Internal papillomatous disease (IPD) is characterized by mucosal papillomas occurring primarily in the oral cavity and cloaca of Neotropical parrots. These lesions can cause considerable morbidity, and in some cases result in mortality. Efforts to demonstrate papillomavirus DNA or proteins in the lesions have been largely unsuccessful. However, increasing evidence suggests that mucosal papillomas may contain psittacid herpesviruses (PsHVs). In this study, PsHV 1 genotype 1, 2, and 3 DNA was found in 100% of mucosal papillomas from 30 Neotropical parrots by PCR using PsHV specific primers. However, Psittacus erithacus papillomavirus and finch papillomavirus DNA were not detected. Additionally, a novel PsHV sequence related to, but phylogenetically distinct from PsHV 1, was identified in 4 African grey parrots (Psittacus erithacus), two of which exhibited papillomas. These findings suggest that mucosal papillomas may develop in parrots latently infected with PsHV. Tumors of the bile and pancreatic ducts have also been observed in parrots with IPD. Other mucosal tumors including carcinomas of the proventriculus and ventriculus may be coincident with bile duct tumors, but cloacal carcinomas usually develop as solitary lesions. To test whether PsHV was associated with these tumors, the fresh tissues from 11 parrots and the formalin-fixed paraffin-embedded (FFPE) tissues of 5 parrots exhibiting mucosal tumors were examined by PCR. All tumors were found to contain PsHV 1 genotype 3 DNA except one bird with a cloacal carcinoma that contained genotype 4. Histologically normal tissues available from six parrots did not contain PsHV DNA. Experiments were performed using the FFPE tissues of 5 parrots with IPD related tumors known to contain PsHV by PCR, to show that the virus was in significantly higher concentration in the neoplastic tissue compared to adjacent histologically normal tissue. Neoplastic and adjacent unaffected cells were dissected from the tissues using laser capture microdissection and the DNA was examined by PCR. In situ hybridization using PsHV specific probes and direct in situ PCR were also performed on the tissues. A strong association was shown between infection by PsHV 1 genotype 3 and birds manifesting IPD related tumors and other neoplasms of the digestive tract.

Darbo uždaviniai buvo: Nustatyti gerklų audinio MMP-2 ir MMP-9 raiškos pokyčius kai yra nepiktybiniai gerklų dariniai (balso klosčių polipai), ikivėžinė gerklų patologija (atsinaujinanti gerklų papilomatozė ir lėtinis hiperplazinis laringitas su keratoze) bei gerklų plokščių ląstelių karcinoma ir įvertinti šių pokyčių diagnozinę vertę. Įvertinti gerklų audinio MMP-2 ir MMP-9 raiškos pokyčių sąsajas su klinikiniais ir morfologiniais nepiktybinių gerklų darinių (balso klosčių polipai), ikivėžinės gerklų patologijos (atsinaujinanti gerklų papilomatozė ir lėtinis hiperplazinis laringitas su keratoze) bei gerklų plokščių ląstelių karcinomos požymiais. Palyginti matrikso metaloproteinazes koduojančių genų MMP-2 (-735 C/T), MMP-2 (-1306 C/T), MMP-9 (-1562 C/T) ir MMP-3 (-1171 5A/6A) polimorfizmo dažnį, kai yra organinės nepiktybinės gerklų ligos bei gerklų plokščių ląstelių karcinoma. Įvertinti matrikso metaloproteinazes koduojančių genų MMP-2 (-735 C/T), MMP-2 (-1306 C/T), MMP-9 (-1562 C/T) ir MMP-3 (-1171 5A/6A) polimorfizmo sąsajas su gerklų plokščių ląstelių karcinoma bei jos klinikiniais ir morfologiniais požymiais. / The goals of the study were as follows: 1. To determine the changes in the expression of the laryngeal tissues MMP-2 and MMP-9 in patients with benign laryngeal tumors (vocal fold polyps), precancerous laryngeal lesions (recurrent respiratory papillomatosis and laryngeal keratosis) and laryngeal squamous cell carcinoma, and assess the diagnostic value of the aforementioned changes. 2. To assess the correlations between the changes in the expression of the laryngeal tissues MMP-2 and MMP-9 and the clinical and morphologic indications of benign laryngeal tumors (vocal fold polyps), precancerous laryngeal lesions (recurrent respiratory papillomatosis and laryngeal keratosis) and laryngeal squamous cell carcinoma. 3. To compare the rate of MMP-2 (-735 C/T), MMP-2 (-1306 C/T), MMP-9 (-1562 C/T) and MMP-3 (-1171 5A/6A) gene polymorphism in cases of non-malignant laryngeal lesions and laryngeal squamous cell carcinoma. 4. To assess the association between MMP-2 (-735 C/T), MMP-2 (-1306 C/T), MMP-9 (-1562 C/T) and MMP-3 (-1171 5A/6A) gene polymorphism, the laryngeal squamous cell carcinoma as well as its clinical and morphologic indications.

La papillomatose respiratoire récurrente (PRR) juvénile est causée par les génotypes 6 et 11 du virus du papillome humain (VPH). Cette maladie est caractérisée par des verrues récurrentes généralement au larynx. La forme sévère peut avoir un impact dévastateur sur la santé et la qualité de vie de l’enfant atteint et de sa famille en raison des conséquences des multiples chirurgies nécessaires et du risque d'obstruction des voies respiratoires. Objectif: Examiner les facteurs de risque associés aux manifestations sévères de la PRR. Méthode: Étude rétrospective des 31 cas diagnostiqués entre janvier 1995 et décembre 2008. Les données démographiques, cliniques, génétiques et virologiques ont été évaluées. Des régressions logistiques furent effectuées afin d'évaluer le rôle des variables indépendantes sur la sévérité de la maladie. Résultats: Nos données suggèrent que les facteurs de risque de sévérité de la PRR seraient associés au genre féminin (Rapport de cotes (RC)=2.60, intervalles de confiance (IC) 95% : 0.44-15.44), au fait d’être premier-né (RC=3.51, IC 95% : 0.17-72.32), à un statut économique faible (RC=5.31, IC 95% : 0.17-164.19), à un jeune âge (RC=0.83, IC 95% : 0.68-1.01), à une charge virale élevée (RC=3.81, IC 95% : 0.23-63.16) et aux condylomes chez la mère pendant la grossesse (RC=12.05, IC 95% : 0.97-149.85). Conclusion: La sévérité de la PRR serait le résultat d'une combinaison de déterminants qui favoriseraient la croissance cellulaire particulièrement chez les jeunes enfants. Des mesures préventives et thérapeutiques visant à restreindre la contamination et la réplication du virus pourraient réduire le fardeau de la maladie. / Juvenile Recurrent Respiratory Papillomatosis is caused by Human Papillomavirus genotypes 6 or 11. It is characterised by recurring warts most commonly in the larynx. The severe form of the disease has a devastating impact on health and life quality of the child and its family because of the consequences of multiple surgical procedures and the constant risk of airway obstruction. Objectives: To study the risk factors associated with severe forms of RRP. Method: We conducted a retrospective case series of the 31 patients diagnosed between January 1995 and December 2008. We analyzed demographic and clinical variables, as well as viral and host factors. Logistic regressions were performed to evaluate the impact of each of independent variables on the disease severity. Results: Our data suggest that risk factors for severe forms of RRP in children could be associated with female gender (OR=2.60, 95% CI: 0.44-15.44), being first-born (OR=3.51 95% CI: 0.17-72.32), lower socio-economic status (OR=5.31, 95% CI: 0.17-164.19), younger age (OR=0.83, 95% CI: 0.68-1.01), high viral load (OR=3.81, 95% CI: 0.230-63.16) and history of condylomas of the mother during pregnancy, (OR=12.05, 95% CI: 0.97-149.85). Conclusion: The severity of the RRP would be the result of a combination of factors which would favor the cellular growth particularly in the young children. Therapeutics and preventive measures to restrict the contamination and the replication of the virus could reduce considerably the burden of this disease.

La papillomatose respiratoire récurrente (PRR) juvénile est causée par les génotypes 6 et 11 du virus du papillome humain (VPH). Cette maladie est caractérisée par des verrues récurrentes généralement au larynx. La forme sévère peut avoir un impact dévastateur sur la santé et la qualité de vie de l’enfant atteint et de sa famille en raison des conséquences des multiples chirurgies nécessaires et du risque d'obstruction des voies respiratoires. Objectif: Examiner les facteurs de risque associés aux manifestations sévères de la PRR. Méthode: Étude rétrospective des 31 cas diagnostiqués entre janvier 1995 et décembre 2008. Les données démographiques, cliniques, génétiques et virologiques ont été évaluées. Des régressions logistiques furent effectuées afin d'évaluer le rôle des variables indépendantes sur la sévérité de la maladie. Résultats: Nos données suggèrent que les facteurs de risque de sévérité de la PRR seraient associés au genre féminin (Rapport de cotes (RC)=2.60, intervalles de confiance (IC) 95% : 0.44-15.44), au fait d’être premier-né (RC=3.51, IC 95% : 0.17-72.32), à un statut économique faible (RC=5.31, IC 95% : 0.17-164.19), à un jeune âge (RC=0.83, IC 95% : 0.68-1.01), à une charge virale élevée (RC=3.81, IC 95% : 0.23-63.16) et aux condylomes chez la mère pendant la grossesse (RC=12.05, IC 95% : 0.97-149.85). Conclusion: La sévérité de la PRR serait le résultat d'une combinaison de déterminants qui favoriseraient la croissance cellulaire particulièrement chez les jeunes enfants. Des mesures préventives et thérapeutiques visant à restreindre la contamination et la réplication du virus pourraient réduire le fardeau de la maladie. / Juvenile Recurrent Respiratory Papillomatosis is caused by Human Papillomavirus genotypes 6 or 11. It is characterised by recurring warts most commonly in the larynx. The severe form of the disease has a devastating impact on health and life quality of the child and its family because of the consequences of multiple surgical procedures and the constant risk of airway obstruction. Objectives: To study the risk factors associated with severe forms of RRP. Method: We conducted a retrospective case series of the 31 patients diagnosed between January 1995 and December 2008. We analyzed demographic and clinical variables, as well as viral and host factors. Logistic regressions were performed to evaluate the impact of each of independent variables on the disease severity. Results: Our data suggest that risk factors for severe forms of RRP in children could be associated with female gender (OR=2.60, 95% CI: 0.44-15.44), being first-born (OR=3.51 95% CI: 0.17-72.32), lower socio-economic status (OR=5.31, 95% CI: 0.17-164.19), younger age (OR=0.83, 95% CI: 0.68-1.01), high viral load (OR=3.81, 95% CI: 0.230-63.16) and history of condylomas of the mother during pregnancy, (OR=12.05, 95% CI: 0.97-149.85). Conclusion: The severity of the RRP would be the result of a combination of factors which would favor the cellular growth particularly in the young children. Therapeutics and preventive measures to restrict the contamination and the replication of the virus could reduce considerably the burden of this disease.