Use of cardiac magnetic resonance imaging and cardiac magnetic resonance feature-tracking strain analysis to characterize and differentiate aortic annular strain patterns in aortic valve regurgitation versus normal aortic valves.

BACKGROUND: Understanding the aortic annulus is important for obtaining reproducible and durable aortic valve (AV) repair and allowing advances for transcatheter AV replacement treatment of aortic regurgitation (AR). Significant limitations exist when using echocardiography and computed tomography-based imaging with feature tracking at the aortic annulus. AIMS: Cardiac magnetic resonance is used to obtain regional longitudinal strain and can be modified to obtain circumferential annular strain at the fibrous and muscular portions of the AV annulus. MATERIALS AND METHODS: Holst et al. use a novel method to characterize and prove that adverse annular deformation occurs at the muscular portion of the AV annulus in patients with AR. In their study, cardiovascular magnetic resonance (CMR) imaging and CMR feature-tracking strain analysis are used to characterize aortic annular regional longitudinal strain (RLS) in humans. RESULTS: The authors convincing show that the direction of muscular annular deformation in patients with AR is opposite to the direction of muscular annular deformation in patients with normal Avs, (median RLS: 4.18% in patients with severe AR vs. -10.41% in well-functioning AVs, p = .024; at RLS muscular annulus). DISCUSSION AND CONCLUSION: The direction of muscular annular deformation in patients with AR is opposite to the direction of muscular annular deformation in patients with normal AVs. This information will have impactful physiological relevance for surgical and percutaneous treatment of aortic valve pathology.

Transforming Growth Factor-ß and Interleukin-1ß Signaling Pathways Converge on the Chemokine CCL20 Promoter.

CCL20 is the only chemokine ligand for the chemokine receptor CCR6, which is expressed by the critical antigen presenting cells, dendritic cells. Increased expression of CCL20 is likely involved in the increased recruitment of dendritic cells observed in fibroinflammatory diseases such as chronic obstructive pulmonary disease (COPD). CCL20 expression is increased by the proinflammatory cytokine IL-1ß. We have determined that IL-1ß-dependent CCL20 expression is also dependent on the multifunctional cytokine TGF-ß. TGF-ß is expressed in a latent form that must be activated to function, and activation is achieved through binding to the integrin αvß8 (itgb8). Here we confirm correlative increases in αvß8 and IL-1ß with CCL20 protein in lung parenchymal lysates of a large cohort of COPD patients. How IL-1ß- and αvß8-mediated TGF-ß activation conspire to increase fibroblast CCL20 expression remains unknown, because these pathways have not been shown to directly interact. We evaluate the 5'-flanking region of CCL20 to determine that IL-1ß-driven CCL20 expression is dependent on αvß8-mediated activation of TGF-ß. We identify a TGF-ß-responsive element (i.e. SMAD) located on an upstream enhancer of the human CCL20 promoter required for efficient IL-1ß-dependent CCL20 expression. By chromatin immunoprecipitation, this upstream enhancer complexes with the p50 subunit of NF-κB on a NF-κB-binding element close to the transcriptional start site of CCL20. These interactions are confirmed by electromobility shift assays in nuclear extracts from human lung fibroblasts. These data define a mechanism by which αvß8-dependent activation of TGF-ß regulates IL-1ß-dependent CCL20 expression in COPD.

Mitral valve prolapse.

Mitral valve prolapse is defined as abnormal bulging of the mitral valve leaflets into the left atrium during ventricular systole. Mitral valve prolapse is a common condition that is a risk factor for mitral regurgitation, congestive heart failure, arrhythmia, and endocarditis. Myxomatous degeneration is the most common cause of mitral prolapse in the United States and Europe, and progression of myxomatous mitral prolapse is the most common cause of mitral regurgitation that requires surgical treatment. Myxomatous degeneration appears to have genetic etiology. The genetics of myxomatous degeneration is complex and not fully worked out; it appears to be heterogeneous with multi-gene, multi-chromosomal autosomal dominance with incomplete penetrance. The molecular disorder of myxomatous degeneration appears to consist of a connective tissue disorder with altered extracellular matrix status and involves the action of matrix metalloproteinase, cysteine endoproteases, and tenomodulin. Treatment of mitral prolapse with regurgitation is complex, and the technological advances that are currently in development will be challenging and controversial.

Interleukin-1beta induces increased transcriptional activation of the transforming growth factor-beta-activating integrin subunit beta8 through altering chromatin architecture.

The integrin αvß8 is a cell surface receptor for the latent domain (LAP) of the multifunctional cytokine TGF-ß. Through its association with LAP, TGF-ß is maintained in a latent form that must be activated to function. Binding to the integrin αvß8 with subsequent metalloproteolytic cleavage of LAP represents a major mechanism of TGF-ß activation in vivo. Altered expression of the integrin ß8 subunit (ITGB8) is found in human chronic obstructive pulmonary disease, cancers, and brain vascular malformations. We have previously shown that the proinflammatory cytokine interleukin-1ß (IL-1ß) increases ITGB8 expression on lung fibroblasts, which increases αvß8-mediated TGF-ß activation in fibrosis and pathologic inflammation. Here we report the mechanism of increased ITGB8 expression by IL-1ß. Our data support a model where the chromatin architecture of the ITGB8 core promoter is altered by nucleosomal repositioning that enhances the interaction of an AP1 complex (containing c-Jun and ATF2). This repositioning is caused by the dissociation of HDAC2 with the ITGB8 core promoter, leading to increased histone H4 acetylation and a loosening of nucleosomal-DNA interactions allowing "opening" of the chromatin structure and increased association of c-Jun and ATF-2. These changes are mediated through NFκB- and p38-dependent pathways. Ultimately, these events culminate in increasing ITGB8 transcription, αvß8 surface expression, and αvß8-mediated TGFß activation.

Transcription of the transforming growth factor beta activating integrin beta8 subunit is regulated by SP3, AP-1, and the p38 pathway.

Integrin alphavbeta8 is a critical regulator of transforming growth factor beta activation in vasculogenesis during development, immune regulation, and endothelial/epithelial-mesenchymal homeostasis. Recent studies have suggested roles for integrin beta8 in the pathogenesis of chronic obstructive pulmonary disease, brain arteriovenous malformations, and select cancers (Araya, J., Cambier, S., Markovics, J. A., Wolters, P., Jablons, D., Hill, A., Finkbeiner, W., Jones, K., Broaddus, V. C., Sheppard, D., Barzcak, A., Xiao, Y., Erle, D. J., and Nishimura, S. L. (2007) J. Clin. Invest. 117, 3551-3562; Su, H., Kim, H., Pawlikowska, L., Kitamura, H., Shen, F., Cambier, S., Markovics, J., Lawton, M. T., Sidney, S., Bollen, A. W., Kwok, P. Y., Reichardt, L., Young, W. L., Yang, G. Y., and Nishimura, S. L. (2010) Am. J. Pathol. 176, 1018-1027; Culhane, A. C., and Quackenbush, J. (2009) Cancer Res. 69, 7480-7485; Cambier, S., Mu, D. Z., O'Connell, D., Boylen, K., Travis, W., Liu, W. H., Broaddus, V. C., and Nishimura, S. L. (2000) Cancer Res. 60, 7084-7093). Here we report the first identification and characterization of the promoter for ITGB8. We show that a SP binding site and a cyclic AMP response element (CRE) in the ITGB8 core promoter are required for its expression and that Sp1, Sp3, and several AP-1 transcription factors form a complex that binds to these sites in a p38-dependent manner. Furthermore, we demonstrate the requirement for Sp3, ATF-2, and p38 for the transcription and protein expression of integrin beta8. Additionally, reduction of SP3 or inhibition of p38 blocks alphavbeta8-mediated transforming growth factor beta activation. These results place integrin beta8 expression and activity under the control of ubiquitous transcription factors in a stress-activated and pro-inflammatory pathway.

Squamous metaplasia amplifies pathologic epithelial-mesenchymal interactions in COPD patients.

Squamous metaplasia (SM) is common in smokers and is associated with airway obstruction in chronic obstructive pulmonary disease (COPD). A major mechanism of airway obstruction in COPD is thickening of the small airway walls. We asked whether SM actively contributes to airway wall thickening through alteration of epithelial-mesenchymal interactions in COPD. Using immunohistochemical staining, airway morphometry, and fibroblast culture of lung samples from COPD patients; genome-wide analysis of an in vitro model of SM; and in vitro modeling of human airway epithelial-mesenchymal interactions, we provide evidence that SM, through the increased secretion of IL-1beta, induces a fibrotic response in adjacent airway fibroblasts. We identify a pivotal role for integrin-mediated TGF-beta activation in amplifying SM and driving IL-1beta-dependent profibrotic mesenchymal responses. Finally, we show that SM correlates with increased severity of COPD and that fibroblast expression of the integrin alpha(v)beta(8), which is the major mediator of airway fibroblast TGF-beta activation, correlated with disease severity and small airway wall thickening in COPD. Our findings have identified TGF-beta as a potential therapeutic target for COPD.

Impact of Aortic Stenosis on Myofiber Stress: Translational Application of Left Ventricle-Aortic Coupling Simulation.

The severity of aortic stenosis (AS) has traditionally been graded by measuring hemodynamic parameters of transvalvular pressure gradient, ejection jet velocity, or estimating valve orifice area. Recent research has highlighted limitations of these criteria at effectively grading AS in presence of left ventricle (LV) dysfunction. We hypothesized that simulations coupling the aorta and LV could provide meaningful insight into myocardial biomechanical derangements that accompany AS. A realistic finite element model of the human heart with a coupled lumped-parameter circulatory system was used to simulate AS. Finite element analysis was performed with Abaqus FEA. An anisotropic hyperelastic model was assigned to LV passive properties, and a time-varying elastance function governed the LV active response. Global LV myofiber peak systolic stress (mean ± standard deviation) was 9.31 ± 10.33 kPa at baseline, 13.13 ± 10.29 kPa for moderate AS, and 16.18 ± 10.59 kPa for severe AS. Mean LV myofiber peak systolic strains were -22.40 ± 8.73%, -22.24 ± 8.91%, and -21.97 ± 9.18%, respectively. Stress was significantly elevated compared to baseline for moderate (p < 0.01) and severe AS (p < 0.001), and when compared to each other (p < 0.01). Ventricular regions that experienced the greatest systolic stress were (severe AS vs. baseline) basal inferior (39.87 vs. 30.02 kPa; p < 0.01), mid-anteroseptal (32.29 vs. 24.79 kPa; p < 0.001), and apex (27.99 vs. 23.52 kPa; p < 0.001). This data serves as a reference for future studies that will incorporate patient-specific ventricular geometries and material parameters, aiming to correlate LV biomechanics to AS severity.

Influence of the emulsion droplet type on the rheological characteristics and microstructure of rennet gels from reconstituted milk.

Rheological and microstructural properties of rennet-induced milk gels containing different fat globules were studied. Recombined milks were prepared by mixing reconstituted low-heat skim milk powder and anhydrous milk fat emulsified with reconstituted skim milk powder (SMP), sodium caseinate (NaCas), whey protein isolate (WPI) or Tween 20. Final elastic modulus of the rennet gels containing WPI- or Tween 20-stabilized fat globules showed significantly lower values compared with those prepared with SMP-emulsified fat globules. SMP-stabilized fat globules interacted with the continuous casein network reinforcing the gel structure. Confocal micrographs supported the rheological data revealing that gels containing SMP-stabilized fat globules formed a tighter network relative to other treatments. Microscopy images also showed some degree of droplet flocculation in the case of gels containing WPI- or Tween 20-stabilized fat globules, and this was most likely the cause of the increase of elastic modulus of these systems. Contrary to reports for acid-induced casein gels, NaCas-stabilized fat globules hindered the formation of rennet gels. These results illustrate that rennet gel structure is affected by droplet-droplet and droplet-casein interactions, which in turn are determined by the composition of the oil-water interface as well as the ionic equilibrium in the reconstituted milk gels.

Rennet Coagulation and Cheesemaking Properties of Thermally Processed Milk: Overview and Recent Developments.

Thermally induced changes in milk proteins and minerals, particularly interactions among caseins and denatured whey proteins, influence important properties of dairy products in both positive and negative ways. Whereas the extensive protein connectivity and increased water-holding capacity resulting from such heat-induced protein modification account for the much desired firmness of acid gels of yogurt, thermal processing, on the other hand, severely impairs clotting and adversely affects the cheesemaking properties of rennet-coagulated cheeses. In technological terms, the principal ongoing challenge in the cheese industry is to take advantage of the water-holding capacity of thermally aggregated whey proteins without compromising the rennetability of cheese milk or the textural and functional attributes of cheese. Including some recent data from the authors' laboratory, this paper will discuss important aspects and current literature on the use of thermally processed milk in the production of rennet-coagulated cheeses and also some of the potential alternatives available for inclusion of whey proteins in cheese, such as the addition of microparticulated whey proteins to cheese milk.

Clinical and six-month angiographic evaluation of coronary arterial graft interrupted anastomoses by use of a self-closing clip device: a multicenter prospective clinical trial.

OBJECTIVES: To evaluate the safety and effectiveness of a self-closing surgical clip with an interrupted technique in left internal thoracic artery to left anterior descending artery bypass grafting. METHODS: Eighty-two patients were enrolled and treated (February 2000 through August 2001) in a prospective, nonrandomized, multicenter trial. Left internal thoracic artery to left anterior descending artery anastomoses were performed in 60 off-pump coronary artery bypasses (73%), 12 conventional coronary artery bypass grafting (15%), and 10 minimally invasive direct coronary artery bypass (12%) procedures. Angiograms (64 to 383 days, mean 200 days) were obtained on 63 patients (77%). Qualitative and quantitative angiographic assessment was performed by an independent core laboratory. RESULTS: The self-closing surgical clip was used for 82 left internal thoracic artery to left anterior descending artery interrupted anastomoses without the requirement for knot tying or primary suture management. Minimum left internal thoracic artery to left anterior descending artery anastomosis time was 3 minutes. There was one perioperative and one late death (both not heart related) and one reexploration for bleeding unrelated to the anastomotic site. FitzGibbon grades were as follows: A (n = 60, 95.2%), B (n = 3, 4.8%) including one kinked left internal thoracic artery, and O (n = 0, 0%). Quantitative analysis (n = 57) showed mean lumen diameters of left internal thoracic artery proximal to the anastomosis of 2.1 mm, at anastomosis of 2.0 mm, and in the left anterior descending artery distal to the anastomosis of 1.9 mm. The average ratio of the anastomosis to the left anterior descending artery diameter was 1.14 (0.45 to 1.93). Anastomotic stenosis as a percentage of average left internal thoracic artery to left anterior descending artery diameter was -2.3%, comparing favorably with results (23% to 24%) reported from the Patency, Outcomes, Economics, Minimally invasive direct coronary artery (POEM) bypass study. CONCLUSIONS: The interrupted technique, facilitated by a self-closing anastomotic clip, yields favorable 6-month angiographic results when compared with other published studies.

Methods for analysis of conjugated linoleic acids and trans-18:1 isomers in dairy fats by using a combination of gas chromatography, silver-ion thin-layer chromatography/gas chromatography, and silver-ion liquid chromatography.

Conjugated linoleic acids (CLA) are octadecadienoic acids (18:2) that have a conjugated double-bond system. Interest in these compounds has expanded since CLA were found to be associated with a number of physiological and pathological responses such as cancer, metastases, atherosclerosis, diabetes, immunity, and body fat/protein composition. The main sources of these conjugated fatty acids are dairy fats. Rumen bacteria convert polyunsaturated fatty acids, especially linoleic and linolenic acids, to CLA and numerous trans- containing mono- and diunsaturated fatty acids. It has been established that an additional route of CLA synthesis in ruminants and monogastric animals, including humans, occurs via delta9 desaturation of the trans-18:1 isomers. To date, a total of 6 positional CLA isomers have been found in dairy fats, each occurring in 4 geometric forms (cis,trans; trans,cis; cis,cis; and trans,trans) for a total of 24. All of these CLA isomers can be resolved only by a combination of gas chromatography (GC), using 100 m highly polar capillary columns, and silver-ion liquid chromatography, using 3 of these 25 cm columns in series. Complete analysis of all the trans-18:1 isomers requires prior isolation of trans monoenes by silver-ion thin-layer chromatography (TLC), followed by GC analysis using the same 100 m capillary columns operated at low temperatures starting from 120 degrees C. These analytical techniques are required to assess the purity of commercial CLA preparations, because their purity will affect the interpretation of any physiological and/or biochemical response obtained. Prior assessment of CLA preparations by TLC is also recommended to determine the presence of any other impurities. The availability of pure CLA isomers will permit the evaluation and analysis of individual CLA isomers for their nutritional and biological activity in model systems, animals, and humans. These techniques are also essential to evaluate dairy fats for their content of specific CLA isomers and to help design experimental diets to increase the level of the desired CLA isomers in dairy fats. These improved techniques are further required to evaluate the CLA profile in monogastric animals fed commercial CLA preparations for CLA enrichment of animal products. This is particularly important because absorption and metabolism will alter the ingested-CLA profile in the animal fed.

Peptide-coated vascular grafts: an in vivo study in sheep.

The data on function and patency of prosthetic vascular grafts in various clinical settings are limited. The purpose of this in vivo study was to compare the function and patency of P15-coated expanded polytetrafluoroethylene (ePTFE) vascular grafts to uncoated ePTFE grafts in sheep. The P15 cell-binding peptide was covalently immobilized onto the surface of ePTFE grafts by a novel atmospheric plasma coating method. We evaluated the amount of neointimal tissue ingrowth present at the arterial and venous sides of the anastomoses and the degree of endothelial cell resurfacing of the luminal surface of the graft. Four P15-coated grafts and two control grafts were implanted as arteriovenous grafts between the femoral artery and vein and the carotid artery and jugular vein in two sheep (n = 6). One animal was euthanized after 14 days and the other after 28 days. The study showed the intimal ingrowth was significantly less. The average intimal thickness of P15-coated grafts (658 microm) was approximately two and a half times less than that of uncoated samples (1657 microm). The newly formed endothelial cell lining was thicker and its coverage was more uniform for P15-coated grafts compared to the uncoated controls.

Foodborne and waterborne pathogenic bacteria in selected Organisation for Economic Cooperation and Development (OECD) countries.

The World Ranking Food Safety Performance reports by Charlebois in 2008 and 2010 importantly stimulated international discussion and encouraged efforts to establish realistic international benchmarks for food safety performance among Organisation for Economic Cooperation and Development (OECD) countries. This paper presents the international incidence of 5 common foodborne pathogens and describes the challenges of comparing international data. Data were compiled from surveillance authorities in the countries, such as the Natl. Notifiable Diseases Surveillance System of Australia; the Canadian Notifiable Diseases Surveillance System; the European Food Safety Authority, EFSA; the Ministry of Health, Labour and Welfare of Japan; New Zealand Food Safety Authority; and the U.S. Center for Disease Control and Prevention. The highest average rates in cases per 100000 people over the 12-y period from 2000 to 2011 for Campylobacter spp. (237.47), Salmonella spp. (67.08), Yersinia spp. (12.09), Verotoxigenic/Shiga toxin producing Escherichia coli (3.38), and Listeria monocytogenes (1.06) corresponded, in order, to New Zealand, Belgium, Finland, Canada, and Denmark. Comparatively, annual average rates for these 5 pathogens showed an increase over the 12-y period in 28%, 17%, 14%, 50%, and 6% of the countries for which data were available. Salmonella spp. showed a decrease in 56% of the countries, while incidence of L. monocytogenes was constant in most countries (94%). Variable protocols for monitoring incidence of pathogens among OECD countries remain. Nevertheless, there is evidence of sufficient standardization of monitoring protocols such as the European Surveillance System, which has contributed to reduce this gap.

Novel mitral repair device for the treatment of severe mitral regurgitation: preclinical ovine acute and chronic implantation model.

OBJECTIVE: A project is now underway to implement a novel percutaneous mitral repair system for severe mitral regurgitation (MR). The initial phase of the project consists of proof-of-concept by testing device characteristics using open surgical implantation. When surgical proof-of-concept of the intended percutaneous design is completed, a second phase of the project will consist of in vivo testing of the percutaneous transseptal system. The device is currently being designed to fold into a 17F catheter system and to unfold within the left atrium where attachment is accomplished using a reversible anchoring system. The purpose of this study was to show functionality of the device in elimination of MR using the open surgical method. METHODS: We have performed surgical prototype device implantation in 5 acute and 7 chronic sheep preparations. We created a P2-flail model of severe (4+) MR in the 12 sheep. Via a minimally invasive left thoracotomy incision and open repair on cardiopulmonary bypass, the device was implanted to determine efficacy of elimination of severe MR. Implantation was considered successful if 4+ regurgitation was converted to 1+ MR or lower. Left ventriculography and epicardial 2-dimensional/3-dimensional echocardiography were used to assess repair; serial 2-dimensional/3-dimensional transthoracic echocardiography was used to assess long-term mitral repair status. RESULTS: Twelve sheep had surgical creation of severe (4+) MR by cutting all chordae to the P2 scallop of the mitral valve; this preparation was tested and was found to produce 100% acute fatality without repair of the mitral valve. Five sheep had acute implantation of the device with elimination of regurgitation in 5/5 sheep. Seven sheep had chronic (1-7 month) implantation of the device. The device was tested in the chronic model for clinical status, residual regurgitation, thrombosis, and histopathology. All sheep had mitigation of MR and survived to the intended date of death. CONCLUSIONS: Proof-of-concept of a novel percutaneous mitral repair device has been completed using an ovine P2-flail severe MR model. The device has characteristics that will allow its use in posterior leaflet degenerative disease and functional/secondary MR. Open, minimally invasive, and robotic surgical implantation of the device can also be developed as an alternative to the percutaneous approach.

Selective targeting of TGF-ß activation to treat fibroinflammatory airway disease.

Airway remodeling, caused by inflammation and fibrosis, is a major component of chronic obstructive pulmonary disease (COPD) and currently has no effective treatment. Transforming growth factor-ß (TGF-ß) has been widely implicated in the pathogenesis of airway remodeling in COPD. TGF-ß is expressed in a latent form that requires activation. The integrin αvß8 (encoded by the itgb8 gene) is a receptor for latent TGF-ß and is essential for its activation. Expression of integrin αvß8 is increased in airway fibroblasts in COPD and thus is an attractive therapeutic target for the treatment of airway remodeling in COPD. We demonstrate that an engineered optimized antibody to human αvß8 (B5) inhibited TGF-ß activation in transgenic mice expressing only human and not mouse ITGB8. The B5 engineered antibody blocked fibroinflammatory responses induced by tobacco smoke, cytokines, and allergens by inhibiting TGF-ß activation. To clarify the mechanism of action of B5, we used hydrodynamic, mutational, and electron microscopic methods to demonstrate that αvß8 predominantly adopts a constitutively active, extended-closed headpiece conformation. Epitope mapping and functional characterization of B5 revealed an allosteric mechanism of action due to locking-in of a low-affinity αvß8 conformation. Collectively, these data demonstrate a new model for integrin function and present a strategy to selectively target the TGF-ß pathway to treat fibroinflammatory airway diseases.

Minimally invasive surgical implantation of the percutaneous left atrial appendage transcatheter occlusion device: initial experience in a canine model.

OBJECTIVE: Atrial fibrillation (AF) is a significant cause of thromboembolism and stroke. Left atrial appendage (LAA) occlusion is associated with a decreased risk of stroke in patients with AF. Percutaneous implantation of the percutaneous LAA transcatheter occlusion (PLAATO) device has shown reduction in stroke risk and decreased need for anticoagulation. A surgical method of PLAATO implantation is proposed for its utility and efficacy in cardiac surgical procedures, where LAA occlusion is indicated as a primary procedure or as an adjunct. We describe a surgical method for PLAATO deployment in an experimental model that simulates various operative scenarios including right minithoracotomy, right video-assisted thoracoscopic, or median sternotomy. METHODS: The PLAATO LAA occlusion device was deployed into the LAA in six dogs using a right minithoracotomy incision and catheter deployment via direct left atrial access. Intracardiac echocardiography and left atrial angiography were used to size, position, and verify proper deployment of the device. RESULTS: Successful PLAATO deployment was achieved in six of the six dogs. One dog required replacement of an undersized device. One dog required replacement of an oversized device. Four dogs required minor repositioning for optimal positioning. Complete flush occlusion of the LAA was achieved in three dogs; the other three dogs had trace leak by LAA angiography. CONCLUSIONS: Experimental surgical implantation of the PLAATO device produces stable and complete occlusion of the LAA. Potential surgical indications for PLAATO implantation include patients with AF where a percutaneous approach to LAA occlusion is contraindicated or in patients with AF who require concurrent cardiac surgery.

Interaction between casein micelles and whey protein/κ-casein complexes during renneting of heat-treated reconstituted skim milk powder and casein micelle/serum mixtures.

Casein micelles were separated from unheated reconstituted skim milk powder (RSMP) and were resuspended in the serum of RSMP that had been heated, with and without dialysis of this serum against unheated RSMP. Using size-exclusion chromatography, it was found that the soluble complexes of whey protein (WP) with κ-casein in the serum of the heated milk bind progressively to unheated casein micelles during renneting, even prior to the onset of clotting. Similar trends were noted when casein micelles from RSMP heated at pH values of 6.7, 7.1, or 6.3, each with different amounts of WP coating the micelles, were renneted in the presence of soluble WP/κ-casein complexes. No matter what was the initial load of micelle-bound WP complexes, all micelle types were capable of binding additional serum protein complexes during renneting. However, it is not clear that this binding of WP/κ-casein complexes to the micellar surface is a direct cause of the impaired rennet clotting of the RSMP.

Mouse and human lung fibroblasts regulate dendritic cell trafficking, airway inflammation, and fibrosis through integrin αvß8-mediated activation of TGF-ß.

The airway is a primary portal of entry for noxious environmental stimuli that can trigger airway remodeling, which contributes significantly to airway obstruction in chronic obstructive pulmonary disease (COPD) and chronic asthma. Important pathologic components of airway remodeling include fibrosis and abnormal innate and adaptive immune responses. The positioning of fibroblasts in interstitial spaces suggests that they could participate in both fibrosis and chemokine regulation of the trafficking of immune cells such as dendritic cells, which are crucial antigen-presenting cells. However, physiological evidence for this dual role for fibroblasts is lacking. Here, in two physiologically relevant models - conditional deletion in mouse fibroblasts of the TGF-ß-activating integrin αvß8 and neutralization of αvß8 in human COPD fibroblasts - we have elucidated a mechanism whereby lung fibroblast chemokine secretion directs dendritic cell trafficking, in a manner that is critically dependent on αvß8-mediated activation of TGF-ß by fibroblasts. Our data therefore indicate that fibroblasts have a crucial role in regulating both fibrotic and immune responses in the lung.