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Eight novel heterocyclic Schiff bases derived from the condensation reactions of indole 3-carboxaldehyde with different l-amino acids (histidine, glutamic acid, aspartic acid, leucine, valine) as well as with some aminophenols, have been synthesized and characterized by various spectroscopic methods (IR, MS, (1)H NMR). Schiff base derivatives of indole 3-carboxaldehyde were labeled with (99m)Tc and radiochemical purity was above 97% which is ascertained by instant thin layer chromatography using different solvent conditions. Stability studies of all the derivatives of indole 3-carboxaldehyde was determined under physiological conditions and were stable for more than 24h. Blood clearance showed a quick wash out from the circulation and biological half life was found to be t((1/2))(F)=1h 15min; t((1/2))(S)=10h 05min. Excellent quality radioimages of tumor bearing mice were recorded showing rapid clearance of background activity, visualization of tumor at 3h and clearance from kidneys of histidine analogue which was further evidenced in biodistribution studies. Antimicrobial activity of these Schiff base compounds was evaluated against Bacillus subtilis, Pseudomonas fluorescence, Staphylococcus aureus, Aspergillus niger, Candida albicans and Trichophyton rubrum.

The potential role of iNOS as a viable therapeutic target for the treatment of asthma was severely hampered by the negative clinical trial results showing that a selective iNOS inhibitor did not affect airway hyperreactivity (AHR) or airway inflammation after allergen challenge in human subjects with asthma. Our laboratory has shown that iNOS inhibition is protective against airway inflammation upon acute, but not chronic, exposures to ovalbumin. More importantly, iNOS inhibition protected against lung fibrosis suggesting that iNOS inhibition may be protective against some aspects of asthma. Here, we show that iNOS inhibition, pharmacologically by L-Nil or by gene knockout, provided an excellent protection against AHR upon an acute, but not chronic, exposure to ovalbumin. Our laboratory also established a reciprocal relationship between iNOS and PARP-1. However, PARP inhibition, protected against inflammation and AHR upon acute and chronic exposure to ovalbumin. It is interesting that PARP-1 inhibition does not completely abrogate expression of iNOS leaving the possibility that the protective effect of PARP inhibition against inflammation and AHR may be associated with reduction but not the complete inhibition of iNOS and associated production of moderate levels of NO. Studies are being conducted to verify this hypothesis and clarify the intricate roles of iNOS in asthma pathogenesis allowing the possibility of being a viable target for the treatment of the disease.

Elevated level of IL-18 is reported in a number of allergic diseases and we earlier reported induced blood IL-18 and esophageal IL-18Rα mRNA in human EoE. Therefore, the current study focuses on the hypothesis that allergen-induced IL-18 and its receptor (IL-18Rα) positive cells may have an important role in EoE pathogenesis. Accordingly, we examined the blood IL-18 and esophageal IL-18Rα mRNA levels in aeroallergen and food allergen induced experimental mouse model of EoE. Herein, we demonstrate that blood IL-18 protein and esophageal IL-18Rα mRNA are induced in the mouse model of EoE and the source of induced esophageal IL-18Rα transcript are iNKT cells. We report that intranasal delivery of rIL-18 induces both mast cells and eosinophilic inflammation in the esophagus. Furthermore, to establish the significance of IL-18 in EoE pathogenesis, we next examined DOX-inducible rtTA-CC10-IL-18 bitransgenic mice that have significantly induced IL-18 protein in the esophagus. Our analysis indicated that DOX exposed IL-18 tg mice develop most of the characteristics of EoE, such as esophageal intraepithelial eosinophilia, increased mast cells and esophageal remodeling including fibrosis. Mechanistically, we show that IL-5 gene-deficient mice and iNKT cell-deficient (CD1d null) mice do not induce EoE in response to IL-18 intranasal challenge. Taken together, we provide evidences that allergen induced IL-18 has a significant role in promoting IL-5 and iNKT dependent EoE pathogenesis.

Two series of N-substituted-2-substituted benzimidazole derivatives, viz. l-benzyl-2-substituted benzimidazole 8-14 and 1-(p-chlorophenyl)-2-substituted benzimidazole 15-21 have been synthesized and tested for their antiviral activities. These compounds have been screened for Tobacco mosaic viruses and Sunhemp rosette viruses and show significant activities.

Background Atopic individuals are predisposed to mounting vigorous T H 2-type immune responses to environmental allergens. The skin is often the first organ that manifests allergic disease and may provide an early entry point for antigen sensitization. Objective We sought to determine whether epicutaneous exposure to the aeroallergen Aspergillus fumigatus induces nasal allergic responses. Furthermore, we aimed to examine the mechanism involved. Methods Wild-type and signal transducer and activator of transcription 6 (STAT6)–deficient mice were exposed to epicutaneous A fumigatus and control antigen ovalbumin. Nasal inflammation and responsiveness to methacholine were monitored. Results Exposure to epicutaneous A fumigatus antigen induced a marked atopic dermatitis-like phenotype in a manner significantly more efficient than epicutaneous ovalbumin. A single A fumigatus intranasal challenge induced clinical nasal responses and hyperresponsiveness to methacholine in the nose as manifested by nasal symptoms, accompanied by allergic airway and nasal inflammation. Mechanistic analysis using gene-targeted mice revealed that the clinical nasal responses and hyperresponsiveness were STAT6-dependent. Although STAT6 was required for changes in nasal responses, it was not required for epicutaneous pathology except eosinophilia. Conclusion Epicutaneous exposure to the aeroallergen A fumigatus potently primes for STAT6-dependent nasal responses. These results draw attention to the cooperative interaction between the nasal tract and skin. Clinical implications The skin is a potent site for antigen sensitization in the development of experimental allergic rhinitis.

Novel bifunctional chelating agents bearing an aromatic rigid backbone have been synthesized and characterized on the basis of spectroscopic techniques. These macrocyclic multidentate chelating agents were conjugated with monoclonal antibody which forms stable complexes with 99mTc with high radiochemical purity.

The purpose of this study was to obtain the convenient, synthetically useful bifunctional chelating agent, 6-(4-isothiocyanatobenzyl)-5,7-dioxo-1,11-(carboxymethyl)-1,4,8,11-tetraazacyclotridecane, and to apply it to stable (99m)Tc-labelling of monoclonal antibodies (mAbs).The chelate was synthesised by reaction of nitrobenzyl malonate and triethylenetetramine followed by alkylation by reacting with bromoacetic acid at pH 10. The amino group was converted to isothiocyanato derivative by reacting with thiophosgene at pH 2.0. Conjugation with mAbs [(anti-carcinoembryonic antigen (CEA) and anti-epidermal growth factor receptor (EGFr)] was performed at pH 8.4 using trisodium phosphate solution by incubating at 37 degrees C for 1 h and subjected to purification on size exclusion chromatography.When radioimmunoconjugates were labelled with (99m)Tc, the specific activity of immunoconjugates was 20-30 mCi/mg of protein and their immunoreactivity exceeded 80%. The stability in serum indicated that the metal remained bound to antibodies. Biodistribution studies in athymic mice grafted with U-87 human glioblastoma multiforme and MDA-MB-468 human breast carcinoma tumours revealed significant localisation of (99m)Tc-labelled antibodies in tumours and reduced accumulation in normal organs.This bifunctional chelating agent is promising for immunoscintigraphy because of good tumour-to-normal organ contrast.

Renal artery stenosis after renal transplant is a dreaded complication. We report two cases of post-renal transplant stenosis in internal iliac artery used for end-to-end anastomosis with the graft renal artery. Both patients were successfully treated by percutaneous transluminal angioplasty with stenting of the internal iliac artery.

The study evaluates the capability of tripalmitin nanoparticles in enhancing the tumor uptake of etoposide, and the influence of administration route on the biodistribution and tumor uptake of etoposide loaded tripalmitin (ETPL) nanoparticles in Dalton's lymphoma tumor bearing mice. ETPL nanoparticles were prepared by melt-emulsification and high pressure homogenization followed by the spray drying of nanodispersion. Characterization of the nanoparticles was done by particle size analysis, zeta potential measurement and scanning electron microscopy. The size of ETPL nanoparticles was 387 nm and possessed negative charge. Etoposide and ETPL nanoparticles were radiolabeled with 99mTc with high labeling efficiency. The labeled complexes showed good in vitro stability in the presence of DTPA/cysteine and serum stability. Etoposide and ETPL nanoparticles were injected by subcutaneous, intravenous or intraperitoneal routes and their biodistribution and tumor uptake were determined. Subcutaneous injection reduced the distribution of ETPL nanoparticles to all the tissues studied whereas after intraperitoneal injection, the distribution of ETPL nanoparticles to tissues was higher than free etoposide. The intravenous injection resulted in lower concentrations of ETPL nanoparticles in the organs of RES compared to free etoposide. ETPL nanoparticles experienced significantly high brain distribution after intraperitoneal injection indicating its potential use in targeting etoposide to brain tumors. After subcutaneous injection, the tissue distribution of ETPL nanoparticles increased with time indicating their accumulation at the injection site for a longer time. The tumor uptake of both etoposide and ETPL nanoparticles was significantly high after subcutaneous injection (P

Re-examination of the leaves of Nyctanthes arbor-tristis led to the isolation and identification of new minor iridoid glucoside, arborside D [1], as its acetyl derivative. The structure of the new compound was determined using spectral methods.

Eosinophil infiltration into the esophagus occurs in a wide range of diseases; however, the underlying pathophysiologic mechanisms involved are largely unknown. We have previously reported that simultaneous delivery of allergen to the lung and gastrointestinal tract induces experimental eosinophilic esophagitis (EE). We aimed to determine whether delivery of a Th2 cytokine (interleukin [IL]-13) to the lung was sufficient for induction of EE.IL-13 was delivered intratracheally to wild-type, signal transducer and activator of transcription (STAT) 6, eotaxin-1, or IL-5-deficient mice. Eosinophil levels and 5'-bromodeoxyuridine (BrdU) incorporation were examined by immunohistochemical staining.Intratracheal delivery of IL-13 induced dose-dependent eosinophil accumulation in the esophagus (but not the stomach). In addition, intratracheal IL-13 induced esophageal epithelial hyperplasia. The ability of IL-13 to induce EE was abolished in STAT6-deficient mice. IL-13-induced EE was nearly completely ablated in IL-5-deficient mice (37.3 +/- 11.6 vs. 3.3 +/- 3.2 eosinophils/mm(2) in wild-type and IL-5-deficient mice, respectively). Additionally, IL-13-induced EE was significantly diminished in eotaxin-1-deficient mice (48.7 +/- 10.3 vs. 14.1 +/- 12.5 eosinophils/mm(2) in wild-type and eotaxin-1-deficient mice, respectively).IL-13 delivery to the lung induces EE by an IL-5, eotaxin-1, and STAT6-dependent mechanism. These results further establish an intimate connection between respiratory and esophageal inflammation.

Objective: Despite recognized hemodynamic derangements during cardiac displacement, most patients appear to tolerate the off-pump procedure well. However, some patients unpredictably become hemodynamically unstable requiring emergency cardiopulmonary bypass or intra-aortic balloon pump support. After an experience of 5,306 multivessel off-pump coronary artery bypasses (OPCABs), this study was undertaken to determine the factors that would identify the patients who were at a higher risk for the procedure. Design: Prospective clinical investigation. Setting: Tertiary care academic cardiac care center. Participants: Five hundred consecutive patients undergoing multivessel OPCAB from September to December 2001. Interventions: Various cardiac and extracardiac factors were charted in prespecified data-entry forms. Multiple logistic regression analysis was done to determine if any identifiable factors were predictors of a higher risk of unacceptable hemodynamic instability during OPCAB. Institution of IABP support or conversion to CPB were the endpoints of the study. Measurements and Main Results: Of the 500 patients studied, significant hemodynamic instability developed in 24 (4.8%) patients. IABP support was instituted in 16 (3.2%) patients, and 8 (1.6%) were converted to CPB. Stepwise logistic regression identified ejection fraction

Asthma, a complex chronic inflammatory pulmonary disorder, is on the rise despite intense ongoing research, underscoring the need for new scientific inquiry. In an effort to provide unbiased insight into the pathogenesis of this disease, we took an empirical approach involving transcript expression profiling of lung tissue from mice with experimental asthma. Employing asthma models induced by different allergens (ovalbumin [OVA] and Aspergillus fumigatus), we found strong induction of trefoil factor-2 (TFF2), a gene involved in epithelial restitution and mucosal secretion in the gastrointestinal tract. Using a combination of pharmacologic delivery and transgenic overexpression, TFF2 was demonstrated to be strongly induced in the lung by interleukin (IL)-4 and IL-13. Notably, TFF2 induction by both OVA and pharmacologic delivery of IL-4 and IL-13 was dependent upon signal transducer and activator of transcription (STAT)6. However, the upregulation of TFF2 by both chronic expression of IL-4 and Aspergillus fumigatus antigen was independent of STAT6. These results establish that TFF2 is an allergen-induced lung gene product differentially regulated by Th2 cytokines and STAT6. Given the important role of trefoil factors in wound healing, epithelial restitution, and maintenance of mucosal integrity in the gastrointestinal tract, these results support a potential role for TFF2, in both the acute and chronic phase of experimental asthma, via separate induction pathways.

Background . Patients with concomitant occlusive disease of coronary and carotid arteries remain at high risk of perioperative stroke and myocardial infarction. Combined coronary artery bypass grafting on cardiopulmonary bypass and carotid endarterectomy has been shown to give good results for this category of patients. In the present study, we analyzed our results of off-pump coronary artery bypass grafting and carotid endarterectomy as a one-stage procedure. Methods . Between January 1997 and December 2000, 82 patients underwent combined off-pump coronary artery bypass grafting and carotid endarterectomy. All patients were evaluated by preoperative carotid duplex scanning and carotid angiography. All patients had more than or equal to 70% carotid artery stenosis. There were 35 asymptomatic patients (42.7%) and 47 symptomatic patients (57.3%). Carotid endarterectomy was performed before coronary artery bypass grafting in all the patients. Results . There were 66 males (80.5%) and 16 females (19.5%) with a mean age of 63 ± 8 years. The average number of grafts was 3.4 ± 0.8. There was no hospital mortality. One patient had perioperative myocardial infarction. None of the patients had stroke. One patient had transient neurologic deficit and 1 patient had temporary 12th nerve dysfunction; both recovered completely. There was no incidence of neck wound infection, although 1 patient developed neck hematoma that required reexploration. At a mean follow-up of 2.2 ± 0.7 years, 1 patient required contralateral carotid endarterectomy and 1 patient died because of cardiac failure. Conclusions . Combined off-pump coronary artery bypass grafting and carotid endarterectomy is a safe and effective procedure in patients with significant concomitant carotid and coronary artery disease.

Eosinophil infiltration into the esophagus occurs in a wide range of diseases; however, the underlying pathophysiological mechanisms involved are largely unknown. We now report that the Th2 cytokine, IL-5, is necessary and sufficient for the induction of eosinophil trafficking to the esophagus. We show that transgenic mice overexpressing IL-5 under the control of a T cell (CD2) or a small intestinal enterocyte (fatty acid-binding protein) promoter have markedly increased eosinophil numbers in the esophagus. For example, esophageal eosinophil levels are 1.9 ± 0.9 and 121 ± 14 eosinophils/mm2 in wild-type and CD2-IL-5-transgenic mice, respectively. Consistent with this effect being mediated by a systemic mechanism, pharmacological administration of IL-5 via a miniosmotic pump in the peritoneal cavity resulted in blood and esophageal eosinophilia. To examine the role of IL-5 in oral Ag-induced esophageal eosinophilia, eosinophilic esophagitis was induced by allergen exposure in IL-5-deficient and wild-type mice. Importantly, IL-5-deficient mice were resistant to eosinophilic esophagitis. Finally, we examined the role of eotaxin when IL-5 was overproduced in vivo. Esophageal eosinophil levels in CD2-IL-5-transgenic mice were found to decrease 15-fold in the absence of the eotaxin gene; however, esophageal eosinophil numbers in eotaxin-deficient IL-5-transgenic mice still remained higher than wild-type mice. In conclusion, these studies demonstrate a central role for IL-5 in inducing eosinophil trafficking to the esophagus.

Eosinophilic esophagitis (EoE) is a recently recognized inflammatory disorder that needs a potential therapeutic strategy. We earlier showed that iNKT cell-deficient mice are protected from allergen-induced EoE. Therefore, we now tested the hypothesis that iNKT cells are induced in the human EoE and is a novel possible target for the treatment of human EoE. Accordingly, we examine number of iNKT cells and eosinophils and expression of iNKT-associated cell surface receptors and chemokines by performing immunofluorescence, qPCR and ELISA in the esophageal biopsies and blood samples of normal subjects (comparison control) and EoE patients. Herein, we show that iNKT cell number, their receptor subcomponents Vα24 and Vβ11 expression, and associated chemokine CXCL16 levels (or expression) are induced significantly in EoE patients compared with normal individuals. In addition, we show that CXCL16 levels (or expression) correlate with the mRNA levels of Vα24 receptor but not well with esophageal eosinophilia in human EoE. Of note, we show that in vivo activation of iNKT cells is sufficient to induce EoE in mice. Furthermore, we show that anti-mCD1d- and anti-hVα24Jα18-neutralizing antibody treatment protects allergen-induced experimental EoE. Taken together, we have shown first time that iNKT cells have a critical pathogenic role in human and experimental EoE. iNKT cell neutralization by humanized anti-CD1d and anti-Vα24Jα18 antibodies might be a novel and potential therapy for human EoE.

Background: IL-13 induces several characteristic features of asthma, including airway eosinophilia, airway hyperresponsiveness, and mucus overproduction; however, the mechanisms involved are largely unknown. Objective: We hypothesized that IL-13–induced inflammatory changes in the lung were dependent in part on IL-5 and eotaxin, two eosinophil-selective cytokines. Methods: Recombinant murine IL-13 was repeatedly administered to the lung by intranasal delivery until the characteristic features of asthma developed. To analyze the role of IL-5 and eotaxin, we subjected eotaxin gene–targeted, IL-5 gene–targeted, eotaxin/IL-5–double-deficient, IL-5 transgenic, and wild-type mice of the Balb/C background to the experimental regime. Results: The induction of IL-13–mediated airway eosinophilia was found to occur independently of eosinophilia in the blood or bone marrow, indicating that IL-13–induced airway inflammation is primarily mediated by local effects of IL-13 in the lung. Eosinophil recruitment into both the lung tissue and bronchoalveolar lavage fluid was markedly attenuated in IL-5–deficient mice in comparison with wild-type controls. Accordingly, IL-13 delivery to IL-5 transgenic mice resulted in a large increase in airway eosinophils in comparison with wild-type mice. Interestingly, IL-13–induced eosinophilia in the bronchoalveolar lavage fluid of eotaxin-deficient mice was not impaired; however, these same mice failed to mount a significant tissue eosinophilia in response to IL-13. Finally, IL-13–induced mucus production was not affected by the presence of IL-5 or eotaxin, suggesting that IL-13–induced mucus secretion is mechanistically dissociated from airway eosinophilia. Conclusion: Selective components of the IL-13–induced asthma phenotype—airway eosinophilia but not mucus secretion—are differentially regulated by IL-5 and eotaxin. IL-5 is required for IL-13 to induce eosinophilia throughout the lung, whereas eotaxin regulates the distribution of airway eosinophils. (J Allergy Clin Immunol 2001;108:594-601.)

Eosinophil infiltration into the esophagus is observed in diverse diseases including gastroesophageal reflux and allergic gastroenteritis, but the processes involved are largely unknown. We now report an original model of experimental esophagitis induced by exposure of mice to respiratory allergen. Allergen-challenged mice develop marked levels of esophageal eosinophils, free eosinophil granules, and epithelial cell hyperplasia, features that mimic the human disorders. Interestingly, exposure of mice to oral or intragastric allergen does not promote eosinophilic esophagitis, indicating that hypersensitivity in the esophagus occurs with simultaneous development of pulmonary inflammation. Furthermore, in the absence of eotaxin, eosinophil recruitment is attenuated, whereas in the absence of IL-5, eosinophil accumulation and epithelial hyperplasia are ablated. These results establish a pathophysiological connection between allergic hypersensitivity responses in the lung and esophagus and demonstrate an etiologic role for inhaled allergens and eosinophils in gastrointestinal inflammation.

A new iridoid diglycoside named koelzioside has been isolated from Scrophularia koelzii . The structure of koelzioside was elucidated by chemical and spectral analysis as 6- O -(2″,3″-di- O -cinnamoyl-4″- O -acetyl)-α- l -rhamnopyranosyl catalpol.

Despite marked advances in the understanding of allergic responses, the mechanisms regulating gastrointestinal allergy are not very well understood. We have developed a model of antigen-induced eosinophil-associated gastrointestinal allergy and characterized the role of eotaxin and IL-5. Challenge of allergen-sensitized mice with oral allergen, in the form of enteric-coated beads, resulted in marked allergen-specific IgG1and IgE, Th2-type (IL-4 and IL-5) cytokine production, and eosinophil accumulation in the blood and small intestine. In the genetic absence of eotaxin, a chemokine constitutively expressed in the gastrointestinal tract, eosinophil recruitment into the small intestine was ablated, and these mice developed enhanced eosinophil accumulation in the blood compared with wild-type mice. Interestingly, in the absence of IL-5, allergen challenge promoted partial eosinophil accumulation into the small intestine and a decline in circulating eosinophil levels. Collectively, these results establish that the accumulation of gastrointestinal eosinophils is antigen induced, can occur independent of IL-5, and provides a molecular mechanism to explain the dichotomy between peripheral blood and tissue eosinophilia. Furthermore, eotaxin is identified as a critical regulator of antigen-induced eosinophilic inflammation in the gastrointestinal tract.

Bleomycin damages DNA and causes lung injury and fibrosis. To determine whether bleomycin is associated with the appearance of DNA damage-inducible proteins, C3H mice received either 0.4 mg bleomycin or normal saline intratracheally and were killed 1 to 14 d later. The lungs were examined for expression of p53, p21(WAF1/PiCl), and proliferating cell nuclear antigen (PCNA) using immunohistochemistry and Western blotting. p53-positive cells first appeared at 5 d after treatment and peaked at 7 d; PCNA-positive cells appeared at 1 d after treatment and peaked at 7 d; and p21-positive cells appeared at 5 d and peaked at 9 d. Western blot analysis confirmed that bleomycin upregulated the DNA damage-inducible proteins in a similar fashion. This is the first evidence that bleomycin causes a p53-dependent response associated with acute injury in the lung.

Protein-A, 42KD cell wall glycoprotein of S. aureus Cowan I enhance mononuclear and polymorphonuclear cell counts in vivo and possesses antitoxic, antitumor, properties. In order to explain the mechanism of its function, the respiratory burst phenomenon in cell and cell free system was studied using lucigenin-dependent chemiluminescence technique. A dose dependent increase in protein A-mediated generation of superoxide radical was observed in resting and PMA stimulated neutrophils. Superoxide dismutase (SOD) was used to confirm the production of superoxide radicals (O2-). To understand the mechanism of protein-A induced O2- generation; NADPH oxidase activity was measured in cell free system using NADPH as a substrate. A significant increase in NADPH oxidase activity was observed in the membrane and post-nuclear supernatant fraction of activated human neutrophils. Cytosolic fraction showed slight enzyme activation. Protein A (SpA)-induced NADPH oxidase activation in the membrane fraction was observed even in the absence of the substrate NADPH. These data indicate that protein A attenuate the NADPH oxidase system to produce O2- radicals.

It has been well documented in the literature that the removal of circulatory immune complexes (CICs) from the host circulation leads to the immunopotentiation as well as generation of antitumor responses in a variety of tumors in rats, cats, dogs and human patients. CICs are the major immunosuppressive factors in tumor bearing host. Protein A (PA) has been extensively used for the removal of these CICs from the sera/plasma of tumor bearers, because PA has the ability to bind with the Fc portion of mammalian immunoglobulins. Previously, we reported for the first time a potent antitumor response by the inoculation of cell free Ehrlich’s ascites fluid adsorbed in vitro over PA containing Staphylococcus aureus Cowan I (SAC) in Ehrlich’s ascites tumor model. However, there was toxicity associated with this form of therapy in terms of early death of treated animals and the depletion of hepatic glutathione pool as well as phase I biotransformation enzyme and increase in glutathione-S-transferase (GST) activities. In the present investigation, tumor bearing animals were treated intraperitoneally (i.p.) on alternate days for 15 days with adsorbed ascites fluid (ad-ASF) (0.1 ml) and glutathione (GSH) (250 mg/kg body weight) separately. We found that GSH supplementation increases mean survival time of GSH and ad-ASF treated mice up to 37.2 days in comparison with 19.9 days for only ad-ASF treated animals, while percent increase in body weight was found to be not affected by the GSH substitution, which remains significantly lower (P

The histological identification of increased eosinophils in the gastrointestinal tract occurs in numerous clinical disorders; however, there is a limited understanding of the mechanisms regulating eosinophil trafficking into this mucosal surface. The results presented in this study characterize the processes regulating eosinophil homing into the gastrointestinal tract at baseline. Eosinophils were found to be present in the lamina propria of 19-day-old embryos and germ-free adult mice at concentrations comparable to those present in non–germ-free adult mice. Furthermore, eosinophil gastrointestinal levels were not altered by increasing circulating eosinophils after pulmonary allergen challenge. Gastrointestinal eosinophil levels were partially reduced in mice deficient in recombinase activating gene-1 (RAG-1), IL-5, or the β common chain (βc), but these reductions paralleled reductions in circulating eosinophils. In contrast, mice deficient in eotaxin had a marked reduction in gastrointestinal eosinophils but normal levels of eosinophils in the hematopoietic compartments. Furthermore, eotaxin was important for regulating eosinophil levels, even in the presence of high levels of IL-5. These investigations demonstrate eosinophil homing into the gastrointestinal tract during embryonic development occurring independently of viable intestinal flora. Furthermore, eotaxin is identified as the primary regulator of eosinophil gastrointestinal homing under homeostatic states, and may therefore have a fundamental role in innate immune responses. J. Clin. Invest. 103:1719–1727 (1999).

Our earlier studies have shown that removal of various blocking factors from the sera of tumor-bearing animals and humans by adsorption over heat-attenuated and formalin-fixed-Staphylococcus aureus Cowan I (SAC) containing Protein A (PA) causes antitumor immune response. It was also shown that this procedure caused regression of a wide variety of established animal and human tumors. In the present investigation, the therapeutic potential of inoculation of ascites fluid adsorbed in vitro over non-viable SAC containing PA has been demonstrated in Ehrlich' s ascites tumor (EAT) in mouse. The antitumor effect was evident by a significant decrease in body weight (p0.001) as well as significant reduction in viability of ascites tumor cells (p0.001) in peritoneal cavity. However, some of the responding animals died earlier than controls, this may be due to the toxicity associated with therapy. The toxic effects were evident in decreased contents of glutathione, and increased activity of glutathione-S-transferase, decreased activity of microsomal enzymes and also in an early death of some of tumor regressed animals. The probable causes of toxicity of the therapy and prospects of reversing these toxic effects are discussed.

Immunocontrast agents used for magnetic resonance imaging require antibodies that preserve the immunoreactivity while containing a high number of chelated paramagnetic ions.Anti-CEA F(ab')2 fragments were coupled to polylysine-Gd-DOTA and polylysine-Gd-DTPA. A paramagnetic load as high as n = 24 to 28 metal ions per antibody was reached.The immunoreactivity of the gadolinium (Gd)-labeled anti-CEA F(ab')2 immunoconjugates was 80% to 85%. Compared with that of commercial chelates, the relaxivity (R1) increase is as follows: Gd-DTPAGd-DOTAGd-H2OPL-Gd-DTPA24-28PL-Gd-DTPA24-28 F(ab')2PL-Gd-DOTA24-28PL-Gd-DOTA24-28 F(ab')2. 1H nuclear magnetic relaxation dispersion data of immunoconjugates showed that the high relaxivity enhancement was the result of a reduction of the molecular tumbling rate. Twenty-four hours after intravenous injection of 50 micrograms (1 mumol Gd/kg) of Gd-labeled immunoconjugates to nude mice grafted with human colorectal carcinoma LS 174T, the tumor uptake was 10% to 15%, resulting in an increase of R1 of up to 15% to 20% versus noninjected mice. No difference was found between PL-Gd-DTPA24-28 F(ab')2 and PL-Gd-DOTA24-28 F(ab')2 immunoconjugates for tumor, liver, and kidney uptake. A high signal intensity of tumor was observed in 50% of the tested mice.

Experimental eosinophilic esophagitis (EE) is a recently described chronic allergic inflammatory disorder whose basic pathogenesis is not well understood. Herein, we focus on the hypothesis that iNKT cell responses are essential for the initiation and progression of EE. We show that iNKT cells and their specific chemokine CXCL16 are induced in human and experimental EE. CXCL16 is primarily derived from esophageal epithelial cells and correlates with the levels of esophageal eosinophils. Notably, iNKT cell-deficient CD1d-null mice are protected from the induction of experimental EE; whereas, NK cell depleted mice exhibit full disease following allergen challenge. Interestingly, in vivo activation of human iNKT cells by the α-galactosylceramide analog PBS57 is sufficient to induce esophageal eosinophilia. Mechanistically, PBS57 exposed human iNKT cells produce IL-5 and IL-13 via STAT5 activation. Taken together, these findings provide evidence that iNKT cells have an essential pathogenic role in the initiation and progression of human and experimental EE, providing new opportunities for therapeutic strategies.

Background: Efficacy of photodynamic therapy can be enhanced by improving uptake, localization, and sub-cellular localization of sensitizers at the sensitive targets. Materials and Methods: Uptake, localization, and photodynamic effects of hematoporphyrin derivative (HpD, Photosan-3; PS-3) and disulfonated aluminum phthalocyanine (AlPcS 2 ) were studied either encapsulated in liposomes or conjugated to a monoclonal antibody to carcinoembryonic antigen (anti-CEA) in a brain glioma cell line, BMG-1. Results: Although the total uptake with encapsulated or conjugated sensitizers was less than the free sensitizers, photodynamic efficiency was higher due to the localization of the sensitizer at the sensitive targets. Biodistribution of intravenously administered technetium ( 99m Tc)-labeled PS-3 analyzed by gamma camera imaging showed maximum accumulation in the liver followed by tumor. Tumor/muscle (T/N) ratio of free PS-3 was higher compared to encapsulated or conjugated PS-3 but the accumulation of PS-3 significantly reduced in brain and cutaneous tissue following modulated delivery. Pharmacokinetics suggested faster accumulation of encapsulated and conjugated PS-3 in the tumor. Conclusion: Localization of sensitizers at sensitive targets and reduced accumulation in normal tissues with liposome encapsulation and antibody conjugation suggest that these two delivery systems can potentially enhance the efficacy of photodynamic treatment.

Quantitative microarray analyses have shown increased expression of interleukin-15 (IL-15) messenger RNA in the esophagus of patients with eosinophilic esophagitis (EoE), a recently recognized allergic disorder with poorly understood pathogenesis.Quantitative polymerase chain reaction and enzyme-linked immunosorbent assay analyses were performed to examine protein and transcript levels in tissue samples from patients with EoE. Tissues from IL-15Ra-deficient and wild-type (control) mice were also examined. Tissue eosinophilia was determined by immunostaining for major basic protein and flow cytometry for cell-surface receptors.Quantitative polymerase chain reaction analyses showed that levels of IL-15 and its receptor IL-15Ra were increased approximately 6- and approximately 10-fold, respectively, in tissues from patients with EoE and approximately 3- and approximately 4-fold, respectively, in mice with allergen-induced EoE. A2-fold increase in serum IL-15 protein levels was also detected in human EoE samples compared with those from healthy individuals. Human IL-15 messenger RNA levels correlated with esophageal eosinophilia (P.001). IL-15Ra-deficient mice were protected from allergen-induced esophageal eosinophilia compared with controls (P.001), even though similar levels of airway eosinophilia were observed in all mice. IL-15 activated STAT5 and CD4(+) T cells to produce cytokines that act on eosinophils. Incubation of primary esophageal epithelial cells from mice and humans with IL-15 caused a dose-dependent increase in the mRNA expression and protein levels of eotaxin-1, -2, and -3.IL-15 mediates in the pathogenesis of EoE. IL-15 activates CD4(+) T cells to produce cytokines that act on eosinophils.

RATIONALE: Eosinophilic esophagitis (EE) is an emerging disease and we previously demonstrated that adaptive T cell immunity is critical in the disease pathogenesis. However, the specific subsets of T cells that promote esophageal eosinophilia are not yet understood. Therefore, we focused our studies to examine the role of CD45RB T subsets, as the pathogenic and regulatory activity of T cells depends on the expression levels of CD45RB. METHODS: A flow cytometric analysis was performed to examine the frequency of CD45RBhigh and CD45RBlow T cell subsets in the mouse esophagus. A real time PCR analysis was performed to examine the pathogenic and anti-inflammatory characteristics of both subsets of T cells. In addition, we adoptively transferred these purified cell subsets in mice with experimental EE and examined their role in the disease pathogenesis. RESULTS: An increase of CD4+CD45RBhigh and a decrease of CD4+CD45RBlow cells were observed in mouse esophagus following the induction of experimental EE. A relatively high transcript level of IL-5 and IL-13 in CD4+CD45RBhigh and IL-10 and TGF-β1 in CD4+CD45RBlow spleen purified cells was observed. Further, our initial experimentation revealed that mice adoptively transferred with 4X105 CD4+CD45RBlow T cells/week were protected from the induction of esophageal eosinophilia compared to control mice that did not received adoptively transferred cells. Mice receiving CD4+CD45RBhigh T cell had no significant change in the number of eosinophils in the esophagus compared to control mice following allergen challenge. CONCLUSIONS: CD4+CD45RBlow T cells have a critical role in down-regulating allergen-induced eosinophil recruitment to the mouse esophagus.

Rationale Aeroallergen induced experimental asthma is accompanied by esophageal eosinophilia, which mimics the pathophysiological changes observed in individuals with eosinophilic esophagitis (EE). Since epidermal allergen sensitization has been recently implicated in the pathogenesis of experimental asthma, we examined the ability of epidermal sensitization to also result in esophageal eosinophilia. Methods We developed a novel model for skin sensitization in which 100μg of Aspergilus fumigatus antigen (Asp) or Saline was topically applied to the shaved upper backs of mice on six occasions over the course of two weeks. One week following sensitization, mice from both groups were given a single intranasal challenge with Asp or Saline (n=4 mice per treatment). Results Epidermal sensitization to Asp caused a 4 to 5-fold increase in serum IgE levels compared to Saline. Additionally, following a single intranasal Asp challenge, epidermal sensitization to Asp resulted in pulmonary inflammation and significantly elevated eosinophils in both bronchoalveolar lavage fluid (BALF) (57.6 ± 8.4×104 vs. 0.2 ± 0.17×104 per lung) and in the esophagus (24.3 ± 7 vs. 4.4 ± 3 per mm2) compared to Saline-challenged mice (p=0.001). Conclusion Epidermal sensitization can prime for aeroallergen induced eosinophilic esophagitis providing a novel link between cutaneous and mucosal allergic responses.

Rationale Epidemiologic studies have suggested that the skin is an early entry point for antigen sensitization as suggested by the frequent occurrence of allergic dermatitis (AD) prior to the development of allergic disorders. We therefore hypothesized that epicutaneous antigen exposure would prime for airway and esophageal allergic responses in an experimental model. Methods AD was induced by applying 100 micrograms of Aspergillus Fumigatus antigen or Vehicle soaked patches on the shaved back of BALB/c mice, at two one week intervals over the course of four weeks. To establish airway and esophageal inflammation, we examined respiratory hyperresponsiveness to methacholine and inflammation in the bone marrow, skin, airways and esophagus. Results Significant skin symptoms including lichenification and excoriations developed in the antigen-sensitized group, whereas no cutaneous changes were observed in the vehicle group (P 4 cells/ml of blood eosinophils, whereas in the vehicle group: 372.8 ± 127.3, 2.8 ± 0.3, 11.4 ± 9.5, respectively (mean ± SEM, p 4 cells/ml and 28.1 ± 6.5cells/mm 2 in the esophagus, whereas 3.1 ± 1.7 and 4.9 ± 1.4 in the vehicle controls (p Conclusions Epicutaneous antigen sensitization followed by a single airway antigen challenge induces systemic responses in the respiratory tract and esophagus. These results highlight the potency of epicutaneous antigen exposure in the development of allergic responses.

Rationale In a mouse model of experimental asthma, we discovered the involvement of gene known as resistin-like molecule (RELM)-alpha and beta that are structurally related group of cytokines implicated in insulin resistance. We report that RELM-alpha and beta have a major role in macrophage activation and airway remodeling in the murine lung. Method The expression of RELM-alpha and beta mRNA was examined following allergen and Th2 cytokine delivery. To understand the in vivo role of RELMs, we intratracheally delivered these cytokines and examined the recruitment of inflammatory cells, collagen deposition and mucus production. Results We found RELM-alpha and beta were induced by respiratory allergen or Th2 cytokine (IL-13) exposure by a STAT 6 dependent mechanism. Interestingly, intratracheal delivery of both RELM cytokines induced dose-dependent accumulation of macrophages in the lung. The macrophages numbers in RELM-alpha and beta treated mice were 14.8 ± 4.2 × 10e4/lung and 11.9 ± 1.5 × 10e4/lung, respectively, in comparison to 3.2 ± 0.6 × 10e4/lung in control mice. Other inflammatory cells including lymphocytes, neutrophils, eosinophils and fibroblast increased significantly to a lesser extent. Furthermore, RELM delivery induced marked perivascular and peribronchial collagen deposition and goblet cell hyperplasia. The collagen deposition in RELM-alpha and beta treated mice were 1.5 ± 0.5 × 10e4/mm2 and 8.8 ± 1.1 × 10e3/um2 compared with 352 ± 53/um2 in control mice. A 23-fold increase in goblet cells were also observed in RELM-alpha treated mice. Conclusion The RELM cytokines are induced in the asthmatic lung, regulated by Th2 cytokines, and are potent inducers of lung remodeling.

A rapid and convenient synthetic methodology for the cyclocondensative transformation of various alkylsulfanylphenylamines with bromoacetyl bromide by supporting on KF-alumina in ionic liquids [bmim][Br] and [bmim][BF 4 ] has been developed to obtain 3-oxo-1,4-benzothiazine in good yields. The product is easily obtained by extraction with ethyl acetate and concentrating under vacuum. Easy recovery of ionic liquid and use in consecutive reactions is also reported.