Chitosan thread in the healing of the cecal wall of rabbits (Oryctolagus cuniculus) submitted to cecorrhaphy.

The development and evaluation of synthesis materials are crucial to reducing the morbidity and magnitude of post-enterorrhaphy surgical complications. Despite the possibility of production, chitosan thread has not yet been used in enterorrhaphy, and its effects on intestinal healing have not been evaluated. Therefore, this study aimed to evaluate the effects of chitosan thread on the intestinal wall repair of rabbits submitted to cecorrhaphy. For this, 42 rabbits were allocated into two groups with 21 animals. One group was submitted to cecorrhaphy with chitosan suture thread (CG) and the other with poliglecaprone suture thread (PG). The occurrence of postoperative complications, the intensity of edema, cellular response, formation of granulation tissue, as well as the deposition and maturation of collagen fibers, and the intensity of vascular endothelial growth factor (VEGF-α) expression, were evaluated during the intestinal wall repair process. The evaluations occurred on the 5th, 15th, and 25th postoperative (PO) days. The animals did not develop peritonitis, but adherence was observed in six animals from CG and seven from PG, with no difference between groups. The polymorphonuclear infiltrate showed higher intensity and higher amount of type III collagen fibers in CG on the 15th PO day. In contrast, a lower amount of type I collagen fibers was observed in CG samples on the 25th PO day. Therefore, the chitosan thread used for cecorrhaphy in rabbits results in minimal postoperative complications, presents biocompatibility, and bioactively assists the tissue repair process of the cecal wall, inducing minimal tissue reaction, stimulating the deposition of type III collagen fibers in the proliferative phase, with sustained VEGF-α expression, but with reduced deposition of type I fibers, indicating a delay in collagen maturation.

Brief topical and intraluminal use of Carolina rinse solution does not attenuate experimental ischemia and reperfusion injury in rabbit jejunum / [O uso tópico breve e intraluminal da solução de Carolina rinse não atenua a injúria de isquemia e reperfusão experimental no jejuno de coelhos ]

Fifteen New Zealand adult rabbits were randomly allocated into three groups: Sham-operated (group A), Ischemia and Reperfusion (group B) and Carolina Rinse Solution (CRS) (group C). Groups B and C were subjected to one hour of ischemia and two hours of reperfusion. In group C, ten minutes before reperfusion, the bowel lumen was filled with CRS, and the segment immersed in CRS. Necrosis and loss of integrity of the villi were visible in groups B and C. Edema of the submucosa and circular muscle was observed in all groups. Hemorrhage was observed in different layers for groups B and C, but group C showed more severe hemorrhage in different layers during reperfusion. All groups showed polymorphonuclear leukocyte infiltration on the base of the mucosa, submucosa, and longitudinal muscle, in addition to polymorphonuclear leukocytes margination in the mucosal and submucosal vessels. Necrosis of enterocytes, muscles, crypts of Lieberkühn and myenteric plexus was observed in groups B and C during reperfusion. Topical and intraluminal Carolina Rinse Solution did not attenuate the effects of ischemia and reperfusion in the small intestine of rabbits.(AU)

Quinze coelhos da raça Nova Zelândia foram alocados em três grupos: instrumentado (grupo A), isquemia e reperfusão (grupo B) e solução de Carolina rinse (CRS) (grupo C). Os grupos B e C foram submetidos a uma hora de isquemia e a duas horas de reperfusão. No grupo C, 10 minutos antes da reperfusão, o segmento isolado foi imerso e teve seu lúmen preenchido com CRS. Os grupos B e C apresentaram necrose e perda progressiva da integridade das vilosidades. Foi observado edema na submucosa e na camada muscular circular em todos os grupos. Nos grupos B e C, foi observada hemorragia em diferentes camadas, mas, no grupo C, a hemorragia foi mais intensa durante a reperfusão. Todos os grupos apresentaram infiltrado de PMN na base da mucosa, na submucosa e na camada muscular longitudinal e marginação de PMN nos vasos da mucosa e da submucosa. Durante a reperfusão, foi observada necrose dos enterócitos, das camadas musculares, das criptas de Lieberkühn e do plexo mioentérico nos grupos B e C. O uso tópico e intraluminal de CRS não atenuou os efeitos da isquemia e da reperfusão no intestino delgado de coelhos.(AU)

Expression of cell cycle inhibitors in canine prostate with proliferative inflammatory atrophy and carcinoma / Expressão de inibidores do ciclo celular na próstata canina com atrofia inflamatória proliferativa e carcinoma

Gene expression of CDKN1A, CDKN1B, and TP53, and immunostaining of p21, p27 and p53 were evaluated to verify the role of these cell cycle inhibitors in canine prostates with proliferative inflammatory atrophy-PIA and prostatic carcinoma-PC. Seventy samples, 15 normal, 30PIA and 25PC. Regarding number of p27 and p53 labeled cells, difference between normal and PIA and PC was observed, as well as between PIA and PC for p53. Immunostaining intensities of p21, p27 and p53 were different when comparing normal tissues to PIA and PC. Sixteen cDNA of canine prostatic FFPE tissue were subjected to RT-PCR and RT-qPCR, four normal, three PIA, and nine PC. CDKN1A mRNA was detected in four PC by RT-PCR, and it was overexpressed when compared to normal by RT-qPCR, in one PIA and six PC. CDKN1B mRNA was detected in three PC by RT-PCR and it was overexpressed in three PC and decreased in one PC. TP53 mRNA was overexpressed in one PIA and three PC. In conclusion, when overexpressed in canine prostate with premalignant and malignant, p21 and p27 play a role controlling cell proliferation, working as a protective factor in the evolution of PIA to PC, and in the PC development, even in the presence of altered p53.(AU)

A expressão gênica de CDKN1A, CDKN1B e TP53, assim como imunomarcação de p21, p27 e p53 foram realizadas a fim de verificar o papel desses inibidores do ciclo celular na próstata canina com atrofia inflamatória proliferativa (PIA) e carcinoma prostático (PC). Foram obtidas70 amostras de próstata canina, sendo 15 de tecido normal, 30 de PIA e 25 de PC. Quanto ao número de células imunomarcadas foi observada diferença entre amostras normais, com PIA e PC para p27 e p53, assim como entre PIA e PC para p53. Para a intensidade de imunomarcação houve diferença entre os tecidos normais e com PIA e PC para p21, p27 e p53. Foram obtidas dezesseis amostras de cDNA a partir de amostras de próstatas caninas embebidas em parafina para a realização da RT-PCR e RT-qPCR, sendo quatro normais, três com PIA, e nove com o PC. O gene CDKN1A foi detectado em quatro das amostras com PC por RT-PCR, e pela RT-qPCR este estava superexpresso em uma PIA e em seis PC quando da comparação com o tecido prostático normal. O CDKN1B foi detectado em três PC por RT-PCR e pela RT-qPCR estava superexpresso em três PC e reduzido em um PC. O TP53 foi detectado em todas as próstatas caninas com PIA e PC por RT-PCR, sendo também superexpresso em uma glândula com PIA e em três com PC. Concluiu-se que p21 e p27 quando superexpressas na próstata canina com lesões pré-malignas (PIA) e malignas (PC) desempenham ação no controle da proliferação celular, possivelmente atuando como fator de proteção na evolução da PIA para PC, e no desenvolvimento do PC, mesmo na presença de p53 alterada. Assim, o próximo passo é avaliar essas proteínas do ciclo celular em casos de PC canino com metástase.(AU)

Parotid gland adenocarcinoma in a cow: case report / Adenocarcinoma de glândula parótida em vaca: relato de caso

A 9-year-old Girolando dairy cow, weighing 400kg, with a history of increased volume in the right parotid region, which extended to the submandibular region, was assisted. Fine needle aspiration cytology was performed, and the cytological findings were consistent with malignant neoplasm of epithelial origin (carcinoma). Because of the unfavorable prognosis, the animal was euthanized and submitted to an anatomopathological examination. Samples of the increased parotid and affected lymph nodes were collected for histopathological evaluation. The microscopic changes were accentuated features of anaplasia, moderate cell proliferation, atypical mitotic figures, and necrosis. Stroma ranged from delicate to scirrhous, and the tumor boundaries were not distinct. These findings substantiated the preliminary histomorphological diagnosis of undifferentiated carcinoma with metastasis in lymph nodes. Immunohistochemical tests were performed with anti-CK Pan (clone AE1AE3), anti-CK HMW (clone 34ßE12), anti-CK19 (clone RCK108), anti-vimentin (clone V9), anti-S100 (polyclonal), and anti-androgen (polyclonal) antibodies. The immunophenotype favored the diagnosis of salivary gland adenocarcinoma. Despite the rareness in cattle, salivary gland adenocarcinoma should be considered in the differential diagnosis of diseases that occur with increased volume in the head, lymphadenopathy, drooling, dysphagia, and progressive weight loss.(AU)

Foi atendida uma vaca da raça Girolando, de nove anos de idade, de aptidão leiteira, pesando aproximadamente 400kg e com histórico de aumento de volume na região parotídea e submandibular direita. Diante do prognóstico desfavorável, o animal foi submetido à eutanásia e encaminhado para exame anatomopatológico. Fragmentos da glândula parótida e dos linfonodos alterados foram colhidos e encaminhados para exame histopatológico. À avaliação microscópica, observaram-se acentuada anaplasia, moderada proliferação celular, figuras de mitose atípicas e focos de necrose. O estroma variava de delicado a esquirroso e os limites do tumor eram imprecisos. Esses achados fundamentaram o diagnóstico de carcinoma indiferenciado com metástase em linfonodos. No exame imuno-histoquímico, foram utilizados anticorpos primários monoclonais anti-CK Pan (clone AE1AE3), anti-CK alto peso molecular (clone 34ßE12), anti-CK19 (clone RCK108), antivimentina (clone V9), anti-S100 (policlonal) e antirreceptor de andrógenos (policlonal). As células neoplásicas apresentaram imunomarcação para todos os anticorpos testados, resultado que favorece o diagnóstico de adenocarcinoma de glândula salivar. Embora raro em bovinos, o adenocarcinoma de glândula salivar deve ser considerado no diagnóstico diferencial de doenças que cursam com aumento de volume na cabeça, linfadenopatia salivação, disfagia e emagrecimento progressivo.(AU)

Metabolomics as a promising tool for early osteoarthritis diagnosis.

Osteoarthritis (OA) is the main cause of disability worldwide, due to progressive articular cartilage loss and degeneration. According to recent research, OA is more than just a degenerative disease due to some metabolic components associated to its pathogenesis. However, no biomarker has been identified to detect this disease at early stages or to track its development. Metabolomics is an emerging field and has the potential to detect many metabolites in a single spectrum using high resolution nuclear magnetic resonance (NMR) techniques or mass spectrometry (MS). NMR is a reproducible and reliable non-destructive analytical method. On the other hand, MS has a lower detection limit and is more destructive, but it is more sensitive. NMR and MS are useful for biological fluids, such as urine, blood plasma, serum, or synovial fluid, and have been used for metabolic profiling in dogs, mice, sheep, and humans. Thus, many metabolites have been listed as possibly associated to OA pathogenesis. The goal of this review is to provide an overview of the studies in animal models and humans, regarding the use of metabolomics as a tool for early osteoarthritis diagnosis. The concept of osteoarthritis as a metabolic disease and the importance of detecting a biomarker for its early diagnosis are highlighted. Then, some studies in plasma and synovial tissues are shown, and finally the application of metabolomics in the evaluation of synovial fluid is described.

Insertion of the LINE-1 element in the C-MYC gene and immunoreactivity of C-MYC, p53, p21 and p27 proteins in different morphological patterns of the canine TVT / Inserção do elemento LINE - 1 no gene C-MYC e imunorreatividade das proteinas C-MYC, p53, p21 e p27 nos diferentes padrões morfológicos do tumor TVT

The canine transmissible venereal tumor (TVT) affects the external genitalia of dogs by the natural transplant of viable tumor cells. Thus, this research aimed to diagnose and characterize TVT morphological patterns, identify the insertion of the LINE-1 element in C-MYC gene, by means of the polymerase chain reaction (PCR), and evaluate the immunohistochemical expression of C-MYC, p53, p21 and p27 proteins. The relationship between C-MYC and p53 proteins and their interference on the expression of p21 and p27 were also studied. For that, 20 samples of naturally occurring TVT were used, subjected to cytopathological, histopathological and immunohistochemical analysis, and to molecular diagnosis of neoplasia. The increased tissue expression and the correlation among C-MYC, p53, p21 and p27 proteins indicate reduction and/or loss of their functionality in the TVT microenvironment, with consequent apoptotic suppression, maintenance of cell growth and progression of neoplasia.(AU)

O tumor venéreo transmissível canino (TVT) afeta a genitália externa de cães pelo transplante natural de células tumorais viáveis. Assim, esta pesquisa teve como objetivo diagnosticar e caracterizar TVT em padrões morfológicos, identificar a inserção do elemento LINE-1 em gene C-MYC, por meio da reação em cadeia da polimerase (PCR), e avaliar a expressão imuno-histoquímica do C-MYC, p53, p21 e p27. A relação entre C-MYC e as proteínas p53 e a sua interferência na expressão de p21 e p27 foram também estudadas. Para isso, foram utilizadas 20 amostras de ocorrência natural de TVT, submetido a exame citopatológico, histopatológica e imuno-histoquímica e ao diagnóstico molecular de neoplasia. A expressão aumentada do tecido e a correlação entre a C-MYC e as proteínas p53, p21 e p27 indicam redução e/ou perda de funcionalidade na TVT em seu microambiente, com consequente supressão apoptótica, manutenção do crescimento celular e progressão da neoplasia.(AU)

Neuropathogenic Capacity of Lentogenic, Mesogenic, and Velogenic Newcastle Disease Virus Strains in Day-Old Chickens.

Strains of Newcastle disease virus (NDV) have different abilities to elicit neurologic signs. To determine the capacity of different NDV strains to replicate and cause lesions in the brain, independently of their peripheral replication, 1-day-old chickens were inoculated in the subdural space with 7 NDV strains of different virulence (4 velogenic, 2 mesogenic, 1 lentogenic). Velogenic strains induced severe necrotizing and heterophilic ventriculitis and meningitis, as well as edema of the neuroparenchyma, and replicated extensively in the nervous tissue by day 2 postinfection, as demonstrated by immunohistochemistry, when all infected birds died. Clinical signs, microscopic lesions, and viral replication were delayed (days 3 and 4 postinfection) with mesogenic strains. Velogenic and mesogenic NDV strains replicated mainly in neurons, and immunolabeling was first detected in surface-oriented areas (periventricular and submeningeal), possibly as a reflection of the inoculation route. The lentogenic NDV strain did not cause death of infected birds; replication was confined to the epithelium of the ependyma and choroid plexuses; and lesions consisted of lymphoid aggregates limited to the choroid plexuses. Results show that extensive NDV replication in the brain is typical of velogenic and mesogenic, but not lentogenic, NDV strains. In addition, this study suggests that differences in the rate of NDV replication in nervous tissue, not differences in neurotropism, differentiate velogenic from mesogenic NDV strains. This study indicates that intracerebral inoculation might be used as an effective method to study the mechanisms of NDV neuropathogenesis.

Pesquisa e atividades de extensão em fitoterapia desenvolvidas pela Rede FitoCerrado: uso racional de plantas medicinais e fitoterápicos por idosos em Uberlândia-MG / Research and extension activities in herbal medicine developed by Rede FitoCerrado: rational use of medicinal plants by the elderly in Uberlândia-MG

Este estudo objetivou realizar atividades de extensão em fitoterapia, a partir de um levantamento de dados sobre as plantas medicinais, fitoterápicos e medicamentos convencionais utilizados por 292 idosos frequentadores de um programa de Atividades Físicas e Recreativas para a Terceira Idade (AFRID), na cidade de Uberlândia-MG, utilizando como instrumento de investigação, um questionário semiestruturado. Dentre os entrevistados verificamos que 88% utilizavam medicamentos prescritos, principalmente para o controle da hipertensão. O uso de plantas medicinais foi relatado por 76,7% dos idosos, sendo as mais citadas: Cymbopogon citratus, Mentha sp., Rosmarinus officinalis, Plectranthus barbatus, Ocimum gratissimum, e Matricaria chamomilla. Dezesseis (5,5%) idosos utilizavam fitoterápicos, principalmente preparados a partir de extratos de Ginkgo biloba, Aesculus hippocastanum e Passiflora incarnata em associação com Crataegus oxyacantha e Salix alba. O uso concomitante de plantas medicinais e fitoterápicos com medicamentos convencionais foi relatado por 86,2% e 81,3% dos idosos, respectivamente. Após a análise dos dados percebemos a necessidade do desenvolvimento de ações educativas para informar e conscientizar os idosos sobre o uso da fitoterapia. Elaboramos uma caderneta e uma cartilha para promoção da difusão dessas informações e o aprimoramento do uso da fitoterapia entre os idosos e, dessa forma, alcançar os profissionais de saúde sobre os riscos e benefícios dessa terapêutica; contribuindo assim para o uso seguro e racional da fitoterapia.

This study aimed to carry out extension activities in herbal medicine from a survey of data on medicinal plants, herbal and conventional medicines used by 292 elderly people who attended a program of physical activity called Physical and Recreational Activities for the Elderly, in Uberlândia-MG, using a semi-structured questionnaire as means of investigation. Among the respondents 88% used prescription drugs, primarily for control of hypertension. The use of medicinal plants was reported by 76.7% of the elderly, being the most cited ones: Cymbopogon citratus, Mentha sp., Rosmarinus officinalis, Plectranthus barbatus, Ocimum gratissimum. and Matricaria chamomilla. Sixteen respondents (5,5%) used herbal medicines, especially those prepared from extracts of Ginkgo biloba, Aesculus hippocastanum, and Passiflora incarnata L. in association with Crataegus oxyacantha L. and Salix alba. The concomitant use of medicinal plants and herbal medicines with conventional drugs was reported by 86.2% and 81.3% of participants, respectively. After analyzing the data, we detected the need to develop educational activities to inform and educate seniors about the use of herbal medicine, encompassing the development a book and a primer for initial dissemination of this information, improving herbal medicine use among the elderly. This course of action would allow a greater knowledge of health professionals about the risks and benefits of this therapy, thereby contributing to the safe and rational use of herbal medicine.

Kinetics of the inflammatory response in subcutaneous cysticercosis induced in mice by Taenia crassiceps.

The larval stage of Taenia crassiceps has been used to study human cysticercosis as these larvae have antigenic similarity to the cysticerci of Taenia solium. The aim of this study was to evaluate the histopathological and immunological changes that followed the inoculation of T. crassiceps cysticerci into the subcutaneous tissue of C57BL/6 mice. Microscopically, granulomas formed of neutrophils and macrophages developed at the sites of inoculation. The serum concentration of the cytokine interferon (IFN)-γ increased throughout the course of infection, while the serum concentration of interleukin-4 increased during the period of transition from the initial phase (7-30 days postinoculation [dpi]) to the late phase (60-90 dpi) of infection. Destruction of the parasite therefore appears to be associated with an increase in IFN-γ, suggesting that a type 1 immune response is important in the control of the parasite.

Óxido nítrico, GSTP-1 e p53: qual o papel desses biomarcadores nas lesões prostáticas do cão? / Nitric oxide, GSTP-1 and p53: what's the role of these biomarkers in dog´s prostate lesions?

Confeccionou-se um microarranjo de tecido (TMA) com 146 amostras de lesões prostáticas caninas. Este continha 17,2 por cento de hiperplasia prostática benigna (HPB), 32,4 por cento de atrofia inflamatória proliferativa (PIA), 2,6 por cento de prostatite, 8,6 por cento de focos de neoplasia intraepitelial prostática (PIN), 29,1 por cento de carcinomas e 9,3 por cento de próstatas normais. Cortes histológicos sequenciais foram feitos e utilizados para reação de imunoistoquímica com os anticorpos primários anti-p-53, anti-NOS-2 e anti-GSTP. Avaliou-se de cada core o escore de células marcadas para cada anticorpo utilizado. Os resultados foram tabulados por grupo diagnóstico e submetidos ao teste Tuckey. Os carcinomas prostáticos do cão e a PIA apresentaram maior número de amostras (41) com mais de 75 por cento das células positivas para NOS-2, demonstrando a influência do estresse oxidativo no desenvolvimento dessas lesões. As próstatas normais e as afecções desta glândula, HPB, PIA, PIN, prostatite e carcinoma, expressaram a proteína GSTP-1, o que conferiu proteção ao tecido prostático canino a danos oxidativos. A proteína p53 estava presente em todas as amostras estudadas, incluindo o tecido prostático normal, porém as lesões prostáticas apresentaram maior número de amostras com escores mais elevados de marcação (escores três e quatro), presente em 95 por cento dos focos de PIA e carcinoma. Concluiu-se que o aumento de expressão de óxido nítrico nas lesões prostáticas no cão e a expressão de GSTP-1 podem ter protegido o tecido prostático canino e que a expressão de p53 foi positiva e uniforme nas próstatas normais e com lesões hiperplásicas e displásicas.

A tissue microarray (TMA) with 149 samples of canine prostatic lesions contained 17.2 percent benign prostatic hyperplasia (BPH), 32.4 percent proliferative inflammatory atrophy (PIA), 2.6 percent prostatitis, 8.6 percent foci prostatic intraepithelial neoplasia (PIN), 29.1 percent carcinomas and 9.3 percent normal prostates. Sequential histological sections were made and used for immunohistochemistry reaction with primary antibodies anti p-53, anti-NOS-2 and anti-GSTP. The score for each antibody employed was evaluated. The results were tabulated by diagnostic group and subjected to Tuckey test. Prostatic carcinomas and PIA had a higher number of samples (41) with over 75 percent of cells positive for NOS-2, demonstrating the influence of oxidative stress in the development of these lesions. The prostates of normal dogs, as well as the disorders of this gland (BPH, PIA, PIN, prostatitis and carcinoma), expressed GSTP-1 protein, which gives protection to the canine prostate tissue against oxidative damage. The p53 protein was present in all samples studied, including normal prostate tissue, but the prostatic lesions had a higher number of samples with higher scores (more than 50 percent of positive cells), in 95 percent of foci of PIA and carcinoma. It was found that an increased expression of NO in prostatic lesions of dogs and that the expression of GSTP-1 can protect the canine prostate tissue, which would contribute to the low frequency of prostate adenocarcinoma in this species. The expression of p53 was positive in all lesions as well as the in normal prostate.

Avaliação histológica da próstata de cães adultos sexualmente intactos / Prostatic histological evaluation in adult not castrated dogs

Avaliou-se histologicamente a próstata de 30 cães adultos e idosos sexualmente intactos que apresentavam ou não sintomatologia clínica de doença prostática, e verificou-se a incidência de possíveis alterações da glândula. Dentre as alterações encontradas, a hiperplasia prostática benigna constituiu o diagnóstico mais comum, 85,6 por cento (n=24), seguida por prostatite crônica, 64,3 por cento (n=18), displasia do epitélio glandular, 42,8 por cento (n=12), atrofia do epitélio glandular, 39,3 por cento (n=11), infiltrado inflamatório focal, 25 por cento (n=7), dilatação glandular focal, 21,4 por cento (n=6), prostatite aguda, 7,1 por cento (n=2), metaplasia escamosa, 3,6 por cento, (n=1), metástase de neoplasia sistêmica, 3,6 por cento (n=1) e abscesso prostático, 3,6 por cento (n=1). Como em muitos casos os cães são assintomáticos, ressalta-se a importância da realização rotineira de exames clínicos específicos, como o toque retal e a ultrassonografia, para o diagnóstico precoce e o tratamento das afecções prostáticas.

The prostates of 30 not castrated old dogs with or without clinical symptoms of prostatic disease were histologically evaluated. It was observed the incidence of possible changes in the gland. Among the changes, benign prostatic hyperplasia (BPH) was the most common diagnosis, accounting for 85.6 percent (n=24), followed by chronic prostatitis, 64.3 percent (n=18), dysplasia of the glandular epithelium, 42.8 percent (n=12), atrophy of the glandular epithelium, 39.3 percent (n=11), focal inflammatory infiltrate, 25 percent (n=7), focal glandular dilation, 21.4 percent (n=6), acute prostatitis, 7.1 percent (n=2), squamous metaplasia, 3.6 percent (n=1), metastasis of systemic neoplasia, 3.6 percent (n=1), and prostatic abscess, 3.6 percent (n=1). Because the lack of symptoms in most of dogs with prostatic changes, the specific clinic exams in routine, as rectal palpation and ultrasonography, are very important to early diagnosis and treatment of dogs with prostatic disease.

Molecular analysis of CYP21A2 can optimize the follow-up of positive results in newborn screening for congenital adrenal hyperplasia.

Neonatal screening for congenital adrenal hyperplasia (CAH) is useful in diagnosing salt wasting form (SW). However, there are difficulties in interpreting positive results in asymptomatic newborns. The main objective is to analyze genotyping as a confirmatory test in children with neonatal positive results. Patients comprised 23 CAH children and 19 asymptomatic infants with persistently elevated 17-hydroxyprogesterone (17OHP) levels. CYP21A2 gene was sequenced and genotypes were grouped according to the enzymatic activity of the less severe allele: A1 null, A2 < 2%, B 3-7%, C > 20%. Twenty-one children with neonatal symptoms and/or 17OHP levels > 80 ng/ml carried A genotypes, except two virilized girls (17OHP < 50 ng/ml) without CAH genotypes. Patients carrying SW genotypes (A1, A2) and low serum sodium levels presented with neonatal 17OHP > 200 ng/ml. Three asymptomatic boys carried simple virilizing genotypes (A2 and B): in two, the symptoms began at 18 months; another two asymptomatic boys had nonclassical genotypes (C). The remaining 14 patients did not present CAH genotypes, and their 17OHP levels were normalized by 14 months of age. Molecular analysis is useful as a confirmatory test of CAH, mainly in boys. It can predict clinical course, identify false-positives and help distinguish between clinical forms of CAH.

Hernioplastia experimental em coelhos por meio de cartilagem auricular bovina conservada em glutaraldeído / Rabbits experimental hernioplasty by means of bovine auricular cartilage preserved in glutaraldehyde

Avaliou-se a viabilidade do emprego da cartilagem auricular bovina conservada em glutaraldeído a 4 por cento na hernioplastia experimental, empregando-se seis coelhos adultos, machos, da raça Nova Zelândia. Para obtenção da hérnia incisional, removeu-se um segmento elíptico de 3cm de comprimento por 1cm de largura, no ponto central à primeira incisão, envolvendo fáscia e tecido muscular, na região da cicatriz umbilical. Dois animais de cada vez foram sacrificados aos 15, 30 e 45 dias após a cirurgia. Nos sacrificados aos 15 dias, observaram-se áreas focais de inflamação, caracterizadas por abscesso e fístula. À microscopia, observou-se área de inflamação e necrose próxima à periferia do implante. Nas amostras colhidas dos animais sacrificados aos 30 dias, não foram evidenciadas alterações clínicas relevantes. Desses, um animal apresentou à microscopia intensa proliferação fibroblástica, moderada neovascularização e células inflamatórias predominantemente mononucleares. Dos sacrificados aos 45 dias, em um ocorreu aderência de alça intestinal ao implante. É possível inferir que o material implantado apresentou satisfatória compatibilidade com o tecido receptor. Conclui-se que o implante de cartilagem auricular bovina conservada na hernioplastia experimental em coelhos apresentou evidências de boa integração tecidual e cicatrização, não havendo eliminação do material implantado.

The auricular cartilage preserved in 4 percent glutaraldehyde was used for experimental hernioplasty in six male, adult, New Zealand rabbits. To create an incision hernia, an elliptic tissue fragment three centimeter-long and one-centimeter wide was removed at a point centrally located from the first incision, embracing fascia and muscle tissue, from the area of umbilical scar. Animals were euthanized, two at a time, 15, 30, or 45 days after surgery. In the animals euthanized after 15 days focal areas of inflammation were observed, characterized by abscesses and fistulas. The histological section showed areas of inflammation and necrosis next to the periphery of the graft. In the animals euthanized after 30 days, there was no evidence of clinical alterations. Microscopic diagnosis of one of these animals showed intense fibroblastic proliferation, moderate neovascularization and inflammatory cells, predominantly mononuclear. One of the animals submitted to euthanasia at 45 days presented at necropsy adherence of bowel to the graft and impaired reconstitution of the parietal peritoneum. It is possible to infer that the grafted material presented satisfactory compatibility with the receptor tissue. Thus, it may be concluded that auricular bovine cartilage grafts preserved in 4 percent glutaraldehyde in experimental hernioplasty in rabbits presented evidence of good tissue integration and healing, with no elimination of the grafted material.

Non-viral lipid-based nanoparticles for targeted cancer systemic gene silencing.

New molecular biology techniques have uncovered the hidden role of genes in cancer. Identification of activated oncogenes, as fundamental genetic differences relative to normal cells, has made it possible to consider such genes as targets for antitumor therapy, namely by applying gene silencing strategies. In this regard, antisense oligonucleotides or small interfering RNAs, constitute promising therapeutic tools. The widespread clinical application of such molecules as modulators of gene expression, is still dependent on several aspects that limit their bioavailability, including: enhanced biological stability, favourable pharmacokinetics, enhanced tumor cell uptake and, consequently, efficient targeted delivery. One of the most promising strategies to overcome the barriers faced by gene silencing molecules, upon systemic administration, involves the use of lipid-based nanoparticles. The first part of this review aims at providing the reader with the molecular mechanism of action of the most important gene silencing molecules used in anticancer therapy. The primary obstacle for translating gene silencing technology from an effective research tool into a feasible therapeutic strategy remains its efficient delivery to the targeted cell type in vivo. Therefore, an overview of different lipid-based strategies for nucleic acid delivery will be presented on the second part. As we learn more about the pharmacokinetics and pharmacodynamics of the carrier and/or of the gene silencing molecules, it will be possible to further improve the delivery strategy that likely in the future will lead to the ideal non-viral particle for targeted cancer systemic gene silencing.