Generation and characterization of induced pluripotent stem cells heterozygous for the Portuguese BRCA2 founder mutation.

Women who inherit heterozygous mutations in the BRCA2 gene have an increased risk of developing cancer, mainly breast and ovarian tumors. A particular BRCA2 mutation (c.156_157insAlu) is exclusively found in families of Portuguese ancestry and is present in approximately 30% of all Portuguese families with hereditary breast and ovarian cancers. We report the generation and characterization of the first iPSC line from a female donor harboring the Portuguese BRCA2 founder mutation. Skin fibroblasts were reprogrammed using a non-integrative Sendai virus. These iPSCs are a valuable tool to study the origin of BRCA2-associated cancer in its earliest phases.

Generation and characterization of induced pluripotent stem cells from a family carrying the BRCA1 mutation c.3612delA.

How BRCA1 germline mutations predispose to cancer remains poorly understood. Induced pluripotent stem cells (iPSCs) represent an emerging model to investigate the molecular mechanisms underlying malignant transformation in primary cells from individuals who are carriers of deleterious mutations in the BRCA1 gene. Here we report the generation and characterization of iPSC lines from a female donor harboring a germline c.3612delA mutation in the BRCA1 gene and her daughter who does not carry the mutation. Skin fibroblasts were reprogrammed using non-integrative Sendai virus and characterized for their pluripotent properties. These iPSCs are a valuable cellular model for personalized pre-clinical research in the context of BRCA1 mutant hereditary cancers.

Atypical ocular Chelonoid herpesvirus manifestations in a captive Loggerhead turtle (Caretta caretta).

A captive loggerhead turtle (Caretta caretta) of unknown sex, 3 years of age, presented with bilateral mucoid secretions, severe chemosis, conjunctival hyperemia, and globe retraction. The animal was evaluated ophthalmologically and systemically, and hematological, microbiological, and conjunctival cytological and biopsy samples were collected for complementary diagnosis. The histopathological examination showed amphophilic intranuclear inclusions associated with severe inflammatory infiltrate. The diagnosis of Chelonid alphaherpesvirus 5 (ChAHV 5) was confirmed with end point PCR. Following systemic treatment with L-lysine, acyclovir and vitamin A, the ocular signs resolved. No amphophilic intranuclear inclusions were seen in a follow-up biopsy 5 months later, and there has been no recurrence of clinical ophthalmic signs during a 4-year follow-up. It is suggested that ChAHV 5 be considered as a differential diagnosis in captive marine turtles that present for conjunctival disease other than fibropapillomatosis.

Tear meniscometry test in wild animals / Teste de meniscometria lacrimal em animais silvestres

ABSTRACT: This study aimed to describe strip meniscometry test values for Amazona aestiva (turquoise-fronted amazon), Ara ararauna (blue-and-yellow macaw) and Caiman latirostris (broad-snouted caiman). Twenty-one Amazona aestiva and 11 Ara ararauna, healthy, adults of unknown sex, and 37 healthy adult Caiman latirostris (27 males and 10 females), were used in this investigation. All animals were manually restrained and I-Tear® Test strip was inserted for 5 s at the edge of the lower tear meniscus, without touching the eyelid or ocular surface, immediately followed by measurement (millimeter per 5 seconds). No differences were reported between left and right eyes for Amazona aestiva (P = 0.824), Ara ararauna (P = 0.262) and Caiman latirostris (P = 0.679). Median and confidence interval were 2.0 (2.25-3.56) mm/5s for Amazona aestiva, 3.0 (2.93-4.15) mm/5s for Ara ararauna and 3.0 (2.55-4.90) mm/5 s for Caiman latirostris. These values for strip meniscometry tear test can be used as parameters for each species and may be helpful for diagnosis and treatment of tear-production disorders in birds and reptiles.

RESUMO: O objetivo deste estudo foi descrever os valores do teste de meniscometria lacrimal para Amazona aestiva (papagaio-verdadeiro), Ara ararauna (arara-canindé) e Caiman latirostris (jacaré-de-papo-amarelo). Vinte e um Amazona aestiva, 11 Ara ararauna adultos, de sexo desconhecido, e 37 Caiman latirostris adultos (27 machos e 10 fêmeas), foram utilizados nesta investigação. Todos os animais foram contidos manualmente e uma tira do I-Tear® test foi inserida por 5 segundos na borda do menisco inferior, sem tocar a pálpebra ou a superfície ocular, e imediatamente mensurada (milímetro por 5 segundos). Não foram encontradas diferenças entre os olhos esquerdos e direitos para Amazona aestiva (P = 0,824), Ara ararauna (P = 0,262) e Caiman latirostris (P = 0,679). Mediana e intervalo de confiança foram de 2,0 (2,25-3,56) mm/5 s para Amazona aestiva, 3,0 (2,93-4,15) mm/5 s para Ara ararauna e 3,0 (2,55-4,90) mm/5 s para Caiman latirostris. Estes valores para o teste de meniscometria podem ser utilizados como parâmetros para estas espécies e podem ser úteis para diagnosticar e tratar distúrbios de produção lacrimal em aves e répteis.

Loss of hierarchical imprinting regulation at the Prader-Willi/Angelman syndrome locus in human iPSCs.

The human chr15q11-q13 imprinted cluster is linked to several disorders, including Prader-Willi (PWS) and Angelman (AS) syndromes. Recently, disease modeling approaches based on induced pluripotent stem cells (iPSCs) have been used to study these syndromes. A concern regarding the use of these cells for imprinted disease modeling is the numerous imprinting defects found in many iPSCs. Here, by reprogramming skin fibroblasts from a control and AS individuals, we generated several iPSC lines and addressed the stability of imprinting status across the PWS/AS domain. We focused on three important regulatory DNA elements which are all differentially methylated regions (DMRs), methylated on the maternal allele: the PWS imprinting center (PWS-IC), which is a germline DMR and the somatic NDN and MKRN3 DMRs, hierarchically controlled by PWS-IC. Normal PWS-IC methylation pattern was maintained in most iPSC lines; however, loss of maternal methylation in one out of five control iPSC lines resulted in a monoallelic to biallelic switch for many imprinted genes in this domain. Surprisingly, MKRN3 DMR was found aberrantly hypermethylated in all control and AS iPSCs, regardless of the methylation status of the PWS-IC master regulator. This suggests a loss of hierarchical control of imprinting at PWS/AS region. We confirmed these results in established iPSC lines derived using different reprogramming procedures. Overall, we show that hierarchy of imprinting control in donor cells might not apply to iPSCs, accounting for their spectrum of imprinting alterations. Such differences in imprinting regulation should be taken into consideration for the use of iPSCs in disease modeling.

Sedative Effects of Intranasal Midazolam Administration in Wild Caught Blue-fronted Amazon (Amazona aestiva) and Orange-winged Amazon (Amazona amazonica) Parrots.

Safe and effective sedation protocols are important for chemical restraint of birds in clinical and diagnostic procedures, such as clinical evaluations, radiographic positioning, and blood collection. These protocols may reduce stress and ease the management of wild-caught birds, which are susceptible to injury or death when exposed to stressful situations. We compare the sedative effect of intranasal midazolam in wild-caught blue-fronted (Amazona aestiva) and orange-winged (Amazona amazonica) Amazon parrots. Ten adult parrots of each species (n = 20), of unknown sex, weighing 0.337 ± 0.04 (blue-fronted) and 0.390 ± 0.03 kg (orange-winged), kg were used. Midazolam (2 mg/kg) was administered intranasally and the total volume of the drug was divided equally between the 2 nostrils. Onset time and total sedation time were assessed. Satisfactory sedation for clinical evaluation was induced in all birds. Onset time and total sedation times were similar in both species: 5.36 ± 1.16 and 25.40 ± 5.72 minutes, respectively, for blue-fronted Amazons and 5.09 ± 0.89 and 27.10 ± 3.73 minutes, respectively, for orange-winged Amazons. A total of 15 animals showed absence of vocalization, with moderate muscle relaxation and wing movement upon handling, and 2 animals presented with lateral recumbence, with intense muscle relaxation and no wing movement, requiring no restraint. Three blue-fronted Amazons had no effective sedation. Intranasally administered midazolam at a dose of 2 mg/kg effectively promoted sedative effects with a short latency time and fast recovery in wild-caught parrots.

Corneal sensitivity in five horse breeds / Sensibilidade corneana em cinco raças de equinos

ABSTRACT: This study aimed to determine and compare corneal sensitivity values in different regions of the cornea in five horse breeds. One hundred and forty five healthy horses, adults of both sexes, of the following breeds - Arabian horse (AH; n=20), Mangalarga Marchador (MM; n=50), Pure Blood Lusitano (PBL; n=35), Quarter Horse (QH; n=20), Brazilian Sport Horse (BSH; n=20)-were investigated. Corneal touch threshold (CTT) was measured with a Cochet-Bonnet esthesiometer in five different corneal regions. Measurements of the median central CTT were: 4.50±0.50cm (AH), 3.50±0.56cm (MM), 3.00±0.25cm (PBL), 2.50±0.44cm (QH) and 2.50±0.00cm (BSH). The central region was the most sensitive and the dorsal region the least sensitive corneal region for all breeds. CTT values differed for corneal regions and horse breeds. The CTT values were different among the corneal regions and the horse breeds. Arabian horses presented higher sensitivity values being the most sensitive in all of the corneal regions.

RESUMO: Objetivou-se com o presente estudo, determinar e comparar valores de sensibilidade corneana em diferentes regiões da córnea em cinco raças de equinos. Foram investigados 145 equinos hígidos, adultos de ambos os sexos, das raças: Puro Sangue Árabe (PSA; n=20), Mangalarga Marchador (MM; n=50), Puro Sangue Lusitano (PBL; n=35), Quarto de Milha (QH; n=20) e Brasileiro de Hipismo (BSH; n=20). O limiar táctil da córnea (LTC) foi mensurado com o auxílio do estesiômetro de Cochet-Bonnet, em cinco diferentes regiões da córnea (central, nasal, ventral, temporal e dorsal). Os valores médios das mensurações do LTC central foram: 4,50±0,50cm (PSA), 3,50±0,56cm (MM), 3,00±0,25cm (PSL), 2,50±0,44cm (QM) e 2,50±0,00cm (BH). A região central foi a mais sensível e a região dorsal a menos sensível para todas as raças. Os valores de LTC diferiram para as regiões da córnea e entre as raças dos cavalos, sendo a raça Puro Sangue Árabe o que apresentou valores maiores de sensibilidade, desta forma sendo o mais sensível em todas as regiões corneanas.

Bullous keratopathy in a yellow-headed caracara (Milvago chimachima) treated with a modified third eyelid flap / Ceratopatia bolhosa em caracará (Milvago chimachima) tratado com flap de terceira pálpebra modificado

A Yellow-headed Caracara (Milvago chimachima) was submitted to the Ophthalmology Service of the Federal University of Bahia with a corneal abnormality. During ophthalmic evaluation the right cornea was stained positively with fluorescein; a blurred bullous lesion, with irregular surface, compatible with the diagnosis of bullous keratopathy was found. This is a rare condition in a bird which was treated successfully with a modified third eyelid flap associated with antibiotic and hyaluronic acid eye drops. The adopted therapeutic proved to be simple to implement and viable for repair of the bullous keratopathy in the Yellow-headed caracara.(AU)

Um gavião carrapateiro (Milvago chimachima) foi encaminhado ao Serviço de Oftalmologia Veterinária da Universidade Federal da Bahia com alteração em córnea. Durante a avaliação oftálmica a prova da fluoresceína foi positiva no olho direito e foi observado lesão bolhosa de aspecto irregular, compatível com o diagnóstico de ceratopatia bolhosa. Esta é uma alteração rara em aves e no presente caso foi tratada através de "flap" modificado de terceira pálpebra associado a utilização de colírio antibiótico e ácido hialurônico. A terapêutica adotada foi de fácil implementação e viável no reparo da ceratopatia bolhosa em gavião carrapateiro.(AU)

Orbital multilobular osteochondrosarcoma in a dog / Osteocondrossarcoma multilobular em órbita de cão

Retrobulbar tumors may arise from any orbital tissue. Some are primary and others may extend from adjacent structures or derived from metastases. Multilobular osteochondrosarcomas are regarded as slowly growing and potentially malignant neoplasms in dogs. This tumor is of rare occurrence on retrobulbar region. This paper aims to report a rare case of female dog, Shih Tzu, spayed, five-year-old, that was referred to the Hospital of Veterinary Medicine, Federal University of Bahia, Brazil, with an increased volume in the orbital space, after enucleation for exophthalmia of unknown etiology. This animal was evaluated clinic and the diagnosis image (radiography and computed tomography) demonstrated a large formation of bone predominantly in the region of the right orbit and zygomatic arch, with invasion and destruction of adjacent bones. Due to her poor quality of life, the dog was submitted to euthanasia and after necropsy. On gross examination, an encapsulated nodular mass with an irregular surface, measuring 5.6 x 4.6 x 3.9 cm, was located in the right lateral skull, occupying the orbit with displacement of the zygomatic arch. The tumor was identified by histopathology as a malignant mesenchymal neoplasm, multilobular osteochondrosarcoma. Thus, differential diagnoses for exophthalmos should include cancers, including the multilobular osteocondrossarcoma.

Tumores retrobulbares podem originar-se de qualquer tecido orbital. Alguns são primários e outros podem estender-se a partir de estruturas adjacentes ou derivados de metástases. O osteocondrosarcoma multilobular é um tumor considerado de crescimento lento, potencialmente maligno e sua ocorrência é considerada rara em região orbital. Este trabalho objetiva descrever um caso de uma cadela, Shih Tzu, castrada, com cinco anos, atendida no Hospital de Medicina Veterinária da Universidade Federal da Bahia, Brasil, com apresentação de aumento de volume no espaço orbital, após enucleação devido a exoftalmia de etiologia desconhecida. O animal foi avaliado clinicamente e por exames de imagem (radiografia e tomografia computadorizada), os quais demonstraram uma grande formação óssea predominantemente na região da órbita direita e arco zigomático, com invasão e destruição dos ossos adjacentes. Devido às más condições de sobrevida apresentada pelo animal, foi indicado o procedimento de eutanásia e por fim, submetido à necropsia. No exame macroscópico, uma massa nodular encapsulada de superfície irregular, medindo 5,6 x 4,6 x 3,9 centímetros, foi localizada no crânio lateral direito, ocupando a órbita com deslocamento do arco zigomático. O tumor foi identificado por exame histopatológico como neoplasia mesenquimal maligna, osteocondrossarcoma multilobular. Desta forma, diagnósticos diferenciais para exoftalmia devem incluir neoplasias, dentre elas o osteocondrossarcoma multilobular.

Avaliaçäo do programa de ginástica especial mediante a variaçäo da freqüênci cardíaca em pacientes internados em enfermaria ortopédica / Evaluation of the special gymnastics program by means of the cardiac frequency variation in patients admitted to an orthopedic infirmary

Visando avaliar o programa de ginástica especial utilizado na enfermaria ortopédica, mediante o comportamento da freqüência cardíaca frente ao exercício, foram selecionados 21 pacientes de idade, que durante o período do estudo realizaram 30 sessöes de ginástica especial. Foi realizado o teste ortostático de Karpaman (1980). Foram analisadas as variáveis: faixa etária, sexo e número de sessöes e calculadas as médias, desvio padräo e suas diferenças estatísticas, foram avaliadas pelo teste t. O teste ortotástico de Karpaman (1980) adaptado foi avaliado a partir de sua escala. Os resultados deste estudo revelaram que a realizaçäo do programa de ginástica especial em pacientes ortopédicos com limitaçäo de movimento pode levar a uma reduçäo significativa da freqüencia cardíaca. A relaçäo entre a reduçäo da freqüencia cardíaca e sexo denotou que as mulheres näo apresentaram reduçäo significativa na freqüência cardíaca. A prova ortostástica adaptada apresentou uma alteraçäo ao final do estudo, demonstrando uma leve melhora na classificaçäo de Karpman (1980), dos sujeitos bradicárdios e desfavorável para a normalidade.