bluuebunny/crossref_metadata_2025_split huggingface dataset:
FC‐27  A spontaneous dog model for in vivo gene therapy of junctional epidermolysis bullosa
Authors: Spirito F. , Capt A. , Guaguere E. , Del Rio M. , Meneguzzi G. 
Published: August 2004

Junctional epidermolysis bullosa (JEB) is a mechanobullous skin disease associated with mutations in the basement membrane components of the dermo‐epidermal junction. This condition is a suitable prototype for proving the feasibility of a gene therapy approach to genodermatoses. As successful genetic treatment requires immunocompetent animal models to study the possible host immune response to the transgene, we have characterized a breed of short‐haired pointers suffering from a mild form of JEB. These animals exhibit skin blistering and erosions following minor trauma. Immunohistochemical analysis of the skin biopsies and immunoprecipitation of spent medium from cultured JEB keratinocytes showed reduced expression and secretion of the α3 chain of laminin 5 (α3, β3, γ2), the major adhesion ligand of basal keratinocytes. The search for genetic mutations detected a homozygous insertional mutation (4818 + 207 ins6.5 kb) in intron 35 of the lama3 gene. Consequently, keratinocytes from dogs with JEB secrete reduced amounts of wild‐type laminin 5, and adhesive properties of the keratinocytes are compromised. Retroviral transfer of wild‐type dog α3 cDNA into JEB keratinocytes enhanced secretion of laminin 5 in the extracellular matrix and restored the adhesion, differentiation and proliferative capacity of the transduced cells. Transplantable fibrin‐based skin equivalents made with transduced JEB keratinocytes and grafted onto SCID mice generated normal cohesive and stratified epithelia and showed stable localized deposition of laminin 5 at the dermo‐epidermal junction. Our results form the basis for preclinical assays of gene therapy on a unique immunocompetent animal model for an inherited skin disease. Funding: DEBRA (UK) Foundation, Association Francaise Contre les Myopathies.
FC‐28  Ultrastructural markers expressed in cultured canine dendritic cells and laminated bodies found in Langerhans cells: similarities and differences
Authors: Ibisch C. , Lieubeau B. , Pradal G. , Bach J. M. 
Published: August 2004

Ultrastructural markers that consist of coated vesicles, regularly laminated bodies, pleiomorphic inclusions and paracrystalline structures have been described in canine cutaneous histiocytoma (CCH), a cutaneous tumour of Langerhans cell origin. These microstructures are also observed in congenital self‐healing histiocytosis, a human Langerhans cell tumour. We have observed for the first time ultrastructural markers in canine dendritic cells (DC) that share some of the features of the microstructures described in CCH. Canine DC were generated in vitro from adherent peripheral blood mononuclear cells. Morphological analyses and ultrastructural and functional studies showed that these cells display typical DC features. After 1 week of in vitro culture of adherent peripheral blood mononuclear cells in the presence of canine IL‐4 and human GM‐CSF, a great proportion of the cells displayed typical cytoplasmic processes (optical and electronic microscopy) and expressed CD14 and MHC class II molecules (cytometric analysis). In addition, allogeneic mixed lymphocyte reactions were performed to assess the ability of these cells to stimulate the proliferation of allogeneic lymphocytes. Compared to monocytes/macrophages, these cells were able to stimulate allogeneic lymphocyte proliferation very strongly, as is reported for DC in other species. Ultrastructural analysis of these DC revealed the presence of cytoplasmic organites, dense granules of variable sizes containing periodic microstructures. These results were obtained for the three different breeds tested. The microstructures previously described in Langerhans cells (e.g. in CCH) share some similarities with periodic microstructures (membrane, laminated aspect, cytoplasmic localization), but not the typical periodic aspect seen in DC. Funding: National Veterinary School of Nantes.
FC‐29  Distribution of γδ‐T lymphocytes in normal canine skin
Authors: Hirt B. C. , Affolter V. K. , Löffler J. F. , Moore P. F. 
Published: August 2004

The exact function of T cells with the γδ‐T‐cell receptor is still poorly understood and their participation in different skin diseases is largely unknown. The goals of this study were to investigate the distribution of γδ‐T cells in normal canine skin and mucocutaneous junctions, and to evaluate their frequency in different body locations. Skin samples were collected from 10 dogs. From each dog, 10 different skin regions were sampled. Two random 30 μm cryosections were used for immunohistochemistry. Each tissue was evaluated for the presence of T cells, αβ‐T cells and γδ‐T cells. The numbers of positive cells per volume of tissue were determined in the epidermis and superficial dermis of each sample using a 42‐point square grid overlay. Accurate thickness of the tissue section was measured with an A2 axis stage micrometer. Results were analysed statistically and 1000 cells/mm3 were used as the threshold value. Independent from body location, the vast majority of γδ‐T cells were observed in the epidermis. Significant occurrences of epidermal γδ‐T cells were found in 50% of samples from the anorectal skin and eyelid, and in 25% of samples from the lip, interdigital skin and pinna. However, <1000 cells/mm3 were observed in the axilla, trunk, inguinal skin, tongue and footpad samples. This study reveals information about the distribution of γδ‐T cells in the skin and, hence, may provide new insights into the prevalence of these cells in skin diseases preferentially developing in certain body locations. Funding: Leukocyte Antigen Biology Laboratory, University of California‐Davis.
FC‐30  Presence of dust mites in the environment of dust mite‐sensitized atopic dogs
Authors: Farmaki R. , Saridomichelakis M. N. , Koutinas A. F. , Papazahariadou M. G. , Leontides L. , Gioulekas D. 
Published: August 2004

Dust mites (DM) are the most common offending aeroallergens in atopic dogs. The aim of this study was to compare the DM load of households with atopic dogs (Group A, n = 8) that had positive intradermal test reactions to Dermatophagoides farinae, D. pteronyssinus, Acarus siro, Lepidoglyphus destructor and/or Tyrophagus putrescentiae to the DM load of households with nonatopic dogs (Group B, n = 4) and of nonpet households (Group C, n = 8). Group A dogs presented with perennial pruritus, were free of pathogenic mites and fleas, did not respond to an elimination diet, and fulfilled the diagnostic criteria of atopic dermatitis. All Group B dogs tested intradermally negative and had no dermatological problems. Dust samples were vacuum collected in a standardized fashion from the human (all groups) and dog mattresses (Groups A and B) or from the couch (Group C) four times, once for each season of the year. The presence of DM was assessed with a commercial test (Acarex test) and stereoscopically. At least one DM was found in all Group A houses. The DM load was not significantly different between the seasons or the three animal groups. The sensitivity of the Acarex test was significantly lower than that of stereoscopic examination (P < 0.001). In conclusion, the environmental DM load was similar between atopic and nonatopic dogs, the presence of dogs in a household didn't increase DM numbers, and stereoscopy was more sensitive than the Acarex test for the detection of DM. Funding: Self‐funded.
FC‐31  Prevalence and characterization of house dust mites and house dust mite allergens collected from the bedding, skin and hair coat of dogs in southwest England
Authors: Jackson A. P. , Shaw S. E. , Hart B. J. , Helps C. R. , Foster A. P. 
Published: August 2004

The house dust mites Dermatophagoides farinae and D. pteronyssinus are commonly used in diagnostic serological and intradermal tests for canine atopic dermatitis in the UK. However, there are few studies that have characterised the exposure of UK pet dogs to these mites. Objectives of this study were to determine the prevalence of mites and to characterise the mite species on skin, hair coat and bedding of pet dogs. A population of nonhospitalized pet dogs for which bedding was available for analysis was recruited irrespective of dermatological status (n = 34). Dust samples (n = 68) were collected from both dogs and their beds using a standardised vacuuming technique. Mites were identified using accepted morphological criteria. House dust mite allergen concentrations were assayed using a standardised ELISA for Der p 1 and Der f 1. Mites were identified in 15 of 68 samples (22%): D. pteronyssinus alone in three of 68, D. pteronyssinus in combination with storage or unclassified mites in five of 68, and storage or unclassified mites alone in seven of 68. Dermatophagoides farinae was not identified in any samples. Der p 1 allergens were detected in 37 of 68 samples (54%), and Der f 1 in four of 68 samples (5.9%). Contrary to studies elsewhere in Europe and North America, these findings support studies of human asthma patients in the UK where exposure to D. pteronyssinus is common, but rare to D. farinae. Given the high prevalence of positive intradermal and serological reactions to D. farinae in atopic dogs, further investigations are warranted to clarify potential cross‐reactivity with other mite allergens. Funding: PetPlan Charitable Trust, Pfizer Animal Health.
FC‐32  Identification of airbourne moulds in stables over a 2‐year period
Authors: Paterson S. , Bacon T. 
Published: August 2004

Fungal moulds are well recognised as a trigger for allergic disease in people. Good correlation is recognized between rising levels of airbourne moulds and allergic reactions to these moulds. This study was designed to evaluate the levels and types of airbourne moulds in different stable types over a 2‐year period. Four stables were chosen within an equine hospital environment. Shavings were used as bedding in all stables throughout the period. The stables were selected as four different constructions with different outlooks and different ventilation, although they were all within the same establishment. Each month throughout the investigation period, a Sabouraud's medium petri dish was mounted vertically on the wall of each stable at a height of 1.5 m and left uncovered for 30 min. The temperature and general weather conditions were recorded. The dish was submitted to the laboratory for culture. A variety of different moulds were identified. These included Penicillium spp., Alternaria spp., Aspergillus spp., Mucor spp. and Candida spp. The different stable constructions grew different numbers of moulds, but the same type on each occasion. Cold icy weather tended to favour growth of Penicillium spp. Aspergillus spp. was identified most commonly when conditions were cold and damp in the autumn. Mould growth was sparse in the spring, but tended to rise in mid‐summer and into the autumn. Funding: Self‐funded.
FC‐33  Expression of IL‐12 receptor β2 gene in atopic dogs
Authors: Kitamiya E. , Momoi Y. , Iwasaki T. 
Published: August 2004

Atopic dermatitis (AD) is very common in dogs, but its pathogenesis is not yet fully understood. It has been suggested that a Th2‐dominant status may be associated with the occurrence of canine AD. IL‐12 is thought to be important for the differentiation of Th1 cells. The IL‐12 receptor β2 (IL‐12Rβ2) gene is considered to play a critical role in signal transduction and is attracting attention as one of the causative genes of AD in humans. The purpose of this study was to investigate the relationship between IL‐12Rβ2 gene expression and canine AD. The canine IL‐12Rβ2 gene was cloned by RT‐PCR and its nucleotide sequences were determined. Canine IL‐12Rβ2 showed 76.8% homology at the amino acid level with human IL‐12Rβ2, and its structural motifs were well conserved. cDNA with a 91 bp deletion including the transmembrane region was also cloned, which consequently produced a frame shift and an early stop codon. The deletion region corresponded to exon 14 of the human IL‐12Rβ2 gene on chromosome 1. The expression of deleted canine IL‐12Rβ2 mRNA in phytohemagglutinin‐stimulated peripheral blood mononuclear cells was examined in seven healthy dogs and 11 AD dogs. Both deleted and intact mRNAs were expressed at constant ratios in healthy and AD dogs. The results indicate that the deletion of the transmembrane region is not associated with the occurrence of AD, and that the expression of the deleted mRNA may be constitutive and produced by alternative splicing. Funding: Self‐funded.
FC‐35  Real‐time evaluation of cytokine and protease expression in flea‐allergic and nonallergic dogs
Authors: Brachelente C. , Wuersch K. , Doherr M. , Reist M. , Peel J. E. , Welle M. 
Published: August 2004

Flea bite allergy is a skin disorder with a complex immunopathogenesis. Th2 cytokines and proteases have been hypothesized to play a role in the pathogenesis. The aims of our study were to evaluate the cytokine and protease expression in skin biopsies and peripheral blood mononuclear cells (PBMC), the histopathological response, and the response to intradermal tests (IDT) in sensitized nonallergic and sensitized allergic dogs. Twenty dogs were exposed to fleas once a day for 4 days. Before flea exposure and after the last flea exposure, an IDT was performed, skin biopsies were taken, and PBMC were isolated. Skin biopsies and PBMC were stimulated with various substances, and real‐time RT‐PCR was performed using primers for tryptase, chymase, IL‐4, IL‐5, IL‐13, TNFα and IFNγ. Intradermal tests and histopathological examination of biopsies revealed much stronger reactions in the allergic group. Before flea exposure, mRNA expression of chymase, TNFα, IFNγ, IL‐5 and IL‐13 in biopsies of allergic dogs was higher compared to nonallergic dogs, whereas in PBMC only the expression of IFNγ was higher. After flea exposure, the difference in mRNA expression between allergic and nonallergic dogs was not as striking. In allergic dogs, stimulation of PBMC with flea antigen resulted in a higher expression of IL‐4 and IL‐13, whereas in biopsies only the expression of IL‐4 was higher. Our results demonstrate that even without the presence of antigen, the pattern of cytokine expression differs between allergic and nonallergic dogs. Furthermore, these results clearly demonstrate that the inflammatory response is much stronger in allergic dogs. Funding: Novartis SA.
FC‐36  The use of immunostimulatory bacterial DNA sequences in allergen‐specific immunotherapy of canine atopic dermatitis
Authors: Mueller R. S. , Veir J. , Fieseler K. V. , Dow S. 
Published: August 2004

The purpose of this study was to evaluate a combination of immunostimulatory bacterial DNA sequences and allergen‐specific immunotherapy for the treatment of canine atopic dermatitis. Seven dogs with nonseasonal atopic dermatitis diagnosed by history, clinical signs and exclusion of differential diagnoses were included. All dogs had been on allergen‐specific immunotherapy for at least 12 months with incomplete responses, were on additional antipruritic therapy and showed residual pruritus. Pruritus was marked by the owner on a visual analogue scale, lesions were determined by a clinician using the Canine Atopic Dermatitis Extent and Severity Index (CADESI), and concurrent medications were recorded before entering the study and after 14 weeks of treatment. Peripheral blood mononuclear cells were isolated and cultured; canine cytokine message for IFNγ, IL‐4, TNF and IL‐10 was quantitated using RT‐PCR. A mixture of allergen extract and liposome‐DNA complexes was injected intradermally at the beginning of the study and after 2, 4, 6, 10 and 14 weeks. CADESI, pruritus and medication scores, and cytokine messages at the beginning and end of the study were compared with a paired t‐test. There were significant improvements in pruritus scores (P = 0.0277). Reductions in medication scores and CADESI were not statistically significant. IL‐4 production decreased significantly (P = 0.0428); decreases in other cytokines were not significant. Although the number of dogs in this pilot study was small, the results warrant further investigation of a combination of immunostimulatory bacterial DNA sequences and allergen‐specific immunotherapy for the treatment of canine atopic dermatitis. Funding: Self‐funded.
FC‐37  Use of recombinant omega interferon therapy in canine atopic dermatitis: a pilot study
Authors: Carlotti D. N. , Madiot G. , Ducret J. , Jasmin P. , Gardey L. 
Published: August 2004

It has been postulated that atopic dogs show a predominant Th2‐type response associated with overexpression of IL‐4 and increased secretion of IgE. In a pilot, open uncontrolled study, 20 dogs with confirmed atopic dermatitis were given Virbagen Omega®, a feline recombinant omega interferon (rfeIFN‐ω), to ascertain whether this type‐1 cytokine could modulate the Th2‐dominated cytokine response. Dogs received rfeIFN‐ω as a monotherapy at the dosage of 1 MU/kg subcutaneously, three times per week for 3 weeks. Flea control and the use of nonmedicated shampoo were maintained during the study. Lesional and pruritus indices for canine atopic dermatitis (LICAD and PICAD, respectively) were assessed before, after treatment (day 21) and 3 weeks post‐treatment (day 42) using a scoring system based on the extent and severity of skin lesions (erythema, excoriation and lichenification scored on 12 anatomic areas) and pruritus (scored on six anatomic areas). Two dogs were excluded, one for an erythematous reaction and another for relapsing folliculitis. After rfeIFNω therapy (day 21), both LICAD and PICAD were significantly improved (P < 0.01). By day 42, mean LICAD and PICAD had decreased by 60 and 51%, respectively, compared to baseline values. Adverse reactions to rfeIFN‐ω were rare and inconsistent. This study suggests that rfeIFN‐ω may be useful in the control of canine atopic dermatitis. However, further work is needed to confirm efficacy and define the optimal dosage through a long‐term randomized controlled trial. Funding: Virbac SA.
FC‐38  Efficacy of conjugated linoleic acid and black currant seed oil in the treatment of canine atopic dermatitis: a double‐blinded, randomized, placebo‐controlled study
Authors: Noli C. , Banni S. 
Published: August 2004

Conjugated linoleic acid (CLA) has shown inhibitory effects on histamine release, eicosanoid production and pruritus in laboratory rodents. Its use in canine atopic dermatitis (AD) has not been reported. The aim of this study was to assess the efficacy of CLA, black currant seed oil (BSO) or a combination of both compared to placebo in dogs with AD. A further aim was to evaluate changes in fatty acid metabolism induced by these treatments. Twenty‐four dogs with AD (no response to food trials and negative intradermal tests for fleas) were randomly allocated to four groups. Each group was treated orally daily for 2 months in a blinded manner with 1 mL/10 kg CLA, 1 mL/10 kg BSO, 1 mL/10 kg CLA + 1 mL/10 kg BSO, or 1 mL/10 kg sugar syrup (placebo). Serum was obtained for polyunsaturated fatty acid analysis on days 0, 30 and 60. Owners assessed pruritus with a visual analogue scale, and veterinarians evaluated lesions with the Canine Atopic Dermatitis Extension and Severity Index on days 0, 30 and 60. The best clinical results occurred with BSO alone and were particularly evident in half of the animals treated. However, global improvement was not significant with any of the treatments. Serum levels of di‐homo‐gamma‐linolenic acid markedly increased in BSO‐treated dogs, which may be related to the clinical improvement. Conjugated linoleic acid at the dosage used in this study does not seem to be helpful in canine AD, whereas BSO may be helpful in a selected population of dogs with AD. Funding: NBF Lanes.
FC‐39  Successful management of canine atopic dermatitis using a plant extract: a randomized, double‐blinded, placebo‐controlled trial
Authors: Nuttall T. J. , Ferguson E. A. , Littlewood J. D. , Grover R. 
Published: August 2004

This study evaluated the efficacy of PhytopicaTM, a proprietary blend of standardised plant extracts, in canine atopic dermatitis (AD). One hundred twenty dogs with perennial AD were recruited on the basis of history and clinical signs, and a positive intradermal allergen test or rFcεRIα serology to perennial allergens. Other pruritic dermatoses were eliminated by antimicrobial treatment, skin scrapings, Sarcoptes serology, flea control and a 6‐week food trial. Exclusion criteria included antimicrobial therapy within 7 days, antihistamines within 14 days, oral/topical glucocorticoids or cyclosporin within 28 days, and parenteral glucocorticoids, essential fatty acids or immunotherapy within 56 days of entry into the study. Dogs [minimum Canine Atopic Dermatitis Extent and Severity Index (CADESI) = 25] were randomly allocated to receive placebo, 100, 200 or 400 mg/kg PhytopicaTM daily for 12 weeks. Their CADESI was assessed every 4 weeks. A modified intention‐to‐treat population was analysed. The mean reductions in CADESI scores at the end of treatment compared to baseline were 4.4% (100 mg/kg; n = 30), 23.4% (200 mg/kg; n = 29, P < 0.01), 8.5% (400 mg/kg; n = 29) and 3.9% (placebo; n = 29). For more severely affected dogs (minimum CADESI ≥ 50 at baseline), there was significant reduction in mean CADESI score (29.3%, P = 0.038) only in the 200 mg/kg treatment group (n = 14). In conclusion, this study demonstrates that PhytopicaTM is an effective nonsteroidal treatment for canine AD. Funding: Phytopharm plc.
FC‐40  Efficacy of combined topical therapy with antiallergic shampoo and lotion for the control of signs associated with atopic dermatitis in dogs
Authors: Reme C. A. , Mondon A. , Calmon J. P. , Poisson L. , Jasmin P. , Carlotti D. N. 
Published: August 2004

This study evaluated the benefit of Allermyl® shampoo and lotion in alleviating signs of atopic dermatitis in dogs. After treatment of previous infections, 35 dogs diagnosed with atopic dermatitis according to Prelaud's criteria were included in the study. Three of the dogs improved on prior elimination diet trials, but signs were not completely controlled. The dogs first underwent a 3‐week antiparasitic therapeutic trial period (permethrin spray once every 7 days and acaricidal medication). Dogs whose lesions and pruritus did not improve entered a 3‐week topical antiallergic treatment period (Allermyl® shampoo and lotion alternately every 3 days). No other treatment was allowed. Erythema, excoriation and lichenification were graded at 12 different sites spanning the whole body surface according to an extent‐severity scale (0–10) to calculate an aggregate lesional index score (LICAD: 0–360). Pruritus was similarly evaluated on six body areas according to a frequency‐intensity scale (0–10) to calculate an aggregate pruritus index score (PICAD: 0–60). Twenty‐nine dogs completed the trial. While LICAD and PICAD did not decrease significantly over the control antiparasitic‐treatment period, both indices decreased significantly after 14 days of Allermyl® therapy (repeated measures ANOVA, P < 0.05). Median (95% confidence interval) lesional and pruritus index reduction were 55% (36–77%) and 58% (42–77%), respectively, within 3 weeks of antiallergic topical treatment. Almost half the dogs (48.3%) were markedly improved (>50% score reduction) with Allermyl®. Obvious clinical benefit (≥30% score reduction) was documented in 75.9% of dogs. Funding: Virbac SA.