Toll-like receptor 9 polymorphisms in brazilian patients with systemic lupus erythematosus: a pilot study / Polimorfismos do receptor toll-like 9 em pacientes brasileiros com lúpus eritematoso sistêmico: um estudo piloto

Toll-like receptor 9 (TLR9) is an important component of the innate immune system and have been associated with several autoimmune diseases, such as Systemic Lupus Erythematosus (SLE). The aim of this study was to investigate polymorphisms in TLR9 gene in a Brazilian SLE patients group and their association with clinical manifestation, particularly Jaccoud’s arthropathy (JA). We analyzed DNA samples from 204 SLE patients, having a subgroup of them presenting JA (n=24). A control group (n=133) from the same city was also included. TLR9 single nucleotide polymorphisms (SNPs) (−1237 C>T and +2848 G>A) were identified by sequencing analysis. The TLR9 gene genotype frequency was similar both in SLE patients and the control group. In the whole SLE population, an association between the homozygosis of allele C at position −1237 with psychosis and anemia (p < 0.01) was found. Likewise, the homozygosis of allele G at position +2848 was associated with a discoid rash (p < 0.05). There was no association between JA and TLR9 polymorphisms. These data show that TLR9 polymorphisms do not seem to be a predisposing factor for SLE in the Brazilian population, and that SNPs are not associated with JA.

O receptor Toll-like 9 (TLR9) é um componente importante do sistema imunológico inato e tem sido associado a várias doenças autoimunes, como o Lúpus Eritematoso Sistêmico (LES). O objetivo deste estudo foi investigar polimorfismos no gene TLR9 em um grupo de pacientes brasileiros com LES e sua associação com a manifestação clínica, particularmente a artropatia de Jaccoud (JA). Foram analisadas amostras de DNA de 204 pacientes com LES, e um subgrupo com JA (n=24). Um grupo de controle (n=133) da mesma cidade também foi incluído. Os polimorfismos de nucleotídeos únicos TLR9 (SNPs) (−1237 C>T e +2848 G>A) foram identificados pela análise de sequenciamento. A frequência do genótipo genético TLR9 foi semelhante tanto em pacientes com LES quanto no grupo controle. Em toda a população de LES, foi encontrada associação entre a homozigose do alelo C na posição −1237 com psicose e anemia (p < 0,01). Da mesma forma, a homozigose do alelo G na posição +2848 foi associada a uma erupção cutânea discoide (p < 0,05). Não houve associação entre polimorfismos JA e TLR9. Esses dados mostram que os polimorfismos TLR9 não parecem ser um fator predisponível para o LES na população brasileira, e que os SNPs não estão associados ao JA.

Toll-like receptor 9 polymorphisms in brazilian patients with systemic lupus erythematosus: a pilot study / Polimorfismos do receptor toll-like 9 em pacientes brasileiros com lúpus eritematoso sistêmico: um estudo piloto

Toll-like receptor 9 (TLR9) is an important component of the innate immune system and have been associated with several autoimmune diseases, such as Systemic Lupus Erythematosus (SLE). The aim of this study was to investigate polymorphisms in TLR9 gene in a Brazilian SLE patients group and their association with clinical manifestation, particularly Jaccouds arthropathy (JA). We analyzed DNA samples from 204 SLE patients, having a subgroup of them presenting JA (n=24). A control group (n=133) from the same city was also included. TLR9 single nucleotide polymorphisms (SNPs) (−1237 C>T and +2848 G>A) were identified by sequencing analysis. The TLR9 gene genotype frequency was similar both in SLE patients and the control group. In the whole SLE population, an association between the homozygosis of allele C at position −1237 with psychosis and anemia (p < 0.01) was found. Likewise, the homozygosis of allele G at position +2848 was associated with a discoid rash (p < 0.05). There was no association between JA and TLR9 polymorphisms. These data show that TLR9 polymorphisms do not seem to be a predisposing factor for SLE in the Brazilian population, and that SNPs are not associated with JA.(AU)

O receptor Toll-like 9 (TLR9) é um componente importante do sistema imunológico inato e tem sido associado a várias doenças autoimunes, como o Lúpus Eritematoso Sistêmico (LES). O objetivo deste estudo foi investigar polimorfismos no gene TLR9 em um grupo de pacientes brasileiros com LES e sua associação com a manifestação clínica, particularmente a artropatia de Jaccoud (JA). Foram analisadas amostras de DNA de 204 pacientes com LES, e um subgrupo com JA (n=24). Um grupo de controle (n=133) da mesma cidade também foi incluído. Os polimorfismos de nucleotídeos únicos TLR9 (SNPs) (−1237 C>T e +2848 G>A) foram identificados pela análise de sequenciamento. A frequência do genótipo genético TLR9 foi semelhante tanto em pacientes com LES quanto no grupo controle. Em toda a população de LES, foi encontrada associação entre a homozigose do alelo C na posição −1237 com psicose e anemia (p < 0,01). Da mesma forma, a homozigose do alelo G na posição +2848 foi associada a uma erupção cutânea discoide (p < 0,05). Não houve associação entre polimorfismos JA e TLR9. Esses dados mostram que os polimorfismos TLR9 não parecem ser um fator predisponível para o LES na população brasileira, e que os SNPs não estão associados ao JA.(AU)

Toll-like receptor 9 polymorphisms in brazilian patients with systemic lupus erythematosus: a pilot study

Abstract Toll-like receptor 9 (TLR9) is an important component of the innate immune system and have been associated with several autoimmune diseases, such as Systemic Lupus Erythematosus (SLE). The aim of this study was to investigate polymorphisms in TLR9 gene in a Brazilian SLE patients group and their association with clinical manifestation, particularly Jaccouds arthropathy (JA). We analyzed DNA samples from 204 SLE patients, having a subgroup of them presenting JA (n=24). A control group (n=133) from the same city was also included. TLR9 single nucleotide polymorphisms (SNPs) (1237 C>T and +2848 G>A) were identified by sequencing analysis. The TLR9 gene genotype frequency was similar both in SLE patients and the control group. In the whole SLE population, an association between the homozygosis of allele C at position 1237 with psychosis and anemia (p 0.01) was found. Likewise, the homozygosis of allele G at position +2848 was associated with a discoid rash (p 0.05). There was no association between JA and TLR9 polymorphisms. These data show that TLR9 polymorphisms do not seem to be a predisposing factor for SLE in the Brazilian population, and that SNPs are not associated with JA.

Resumo O receptor Toll-like 9 (TLR9) é um componente importante do sistema imunológico inato e tem sido associado a várias doenças autoimunes, como o Lúpus Eritematoso Sistêmico (LES). O objetivo deste estudo foi investigar polimorfismos no gene TLR9 em um grupo de pacientes brasileiros com LES e sua associação com a manifestação clínica, particularmente a artropatia de Jaccoud (JA). Foram analisadas amostras de DNA de 204 pacientes com LES, e um subgrupo com JA (n=24). Um grupo de controle (n=133) da mesma cidade também foi incluído. Os polimorfismos de nucleotídeos únicos TLR9 (SNPs) (1237 C>T e +2848 G>A) foram identificados pela análise de sequenciamento. A frequência do genótipo genético TLR9 foi semelhante tanto em pacientes com LES quanto no grupo controle. Em toda a população de LES, foi encontrada associação entre a homozigose do alelo C na posição 1237 com psicose e anemia (p 0,01). Da mesma forma, a homozigose do alelo G na posição +2848 foi associada a uma erupção cutânea discoide (p 0,05). Não houve associação entre polimorfismos JA e TLR9. Esses dados mostram que os polimorfismos TLR9 não parecem ser um fator predisponível para o LES na população brasileira, e que os SNPs não estão associados ao JA.

Toll-like receptor 9 polymorphisms in brazilian patients with systemic lupus erythematosus: a pilot study / Polimorfismos do receptor toll-like 9 em pacientes brasileiros com lúpus eritematoso sistêmico: um estudo piloto

Abstract Toll-like receptor 9 (TLR9) is an important component of the innate immune system and have been associated with several autoimmune diseases, such as Systemic Lupus Erythematosus (SLE). The aim of this study was to investigate polymorphisms in TLR9 gene in a Brazilian SLE patients group and their association with clinical manifestation, particularly Jaccoud's arthropathy (JA). We analyzed DNA samples from 204 SLE patients, having a subgroup of them presenting JA (n=24). A control group (n=133) from the same city was also included. TLR9 single nucleotide polymorphisms (SNPs) (−1237 C>T and +2848 G>A) were identified by sequencing analysis. The TLR9 gene genotype frequency was similar both in SLE patients and the control group. In the whole SLE population, an association between the homozygosis of allele C at position −1237 with psychosis and anemia (p < 0.01) was found. Likewise, the homozygosis of allele G at position +2848 was associated with a discoid rash (p < 0.05). There was no association between JA and TLR9 polymorphisms. These data show that TLR9 polymorphisms do not seem to be a predisposing factor for SLE in the Brazilian population, and that SNPs are not associated with JA.

Resumo O receptor Toll-like 9 (TLR9) é um componente importante do sistema imunológico inato e tem sido associado a várias doenças autoimunes, como o Lúpus Eritematoso Sistêmico (LES). O objetivo deste estudo foi investigar polimorfismos no gene TLR9 em um grupo de pacientes brasileiros com LES e sua associação com a manifestação clínica, particularmente a artropatia de Jaccoud (JA). Foram analisadas amostras de DNA de 204 pacientes com LES, e um subgrupo com JA (n=24). Um grupo de controle (n=133) da mesma cidade também foi incluído. Os polimorfismos de nucleotídeos únicos TLR9 (SNPs) (−1237 C>T e +2848 G>A) foram identificados pela análise de sequenciamento. A frequência do genótipo genético TLR9 foi semelhante tanto em pacientes com LES quanto no grupo controle. Em toda a população de LES, foi encontrada associação entre a homozigose do alelo C na posição −1237 com psicose e anemia (p < 0,01). Da mesma forma, a homozigose do alelo G na posição +2848 foi associada a uma erupção cutânea discoide (p < 0,05). Não houve associação entre polimorfismos JA e TLR9. Esses dados mostram que os polimorfismos TLR9 não parecem ser um fator predisponível para o LES na população brasileira, e que os SNPs não estão associados ao JA.

Discoid lupus erythematosus in dog / Lúpus eritematoso discoide em cão

Lupus erythematosus complex is an immune-mediated dermatological disease, mainly represented by the generalized and discoid forms. The last form described is milder, as it is limited to the appearance of lesions, usually on the face and in mucocutaneous regions. Its pathophysiology is considered multifactorial, however, continuous exposure to ultraviolet radiation seems to be very relevant to trigger and/or worsen clinical manifestations. DeÞ nitive diagnosis is obtained by histopathological analysis, and treatment is mainly based on immunosuppression and protection against ultraviolet radiation. The objective of this study was to report the case and clinical evolution of a mixed breed bitch, diagnosed with discoid lupus erythematosus. The bitch presented moderate desquamation, crusts and depigmentation restricted to the nasal plane. No other clinical or laboratory Þ ndings were evidenced in the screening tests. Upon conÞ rmation by histopathology, the initial therapy was started with oral prednisolone. The owners were also instructed to avoid exposure to sunlight, as well as to use topical protectors against ultraviolet radiation. The patient presented good response to therapy, showing remission of signs. Other sporadic recurrences were observed later, however, they were controlled only with topical corticosteroids, but always reinforcing the other precautions of environmental management. This work also addressed the risks, beneÞ ts and need to institute ongoing care to control discoid lupus erythematosus. Therapeutic success can vary among patients, as the intensity of the disease can be manifested in varying degrees. Therefore, in those individuals in which the condition is mild, it may be advantageous to opt for more conservative therapies in order to avoid side effects.(AU)

O complexo lúpus eritematoso é uma enfermidade dermatológica imunomediada, sendo principalmente representado pela forma generalizada e discoide. A última descrita é mais branda, pois se limita ao aparecimento de lesões geralmente em face e em regiões muco-cutâneas. Sua Þ - siopatogenia é considerada multifatorial, entretanto, a exposição contínua à radiação ultravioleta parece ser muito relevante para desencadear e/ou agravar as manifestações clínicas. O diagnóstico deÞ nitivo é obtido pela análise histopatológica, e o tratamento se baseia principalmente na imunossupressão e proteção contra a radiação ultravioleta. O objetivo deste trabalho foi relatar o caso e evolução clínica de uma cadela sem raça deÞ nida, diagnosticada com lúpus eritematoso discoide. A mesma apresentou moderada descamação, crostas e despigmentação restritas ao plano nasal. Nenhum outro achado clínico ou laboratorial foi evidenciado nos demais exames de triagem. Após conÞ rmado pela histopatologia, a terapia inicial foi instituída a partir da prednisolona por via oral. Os tutores também foram orientados a evitar exposição a luz solar, bem como, fazer a utilização de protetores tópicos contra a radiação ultravioleta. A paciente teve boa resposta à terapia, apresentando remissão dos sinais. Após este episódio, outras recidivas esporádicas foram observadas, entretanto, controladas apenas com corticoides tópicos, mas sempre reforçando os demais cuidados com o manejo ambiental. Este trabalho também abordou os riscos, benefícios e necessidade de instituir o cuidado contínuo para controle do lúpus eritematoso discoide. O sucesso terapêutico pode variar entre os pacientes, uma vez que a intensidade da doença pode ser manifestada em vários graus. Portanto, naqueles indivíduos em que o quadro é brando, pode ser vantajoso optar por terapias mais conservadoras, a Þ m de evitar seus efeitos colaterais.(AU)

Treatment of discoid lupus erythematosus in a dog with human intravenous immunoglobulin

Background: Discoid lupus erythematosus (DLE) is a common canine autoimmune skin disease, in which systemic manifestations are absent. Skin Lesions are usually present on the nasal planum, and characterised by erythema, depigmentation, erosion, ulceration, and crusting. The diagnosis is based on histopathological results, which should demonstrate lymphoplasmacytic lichenoid-interface dermatitis. Human intravenous immunoglobulin (hIVIg) has been used in veterinary medicine to treat cutaneous diseases including erythema multiforme, PF, and severe adverse cutaneous drug reactions. In human medicine, it has been effective to treat DLE. This report firstly describes the clinical response to hIVIg in a dog with DLE resistant to common immunosuppressive drugs. Case: A 5-year-old, intact female Shih Tzu presented with a 1-month history of slowly progressive black crusting on the nasal planum, chin, and claw. Based on the results of a dermatologic examination, superficial pyoderma was diagnosed. The skin lesions did not improve during and after anti-infective treatment. After removing the crusts, a skin biopsy was obtained from the muzzle. Histopathology of lesional skin biopsy specimens revealed lymphoplasmacytic interface dermatitis at the dermoepidermal junction. Microscopic examination also revealed vacuolar changes and pigmentary incontinence of the basal layer as a lichenoid tissue reaction. No mites or fungi were detected on the skin section. The absence of acantholytic cells excluded pemphigus foliaceus, which is also characterised by the lesions of the nasal planum. Based on the distribution of the lesions, histopathology and exclusion of other dermatoses, the dog was diagnosed with DLE. The skin lesions temporarily improved after treatment with prednisolone (2 mg/kg PO q12h). However, after tapering the dose of prednisolone, new black crusts developed on the nasal planum and claw. Although the dog was successively treated with other immunosuppressive drugs, including azathioprine, cyclosporin with dexamethasone, and mycophenolate mofetil, black crusts still remained. Due to the low efficacy of these immunosuppressive drugs, hIVIg was administered at 0.5 g/kg once daily for 4 days, for a total dose of 2 g/kg. During hIVIg administration, the crusted lesions gradually improved. After the hIVIg administration, the dog was treated with prednisolone (1 mg/kg PO q12h). The lesions were almost in complete remission at 21 days after an additional application of prednisolone. The skin lesions did not recur, and the treatment was eventually discontinued after 6 weeks of additional prednisolone application. Discussion: The standard treatment of canine DLE includes glucocorticoids, and second-line immunosuppressive drugs, such as azathioprine and cyclosporine, are usually added in cases resistant to steroids. This case suggests that hIVIg may be beneficial as an adjunctive treatment option for canine DLE, especially when the application of standard immunosuppressive drugs is limited due to adverse effects or low efficacy. There is evidence from several studies that the steroid-sparing effect of hIVIg is significant in human patients. In the current case, the effective dose of prednisolone was reduced to 2 mg/kg/day after hIVIg administration, and prednisolone therapy was finally discontinued completely. The hIVIg appears to lower the daily steroid dose requirement in this dog.

Lúpus eritematoso discoide em cão / Dischoid erythematosus lupus in dog

O Lúpus Eritematoso Discoide (LED) é uma doença autoimune que causa alterações no sistema tegumentar dos animais. Ela acomete cães, gatos, equinos e humanos. Sua etiologia ainda não é bem elucidada, mas acredita-se que fatores genéticos, hormonais e ambientais possam possivelmente desencadeá-la. Essa enfermidade é caracterizada por despigmentação, eritema e descamação nas áreas acometidas, podendo em casos crônicos úlceras serem observadas. Seu diagnóstico baseia-se nos resultados obtidos na anamnese, histórico, exame físico e no exame histopatológico. Por se tratar de uma doença autoimune não há cura definitiva, ocorre apenas o controle, que para tal, pode-se utilizar drogas imunossupressoras e restrição solar. Objetiva-se com presente trabalho relatar um caso de Lúpus Eritematoso Discoide em um cão. No caso relatado o animal apresentava lesões com aspecto eritematoso, descamativo e ulcerado, na região nasal, nos lábios e gengiva, região perianal e na região abdominal caudal. O diagnóstico confirmatório da doença foi realizado por meio do exame histopatológico. Como tratamento foi instituído a corticoterapia a base de prednisolona na dose de 2mg/kg, BID, por 10 dias, posteriormente a dose foi reduzida para 1mg/kg, BID, por mais 10 dias e depois 1mg/kg, em dias alternados, também foi recomendado a restrição solar. O animal respondeu positivamente ao tratamento instituído com melhora significativa dos sinais clínicos apresentados. Contudo, deve-se salientar que a terapêutica instituída com objetivo de controle, pois conforme já foi mencionado não há cura definitiva para a enfermidade.

Discoid Lupus Erythematosus (LED) is an autoimmune disease that causes changes in the integumentary system of animals. It affects dogs, cats, horses and humans. Its etiology is still not well understood, but it is believed that genetic, hormonal and environmental factors may possibly trigger it. This disease is characterized by depigmentation, erythema and desquamation in the affected areas, and in chronic cases ulcers may be observed. Its diagnosis is based on the results obtained in the anamnesis, history, physical examination and histopathological examination. Because it is an autoimmune disease, there is no definitive cure, only control occurs, for which, immunosuppressive drugs and solar restriction can be used. The objective of this work is to report a case of Discoid Lupus Erythematosus in a dog. In the case reported, the animal presented lesions with an erythematous, scaly and ulcerated aspect, in the nasal region, on the lips and gums, perianal region and in the caudal abdominal region. The confirmatory diagnosis of the disease was performed by means of histopathological examination. As treatment, prednisolone-based corticosteroids were administered at a dose of 2mg / kg, BID, for 10 days, then the dose was reduced to 1mg / kg, BID, for another 10 days and then 1mg / kg, every other day, as well. solar restriction was recommended. The animal responded positively to the treatment instituted with a significant improvement in the clinical signs presented. However, it should be noted that the therapy instituted for the purpose of control, as as already mentioned, there is no definitive cure for the disease.

Lúpus eritematoso discoide em cão / Dischoid erythematosus lupus in dog

O Lúpus Eritematoso Discoide (LED) é uma doença autoimune que causa alterações no sistema tegumentar dos animais. Ela acomete cães, gatos, equinos e humanos. Sua etiologia ainda não é bem elucidada, mas acredita-se que fatores genéticos, hormonais e ambientais possam possivelmente desencadeá-la. Essa enfermidade é caracterizada por despigmentação, eritema e descamação nas áreas acometidas, podendo em casos crônicos úlceras serem observadas. Seu diagnóstico baseia-se nos resultados obtidos na anamnese, histórico, exame físico e no exame histopatológico. Por se tratar de uma doença autoimune não há cura definitiva, ocorre apenas o controle, que para tal, pode-se utilizar drogas imunossupressoras e restrição solar. Objetiva-se com presente trabalho relatar um caso de Lúpus Eritematoso Discoide em um cão. No caso relatado o animal apresentava lesões com aspecto eritematoso, descamativo e ulcerado, na região nasal, nos lábios e gengiva, região perianal e na região abdominal caudal. O diagnóstico confirmatório da doença foi realizado por meio do exame histopatológico. Como tratamento foi instituído a corticoterapia a base de prednisolona na dose de 2mg/kg, BID, por 10 dias, posteriormente a dose foi reduzida para 1mg/kg, BID, por mais 10 dias e depois 1mg/kg, em dias alternados, também foi recomendado a restrição solar. O animal respondeu positivamente ao tratamento instituído com melhora significativa dos sinais clínicos apresentados. Contudo, deve-se salientar que a terapêutica instituída com objetivo de controle, pois conforme já foi mencionado não há cura definitiva para a enfermidade.(AU)

Discoid Lupus Erythematosus (LED) is an autoimmune disease that causes changes in the integumentary system of animals. It affects dogs, cats, horses and humans. Its etiology is still not well understood, but it is believed that genetic, hormonal and environmental factors may possibly trigger it. This disease is characterized by depigmentation, erythema and desquamation in the affected areas, and in chronic cases ulcers may be observed. Its diagnosis is based on the results obtained in the anamnesis, history, physical examination and histopathological examination. Because it is an autoimmune disease, there is no definitive cure, only control occurs, for which, immunosuppressive drugs and solar restriction can be used. The objective of this work is to report a case of Discoid Lupus Erythematosus in a dog. In the case reported, the animal presented lesions with an erythematous, scaly and ulcerated aspect, in the nasal region, on the lips and gums, perianal region and in the caudal abdominal region. The confirmatory diagnosis of the disease was performed by means of histopathological examination. As treatment, prednisolone-based corticosteroids were administered at a dose of 2mg / kg, BID, for 10 days, then the dose was reduced to 1mg / kg, BID, for another 10 days and then 1mg / kg, every other day, as well. solar restriction was recommended. The animal responded positively to the treatment instituted with a significant improvement in the clinical signs presented. However, it should be noted that the therapy instituted for the purpose of control, as as already mentioned, there is no definitive cure for the disease.(AU)

Correlation between histopathological findings and results of direct immunofluorescence in the diagnosis of canine localized cutaneous chronic lupus erythematosus

Background: Localized chronic cutaneous lupus erythematosus (CCLE), also known as discoid lupus erythematosus, is one of the most prevalent canine autoimmune skin diseases. Histopathology is considered the gold standard for the diagnosis of CCLE and the accuracy of which can be increased by use of direct immunofluorescence (DIF). This study aimed to investigate the fluorescence pattern revealed by DIF in cases of canine localized CCLE and to establish and compare its effectiveness with that obtained from histopathology.Materials, Methods & Results: Eleven dogs suspicious to localized CCLE, i.e., those animals that presented mucocutaneous lesions as erythema, leucoderma, erosive-ulcerative lesions, and loss of nasal planum architecture and its transition to the haired skin underwent medical physical and laboratory examinations (blood count, platelet count, determination of alanine transaminase, alkaline phosphatase, total protein, serum albumin, urea, creatinine). Only those animals that proved to be normal across both the physical and laboratorial evaluation were included in Group I. Animals belonging to this group were submitted to general anesthesia to biopsy two samples of lesioned skin from nasal planum to histopathologic examination and DIF test. Five dogs with no skin lesions were included in Group II as negative control to the DIF assay. Two samples of no lesioned skin from nasal planum were biopsied to histopathologic and DIF evaluation. The kappa (k) coefficient was used to determine the degree of agreement and reliability of the results of both tests. A P-value < 5% was considered to be statistically significant. In Group I, all animals were normal across both the physical and laboratorial evaluation. A diagnosis of canine CCLE was established in 81.8% (9/11) of the animals based on histopathology analysis.[...]

Correlation between histopathological findings and results of direct immunofluorescence in the diagnosis of canine localized cutaneous chronic lupus erythematosus

Background: Localized chronic cutaneous lupus erythematosus (CCLE), also known as discoid lupus erythematosus, is one of the most prevalent canine autoimmune skin diseases. Histopathology is considered the gold standard for the diagnosis of CCLE and the accuracy of which can be increased by use of direct immunofluorescence (DIF). This study aimed to investigate the fluorescence pattern revealed by DIF in cases of canine localized CCLE and to establish and compare its effectiveness with that obtained from histopathology.Materials, Methods & Results: Eleven dogs suspicious to localized CCLE, i.e., those animals that presented mucocutaneous lesions as erythema, leucoderma, erosive-ulcerative lesions, and loss of nasal planum architecture and its transition to the haired skin underwent medical physical and laboratory examinations (blood count, platelet count, determination of alanine transaminase, alkaline phosphatase, total protein, serum albumin, urea, creatinine). Only those animals that proved to be normal across both the physical and laboratorial evaluation were included in Group I. Animals belonging to this group were submitted to general anesthesia to biopsy two samples of lesioned skin from nasal planum to histopathologic examination and DIF test. Five dogs with no skin lesions were included in Group II as negative control to the DIF assay. Two samples of no lesioned skin from nasal planum were biopsied to histopathologic and DIF evaluation. The kappa (k) coefficient was used to determine the degree of agreement and reliability of the results of both tests. A P-value < 5% was considered to be statistically significant. In Group I, all animals were normal across both the physical and laboratorial evaluation. A diagnosis of canine CCLE was established in 81.8% (9/11) of the animals based on histopathology analysis.[...](AU)

Correlation between Histopathological Findings and Results of Direct Immunofluorescence in the Diagnosis of Canine Localized Cutaneous Chronic Lupus Erythematosus

Background: Localized chronic cutaneous lupus erythematosus (CCLE), also known as discoid lupus erythematosus, is one of the most prevalent canine autoimmune skin diseases. Histopathology is considered the gold standard for the diagnosis of CCLE and the accuracy of which can be increased by use of direct immunofluorescence (DIF). This study aimed to investigate the fluorescence pattern revealed by DIF in cases of canine localized CCLE and to establish and compare its effectiveness with that obtained from histopathology.Materials, Methods & Results: Eleven dogs suspicious to localized CCLE, i.e., those animals that presented mucocutaneous lesions as erythema, leucoderma, erosive-ulcerative lesions, and loss of nasal planum architecture and its transition to the haired skin underwent medical physical and laboratory examinations (blood count, platelet count, determination of alanine transaminase, alkaline phosphatase, total protein, serum albumin, urea, creatinine). Only those animals that proved to be normal across both the physical and laboratorial evaluation were included in Group I. Animals belonging to this group were submitted to general anesthesia to biopsy two samples of lesioned skin from nasal planum to histopathologic examination and DIF test. Five dogs with no skin lesions were included in Group II as negative control to the DIF assay. Two samples of no lesioned sk

HISTOPATHOLOGICAL EVALUATION OF SELECTED PALPEBRAL DISEASES IN DOGS: INVESTIGATION OF THE INTERSECTION BETWEEN VETERINARY DERMATOPATHOLOGY AND OPTHALMOLOGY

Os estudos, investigações e publicações sobre as alterações clínicas e histopatológicas das doenças que afetam as pálpebras de cães são muito escassos. Isso provavelmente se deve ao fato de as pálpebras serem estruturas que pertencem à área de atuação comum a duas especialidades médicas: a dermatologia e a oftalmologia, não se encaixando completamente em nenhuma delas. Tanto os dermatologistas não se sentem confortáveis em tratar de uma área que pertence ao olho; quanto os oftalmologistas, hesitam em abordar uma região tão ligada à dermatologia. Certamente o diagnóstico e consequente tratamento exitoso das afecções palpebrais exige conhecimentos sólidos de cada umas dessas especialidades. Assim sendo, estudos mais aprofundados e específicos das alterações patológicas palpebrais e que incluam conhecimentos e experiências das duas especialidades têm grande relevância no desenvolvimento da clínica veterinária de pequenos animais. Além disso, é importante que aqueles responsáveis pelo diagnóstico histopatológicos das alterações palpebrais, os patologistas, sejam apresentados a essa área de intersecção de especialidades e, uma vez habituados e conhecedores dela, tornem-se aliados dos oftalmologistas e dermatologistas no diagnóstico das afecções palpebrais. A presente tese de doutorado está dividida em dois capítulos. O primeiro capítulo trata-se de um estudo retrospectivo de 290 casos de adenomas e epiteliomas da glândula tarsal em cães. Nele determina-se a frequência destas neoplasias palpebrais em dois laboratórios de Patologia Veterinária que recebem amostras de todo o Brasil. Além disso, discutem-se os aspectos histopatológicos dessas duas neoplasias e propõe-se uma nova forma de diferenciação entre elas. Este estudo deu origem a um artigo publicado no periódico Brazilian Journal of Veterinary Pathology. O segundo capítulo avalia os achados histopatológicos e imuno-histoquímicos das lesões despigmentantes das pálpebras de cães. O estudo sugere que a despigmentação palpebral acompanhada do padrão histopatológico de dermatite de interface é uma manifestação localizada do lúpus eritematoso discoide. Além disso, o demonstra que o lúpus eritematoso discoide localizado pode ser restrito às pálpebras de cães e propõe o termo blefarite lupoide para essa manifestação. O estudo também avaliou a expressão da proteína p53 nas pálpebras despigmentadas com dermatite de interface. A correlação entre a expressão da proteína p53 com a presença de dermatite de interface em pálpebras despigmentadas pode ajudar no entendimento dos mecanismos das dermatopatias que levam à despigmentação, resultando em diagnósticos e prognósticos mais precisos. Esse artigo será publicado em periódico direcionado a clínicos veterinários dermatologistas ou oftalmologistas.

Studies, investigations and publications about clinical and histopathological alterations of diseases affecting the eyelids in dogs are very scarce. This is probably because the eyelids are structures belonging to an area common to two medical fields: dermatology and ophthalmology, and do not totally fit into neither. Dermatologists usually do not feel confident to treat an area belonging to the eye; and ophthalmologists hesitate to approach an area so connected to dermatology. Certainly, the diagnosis and consequent successful treatment of palpebral conditions demand solid knowledge from each of these specialties. Therefore, it is clear the need of deeper and more specific studies of palpebral diseases including knowledge and experience from both specialties. Besides, it is important that the pathologists themselves should be more familiar with this intersection area of specialties and make the connection between ophthalmologists and dermatologists in the diagnosis of eyelid diseases. The present doctorate thesis was divided into two chapters. The first one is about a retrospective study of 290 cases of adenomas and epitheliomas of the tarsal gland in dogs. In this chapter, the frequency of these eyelid neoplasias was determined in two Veterinary Pathology laboratories that receive samples from all over Brazil. Besides, the histopathological aspects of both neoplasias have been discussed and a new form of differentiation between them has been proposed. This study gave origin to an article published in the Brazilian Journal of Veterinary Pathology.The second chapter evaluates the histopathological and immuno-histochemical findings on the depigmenting lesions in eyelids of dogs. The study suggests that depigmented eyelids with interface dermatitis are, in fact, a clinical manifestation of localized discoid lupus erythematosus. Besides, the study demonstrates that localized discoid lupus erythematosus can be restricted to the eyelids of dogs and proposes the term lupoid blepharitis for those cases. The expression of p53 protein in depigmented eyelids with interface dermatitis also was investigated. The results of this study may help to better understanding of the mechanisms behind the disorders that lead to depigmentation, resulting in more precise diagnosis and prognosis. The manuscript will be published in a periodical with the focus directed to veterinary clinicians specialized in dermatology or ophthalmology.

DIAGNÓSTICO DAS DOENÇAS DERMATOLÓGICAS EM PEQUENOS ANIMAIS: PRINCIPAIS DESAFIOS.

As doenças dermatológicas em pequenos animais podem ser consideradas um desafio para tutores, clínicos e patologistas. O objetivo desse estudo foi a pesquisa de condições que interferem no diagnóstico das dermatopatias não neoplásicas em cães e gatos. Esse trabalho foi dividido em três partes. Na primeira e na segunda parte do trabalho foram aplicados dois questionários, o primeiro a médicos veterinários de onze cidades do estado do Paraná e o segundo a tutores de pequenos animais com doença dermatológica em um hospital veterinário. Na terceira parte do trabalho foi realizada a avaliação dos aspectos histopatológicos que possam ser utilizados para a diferenciação das dermatopatias não neoplásicas em cães e gatos, fazendo sua correlação com os achados clínicos. Na primeira etapa do trabalho foi observado que a principal dificuldade encontrada na rotina dermatológica foi comprometimento do tutor do animal (54,95%). Recidivas de até 30% foram relatadas por 40% dos clínicos. Houve predomínio de tutores de classe média nos atendimentos clínicos (56%) e a maioria dos clínicos (84%) já utilizou o exame histopatológico e espera do mesmo a obtenção de diagnóstico (73,44%) e maior rapidez para o resultado (27,91%). Como principal sugestão na obtenção do diagnóstico foi a realização de exames complementares (48,33%). Na segunda etapa do trabalho foi observado que a maioria dos tutores buscou tratamento em até um mês (54%) do início da doença, entretanto 16% aguardaram dois meses e 30%, mais de seis meses para iniciarem um tratamento. Apenas 38% dos tutores relataram melhora do animal após o tratamento. Os tutores que não realizaram o tratamento (12%) justificaram o fato principalmente devido ao tratamento ser muito caro (33,33%) e por acharem que não estava melhorando (33,33%). A maioria dos tutores (60%) relata que não foi solicitada a realização de qualquer tipo de exame complementar antes do início do tratamento. Na terceira etapa da pesquisa, 40 animais foram biopsiados, e destes 95% eram cães e 5% eram gatos. Nos exames realizados em cães, 45% eram dermatopatias alérgicas, 12,5% eram imunomediadas, 7,5% eram endócrinas e 5% eram parasitárias. Outras dermatopatias totalizaram em 10% e os casos inconclusivos chegaram a 15%. Entre os cães alérgicos 38,89% eram sem raça definida. Segundo os tutores a doença alérgica iniciou entre menos de 1 ano a 3 anos de idade (66,67%) do animal e as primeiras lesões que surgiram eram principalmente eritematosas (27,78%), formavam crostas na pele (27,78%) em região ventral (44,44%) e membros (27,78%). Apenas 44,44% dos cães alérgicos chegaram ao diagnóstico definitivo, em 27,78% pacientes foram realizados protocolos diferentes e 27,78% dos cães não retornaram a consulta clínica. Entre os animas que chegaram ao diagnóstico, 50% foram compatíveis com hipersensibilidade alimentar, 37,5% com dermatite alérgica a picada de pulga e 12,5% com dermatite de contato. O principal ponto avaliado na histopatologia dos animais alérgicos foi o padrão inflamatório. As dermatopatias imunomediadas encontradas foi o lúpus eritematoso discóide (lesão em plano nasal e dorsal do focinho) e o pênfigo foliáceo (tronco, membros, cabeça e coxim). No exame histopatológico foi visualizado infiltrado inflamatório linfoplasmocitário em faixa e pústula eosinofília, respectivamente para essas doenças. A dermatopatia endócrina encontrada foi o hipotireoidismo (alopecia e hiperpigmentação generalizada) com predomínio de folículos telogênicos no exame histopatológico. Nos animais com dermatopatia parasitária (lesões em abdome, membros, dígitos, períneo, tórax e cauda) foi observado foliculite linfoplasmocitária estruturas intrafoliculares compatíveis com Demodex sp. no exame histopatológico. As outras dermatopatias encontradas foram a alopecia por diluição de cor, furunculose interdigital, acne canina e piodermite dos calos em um mesmo animal, Acanthosis Nigricans do Dachshund e reação a corpo estranho (granuloma causado por ceratina). As doenças dos felinos foram dermatite atópica (lesões em cabeça e bilateral flanco) e criptococose cutânea (lesões em face e base da orelha). Na histopatologia havia dermatite ulcerativa mastocitária e eosinofílica perivascular a intersticial e dermatite ulcerativa e piogranulomatosa com estruturas leveduriformes intralesionais, respectivamente. Concluímos que a dificuldade encontrada no diagnóstico das dermatopatias é por falta de padronização e/ou conhecimentos do médico veterinário e falha na comunicação entre o patologista, o clínico e o tutor do animal.

Dermatological diseases in small animals can be considered a challenge for tutors, clinicians and pathologists. The objective of this study was to investigate conditions that interfere in the diagnosis of non-neoplastic dermatopathies in dogs and cats. This work was divided into three parts. In the first and second part of the study, two questionnaires were applied, the first to veterinarian physician from 11 cities in the state of Paraná and the second to tutors of small animals with dermatological disease in a veterinary hospital. In the third part of the study, the evaluation of the histopathological aspects that could be used for the differentiation of non-neoplastic dermatopathies in dogs and cats was carried out, correlating with the clinical findings. In the first stage of the study, it was observed that the main difficulty found in the dermatological routine was commitment of the animal guardian (54.95%). Relapses of up to 30% were reported by 40% of clinicians. There was a predominance of middle-class tutors in clinical consultations (56%) and the majority of clinicians (84%) had already used the histopathological examination and expected diagnosis (73.44%) and faster results (27.91%). The main suggestion in the diagnosis was the performance of complementary tests (48.33%). In the second stage of the study, it was observed that most of the tutors sought treatment in up to one month (54%) of the onset of the disease, however, 16% waited two months and 30%, more than six months to start treatment. Only 38% of tutors reported improvement of the animal after treatment. The tutors who did not perform the treatment (12%) justified the fact mainly because the treatment was very expensive (33.33%) and because they thought it was not improving (33.33%). Most tutors (60%) report that no further type of examination was required prior to initiation of treatment. In the third stage of the research, 40 animals were biopsied, and these 95% were dogs and 5% were cats. In the examinations performed in dogs, 45% were allergic dermatopathies, 12.5% were immunomediated, 7.5% were endocrine and 5% were parasitic. Other dermatopathies totaled 10% and the inconclusive cases reached 15%. Among the allergic dogs 38.89% were undefined. According to the tutors the allergic disease started between less than 1 year to 3 years of age (66.67%) of the animal and the first lesions that appeared were mainly erythematous (27.78%), formed skin crusts (27.78% ) in the ventral region (44.44%) and limbs (27.78%). Only 44.44% of the allergic dogs arrived at the definitive diagnosis, 27.78% of the patients had different protocols, and 27.78% of the dogs did not return to the clinic. Among the groups that arrived at the diagnosis, 50% were compatible with food hypersensitivity, 37.5% with allergic dermatitis to flea bite and 12.5% with contact dermatitis. The main point evaluated in the histopathology of allergic animals was the inflammatory pattern. The immune-mediated skin diseases were discoid lupus erythematosus (lesion in the nasal and dorsal plane of the muzzle) and pemphigus foliaceus (trunk, limbs, head and cushion). In the histopathological examination, inflammatory lymphoplasmacytic infiltrate in the band and pustule eosinophilia were visualized, respectively, for these diseases. The endocrine dermatopathy found was hypothyroidism (alopecia and generalized hyperpigmentation) with a predominance of telogenic follicles in histopathological examination. In animals with parasitic dermatopathy (lesions in abdomen, limbs, digits, perineum, thorax and tail) we observed lymphoplasmacytic folliculitis intrafollicular structures compatible with Demodex sp. Histopathological examination. The other dermatopathies found were color-diluted alopecia, interdigital furunculosis, canine acne and callus pyoderma in the same animal, Dachshund's Acanthosis Nigricans and foreign body reaction (granuloma caused by keratin). Feline diseases were atopic dermatitis (lesions on the head and bilateral flank) and cutaneous cryptococcosis (lesions on the face and base of the ear). In the histopathology there was ulcerative mastocytoma and eosinophilic perivascular to interstitial dermatitis and ulcerative and piogranulomatous dermatitis with intralesional yeast structures, respectively. We conclude that the difficulty found in the diagnosis of dermatopathies is due to the lack of standardization and / or knowledge of the veterinarian and the lack of communication between the pathologist, clinician and animal guardian.

Lúpus eritematoso discóide relato de caso / Discoid lupus erythematosus - a case report

O lúpus eritematoso é uma afecção imunomediada de etiologia desconhecida que pode apresentar-sesob duas formas: uma benigna, com lesões cutâneas bem localizadas e denominada Lúpus EritematosoDiscóide (LED) e outra multissistêmica, afetando vários segmentos do organismo, denominada LúpusEritematoso Sistêmico (LES). O presente trabalho tem por objetivo apresentar um relato de caso de umcão da raça boxer, com diagnóstico de lúpus eritematoso discoide (LED). (AU)

Systemic lupus erythematosus is an immune-mediated disease of un known etiology which may presentin two ways: benign skin lesionswell-located, called Discoid Lupus Erythematosus (LED) andmultisystem involvement affecting various segments of the body, called Systemic Lupus Erythematosus(LES). This paper aims to present a case of a dog breed boxer diagnosed with discoid lupus erythematosus (LED). (AU)

Lúpus eritematoso discóide – relato de caso / Discoid lupus erythematosus - a case report

O lúpus eritematoso é uma afecção imunomediada de etiologia desconhecida que pode apresentar-sesob duas formas: uma benigna, com lesões cutâneas bem localizadas e denominada Lúpus EritematosoDiscóide (LED) e outra multissistêmica, afetando vários segmentos do organismo, denominada LúpusEritematoso Sistêmico (LES). O presente trabalho tem por objetivo apresentar um relato de caso de umcão da raça boxer, com diagnóstico de lúpus eritematoso discoide (LED).

Systemic lupus erythematosus is an immune-mediated disease of un known etiology which may presentin two ways: benign skin lesionswell-located, called Discoid Lupus Erythematosus (LED) andmultisystem involvement affecting various segments of the body, called Systemic Lupus Erythematosus(LES). This paper aims to present a case of a dog breed boxer diagnosed with discoid lupus erythematosus (LED).

Correlation between Histopathological Findings and Results of Direct Immunofluorescence in the Diagnosis of Canine Localized Cutaneous Chronic Lupus Erythematosus

Background: Localized chronic cutaneous lupus erythematosus (CCLE), also known as discoid lupus erythematosus, is one of the most prevalent canine autoimmune skin diseases. Histopathology is considered the gold standard for the diagnosis of CCLE and the accuracy of which can be increased by use of direct immunofluorescence (DIF). This study aimed to investigate the fluorescence pattern revealed by DIF in cases of canine localized CCLE and to establish and compare its effectiveness with that obtained from histopathology.Materials, Methods & Results: Eleven dogs suspicious to localized CCLE, i.e., those animals that presented mucocutaneous lesions as erythema, leucoderma, erosive-ulcerative lesions, and loss of nasal planum architecture and its transition to the haired skin underwent medical physical and laboratory examinations (blood count, platelet count, determination of alanine transaminase, alkaline phosphatase, total protein, serum albumin, urea, creatinine). Only those animals that proved to be normal across both the physical and laboratorial evaluation were included in Group I. Animals belonging to this group were submitted to general anesthesia to biopsy two samples of lesioned skin from nasal planum to histopathologic examination and DIF test. Five dogs with no skin lesions were included in Group II as negative control to the DIF assay. Two samples of no lesioned sk

Correlation between Histopathological Findings and Results of Direct Immunofluorescence in the Diagnosis of Canine Localized Cutaneous Chronic Lupus Erythematosus

Background: Localized chronic cutaneous lupus erythematosus (CCLE), also known as discoid lupus erythematosus, is one of the most prevalent canine autoimmune skin diseases. Histopathology is considered the gold standard for the diagnosis of CCLE and the accuracy of which can be increased by use of direct immunofluorescence (DIF). This study aimed to investigate the fluorescence pattern revealed by DIF in cases of canine localized CCLE and to establish and compare its effectiveness with that obtained from histopathology.Materials, Methods & Results: Eleven dogs suspicious to localized CCLE, i.e., those animals that presented mucocutaneous lesions as erythema, leucoderma, erosive-ulcerative lesions, and loss of nasal planum architecture and its transition to the haired skin underwent medical physical and laboratory examinations (blood count, platelet count, determination of alanine transaminase, alkaline phosphatase, total protein, serum albumin, urea, creatinine). Only those animals that proved to be normal across both the physical and laboratorial evaluation were included in Group I. Animals belonging to this group were submitted to general anesthesia to biopsy two samples of lesioned skin from nasal planum to histopathologic examination and DIF test. Five dogs with no skin lesions were included in Group II as negative control to the DIF assay. Two samples of no lesioned sk

Correlation between Histopathological Findings and Results of Direct Immunofluorescence in the Diagnosis of Canine Localized Cutaneous Chronic Lupus Erythematosus

Background: Localized chronic cutaneous lupus erythematosus (CCLE), also known as discoid lupus erythematosus, is one of the most prevalent canine autoimmune skin diseases. Histopathology is considered the gold standard for the diagnosis of CCLE and the accuracy of which can be increased by use of direct immunofluorescence (DIF). This study aimed to investigate the fluorescence pattern revealed by DIF in cases of canine localized CCLE and to establish and compare its effectiveness with that obtained from histopathology.Materials, Methods & Results: Eleven dogs suspicious to localized CCLE, i.e., those animals that presented mucocutaneous lesions as erythema, leucoderma, erosive-ulcerative lesions, and loss of nasal planum architecture and its transition to the haired skin underwent medical physical and laboratory examinations (blood count, platelet count, determination of alanine transaminase, alkaline phosphatase, total protein, serum albumin, urea, creatinine). Only those animals that proved to be normal across both the physical and laboratorial evaluation were included in Group I. Animals belonging to this group were submitted to general anesthesia to biopsy two samples of lesioned skin from nasal planum to histopathologic examination and DIF test. Five dogs with no skin lesions were included in Group II as negative control to the DIF assay. Two samples of no lesioned sk

Correlation between Histopathological Findings and Results of Direct Immunofluorescence in the Diagnosis of Canine Localized Cutaneous Chronic Lupus Erythematosus

Background: Localized chronic cutaneous lupus erythematosus (CCLE), also known as discoid lupus erythematosus, is one of the most prevalent canine autoimmune skin diseases. Histopathology is considered the gold standard for the diagnosis of CCLE and the accuracy of which can be increased by use of direct immunofluorescence (DIF). This study aimed to investigate the fluorescence pattern revealed by DIF in cases of canine localized CCLE and to establish and compare its effectiveness with that obtained from histopathology.Materials, Methods & Results: Eleven dogs suspicious to localized CCLE, i.e., those animals that presented mucocutaneous lesions as erythema, leucoderma, erosive-ulcerative lesions, and loss of nasal planum architecture and its transition to the haired skin underwent medical physical and laboratory examinations (blood count, platelet count, determination of alanine transaminase, alkaline phosphatase, total protein, serum albumin, urea, creatinine). Only those animals that proved to be normal across both the physical and laboratorial evaluation were included in Group I. Animals belonging to this group were submitted to general anesthesia to biopsy two samples of lesioned skin from nasal planum to histopathologic examination and DIF test. Five dogs with no skin lesions were included in Group II as negative control to the DIF assay. Two samples of no lesioned sk

Correlation between Histopathological Findings and Results of Direct Immunofluorescence in the Diagnosis of Canine Localized Cutaneous Chronic Lupus Erythematosus

Background: Localized chronic cutaneous lupus erythematosus (CCLE), also known as discoid lupus erythematosus, is one of the most prevalent canine autoimmune skin diseases. Histopathology is considered the gold standard for the diagnosis of CCLE and the accuracy of which can be increased by use of direct immunofluorescence (DIF). This study aimed to investigate the fluorescence pattern revealed by DIF in cases of canine localized CCLE and to establish and compare its effectiveness with that obtained from histopathology.Materials, Methods & Results: Eleven dogs suspicious to localized CCLE, i.e., those animals that presented mucocutaneous lesions as erythema, leucoderma, erosive-ulcerative lesions, and loss of nasal planum architecture and its transition to the haired skin underwent medical physical and laboratory examinations (blood count, platelet count, determination of alanine transaminase, alkaline phosphatase, total protein, serum albumin, urea, creatinine). Only those animals that proved to be normal across both the physical and laboratorial evaluation were included in Group I. Animals belonging to this group were submitted to general anesthesia to biopsy two samples of lesioned skin from nasal planum to histopathologic examination and DIF test. Five dogs with no skin lesions were included in Group II as negative control to the DIF assay. Two samples of no lesioned sk