contrast, the protein levels were not changed by cycloheximide among these groups. Apicidin and n-butyrate induced the continuous phosphorylation of eIF-2α for up to 8 days. Conclusions: The decrease in the level of FIP1L1-PDGFRα protein by continuous inhibition of HDAC may be due to the decrease in the translation rate of FIP1L1-PDGFRA.
The Synthetic PPARγ Agonist Troglitazone Inhibits Eotaxin-Enhanced Eosinophil Adhesion to ICAM-1-Coated Plates
Authors: Hirasawa Hiroyuki , Chiba Takahito , Ueki Shigeharu , Kamada Yumiko , Ito Wataru , Takeda Masahide , Fujita Miyoshi , Kato Hikari , Kayaba Hiroyuki , Chihara Junichi
Published: May 2008

Accumulation and activation of eosinophils in tissue are critical events in the allergic inflammatory response and adhesion molecules play important roles in this process. We previously demonstrated that human eosinophils expressed a nuclear receptor, peroxisome proliferator-activated receptor γ (PPARγ), and that stimulation with a PPARγ agonist attenuated cytokine/chemokine-induced eosinophil activation, such as survival, chemotaxis and degranulation. In the present study, we investigated the effect of troglitazone, a synthetic PPARγ agonist, on adherence to intercellular adhesion molecule-1 (ICAM-1). Eosinophils were purified from human peripheral blood, and the functional adherence to recombinant soluble ICAM-1-coated plates was examined. We found that in the presence of eotaxin, troglitazone inhibited eosinophil adherence in a concentration-dependent manner. This novel activity appears to be associated with modulation of qualitative change of integrins in response to eotaxin, because quantitative reduction of CD11a, CD11b and CD18 expression by troglitazone was not observed using flow cytometry. The PPARγ agonist troglitazone has a potent inhibitory effect on eosinophil adhesion to ICAM-1, and this may be a therapeutic modality for the treatment of eosinophil-related diseases including bronchial asthma.
Neutrophil Proteases Activate Eosinophil Function in vitro
Authors: Hiraguchi Yukiko , Nagao Mizuho , Hosoki Koa , Tokuda Reiko , Fujisawa Takao
Published: May 2008

Background: Recent evidence suggests that both neutrophilic and eosinophilic inflammation persist in the airways of patients with severe asthma. Mechanisms for interaction between neutrophils and eosinophils are still to be understood. Since eosinophils express protease-activated receptor 2, neutrophil-derived serine proteases may activate eosinophils. Objective: We investigated the effect of neutrophil serine proteases on eosinophil effector functions. Methods: Peripheral blood eosinophils were stimulated with elastase, cathepsin G and proteinase 3. Superoxide generation was quantitated with the cytochrome C reduction method. A panel of cytokines and chemokines in the culture supernatants were measured with a multiplex beads array system. Effects of an elastase inhibitor, sivelestat, and a serine protease inhibitor, PMSF, on the protease-induced reactions were also tested. Results: Neutrophil proteases significantly induced superoxide production from eosinophils. Elastase was the most potent among them. Sivelestat and PMSF inhibited the reaction. The proteases induced production of IL-6, IL-8, TNF-α and GRO-α, that have a possible connection with neutrophilic inflammation. Conclusion: Neutrophil proteases activate eosinophils to produce superoxide, proinflammatory cytokines and neutrophilotactic chemokines and may further aggravate airway inflammation in patients with severe asthma.
Inhibitory Effect of Budesonide Alone and in Combination with Formoterol on IL-5 and RANTES Production from Mononuclear Cells
Authors: Soma Tomoyuki , Takaku Yotaro , Kobayashi Takehito , Hagiwara Koichi , Kanazawa Minoru , Uematsu Kazutsugu , Nagata Makoto
Published: May 2008

Background:The use of a budesonide (BUD)/formoterol (FOR) combination (Symbicort®) for both maintenance and reliever therapy reduces the exacerbation of asthma better than the traditional fixed-dose regimens. In the early stage of exacerbation, the combination therapy could intervene with the development of subsequent airway inflammation. The objective of the study was to evaluate whether in the early stage of cell activation the use of BUD/FOR combination would modify the adhesion of eosinophils or production of cytokines from mononuclear cells. Methods:Human peripheral blood eosinophils, pretreated with BUD (0.1 µM), FOR (0.1 µM) or BUD/FOR combination for 30 min at 37°C, were stimulated with IL-5, leukotriene D4 or phorbol myristate acetate, and eosinophil adhesion was evaluated using an eosinophil peroxidase assay. BUD (0.1 µM), FOR (0.1 µM) or BUD/FOR combination was added to the peripheral blood mononuclear cells stimulated with ionomycin plus phorbol myristate acetate. Concentrations of IL-5 and RANTES in the cell supernatant were measured using ELISA. Results:BUD, FOR or BUD/FOR combination did not modify the eosinophil adhesion induced by any stimulators. In contrast, BUD or BUD/FOR combination significantly reduced the productions of IL-5 and RANTES in peripheral blood mononuclear cells (BUD: p < 0.0001, p = 0.027 vs. control; BUD/FOR: p < 0.0001, p = 0.031 vs. control) when the drugs were added 15 min, but not 4 h, following cell stimulation. Conclusion: In the early stage of exacerbation of asthma, inhaled BUD may suppress the progression of the allergic inflammatory cascade by inhibiting the mononuclear cells. These results also underline the importance of using BUD in rescue inhaler.
Immunohistochemical Localization of Transient Receptor Potential Vanilloid Subtype 1 in the Trachea of Ovalbumin-Sensitized Guinea Pigs
Authors: Watanabe Naoto , Horie Syunji , Spina Domenico , Michael Gregory J. , Page Clive P. , Priestley John V.
Published: May 2008

Background: We previously found many transient receptor potential vanilloid receptor subtype 1 (TRPV1) axons in the tracheal smooth muscle and epithelium of the guinea pig airway. One report indicates that the number of TRPV1 axons is significantly increased in patients with cough variant asthma. Aim: To determine whether the distribution of TRPV1 in the airways is altered in guinea pigs with an allergic phenotype. Methods: Ten guinea pigs were assigned to 2 groups in a double-blind study. Five animals were sensitized with ovalbumin and the other 5 underwent sham sensitization. Cryopreserved sections (30 µm) of tracheal tissues removed from each animal were stained with polyclonal serum rabbit anti-TRPV1 antibody (1:30,000) and examined by confocal microscopy. Results: Axons immunoreactive to TRPV1 localized to fine axons within the epithelium and around areas of smooth muscle, were more densely stained and frequent in the ovalbumin than in the sham group. Conclusion: The number of TRPV1-immunoreactive axons in the trachea increases under allergic inflammatory conditions.
A Contraction Assay System Using Established Human Bronchial Smooth Muscle Cells
Authors: Kitamura Noriko , Kaminuma Osamu , Kobayashi Noriaki , Mori Akio
Published: May 2008

Background: To further understand the mechanisms of airway obstruction in asthma, it is crucial to investigate contractile responses of human airway smooth muscle. An in vitro assay system employing collagen gels embedded with well-established bronchial smooth muscle cells of human origin was explored in the present study. Methods: Commercially available cultured human bronchial smooth muscle cells were embedded into a collagen gel. Well-known constrictors, histamine and methacholine, were added to the gel. The gel images were captured by an image analyzer, and contractile responses were evaluated. Results: Histamine and methacholine induced contraction of the gels in a dose-dependent manner. Pyrilamine, an H1 receptor antagonist, inhibited gel contraction in an agonist-specific manner. Conclusion: Our contraction assay system, employing widely distributed cultured cells, was highly reproducible and precise. It may go a long way toward understanding mechanisms of asthmatic responses and evaluation of antiasthma drugs.
Induction of Apoptosis in Human Basophils by Anti-Fas Antibody Treatment in vitro
Authors: Matsumoto Kenji , Maeda Akihiko , Bochner Bruce S. , Wakiguchi Hiroshi , Saito Hirohisa
Published: May 2008

Background: Basophils are thought to play an important role in the pathogenesis of allergic inflammation; however, the factors associated with basophil death are not fully understood. Fas (CD95) is a member of the TNF receptor superfamily and is known to induce apoptosis in activated T cells, neutrophils and eosinophils. In the present study, the expression and function of Fas in human basophils were investigated in vitro. Methods: Human cultured basophils were obtained by culturing cord blood-derived CD34+ cells in the presence of 2.5 ng/ml of IL-3 for 5–6 weeks. The expression of Fas was measured using flow cytometry. Cell viability and morphological changes after the incubation of basophils with anti-Fas mAb (clone CH11, IgM) in the presence of 1 ng/ml of IL-3 were measured using the trypan blue dye exclusion test and light microscopy, respectively. Results: Human cultured basophils constitutively and significantly expressed Fas on their cell surfaces. Treatment with anti-Fas monoclonal antibody (mAb) significantly reduced basophil viability in a time- and dose-dependent manner. When basophils were incubated with 10 ng/ml of anti-Fas mAb or control for 72 h, the basophil viability was 27.3 ± 8.8% and 89.3 ± 5.2%, respectively (p < 0.01). Anti-Fas mAb-treated basophils were shrunken and exhibited condensed nuclei, consistent with apoptosis. Conclusions: Our findings indicate that human basophils express functional Fas on their cell surfaces, and signaling via Fas may regulate basophil survival in vivo.
Allergen-Induced Basophil CD203c Expression as a Biomarker for Rush Immunotherapy in Patients with Japanese Cedar Pollinosis
Authors: Nagao Mizuho , Hiraguchi Yukiko , Hosoki Koa , Tokuda Reiko , Usui Tomoko , Masuda Sawako , Yamaguchi Masao , Fujisawa Takao
Published: May 2008

Background: Rush immunotherapy (RIT) can confer rapid clinical benefit on patients with allergic rhinitis or asthma. However, biomarkers representing mechanisms for the efficacy of RIT are still to be established. CD203c is a basophil activation marker known to be upregulated by cross-linking of the FcΕRIα receptor and may serve as a useful marker. Objective: We sought to investigate the changes in allergen-induced CD203c expression in patients with Japanese cedar pollen (JCP) pollinosis who received RIT. Methods: Nine patients treated with RIT were enrolled in the study. Whole blood was incubated with various concentrations of JCP extract. CD203c expression on basophils was quantitated by means of flow cytometry. JCP-specific IgG4 levels in sera were measured with ELISA. Basophil histamine release, CAP-RAST to JCP (JCP-IgE) and total IgE were also examined. The biomarkers listed above were evaluated before and sequentially after RIT. Symptom and quality of life scores were obtained during pre- and posttreatment pollen seasons. Results: All patients showed significant improvement in symptom and quality of life scores after RIT. Serum JCP-specific IgG4 titers were significantly elevated at 1 month and remained at high levels 12 months after the treatment. Stimulation with JCP extract induced enhancement of basophil CD203c expression in a concentration-dependent manner except for 2 subjects in whom no increase in CD203c by an anti-IgE antibody was observed (nonresponders). Significant reductions in the responses were observed in 4 subjects after RIT (reduction in CD203c expression, RCE) whereas no changes were seen in 3 subjects (non-RCE). RCE subjects were older than non-RCE counterparts, with mean ages of 20 and 12 years, respectively. No significant changes in JCP-specific IgE and total IgE levels were seen before and after RIT. Conclusion: Allergen-induced CD203c expression in basophils may represent, at least in part, the cellular mechanism for the therapeutic responses to RIT for JCP pollinosis. However, further larger-scale studies to confirm the utility of the test are necessary.
Blocking Antibody Is Generated in Allergic Rhinitis Patients during Specific Immunotherapy Using Standardized Japanese Cedar Pollen Extract
Authors: Kawakami Ayako , Koketsu Rikiya , Suzukawa Maho , Nagao Mizuho , Hiraguchi Yukiko , Tokuda Reiko , Fujisawa Takao , Nagase Hiroyuki , Ohta Ken , Yamamoto Kazuhiko , Yamaguchi Masao
Published: May 2008

Background: Japanese cedar pollen is by far the most important cause of allergic rhinitis in Japan. In this study, we assessed the induction of blocking antibody during specific immunotherapy (SIT) using a recently standardized allergen extract from Japanese cedar pollen. Methods: Basophils from nonallergic subjects were passively sensitized with serum samples prepared from pollinosis patients before and after SIT; all patients showed good clinical efficacy. The cells were then stimulated with the standardized allergen, and histamine release was measured. In most experiments, the basophil stimulation buffer contained 1% serum. Results: Pollinosis patients’ sera obtained both before and after SIT showed essentially similar sensitizing capacity for basophils. Basophil degranulation in response to a relatively low concentration of pollen extract was effectively suppressed by addition of post-SIT serum samples, indicating the presence of blocking antibody. The blocking antibody was IgG, and its potency varied widely among the donor patients. Conclusions: The standardized allergen extract from Japanese cedar pollen is useful not only for clinical application in SIT, but also for testing for induction of blocking antibody during SIT.
HMG-CoA Reductase Inhibitor Simvastatin Inhibits Proinflammatory Cytokine Production from Murine Mast Cells
Authors: Kagami Shin-ichiro , Kanari Hiroko , Suto Akira , Fujiwara Michio , Ikeda Kei , Hirose Koichi , Watanabe Norihiko , Iwamoto Itsuo , Nakajima Hiroshi
Published: May 2008

Background: Statins inhibit 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase, a key rate-limiting enzyme in the mevalonate pathway. Accumulating data suggest that statins exhibit anti-inflammatory effects on a number of experimental models including experimental autoimmune encephalomyelitis and antigen-induced allergic airway inflammation. However, the mechanism underlying the anti-inflammatory effect of statins is still largely unknown. In this study, we examined the effect of a representative statin, simvastatin, on proinflammatory cytokine production from murine mast cells. Methods: Bone marrow-derived mast cells (BMMCs) were stimulated with lipopolysaccharide (LPS) in the presence or absence of simvastatin, and TNF-α and IL-6 production from BMMCs was evaluated at mRNA and protein levels. The effect of simvastatin on the expression of tristetraprolin, an RNA-binding protein that promotes decay of TNF-α mRNA, was evaluated. Results: Incubation of BMMCs with simvastatin resulted in the inhibition of LPS-induced TNF-α production at both mRNA and protein levels. Simvastatin also inhibited IL-6 production from LPS-stimulated BMMCs. However, simvastatin did not enhance the expression of tristetraprolin. Conclusions: Simvastatin inhibits the production of TNF-α and IL-6 from activated mast cells in part by inhibiting de novo synthesis of their transcripts and the inhibition may account for the anti-inflammatory effect of simvastatin.