A small scale study to evaluate the efficacy of microneedling in the presence or absence of platelet-rich plasma in the treatment of post-clipping alopecia in dogs.

BACKGROUND: Post-clipping alopecia often has a clinically poor response to therapy and prolonged alopecia is a source of anxiety for some owners. In humans and dogs, superficial microtrauma via a microneedling (MN) device induces mechanical stimulation of the hair follicle with resultant hair regrowth. Human studies suggest that concurrent application of platelet-rich plasma (PRP) with MN induces more rapid regrowth of better-quality hair than microneedling alone. HYPOTHESIS: Microneedling with PRP will induce more rapid regrowth of better quality hair. ANIMALS: Four unrelated client-owned dogs diagnosed with post-clipping alopecia. METHODS AND MATERIALS: This was a prospective study. The affected site was divided in half, with the first half treated with MN alone and the second half treated with MN + PRP. Hair regrowth was assessed by clinician and owner using a hair growth assessment scale (HGAS) at one, three, six and 12 months. RESULTS: At three months, all dogs had improved and three exhibited greater hair regrowth on the MN + PRP side. A similar response was noted bilaterally in three dogs, which had improved by 76-100% at six months and remained unchanged at 12 months. One dog improved by < 26% at six months, but had> 50% re-growth by 12 months. The small sample size precluded statistical analysis. CONCLUSIONS AND CLINICAL IMPORTANCE: In dogs with post-clipping alopecia, MN + PRP appeared to induce more rapid hair regrowth than MN; however, overall results were visibly equivalent by six months regardless of method. Both MN and MN + PRP proved successful for treating post-clipping alopecia.

Sodium Hypochlorite/Salicylic Acid Shampoo for Treatment of Canine Staphylococcal Pyoderma.

The emergence of methicillin-resistant Staphylococcus pseudintermedius has increased the interest in topical therapy for treating canine pyoderma. Shampooing with chlorhexidine followed by dilute bleach rinses are often recommended, but household bleach can dry the skin and is unpleasant to use. A shampoo formulated with sodium hypochlorite and salicylic acid was evaluated as sole therapy for dogs with superficial pyoderma associated with S. pseudintermedius, including methicillin-resistant strains. Client-owned dogs were recruited based on positive culture for methicillin-resistant staphylococci or prior failure of pyoderma to respond to antibiotics. This prospective, open-label pilot study assessed the efficacy of the shampoo when used three times weekly for 4 wk. Dogs were evaluated at baseline and at 2 and 4 wk by cytology, clinical examination, and owner assessment. Digital images were also obtained. Baseline bacterial counts, clinical assessments and owner scores were significantly improved at 2 and 4 wk. Clients completing the study reported excellent lathering and dispersion, reduction in odor, and brightening of white and light coats. No owners reported skin dryness or other adverse events during the study. We conclude that this shampoo containing sodium hypochlorite in a vehicle that avoids skin drying is an effective treatment for canine pyoderma.

Agreement between allergen-specific IgE assays and ensuing immunotherapy recommendations from four commercial laboratories in the USA.

BACKGROUND: Canine allergen-specific IgE assays in the USA are not subjected to an independent laboratory reliability monitoring programme. HYPOTHESIS/OBJECTIVES: The aim of this study was to evaluate the agreement of diagnostic results and treatment recommendations of four serum IgE assays commercially available in the USA. METHODS: Replicate serum samples from 10 atopic dogs were submitted to each of four laboratories for allergen-specific IgE assays (ACTT(®) , VARL Liquid Gold, ALLERCEPT(®) and Greer(®) Aller-g-complete(®) ). The interlaboratory agreement of standard, regional panels and ensuing treatment recommendations were analysed with the kappa statistic (κ) to account for agreement that might occur merely by chance. Six comparisons of pairs of laboratories and overall agreement among laboratories were analysed for ungrouped allergens (as tested) and also with allergens grouped according to reported cross-reactivity and taxonomy. RESULTS: The overall chance-corrected agreement of the positive/negative test results for ungrouped and grouped allergens was slight (κ = 0.14 and 0.13, respectively). Subset analysis of the laboratory pair with the highest level of diagnostic agreement (κ = 0.36) found slight agreement (κ = 0.13) for ungrouped plants and fungi, but substantial agreement (κ = 0.71) for ungrouped mites. The overall agreement of the treatment recommendations was slight (κ = 0.11). Altogether, 85.1% of ungrouped allergen treatment recommendations were unique to one laboratory or another. CONCLUSIONS AND CLINICAL IMPORTANCE: Our study indicated that the choice of IgE assay may have a major influence on the positive/negative results and ensuing treatment recommendations.

Update on equine allergies.

Horses develop many skin and respiratory disorders that have been attributed to allergy. These disorders include pruritic skin diseases, recurrent urticaria, allergic rhinoconjunctivitis, and reactive airway disease. Allergen-specific IgE has been detected in these horses, and allergen-specific immunotherapy is used to ameliorate clinical signs. The best understood atopic disease in horses is insect hypersensitivity, but the goal of effective treatment with allergen-specific immunotherapy remains elusive. In this review, updates in pathogenesis of allergic states and a brief mention of the new data on what is known in humans and dogs and how that relates to equine allergic disorders are discussed.

Use of modified ciclosporin in the management of feline pemphigus foliaceus: a retrospective analysis.

BACKGROUND: Glucocorticoids as sole therapy for pemphigus foliaceus (PF) in cats are not always successful, and it is common to need additional immunomodulating agents to manage the disease. HYPOTHESIS/OBJECTIVES: This retrospective study evaluated the use of modified ciclosporin as an adjuvant or sole immunomodulating drug in cats with PF and compared their response to PF cats managed with chlorambucil. ANIMALS: Fifteen client-owned cats diagnosed with PF that received ciclosporin and/or chlorambucil as part of their treatment and had adequate follow-up to assess treatment response were evaluated. METHODS: Records were reviewed from feline PF patients presented between the years of 1999 and 2009. Cats were divided into two treatment groups: those treated with ciclosporin and those treated with chlorambucil. Most cats in both groups also received concurrent systemic glucocorticoids. Each group contained six patients. Three cats were treated with both medications and are discussed separately. Time to disease remission, remission-inducing glucocorticoid dose, maintenance or final glucocorticoid dose, disease response and adverse effects were assessed. RESULTS: There was no significant difference in remission times or disease response between groups. All six patients maintained with ciclosporin for PF management were weaned off systemic glucocorticoids, while glucocorticoid therapy was stopped in only one of the six cats receiving chlorambucil. CONCLUSIONS AND CLINICAL IMPORTANCE: Modified ciclosporin is effective in the management of feline pemphigus foliaceus and is glucocorticoid sparing.

Update on pathogenesis, diagnosis, and treatment of atopic dermatitis in dogs.

Improved understanding of the pathogenesis of atopic dermatitis in dogs has led to more effective treatment plans, including skin barrier repair and new targeted treatments for management of allergy-associated itch and inflammation. The intent of this review article is to provide an update on the etiologic rationale behind current recommendations that emphasize a multimodal approach for the management of atopic dermatitis in dogs. Increasing knowledge of this complex disease process will help direct future treatment options.

Phosphatidylserine-dependent ingestion of apoptotic cells promotes TGF-beta1 secretion and the resolution of inflammation.

Ingestion of apoptotic cells in vitro by macrophages induces TGF-beta1 secretion, resulting in an anti-inflammatory effect and suppression of proinflammatory mediators. Here, we show in vivo that direct instillation of apoptotic cells enhanced the resolution of acute inflammation. This enhancement appeared to require phosphatidylserine (PS) on the apoptotic cells and local induction of TGF-beta1. Working with thioglycollate-stimulated peritonea or LPS-stimulated lungs, we examined the effect of apoptotic cell uptake on TGF-beta1 induction. Viable or opsonized apoptotic human Jurkat T cells, or apoptotic PLB-985 cells, human monomyelocytes that do not express PS during apoptosis, failed to induce TGF-beta1. PS liposomes, or PS directly transferred onto the PLB-985 surface membranes, restored the TGF-beta1 induction. Apoptotic cell instillation into LPS-stimulated lungs reduced proinflammatory chemokine levels in the bronchoalveolar lavage fluid (BALF). Additionally, total inflammatory cell counts in the BALF were markedly reduced 1-5 days after apoptotic cell instillation, an effect that could be reversed by opsonization or coinstillation of TGF-beta1 neutralizing antibody. This reduction resulted from early decrease in neutrophils and later decreases in lymphocytes and macrophages. In conclusion, apoptotic cell recognition and clearance, via exposure of PS and ligation of its receptor, induce TGF-beta1 secretion, resulting in accelerated resolution of inflammation.

Elastase-mediated phosphatidylserine receptor cleavage impairs apoptotic cell clearance in cystic fibrosis and bronchiectasis.

Cystic fibrosis is characterized by an early and sustained influx of inflammatory cells into the airways and by release of proteases. Resolution of inflammation is normally associated with the orderly removal of dying apoptotic inflammatory cells through cell recognition receptors, such as the phosphatidylserine receptor, CD36, and alpha v integrins. Accordingly, removal of apoptotic inflammatory cells may be impaired in persistent inflammatory responses such as that seen in cystic fibrosis airways. Examination of sputa from cystic fibrosis and non-cystic fibrosis bronchiectasis patients demonstrated an abundance of apoptotic cells, in excess of that seen in patients with chronic bronchitis. In vitro, cystic fibrosis and bronchiectasis airway fluid directly inhibited apoptotic cell removal by alveolar macrophages in a neutrophil elastase-dependent manner, suggesting that elastase may impair apoptotic cell clearance in vivo. Flow cytometry demonstrated that neutrophil elastase cleaved the phosphatidylserine receptor, but not CD36 or CD32 (Fc gamma RII). Cleavage of the phosphatidylserine receptor by neutrophil elastase specifically disrupted phagocytosis of apoptotic cells, implying a potential mechanism for delayed apoptotic cell clearance in vivo. Therefore, defective airway clearance of apoptotic cells in cystic fibrosis and bronchiectasis may be due to elastase-mediated cleavage of phosphatidylserine receptor on phagocytes and may contribute to ongoing airway inflammation.

Life after corpse engulfment: phagocytosis of apoptotic cells leads to VEGF secretion and cell growth.

Removal of apoptotic cells by neighboring viable cells or professional phagocytes is essential for the maintenance of tissue homeostastis. Here we show that the phagocytosis of apoptotic Jurkat T cells by mouse epithelial cells (HC-11) and peritoneal macrophages leads to the secretion of growth and survival factors. We characterized VEGF as one of these factors which subsequently promote the proliferation of endothelial cells. Further we demonstrate that the phagocytosis of apoptotic bodies inhibits both spontanous and UV-irradiation-induced apoptosis in endothelial and epithelial cells. These effects were not observed when phagocytes had been exposed to viable or necrotic Jurkat T cells. We conclude that phagocytosis of apoptotic cells leads to secretion of growth and survival factors by phagocytes that represents a new form of life-promoting cell-cell interaction.

The role of the macrophage in apoptosis: hunter, gatherer, and regulator.

Clearance of cellular corpses is a critical feature of apoptosis in vivo during development, tissue homeostasis, and resolution of inflammation. As the professional phagocytes of the body, macrophages play a key role in this process. By recognizing emerging signals using several different receptors, macrophages engulf apoptotic cells swiftly and efficiently. In addition, the binding of apoptotic cells profoundly down-regulates the ability of the macrophage to produce inflammatory mediators by inducing the release of antiinflammatory mediators. Finally, macrophages may actually induce cell death in specific cells during embryogenesis. Abnormalities of apoptotic cell clearance may contribute to the pathogenesis of chronic inflammatory diseases, including those of autoimmune etiology. It is also possible that certain malignant tumor cells co-opt the mechanisms for apoptotic cell clearance to avoid immune surveillance by subverting macrophage and dendritic cell responses.

Beryllium induces apoptosis in human lung macrophages.

The link between metal-induced apoptosis and granulomatous inflammation in human disease pathogenesis is not established. The presence of TUNEL positive nuclei in chronic beryllium disease (CBD) pulmonary granulomas suggested the possibility that beryllium (Be) could induce apoptosis in adherent macrophages from CBD bronchoalveolar (BAL) cells. Apoptosis was measured in un-stimulated and Be-stimulated BAL adherent macrophages from CBD (n = 21) and Be-sensitized (BeS, n = 16) subjects. Be-stimulated CBD and BeS macrophages underwent caspase-dependent nuclear fragmentation, cytoplasmic membrane blebbing and CD14 loss. Be-stimulated adherent macrophage apoptosis was not due to TNF-alpha production. Apoptosis, CD14 loss and TNF-alpha production were not observed in unstimulated BAL macrophages. Thus, Be-stimulated BAL adherent macrophage apoptosis occurred whether cells were derived from patients with granulomatous inflammation or not, was caspase-mediated and occurred independent of TNF-alpha levels. We conclude that Be-induced human lung adherent macrophage apoptosis could contribute to the host's continued re-exposure to Be resulting in chronic granulomatous inflammation.

Of Horses and Men: Urticaria.

Abstract- Urticarial eruptions, with or without pruritus, are common lesions in horses. The pathogenesis of these lesions can include immunological and other mechanisms. Research in the human field suggests that the mast cell co-ordinates the urticarial response by releasing a complex array of inflammatory mediators. Other cells, including the neutrophil, the eosinophil and the macrophage, may also play a role in the development of wheals. Elucidation of the role of many of these cells and mediators in the evolution of urticaria is only just beginning. Successful treatment of this dermatological disorder requires a careful search for the underlying cause; however, this can be difficult to identify in many cases. The purpose of this review is to discuss the potential factors leading to urticaria in horses and to describe a standard diagnostic approach for determination of the underlying cause. Résumé- Des eruptions urticariennes avec ou sans prurit sont des lésions communes chez le cheval. La pathogénie de ces lésions peut être en rapport avec des mécanismes immunologiques et autres. Les recherches dans le domaine humain suggèrent que les mastocytes coordonnent la réponse urticarienne en libérant un ensemble complexe de médiateurs de l'inflammation. D'autres cellules, comprenant les neutrophiles, les éosinophiles et les macrophages sont aussi susceptibles de jouer un rôle dans le développment des plaques ortiées. La compréhension du rôle de plusieurs de ces cellules et de ces médiateurs dans l'evolution de l'urticaire ne fait que commencer. La réussite du traitement de ce trouble cutané demande une recherche minutieuse de la cause sous-jacente; toutefois, elle peut être difficile à identifier dans de nombreux cas. Le propos de cette revue est de discuter des facteurs potentiels conduisant à l'urticaire chez le cheval et de décrire une approche diagnostique standard afin de déterminer la cause sous-jacente. Zusammenfassung- Häufig treten bei Pferden bei Urticaria Eruptionen auf, die mit oder ohne Pruritus verlaufen können. Die Pathogenese dieser Veränderungen umfasst immunologische und andere Mechanismen. Untersuchungen in der Humanmedizin weisen darauf hin, dass Mastzellen die Reaktionen bei Urticaria koordinieren, indem sie einen ganzen Komplex von Entzündungsmediatoren ausschütten. Andere Zellen, auch Neutrophile, Eosinophile und Makrophagen, sind vermutlich ebenfalls an dieser Entwicklung beteiligt. Die Rolle von zahlreichen Zellen und Entzündungsmediatoren bei der Entstehung einer Urticaria ist erst in Ansätzen bekannt. Voraussetzung für die erfolgreiche Behandlung dieser Hauterkrenkung ist eine sorgfältige Suche nach der eigentlichen Ursache, die sich aber oft schwierig gestaltet. Dieser Artikel soll die möglichen Ursachen für eine Utricaria beim Pferd und die Standarddiagnostik bei der Bestimmung der eigentlichen Ursache aufzeigen. Resumen Las erupciones urticariformes, con o sin prurito, son lesiones frecuentes en el caballo. La patogenia de estas lesiones incluyen tanto mecanismos inmunológicos como mecanismos no inmunológicos. Diferentes investigaciones realizadas en el campo de la patología humana sugieren que la célula cebada coordina la respuesta urticariforme liberando gran cantidad de mediadores inflamatorios. Otras células, entre ellas el neutrófilo el eosinófilo y el macrófago, también juegan un papel importante en el desarrollo de los habones. En la actualidad se empieza a conocer el papel que estas células juegan en la evolución de la urticaria. Un tratamiento efectivo de esta enfermedad depende de la detección de la causa desencadenante, aunque en muchos casos esto no es posible. El objetivo de esta revisión es discutir los factores que pueden conducir a la urticaria en el caballo y describir el protocolo diagnóstico adecuado para la detección del agente desencadenante.

Epithelial cells remove apoptotic epithelial cells during post-lactation involution of the mouse mammary gland.

Following the cessation of lactation, the mammary gland undergoes a physiologic process of tissue remodeling called involution in which glandular structures are lost, leaving an adipose tissue compartment that takes up a much larger proportion of the tissue. A quantitative morphometric analysis was undertaken to determine the mechanisms for clearance of the epithelial cells during this process. The involution process was set in motion by removal of pups from 14-day lactating C57BL/6 mice. Within hours, milk-secreting epithelial cells were shed into the glandular lumen. These cells became apoptotic, exhibiting exposure of phosphatidylserine residues on their surfaces, activation of effector caspase-3, staining for caspase-cleaved keratin 18, loss of internal organellar structure, and nuclear breakdown, but minimal blebbing or generation of apoptotic bodies. Clearance of residual milk and the shed epithelial cells was rapid, with most of the removal occurring in the first 72 h. Intact apoptotic epithelial cells were engulfed in large numbers by residual viable epithelial cells into spacious efferosomes. This process led to essentially complete involution within 4 days, at which point estrous cycling recommenced. Macrophages and other inflammatory cells did not contribute to the clearance of either residual milk or apoptotic cells, which appeared to be due entirely to the epithelium itself.

Interaction between phosphatidylserine and the phosphatidylserine receptor inhibits immune responses in vivo.

Phosphatidylserine (PS) on apoptotic cells promotes their uptake and induces anti-inflammatory responses in phagocytes, including TGF-beta release. Little is known regarding the effects of PS on adaptive immune responses. We therefore investigated the effects of PS-containing liposomes on immune responses in mice in vivo. PS liposomes specifically inhibited responses to Ags as determined by decreased draining lymph node tissue mass, with reduced numbers of total leukocytes and Ag-specific CD4(+) T cells. There was also a decrease in formation and size of germinal centers in spleen and lymph nodes, accompanied by decreased levels of Ag-specific IgG in blood. Many of these effects were mimicked by an agonistic Ab-specific for the PS receptor. TGF-beta appears to play a critical role in this inhibition, as the inhibitory effects of PS were reversed by in vivo administration of anti-TGF-beta Ab. PS-containing liposomes did not appear to directly inhibit dendritic cell maturation in vitro in response to a variety of stimuli, nor did it prevent their migration to regional lymph nodes in vivo, suggesting that the inhibitory effects may have resulted from complicated interactions between tissue cells and dendritic cells, subsequently inhibiting their ability to productively activate T lymphocytes.

Defective apoptotic cell phagocytosis attenuates prostaglandin E2 and 15-hydroxyeicosatetraenoic acid in severe asthma alveolar macrophages.

RATIONALE: Clearance of apoptotic cells is crucial to the resolution of inflammation and development of fibrosis, but the process is not well understood in normal or diseased human lungs. OBJECTIVES: To determine phagocytosis of apoptotic cells by primary human alveolar macrophages and whether defects in uptake of apoptotic cells are associated with decreases in antiinflammatory/antifibrotic mediators. METHODS: Human bronchoalveolar lavage macrophages (AMphis) from normal control subjects and subjects with mild-moderate or severe asthma were examined in vitro for phagocytosis of apoptotic human T-cell line Jurkats and secretion of inflammatory mediators. MEASUREMENTS AND MAIN RESULTS: AMphis from normal subjects and patients with mild-moderate asthma were able to phagocytose apoptotic cells in response to LPS, resulting in an induction of the antifibrotic and/or antiinflammatory eicosanoids, prostaglandin E2 (PGE2) and 15-hydroxyeicosatetraenoic acid (HETE). In contrast, AMphis from patients with severe asthma had defective LPS-stimulated uptake of apoptotic cells, with associated failure to induce PGE2 and 15-HETE. In addition, LPS-stimulated basal levels of tumor necrosis factor alpha and granulocyte-macrophage colony-stimulating factor were reduced in all patients with asthma, whereas PGE2 and 15-HETE were reduced only in patients with severe asthma. Dexamethasone enhanced specific uptake of apoptotic cells in all subjects, while suppressing inflammatory mediator secretion. CONCLUSIONS: A decrease in AMphis LPS-responsiveness in severe asthma is manifested by defective apoptotic cell uptake and reduces secretion of inflammatory mediators. This may contribute to the chronicity of inflammation and remodeling in lungs of patients with asthma.