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In this study we aim to evaluate the assessment of bronchial pathologies in a murine model of lung transplantation with grating-based X-ray interferometry in vivo. Imaging was performed using a dedicated grating-based small-animal X-ray dark-field and phase-contrast scanner. While the contrast modality of the dark-field signal already showed several promising applications for diagnosing various types of pulmonary diseases, the phase-shifting contrast mechanism of the phase contrast has not yet been evaluated in vivo. For this purpose, qualitative analysis of phase-contrast images was performed and revealed pathologies due to previous lung transplantation, such as unilateral bronchial stenosis or bronchial truncation. Dependent lung parenchyma showed a strong loss in dark-field and absorption signal intensity, possibly caused by several post transplantational pathologies such as atelectasis, pleural effusion, or pulmonary infiltrates. With this study, we are able to show that bronchial pathologies can be visualized in vivo using conventional X-ray imaging when phase-contrast information is analysed. Absorption and dark-field images can be used to quantify the severity of lack of ventilation in the affected lung. more...

Purpose Lung transplant shows poor long-term survival due to chronic lung allograft dysfunction (CLAD). VEGF-C/VEGFR3 signaling may regulate lymphatic vessel activation and initiate alloimmunity after transplantation. However, its role in CLAD development remains unclear. Methods The CLAD model was established in mouse orthotopic left lung transplantation with a single mismatch (HLA-A2 knock-in C57BL/6 to C57BL/6). The allografts were collected at 14, 28 and 56d after surgery. Syngeneic transplantations were used as control. We analyzed the change of lymphatic density, lymphatic phenotype and the activation of pro-lymphangiogenic genes during CLAD development. Results We found increased expression of PROX-1 7d after transplantation in CLAD model. We further observed a significant increase of lymphatic vessel density 56d after transplantation in CLAD model. The trend of increased lymphatic activation reached no significance (Figure 1). Conclusion Our results show that lung transplantation leads to the rapid activation of pro-lymphangiogenic phenotype with consequent increase of lymphatic vessel density in CLAD model. The early activation of lymphangiogensis during the CLAD development may stimulate chronic alloimmune response. VEGF-C/VEGFR3 inhibition during lung transplant might therefore prevent CLAD. more...

Background: We have previously shown that the allergic sensitization to ovalbumin does not represent a superantigen-like immune response. In gene-targeted mice (ΔD-iD) with a single modified Diversity gene segment (DH) of the immunoglobulin heavy chain, enriched for charged amino acids, the asthma phenotype in a murine model was markedly alleviated compared to wild-type animals. Objective: We now sought to determine whether the confinement to a single DH gene segment alone leads to a reduced allergic phenotype. Methods: We examined another gene-targeted mouse strain (ΔD-DFL) with a single DH gene segment which encodes for neutral amino acids, thus reflecting the preferential repertoire in wild-type mice. Mice were sensitized intraperitoneally to ovalbumin. Results: Despite the constraint to a single DH gene segment, ΔD-DFL mice mounted high total and allergen-specific IgG1 and IgE serum levels after sensitization to ovalbumin. The affinity constants of allergen-specific IgG1 antibodies did not differ between ΔD-DFL and wild type. Following challenge with aerosolized allergen, a marked local TH2 cytokine response and an eosinophilic airway inflammation developed. Quantitative histology revealed increased mucus production and intense goblet cell metaplasia which were identical to those in wild type. Moreover, ΔD-DFL mice developed an airway hyperreactivity to methacholine and to the specific allergen, which both did not differ from those in wild-type animals. Conclusion: A single DH gene segment is sufficient for the establishment of the asthma phenotype in a murine model of allergic airway inflammation. Thus, the allergic phenotype depends on the amino acid composition and not on the diversity of the classical antigen-binding site. more...

The lung epithelia facilitate wound closure by secretion of various cytokines and growth factors. Nerve growth factor (NGF) has been well described in airway inflammation; however, its likely role in lung repair has not been examined thus far. To investigate the repair function of NGF, experiments were performed in vitro using cultured alveolar epithelial cells and in vivo using a naphthalene-induced model of Clara epithelial cell injury. Both in vitro and in vivo experiments revealed airway epithelial cell proliferation following injury to be dependent on NGF and the expression of its receptor, tropomyosin-receptor-kinase A. Additionally, NGF also augmented in vitro migration of alveolar type II cells. In vivo, transgenic mice over-expressing NGF in Clara cells (NGFtg) did not reveal any proliferation or alteration in Clara cell phenotype. However, following Clara cell specific injury, proliferation was increased in NGFtg and impaired upon inhibition of NGF. Furthermore, NGF also promoted the expression of collagen I and fibronectin in vitro and in vivo during repair, where significantly higher levels were measured in re-epithelialising NGFtg mice. Our study demonstrates that NGF promotes the proliferation of lung epithelium in vitro and the renewal of Clara cells following lung injury in vivo. more...

Introduction The most widely used protocol for the induction of experimental allergic airway inflammation in mice involves sensitization by intraperitoneal (i.p.) injections of the antigen ovalbumin (OVA) used in conjunction with the adjuvant aluminium hydroxide (alum). Although adjuvants are frequently used, there are questions regarding the necessity of alum for murine asthma studies due to the non-physiological nature of this chemical. Objective The objective of this study was to compare experimental asthma phenotypes between adjuvant and adjuvant-free protocols of murine allergic airway inflammation in an attempt to develop a standardized alternative to adjuvant use. Method An adjuvant-free OVA model of experimental asthma was investigated in BALB/c mice using i.p. or subcutaneous (s.c.) sensitization routes. For the s.c. sensitization, β-galactosidase (β-gal) was also tested as an antigen. In addition, OVA adjuvant and adjuvant-free sensitization protocols were compared in BALB/c and C57BL/6 mice. Open-field testing was performed to assess the effect of alum on mouse behaviour. Results Comparison of adjuvant vs. adjuvant-free and i.p. vs. s.c. protocols revealed that both adjuvant use and route of antigen application significantly influenced OVA-specific antibody production. Comparison of adjuvant and adjuvant-free protocols in this study clearly demonstrated the non-requirement of alum for the induction of acute allergic airway inflammation, as both protocols induce a similar disease phenotype. BALB/c mice were significantly more susceptible than C57BL/6 mice to sensitization. Using the improved s.c. adjuvant-free protocol, it was demonstrated that alternative antigens such as β-gal can also be utilized. Behavioural studies indicated severe distress in mice treated with alum. Conclusion The OVA s.c. adjuvant-free protocol used in this study generates a phenotype comparable to the benchmark adjuvant protocol widely used in the literature. The adjuvant-free alternative avoids the added complication of non-physiological adjuvants that may interfere with asthma treatment or prevention strategies. more...

Aldose reductase (AR) has been implicated in the etiology of diabetic complications. Under diabetic conditions, the elevated vascular glucose level causes an increased flux through the polyol pathway, which induces functional and morphological changes associated with secondary diabetic complications such as cataract, neuropathy, and nephrop?athy. Oxidative stress, antioxidants, and the polyol pathway have recently been found to be linked in pathological states. A large number of structurally different compounds have been studied as potent in vitro AR inhibitors (ARIs). However, with few exceptions, these compounds did not show clinical benefit, and some even produced serious side effects. In view of the ARI activity of certain indole derivative compounds and antioxidant properties of melatonin, we investigated some indole-based analogs of melatonin derivatives. Antioxidant and ARI activity tests were applied to nine indole derivatives that are substituted at the third and fifth positions. Also, the relationship between ARI and antioxidant enzyme activity is discussed. more...

Summary Background Non-selective cation influx through canonical transient receptor potential channels (TRPCs) is thought to be an important event leading to airway inflammation. TRPC6 is highly expressed in the lung, but its role in allergic processes is still poorly understood. Objective The purpose of this study was to evaluate the role of TRPC6 in airway hyperresponsiveness (AHR) and allergic inflammation of the lung. Methods Methacholine-induced AHR was assessed by head-out body plethysmography of wild type (WT) and TRPC6−/− mice. Experimental airway inflammation was induced by intraperitoneal ovalbumin (OVA) sensitization, followed by OVA aerosol challenges. Allergic inflammation and mucus production were analysed 24 h after the last allergen challenge. Results Methacholine-induced AHR and agonist-induced contractility of tracheal rings were increased in TRPC6−/− mice compared with WT mice, most probably due to compensatory up-regulation of TRPC3 in airway smooth muscle cells. Most interestingly, when compared with WT mice, TRPC6−/− mice exhibited reduced allergic responses after allergen challenge as evidenced by a decrease in airway eosinophilia and blood IgE levels, as well as decreased levels of T-helper type 2 (Th2) cytokines (IL-5, IL-13) in the bronchoalveolar lavage. However, lung mucus production after allergen challenge was not altered by TRPC6 deficiency. Conclusions TRPC6 deficiency inhibits specific allergic immune responses, pointing to an important immunological function of this cation channel in Th2 cells, eosinophils, mast cells and B cells. more...

PurposeTo elucidate whether the gene polymorphisms of glutathione S-transferase (GST) M1, T1, and PI are associated with the development of exudative age-related macular degeneration.MethodsThe authors genotyped 35 white patients with exudative age-related macular degeneration and 159 healthy controls. Genomic DNA from peripheral blood was examined using polymerase chain reaction and defined for the genetic polymorphisms of GST.ResultsNo association was observed between GSTM1, GSTT1, and GSTP1 polymorphisms and age-related macular degeneration risk (p>0.05). The frequencies of the combination of the GSTM1 (null) and GSTP1 (mutant), GSTM1 (null), and GSTT1 (null) genotype polymorphisms in patients with exudative age-related macular degeneration differed greatly from those of the control group (p=0.001 OR [95% CI]: 7.70 [2.28–25.98] and p=0.007 OR [95% CI]: 3.88 [1.51–10.02], respectively).ConclusionsThe present study suggests that the GSTM1 (null) and GSTT1 (null), GSTM1 (null), and GSTP1 (mutant) combinations may be a genetic risk factor for the development of exudative age-related macular degeneration. However, the potential role of GST polymorphisms as a marker of susceptibility to age-related macular degeneration needs further studies in a larger number of patients. more...

BACKGROUND: Increased mucus production is a critical factor impairing lung function in patients suffering from bronchial asthma, the most common chronic inflammatory lung disease worldwide. OBJECTIVE: This study aimed at investigating whether goblet cell (GC) metaplasia and mucus production are differentially regulated in proximal and distal airways. METHODS: Female Balb/c mice were sensitized to ovalbumin (OVA) and challenged with an OVA-aerosol on two consecutive days for one week (acute) or twelve weeks (chronic). Real-time RT-PCR analysis was applied on microdissected airways. RESULTS: In acutely and chronically OVA-challenged mice GC metaplasia and mucus production was observed in proximal but not in distal airways. In contrast, inflammation reflected by the infiltration of eosinophils and expression of the TH2-type cytokines IL-4 and IL-13 was increased in both, proximal and distal airways. Abundance of IL-13Rα1 was lower in distal airways of healthy control mice. Under acute and chronic OVA-exposure, activation of IL-13Rα1-dependent signaling cascade, reflected by Spdef and Foxo3A transcription factors, was attenuated in distal compared to proximal airways. CONCLUSION & CLINICAL RELEVANCE: These data indicate that distal airways might be less sensitive to IL-13 induced GC metaplasia and mucus production through lower expression of IL-13Rα1 and attenuated activation of downstream signaling. This might represent a protective strategy in order to prevent mucus plugging of distal airways and thus, impaired ventilation of attached alveoli. more...

In the present study, the CAMP-factor (cfb) gene of streptococci of serological group B (Streptococcus agalactiae) and the CAMP-factor (cfu) gene of S. uberis could be amplified by polymerase chain reaction. A cfb specific amplicon could be observed for all 128 phenotypically CAMP-positive S. agalactiae, for the phenotypically CAMP-negative S. agalactiae strain 74-360, and for 2 S. difficile reference strains. A cfu specific amplicon could be observed for all 7 phenotypically CAMP-positive S. uberis. Four S. agalactiae strains isolated from 4 cows with mastitis appeared to be phenotypically CAMP-negative and negative in the cfb gene PCR. The CAMP-positive and CAMP-negative isolates, including both S. difficile, could be identified as S. agalactiae by amplification of a S. agalactiae specific part of the V2 region of the 16S rRNA and a species-specific part of the 16S-23S rRNA intergenic spacer region. Amplification of an internal fragment of the cfb gene with a reduced annealing temperature yielded positive reactions not only for CAMP-positive S. agalactiae, but also for phenotypically CAMP-positive S. pyogenes (n = 4), S. canis (n = 28), and S. uberis (n = 7), indicating a close relation of the CAMP genes of these 4 species. The relation could be further demonstrated by sequencing the internal fragment of the CAMP-factor (cfg) gene of S. canis and comparing the sequence with those of S. agalactiae, S. pyogenes, and S. uberis.Key words: CAMP factor, cfb, cfu, S. canis. more...

Background—When bound to mast cell FceRI, IgE serves as antigen receptor for allergic reactions, permitting specific identification of the allergen. Although the core of the classic antigen-binding site is heavy chain complementarity determining region 3 (CDR-H3), recent studies suggest that allergens might also bind IgE in a superantigen-like fashion outside the classic antigen-binding site. Objective—We sought to evaluate the contribution of the classic CDR-H3-centric antigenbinding site to the development of an allergic phenotype. Methods—Using a murine model of experimental asthma, we characterized a gene targeted mouse strain expressing an altered range of CDR-H3s (ΔD-iD mice) in response to the hydrophobic allergen ovalbumin. Mutant and wildtype (wt) mice were sensitized intraperitoneally with ovalbumin; non-sensitized mice served as controls. Results—We found the composition of the classic CDR-H3-centric antigen-binding site to be critical for the development of characteristic aspects of allergic asthma: (i) Compared to wt animals, ΔD-iD mice showed a significantly less pronounced ovalbumin-induced rise in allergenspecific IgE levels and hence in total serum IgE levels. (ii) In addition, ΔD-iD mice demonstrated a significant reduction in eosinophilic airway inflammation, as well as in IL-4, IL-5, and IL-13 levels in BAL fluids. Conclusion—Allergic sensitization and airway inflammation depend on the composition of the predominant CDR-H3 repertoire, suggesting that the classic CDR-H3-centric antigen-binding site plays a crucial role in creating the immunological interface between allergen and IgE. Our results further emphasize a central role of IgE, not only in mediating but also in regulating the allergic immune response. more...

Summary Background A subset of food-allergic patients does not only respond clinically with symptoms in the gastro-intestinal tract but also with asthmatic reactions. Objective The aim of this study was to analyse whether CD4+ T cells from mice with intestinal immediate-hypersensitivity reactions to food allergen are involved in the development of experimental asthma. Methods BALB/c mice were intraperitoneally sensitized to ovalbumin (OVA), followed by repeated intra-gastric (i.g.) OVA challenges. Control animals were either sham-sensitized or sham-challenged with phosphate-buffered saline (PBS). Duodenum, jejunum, ileum and colon were histologically examined. CD4+ T cells from mesenteric lymph nodes were transferred from various donor groups into recipient mice that received either OVA or PBS aerosol challenges. Recipients were analysed by measurements of lung function using head-out body-plethysmography and examination of broncho-alveolar lavage and lung histology. Results The highest levels of OVA-specific IgE antibody levels were detected in OVA-sensitized and OVA-challenged mice. Throughout the lower intestinal tract, a marked infiltration with eosinophils was observed, and goblet cell numbers as well as goblet cell area were significantly increased. The villus/crypt ratio was decreased compared with controls. The transfer of CD4+ T cells from mesenteric lymph nodes of OVA-sensitized and OVA-challenged mice triggered airway hyperreactivity and eosinophilic airway inflammation in recipients aerosol challenged with OVA, but not with PBS. Conclusion We conclude that CD4+ T cells from mesenteric lymph nodes of mice with allergen-induced immediate-type hypersensitivity reactions in the gut are able to transfer the phenotype of experimental asthma. more...

ÖZET Bu araştırma, Şanlıurfa İli kent merkezinde Cutaneous Leishmaniasis (CL) veya diğer adıyla Şark Çıbanı hastalığının prevalansını tahmin edebilmek, halkın bu hastalık için sağlık kuruluşlarından ne ölçüde yararlandığım saptayabilmek, halkın hastalık ile ilgili bilgi düzeylerini ortaya çıkarabilmek ve Şark Çıbanı hastalığının bulaşması ile ilgili risk faktörlerini tanımlayabilmek için planlanmıştır. Araştırmanın veri toplama aşaması, Şanlıurfa İli'nde Nisan-Mayıs 1998 tarihleri arasında, seçilmiş örneklem grubunda (397 hanede yaşayan 2432 kişide) gerçekleştirilmiştir. Hanelerin seçiminde araştırma bölgesi mahallelerin nüfusu oranında rasgele örnekleme yöntemi kullanılmıştır. Hanelerdeki kişilerle yapılan yüz yüze görüşmeler ve kişilerin muayeneleri sonucu, Cutaneous Leishmaniasis hastalığının prevalansı % 10,44 olarak bulunmuştur. Araştırma sırasında tespit edilen vakaların % 42,13'ü, teşhis için ya hiçbir yere başvurmadığı veya teşhis için yararlı olmayan yardım yollarına başvurduğu, son bir yıl içinde gelişen vakalara bakıldığında bu oranın, % 19,25 'e düştüğü görülmüştür. Araştırma kapsamında kişilerin CL ile ilgili bilgi düzeylerinin de tespiti için her haneden bir kişi ile görüşme yapılmıştır. Görüşme yapılan kişilerin % 85,39'u Şark Çıbanı hastalığım duyduğu anlaşılmıştır. Hastalığı hiç duymayanların ailelerinde de hiç vakaya rastlanmamıştır. Görüşülen kişilerin % 16,22'si hastalığın nedenini, % 5,90'ı hastalığın bulaşma şeklini, % 7,66'sı hastalıktan korunma yollarını tam olarak bildiği, ayrıca % 70,5 1 `i hastalığın tedavisinin nerede yapıldığını, % 58,12'sinin tedavinin nasıl yapıldığını bildiği saptanmıştır. Ailede Şark Çıbanı olgusunun bulunmasının kişilerin bilgi düzeyi üzerine olumlu etki yaptığı görülmüştür. Yapılan araştırma sonunda müstakil ve briketten inşa edilen evlerin, ev dışındaki tuvaletlerin, fosseptik çukurlu evlerin, vektör üremesini ve hastalığın bulaşmasını kolaylaştırdığı tespit edilmiştir. Ayrıca bölgede çok sayıda bulunan kötü sağlık şartlarındaki hayvan barınakları ve gübreliklerin de tatarcık üremesini kolaylaştırdığı bulunmuştur. Bu sonuçlar, Cutaneous Leishmaniasis hastalığının, araştırma bölgesindeki her on kişiden birini etkilediğini, hastalığın kötü çevre koşullan ve konut özellikleri ile ilgili olduğunu göstermiştir. Ayrıca Sağlık Müdürlüğü' nün hastalık ile ilgili yeterince güvenilir veri toplayamadığını, bölgede yaşayan kişilerin hastalığı duymalarına rağmen, yeterince bilgili ve duyarlı olmadıklarım ortaya çıkarmıştır. Anahtar Kelimeler : Cutaneous Leishmaniasis, Şanlıurfa, Prevalans, Risk Faktörleri, Bilgi düzeyi. IV ABSTRACT This research is planned to guess the prevalence of CL disease in Sanliurfa, to determine in what frequency the society benefit from health centers for that disease, to determine the society's knowledge level about disease and to define risk factors about transmission of CL. This research is done on 2432 people who are living in 397 houses, between April-May 1998 in Sanliurfa. In ratio of population, randomized sampling method is used to choose the houses in research area. As a results of the interwievs done with the people facely in the houses and examining the people, the prevalence of CL disease is found % 10,44. During the research, it's seen that they applied nowhere for diagnosis or they applied useless assistance for diagnosis, and in last one year, this ratio was decreased % 19,25. During the research, the interwievis done with one person from each family to determine the level of knowledge about disease. % 85,39 of the interwieved people are known about CL. There was no case in the families who are not aware of the disease. It's seen that % 16,22 of the interwieved people know the cause of the disease, % 5,90 of them know the way of transmission, % 7,66 of them know ways of the prevention about CL, % 70,51 of them know how the treatment is done. It's seen that presence of Cutaneous Leishmaniasis in the family affect the knowledge of the people positively. It's determined in the research that houses with garden and brick houses, outdoor touilets and houses with cesspool ease the vector breeding and transmission of the disease. It's determined that, shelters which are bad health conditions ease the breeding of sandfly. These results show that CL affect one in each ten people, and the disease is related with bad environmental conditions and with house charesteristics. It's also seen that Provincial Health Directorate can't collect safe datas and although the people who live in region hear the disease, they don't have enough knowledge and they are not sensitive. Key words : Cutaneous Leishmaniasis, Sanliurfa, Prevalence, Risk factors, Knwledge Level. 116 more...

ARKIVOC: vol. 2008, no. 2, (issn) 1551-7012, (dlps) 5550190.0009.210, This work is licensed under a Creative Commons Attribution-NonCommercial 3.0 License. Please contact mpub-help@umich.edu to use this work in a way not covered by the license. more...