To determine the spectrum of ocular surface squamous cell neoplasia (OSSN) in young patients presenting at the St John Eye hospital. The objectives were: (1) documentation of the occurrence of OSSN in young patients at St John Eye hospital (2) to determine the possible role of HIV as a risk factor and the correlation of disease severity with CD4 count.

Afhandling - Copenhagen, 1959. / Summary in English and Danish.

Afhandling - Copenhagen, 1959. / Summary in English and Danish.

Diss. (sammanfattning)--Stockholm : Karolinska institutet, 2009.

Background: Conjunctival pedicle flaps are one of the most frequently employed surgical interventions used to address a variety of sight threatening corneal diseases in veterinary ophthalmic practice. Securing the conjunctiva to the cornea is typically achieved through suturing, which is technically challenging and can result in prolonged surgical times, increased corneal edema, increased scar tissue, foreign body reaction, suture abscess and dehiscence. In human ophthalmology, a number of sutureless techniques to affix ocular tissues are being explored. Specifically, these approaches include synthetic tissue adhesives, bioadhesives, and hydrogel sealants. The proposed advantages of adhesives over suture, include reduced operative times, watertight seals, decreased foreign-body sensation and inflammation, faster healing times and tissue regeneration with original architecture restoration. Objective. To evaluate the maximum tensile force a conjunctival pedicle flap is able to withstand with respect to different fixation methods, i.e., Tisseel® fibrin glue, ethyl cyanoacrylate adhesive, ReSure® hydrogel sealant, or 8-0 VICRYL suture. Animals Studied. Ex-vivo porcine globes Procedures. Following a 500-micron restricted depth lamellar keratectomy, conjunctival pedicle flaps were dissected and secured to corneal defects with either the bioadhesive Tisseel®, or the synthetic adhesives ReSure®, ethyl cyanoacrylate, or 8-0 VICRYL® suture. Harvested corneoconjunctival flap interfaces were clamped to an accelerometer and potentiometer device, and loaded under video surveillance until the point of failure. Peak load at failure was determined for each test and used to compare between sample types. Results. 40 flaps underwent tensile force testing, with 6 being omitted for dehiscence prior to tensile testing. Of the 34 tests included in analysis, 10 conjunctival flaps were secured with suture, 10 with cyanoacrylate, 8 with ReSure® hydrogel sealant, and 6 with Tisseel® fibrin glue. A significant increase in maximum withstood tensile force was recorded between sutured flap fixation when compared with cyanoacrylate glue (p=0.02474), ReSure® hydrogel sealant (p= 0.00000), and Tisseel® fibrin glue (p= 0.00002). Cyanoacrylate fixation was significantly stronger when compared with ReSure® hydrogel sealant and Tisseel fibrin glue (p=0.01194 and 0.01798 respectively). There was no significant difference in adhesion strength between ReSure® hydrogel sealant and Tisseel® fibrin glue (p=0.95675). Conclusions. Conjunctival pedicle flap fixation using 8-0 VICRYL® suture fixation was able to withstand significantly greater maximum tensile force application in comparison with the ReSure®, Tisseel®, or cyanoacrylate adhesives. / Master of Science / Conjunctival pedicle flaps are one of the most frequently employed surgical interventions to address sight threatening corneal disorders in companion animals. Due to its redundant nature and close proximity to the corneal surface, conjunctival tissue is readily available for grafting to the cornea. It is surgically dissected to appropriate size and repositioned over the corneal defect where it effectively aids in healing through direct provision of structural support and indirectly via its rich blood supply. Securing the conjunctiva to cornea is typically achieved through suturing, which is technically challenging and can result in prolonged surgical times, increased corneal edema, increased scar tissue, foreign body reaction, abscess and dehiscence. In human ophthalmology, a number of sutureless techniques to affix ocular tissues are being explored. Specifically, these approaches include synthetic tissue adhesives, bioadhesives, and hydrogel sealants. The proposed advantages of tissue adhesives over suture, include reduced operation times, watertight closures, decreased foreign-body reaction and inflammatory response, faster healing times and increased ability to induce regeneration of the original tissue architecture. The purpose of this study was to evaluate the maximum tensile force a corneoconjunctial pedicle flap is able to withstand with respect to four different fixation methods, 40 ex-vivo porcine globes underwent conjunctival pedicle flap procedures. Each pedicle flap was secured to cornea with either 8-0 Vicryl® suture, Tisseel®, ethyl cyanoacrylate, or ReSure®. After harvesting from the globe, the corneoconjunctival unions were clamped to an accelerometer and potentiometer device, and loaded under video surveillance until the point of failure. The peak load was determined for each test and used to compare between sample types.

A leishmaniose visceral canina é uma zoonose, no Brasil causado pelo protozoário Leishmania chagasi (syn. L. infantum). É endêmica em 88 países, os quais compreendem regiões tropicais e subtropicais do Velho e do Novo Mundo, com incidência estimada em 2 milhões de casos por ano. No ambiente urbano, o cão doméstico é considerado o principal reservatório do parasito. A transmissão da doença ocorre através da picada do vetor, díptero flebotomíneo, da espécie Lutzomyia longipalpis. O diagnóstico pode ser feito através de métodos diretos, como esfregaço preparado com os diferentes órgãos linfoides, Reação em Cadeia da Polimerase (PCR), cultivo in vitro do parasito, ou por métodos sorológicos, como ELISA (Ensaio Imunoenzimático) e RIFI (Reação de Imunofluorescência Indireta). O controle epidemiológico da doença humana envolve o tratamento sistemático dos casos humanos, borrifação de inseticida em região domiciliar e peridomiciliar e eliminação de cães soropositivos, ponto mais controverso do programa de controle. O objetivo do presente trabalho foi avaliar a técnica não invasiva do suabe conjuntival na identificação por PCR de leishmaniose canina. As amostras, de sangue e suabe conjuntival, foram coletadas durante inquérito epidemiológico realizado na cidade de Ilha Solteira - SP. A prevalência de animais positivos em pelo menos um dos três testes (RIFI, PCR de sangue, e PCR de suabe conjuntival) foi de 28,17% (60/213). No teste sorológico RIFI, um total de 13,6% (29/213) dos cães reagiram positivamente. Na PCR do suabe conjuntival, 13,1% (28/213) dos cães foram positivos, e na PCR de sangue total também obtivemos 13,1% (28/213) positivos. Comparando os animais testados pela RIFI e pela PCR de suabe conjuntival, 7,9% (17/213) animais foram positivos em ambos os testes, enquanto 81,2% (173/213) animais foram negativos em ambos os testes (PCR-SC e RIFI). Dos animais testados para PCR-SC, 5,1% (11/213) foram positivos apenas neste teste. Na sorologia, 5,6% (12/213) reagiram positivamente apenas pela RIFI. A sensibilidade e especificidade da PCR-SC foram respectivamente 58,6% e 94,0%, valor preditivo positivo de 61,0%, valor preditivo negativo de 94,0%, e o índice kappa 0,53, o qual demonstra moderada concordância entre os testes. Ao se comparar a RIFI com a PCR-SG, dos animais testados, 3,3% (7/213) foram positivos nos dois testes simultaneamente, 13,6% (29/213) foram positivos apenas na RIFI, logo, 9,9% (21/213) foram positivos apenas na PCR-SG. Foram negativos nos dois testes 76,5% (163/231). A PCR-SG demonstrou sensibilidade de 24,1%, especificidade de 88,5%, valor preditivo positivo de 25,0% e valor preditivo negativo de 88,0% e índice kappa de 0,13, quando comparado à RIFI, indicando uma fraca concordância. Além disso, foram positivos em todos os três testes 2,35% (5/213), e apenas 0,47% (1/213) foi positivo nos testes de PCR-SG e PCR-SC. Os resultados demonstraram que o suabe conjuntival é uma boa alternativa, fácil e pouco invasiva, que pode ser utilizada para auxiliar inquéritos e estudos epidemiológicos da Leishmaniose Visceral Canina. / Canine visceral leishmaniasis is a zoonosis in Brazil caused by the protozoan Leishmania chagasi (syn. L. infantum). It is endemic in 88 countries, which include tropical and subtropical regions of the Old and New World, with an estimated incidence of 2 million cases per year. In the urban environment, the domestic dog is the main reservoir of the parasite. Disease transmission occurs through the bite of the vector, sandfly, Lutzomyia longipalpis species. The diagnosis by direct methods can be made, such as smear prepared with different lymphoid organs, Polymerase Chain Reaction (PCR) in vitro culture of the parasite, or by serological methods such as ELISA (enzyme immunoassay) and IFAT (Indirect Immunofluorescence Test). The epidemiological control of human disease involves the systematic treatment of human cases, insecticide spraying in domiciliary area and elimination of seropositive dogs, most controversial point of the control program. The objective of this study was evaluate the noninvasive technique of conjunctival swab for canine leishmaniasis PCR identification. Were collected Samples of blood and conjunctival swab during an epidemiological survey conducted in the city of Ilha Solteira SP. The prevalence of positive animals in at least one of the three test (IFAT, blood PCR, and swab PCR) was 28.17% (60/213). In IFAT serological test, 13.6% (29/213) of the dogs responded positively. In conjunctival swabs PCR, 13.1% (28/213) dogs were positive, and PCR of whole blood obtained 13.1% (28/213) positive. Comparing the animals tested by IFAT and conjunctival swab PCR, 7.9% (17/213) were positive in both tests, while 81.2% (173/213) animals were negative in both tests. Of the tested animals for swab PCR, 5.1% (11/213) were positive only in this test. In serology, 5.6% (12/213) reacted positively only by IFAT. The sensitivity and specificity of swab PCR were respectively 58.6% and 94.0%, positive predictive value 61.0%, negative predictive value of 94.0%, and kappa 0.53, which demonstrates moderate agreement between tests. Comparing the IFAT with blood PCR, the animals tested, 3.3% (7/213) were positive in both tests simultaneously, 13.6% (29/213) were positive only by IFAT, so, 9, 86% (21/213) were positive only by blood PCR. Were negative in both tests 76.5% (163/213). The blood PCR demonstrated a sensitivity of 24.1%, specificity 88.5%, positive predictive value 25% and negative predictive value of 88% and coefficient of agreement (kappa) of 0.13, compared to IFAT, indicating poor agreement. Furthermore, were positive in all three tests 2.35% (5/213), and only 0.47% (1/213) were positive in the PCR-SC and PCR-SG test. The results showed that conjunctival swab is a good alternative, easier and less invasive that can be used to aid investigations and epidemiological studies of Canine Visceral Leishmaniasis.

Cães, gatos e equinos domésticos podem ser importantes reservatórios de agentes infecciosos potencialmente zoonóticos, podendo-se destacar entre eles as leishmanioses. Por esse motivo, torna-se necessário aprimorar o conhecimento sobre essas enfermidades, estudando mais detalhadamente sua ecoepidemiologia, importante fonte de informações para a tomada de decisões com relação ao controle das mesmas. O presente estudo objetivou verificar a ocorrência de Leishmania spp. em cães, gatos e equinos procedentes de regiões endêmicas e não endêmicas do estado de São Paulo, por meio dos testes diagnósticos Imunofluorescência Indireta (RIFI) e Reação em cadeia pela Polimerase (PCR) e objetivou também comprovar a eficácia da PCR realizada com DNA extraído de amostras de suabe da conjuntiva ocular de gatos e equinos. Foram encontrados cães infectados por Leishmania spp. em Itapecerica da Serra, São Lourenço da Serra e São Paulo, com freqüências de 3,12% (1/32) (PCR de suabe), 10% (1/10) (PCR de suabe) e 1,72% (02/116) (PCR de suabe) / 6,89% (08/116) (PCR de sangue), respectivamente. As amostras analisadas dos gatos provenientes dos municípios de Pirassununga e São Lourenço da Serra, foram positivas para Leishmania spp., com frequências de 28,57% (02/07) (PCR de suabe) / 28,57% (02/07) (RIFI) e 33,33% (1/3) (RIFI) respectivamente. Equinos infectados por Leishmania spp. foram verificados nas cidades de Bragança Paulista e Ilha Solteira, com frequências de 100% (14/14) (PCR de sangue) e 90% (36/40) (PCR de suabe)/ 100% (40/40) (PCR de sangue)/ RIFI 2,5% (01/40) respectivamente. Os resultados demonstram a presença de Leishmania spp infectando cães, gatos e equinos no Estado de São Paulo e que o suabe de conjuntiva ocular foi capaz de detectar gatos e equinos infectados. A coleta de amostras da conjuntiva ocular mostrou-se de fácil execução em felinos, mas nem tanto em equinos, quando comparado à coleta de sangue. / Domestic dogs, cats and horses can be important reservoirs of potential zoonotic agents, as leishmaniasis diseases. Therefore it is necessary improving knowledge on these diseases by studying the ecoepidemiology of leishmaniasis, which is an important source to make decisions on the disease control. The present study aimed to verify the occurrence of Leishmania spp. in dogs, cats and horses from endemic and non-endemic regions from São Paulo State using diagnostics tests as Indirect Immunofluorescence Test (RIFI) and Polymerase Chain Reaction (PCR), and to study the efficacy of the PCR protocol on DNA extracted from ocular conjunctival swab samples from cats, dogs and horses. Leishmania spp. was found in dogs from Itapecerica da Serra, São Lourenço da Serra and São Paulo with frequencies of 3,12% (1/32) (swab PCR), 10%(1/10) (swab PCR) and 1,72% (02/116) (swab PC)/ 6,89% (08/116) (blood PCR) respectively. The cats samples analyzed from Pirassununga and São Lourenço da Serra were positive for Leishmania spp. showing frequencies of 28,57%(02/07) (swab PCR)/ 28,57%(02/07) (RIFI) and 33,33%(1/3) (RIFI) respectively. Leishmania spp. infected horses were detected in Bragança Paulista and Ilha Solteira cities with frequencies of 100% (14/14) (blood PCR) and 90% (36/40) (swab PCR)/ 100% (40/40) (blood PCR)/ RIFI 2,5% (01/40) respectively. Results demonstrated the presence of Leishmania spp infection in dogs, cats and horses from São Paulo and that is possible detect Leishmania spp. DNA in ocular conjunctival swab from infected cats and horses. Colect samples from ocular conjuntiva is more easily made in felines than horses, when compared to blood sample collection.

Hund, bakterieller/ zytologischer Konjunktivalstatus, Konjunktivitis, Klinikaufenthalt

A leishmaniose visceral canina é uma zoonose, no Brasil causado pelo protozoário Leishmania chagasi (syn. L. infantum). É endêmica em 88 países, os quais compreendem regiões tropicais e subtropicais do Velho e do Novo Mundo, com incidência estimada em 2 milhões de casos por ano. No ambiente urbano, o cão doméstico é considerado o principal reservatório do parasito. A transmissão da doença ocorre através da picada do vetor, díptero flebotomíneo, da espécie Lutzomyia longipalpis. O diagnóstico pode ser feito através de métodos diretos, como esfregaço preparado com os diferentes órgãos linfoides, Reação em Cadeia da Polimerase (PCR), cultivo in vitro do parasito, ou por métodos sorológicos, como ELISA (Ensaio Imunoenzimático) e RIFI (Reação de Imunofluorescência Indireta). O controle epidemiológico da doença humana envolve o tratamento sistemático dos casos humanos, borrifação de inseticida em região domiciliar e peridomiciliar e eliminação de cães soropositivos, ponto mais controverso do programa de controle. O objetivo do presente trabalho foi avaliar a técnica não invasiva do suabe conjuntival na identificação por PCR de leishmaniose canina. As amostras, de sangue e suabe conjuntival, foram coletadas durante inquérito epidemiológico realizado na cidade de Ilha Solteira - SP. A prevalência de animais positivos em pelo menos um dos três testes (RIFI, PCR de sangue, e PCR de suabe conjuntival) foi de 28,17% (60/213). No teste sorológico RIFI, um total de 13,6% (29/213) dos cães reagiram positivamente. Na PCR do suabe conjuntival, 13,1% (28/213) dos cães foram positivos, e na PCR de sangue total também obtivemos 13,1% (28/213) positivos. Comparando os animais testados pela RIFI e pela PCR de suabe conjuntival, 7,9% (17/213) animais foram positivos em ambos os testes, enquanto 81,2% (173/213) animais foram negativos em ambos os testes (PCR-SC e RIFI). Dos animais testados para PCR-SC, 5,1% (11/213) foram positivos apenas neste teste. Na sorologia, 5,6% (12/213) reagiram positivamente apenas pela RIFI. A sensibilidade e especificidade da PCR-SC foram respectivamente 58,6% e 94,0%, valor preditivo positivo de 61,0%, valor preditivo negativo de 94,0%, e o índice kappa 0,53, o qual demonstra moderada concordância entre os testes. Ao se comparar a RIFI com a PCR-SG, dos animais testados, 3,3% (7/213) foram positivos nos dois testes simultaneamente, 13,6% (29/213) foram positivos apenas na RIFI, logo, 9,9% (21/213) foram positivos apenas na PCR-SG. Foram negativos nos dois testes 76,5% (163/231). A PCR-SG demonstrou sensibilidade de 24,1%, especificidade de 88,5%, valor preditivo positivo de 25,0% e valor preditivo negativo de 88,0% e índice kappa de 0,13, quando comparado à RIFI, indicando uma fraca concordância. Além disso, foram positivos em todos os três testes 2,35% (5/213), e apenas 0,47% (1/213) foi positivo nos testes de PCR-SG e PCR-SC. Os resultados demonstraram que o suabe conjuntival é uma boa alternativa, fácil e pouco invasiva, que pode ser utilizada para auxiliar inquéritos e estudos epidemiológicos da Leishmaniose Visceral Canina. / Canine visceral leishmaniasis is a zoonosis in Brazil caused by the protozoan Leishmania chagasi (syn. L. infantum). It is endemic in 88 countries, which include tropical and subtropical regions of the Old and New World, with an estimated incidence of 2 million cases per year. In the urban environment, the domestic dog is the main reservoir of the parasite. Disease transmission occurs through the bite of the vector, sandfly, Lutzomyia longipalpis species. The diagnosis by direct methods can be made, such as smear prepared with different lymphoid organs, Polymerase Chain Reaction (PCR) in vitro culture of the parasite, or by serological methods such as ELISA (enzyme immunoassay) and IFAT (Indirect Immunofluorescence Test). The epidemiological control of human disease involves the systematic treatment of human cases, insecticide spraying in domiciliary area and elimination of seropositive dogs, most controversial point of the control program. The objective of this study was evaluate the noninvasive technique of conjunctival swab for canine leishmaniasis PCR identification. Were collected Samples of blood and conjunctival swab during an epidemiological survey conducted in the city of Ilha Solteira SP. The prevalence of positive animals in at least one of the three test (IFAT, blood PCR, and swab PCR) was 28.17% (60/213). In IFAT serological test, 13.6% (29/213) of the dogs responded positively. In conjunctival swabs PCR, 13.1% (28/213) dogs were positive, and PCR of whole blood obtained 13.1% (28/213) positive. Comparing the animals tested by IFAT and conjunctival swab PCR, 7.9% (17/213) were positive in both tests, while 81.2% (173/213) animals were negative in both tests. Of the tested animals for swab PCR, 5.1% (11/213) were positive only in this test. In serology, 5.6% (12/213) reacted positively only by IFAT. The sensitivity and specificity of swab PCR were respectively 58.6% and 94.0%, positive predictive value 61.0%, negative predictive value of 94.0%, and kappa 0.53, which demonstrates moderate agreement between tests. Comparing the IFAT with blood PCR, the animals tested, 3.3% (7/213) were positive in both tests simultaneously, 13.6% (29/213) were positive only by IFAT, so, 9, 86% (21/213) were positive only by blood PCR. Were negative in both tests 76.5% (163/213). The blood PCR demonstrated a sensitivity of 24.1%, specificity 88.5%, positive predictive value 25% and negative predictive value of 88% and coefficient of agreement (kappa) of 0.13, compared to IFAT, indicating poor agreement. Furthermore, were positive in all three tests 2.35% (5/213), and only 0.47% (1/213) were positive in the PCR-SC and PCR-SG test. The results showed that conjunctival swab is a good alternative, easier and less invasive that can be used to aid investigations and epidemiological studies of Canine Visceral Leishmaniasis.

Cães, gatos e equinos domésticos podem ser importantes reservatórios de agentes infecciosos potencialmente zoonóticos, podendo-se destacar entre eles as leishmanioses. Por esse motivo, torna-se necessário aprimorar o conhecimento sobre essas enfermidades, estudando mais detalhadamente sua ecoepidemiologia, importante fonte de informações para a tomada de decisões com relação ao controle das mesmas. O presente estudo objetivou verificar a ocorrência de Leishmania spp. em cães, gatos e equinos procedentes de regiões endêmicas e não endêmicas do estado de São Paulo, por meio dos testes diagnósticos Imunofluorescência Indireta (RIFI) e Reação em cadeia pela Polimerase (PCR) e objetivou também comprovar a eficácia da PCR realizada com DNA extraído de amostras de suabe da conjuntiva ocular de gatos e equinos. Foram encontrados cães infectados por Leishmania spp. em Itapecerica da Serra, São Lourenço da Serra e São Paulo, com freqüências de 3,12% (1/32) (PCR de suabe), 10% (1/10) (PCR de suabe) e 1,72% (02/116) (PCR de suabe) / 6,89% (08/116) (PCR de sangue), respectivamente. As amostras analisadas dos gatos provenientes dos municípios de Pirassununga e São Lourenço da Serra, foram positivas para Leishmania spp., com frequências de 28,57% (02/07) (PCR de suabe) / 28,57% (02/07) (RIFI) e 33,33% (1/3) (RIFI) respectivamente. Equinos infectados por Leishmania spp. foram verificados nas cidades de Bragança Paulista e Ilha Solteira, com frequências de 100% (14/14) (PCR de sangue) e 90% (36/40) (PCR de suabe)/ 100% (40/40) (PCR de sangue)/ RIFI 2,5% (01/40) respectivamente. Os resultados demonstram a presença de Leishmania spp infectando cães, gatos e equinos no Estado de São Paulo e que o suabe de conjuntiva ocular foi capaz de detectar gatos e equinos infectados. A coleta de amostras da conjuntiva ocular mostrou-se de fácil execução em felinos, mas nem tanto em equinos, quando comparado à coleta de sangue. / Domestic dogs, cats and horses can be important reservoirs of potential zoonotic agents, as leishmaniasis diseases. Therefore it is necessary improving knowledge on these diseases by studying the ecoepidemiology of leishmaniasis, which is an important source to make decisions on the disease control. The present study aimed to verify the occurrence of Leishmania spp. in dogs, cats and horses from endemic and non-endemic regions from São Paulo State using diagnostics tests as Indirect Immunofluorescence Test (RIFI) and Polymerase Chain Reaction (PCR), and to study the efficacy of the PCR protocol on DNA extracted from ocular conjunctival swab samples from cats, dogs and horses. Leishmania spp. was found in dogs from Itapecerica da Serra, São Lourenço da Serra and São Paulo with frequencies of 3,12% (1/32) (swab PCR), 10%(1/10) (swab PCR) and 1,72% (02/116) (swab PC)/ 6,89% (08/116) (blood PCR) respectively. The cats samples analyzed from Pirassununga and São Lourenço da Serra were positive for Leishmania spp. showing frequencies of 28,57%(02/07) (swab PCR)/ 28,57%(02/07) (RIFI) and 33,33%(1/3) (RIFI) respectively. Leishmania spp. infected horses were detected in Bragança Paulista and Ilha Solteira cities with frequencies of 100% (14/14) (blood PCR) and 90% (36/40) (swab PCR)/ 100% (40/40) (blood PCR)/ RIFI 2,5% (01/40) respectively. Results demonstrated the presence of Leishmania spp infection in dogs, cats and horses from São Paulo and that is possible detect Leishmania spp. DNA in ocular conjunctival swab from infected cats and horses. Colect samples from ocular conjuntiva is more easily made in felines than horses, when compared to blood sample collection.