Image processing methodology for assessment of drilling induced damage in CFRP

Dissertação para obtenção do Grau de Mestre em Engenharia Mecânica

DNA damage induced by acrylamide: roe of genetic polymorphisms in DNA damage levels

RESUMO:Em 1994 a acrilamida (AA) foi classificada pela IARC como um provável cancerígeno para o homem. Para além da utilização de AA em numerosas aplicações industriais, a AA está também presente numa grande variedade de alimentos ricos em amido e processados a temperaturas elevadas. Esta exposição através da ingestão de produtos alimentares despoletou elevadas preocupações ao nível do risco para a saúde pública e poderá implicar um risco adicional para o aparecimento de cancro. A glicidamida (GA), o metabolito epóxido formado a partir da oxidação da AA provavelmente através do citocromo P450 2E1, é considerada por vários estudos, o principal responsável pela carcinogenicidade da AA. Actualmente existe uma escassez de resultados relativamente aos mecanismos de genotoxicidade da AA e GA em células de mamífero. Por este motivo, o objectivo deste estudo centra-se na avaliação das consequências genéticas da exposição à AA e GA, recorrendo-se para tal ao uso de células de mamífero como modelo. Tendo como base este objectivo avaliou-se a citotoxicidade da AA e GA, através do ensaio do MTT, e realizaram-se dois testes citogenéticos, o teste das aberrações cromossómicas (CAs) e o teste da troca de cromátides irmãs (SCEs), de modo a avaliar as lesões de DNA induzidas por estes compostos em células de hamster Chinês V79. Os resultados globalmente mostraram que a GA é mais citotóxica e clastogénica do que a AA. No âmbito deste trabalho, foi também efectuada a quantificação de aductos específicos de DNA, nomeadamente N7-(2-carbamoil-2-hidroxietil)guanina (N7-GA-Gua) e N3-(2-carbamoil-2-hidroxietil)adenina (N3-GA-Ade). Os resultados obtidos permitem afirmar que os níveis de N7-GA-Gua e a concentração de GA apresentam uma relação linear dose-resposta. Foi também identificada uma óptima correlação entre os níveis de N7-GA-Gua e a frequência de troca de cromátides irmãs. Adicionalmente, e de forma a compreender os mecanismos de toxicidade da AA, estudaram-se os mecanismos dependentes da modulação do glutationo reduzido (GSH), nomeadamente da butionina sulfoximina (BSO), um inibidor da síntese de GSH, do GSH-monoetil estér (GSH-EE), um composto permeável nas células e que é intra-celularmente hidrolisado a GSH e ainda do GSH adicionado exogenamente ao meio de cultura, em células V79. Os resultados obtidos reforçaram o papel da modulação do GSH nos efeitos de citotoxicidade e clastogenicidade da AA. Para além dos estudos efetuados com células V79, procedeu-se também à determinação da frequência de SCEs, à quantificação de aductos específicos de DNA, bem como ao ensaio do cometa alcalino em amostras de dadores saudáveis expostos à AA e GA. Tanto os resultados obtidos através do ensaio das SCE, como pela quantificação de aductos específicos de DNA, ambos efectuados em linfócitos estimulados, originaram resultados comparáveis aos obtidos anteriormente para as células V79, reforçando a ideia de que a GA é bastante mais genotóxica do que a AA. Por outro lado, os resultados obtidos pelo ensaio do cometa para exposição à AA e GA mostraram que apenas esta última aumenta o nível das lesões de DNA. Outro objectivo deste trabalho, foi a identificação de possíveis associações existentes entre as lesões de DNA, quantificadas através do ensaio das SCEs e do cometa, e biomarcadores de susceptibilidade, tendo em conta os polimorfismos genéticos individuais envolvidos na destoxificação e nas vias de reparação do DNA (BER, NER, HRR e NHEJ) em linfócitos expostos à GA. Tal permitiu identificar associações entre os níveis de lesão de DNA determinados através do ensaio das SCEs, e os polimorfismos genéticos estudados, apontando para uma possível associação entre o GSTP1 (Ile105Val) e GSTA2 (Glu210Ala) e a frequência de SCEs. Por outro lado, os resultados obtidos através do ensaio do cometa sugerem uma associação entre as lesões de DNA e polimorfismos da via BER (MUTYH Gln335His e XRCC1 Gln39Arg) e da via NER (XPC Ala499val e Lys939Gln), considerando os genes isoladamente ou combinados. Estes estudos contribuem para um melhor entendimento da genotoxicidade e carcinogenicidade da AA e GA em células de mamífero, bem como da variabilidade da susceptibilidade individual na destoxificação e reparação de lesões de DNA provocadas pela exposição a estes xenobióticos alimentares. ----------- ABSTRACT:Acrylamide (AA) has been classified as a probable human carcinogen by IARC. Besides being used in numerous industrial applications, AA is also present in a variety of starchy cooked foods. This AA exposure scenario raised concerns about risk in human health and suggests that the oral consumption of AA is an additional risk factor for cancer. A considerable number of findings strongly suggest that the reactive metabolite glycidamide (GA), an epoxide generated presumably by cytochrome P450 2E1, plays a central role in AA carcinogenesis. Until now there are a scarcity of results concerning the mechanisms of genotoxicity of AA and GA in mammalian cells. In view of that, the study described in this thesis aims to unveil the genetic consequences of AA and GA exposure using mammalian cells as a model system. With this aim we evaluated the cytotoxicity of AA and GA using the MTT assay and subsequently performed two cytogenetic end-points: chromosomal aberrations (CAs) and sister chromatid exchanges (SCEs), in order to evaluate DNA damage induced by these compounds in V79 Chinese hamster cell line. The results showed that GA was more cytotoxic and clastogenic than AA. Within the scope of this thesis the quantification of specific DNA adducts were also performed, namely N7-(2-carbamoyl-2-hydroxyethyl)guanine (N7-GA-Gua) and N3-(2-carbamoyl-2-hydroxyethyl)adenine (N3-GA-Ade). Interestingly, the GA concentration and the levels of N7-GA-Gua presented a linear dose-response relationship. Further, a very good correlation between the levels of N7-GA-Gua and the extent of SCEs were observed. In order to understand the mechanisms of AA-induced toxicity, the modulation of reduced glutathione (GSH)-dependent mechanisms were studied, namely the evaluation of the effect of buthionine sulfoximine (BSO), an effective inhibitor of GSH synthesis, of GSH-monoethyl ester (GSH-EE), a cell permeable compound that is intracellularly hydrolysed to GSH and also of GSH endogenously added to culture medium,z in V79 cell line. The overall results reinforced the role of GSH in the modulation of the cytotoxic and clastogenic effects induced by AA.Complementary to the studies performed in V79 cells, SCEs, specific DNA-adducts and alkaline comet assay in lymphocytes from healthy donors exposed to AA and GA were also evaluated. Both, the frequency of SCE and the quantification of specific GA DNA adducts, produced comparable results with those obtained in V79 cell line, reinforcing the idea that GA is far more genotoxic than AA. Further, the DNA damaging potential of AA and GA in whole blood leukocytes evaluated by the alkaline comet assay, showed that GA, but not AA, increases DNA damage. Additionally, this study aimed to identify associations between DNA damage and biomarkers of susceptibility, concerning individual genetic polymorphisms involved in detoxification and DNA repair pathways (BER, NER, HRR and NHEJ) on the GA-induced genotoxicity assessed by the SCE assay and by the alkaline comet assay. The extent of DNA damage determined by the levels of SCEs induced by GA seems to be modulated by GSTP1 (Ile105Val) and GSTA2 (Glu210Ala) genotypes. Moreover, the results obtained from the comet assay suggested associations between DNA damage and polymorphisms of BER (MUTYH Gln335His and XRCC1 Gln399Arg) and NER (XPC Ala499Val and Lys939Gln) genes, either alone or in combination. The overall results from this study contribute to a better understanding of the genotoxicity and carcinogenicity of AA and GA in mammalian cells, as well as the knowledge about the variability in individual susceptibility involved in detoxification and repair of DNA damage due to these dietary xenobiotics.

Carbon monoxide, autophagy and cytoprotection in response to cerebral ischemia

Dissertação para obtenção do Grau de Mestre em Genética Molecular e Biomedicina

Dissecting neuronal dysfunction and microglia/motoneurons cross-talk in ALS: an immunofluorescence directed study

Dissertação para obtenção do Grau de Mestre em Genética Molecular e Biomedicina

Despopulação neuronal pancreática em chagásicos crônicos

Revendo a literatura não encontramos estudos anatômicos dos gânglios intrapancreáticos na forma crônica da doença de Chagas; lesões dos mesmos poderiam explicar, ao menos em parte, os distúrbios funcionais do pâncreas exócrino e endócrino descritos nesta forma da doença. Decidimos então analisar morfologicamente tais gânglios. Para isso, estudamos segmentos transversais da cabeça, corpo e cauda do pâncreas de doze chagásicos crônicos, com idade média de 46,5 ± 9,1 anos, e quatorze controles, com idade média de 41,2 ± 11,0 anos. Os segmentos foram processados histologicamente e seccionados de forma seriada até o esgotamento, analisando-se os cortes múltiplos de sete. Para análise estatística, usamos o teste não-paramétrico de Mann-Whitney. Na cabeça do pâncreas, a contagem de neurônios teve média de 57,3 ± 50,8 para o grupo chagásico e 117,5 ± 99,0 para o grupo controle (p < 0,05); no corpo, 25,9 ± 19,4 para o grupo chagásico e 54,7 ± 47,8 para o controle (p < 0,05); na cauda, 23,4 ± 16,3 para o chagásico e 54,1 ± 29,2 para o controle (p < 0,01), sendo a contagem total de 106,6 ± 71,1 para o chagásico e 226,3 ± 156,5 para o controle (p < 0,01). Nossos achados nos permitiram concluir que: a) ocorreu despopulação neuronal estatisticamente significante no grupo chagásico em relação ao controle, em cada segmento pancreático analisado, bem como no órgão como um todo; b) 50% dos chagásicos tiveram número total de neurônios inferior ao menor número dos controles (80); c) 75% e 91,6% dos chagásicos tiveram número de neurônios inferior, respectivamente, à mediana (171) e à média (226) do grupo controle; d) assim, a despopulação neuronal pancreática foi frequente, porém não constante; e) o fator idade não pareceu ter sido o responsável pela despopulação neuronal dos chagásicos.

Despopulação neuronal cardíaca em hamsters (Mesocricetus auratus) cronicamente infectados com o Trypanosoma cruzi

Com o objetivo de se obter um modelo experimental que permitisse estabelecer a despopulação (desnervação) neuronal cardíaca procurou-se pesquisar o comportamento do sistema nervoso intracardíaco em hamsters cronicamente infectados com o T. cruzi. Para tal fim, realizaram-se contagens dos neurônios do plexo nervoso autonômico intracardíaco em hamsters inoculados com 35.000 formas sangüíneas de três cepas diferentes, sacrificados 5, 8 e 10 meses depois da infecção. Demonstrou-se, pela primeira vez, destruição neuronal significativa num modelo experimental, similar à que ocorre na doença de Chagas humana.

Migration and differentiation of neuronal precursors in the postnatal brain: insights from the subventricular zone and cerebellum

Fundação para a Ciência e a Tecnologia - SFRH/BD/42848/2008, através do Programa MIT_Portugal em Sistemas de Bioengenharia; projectos PTDC/SAUNEU/104415/2008 e Projecto ref. 96542 da Fundação Caloust Gulbenkian

Geospatial assessment of pest-induced forest damage through the use of UAV-bases NIR imaging and Gi-technology

Dissertation submitted in partial fulfillment of the requirements for the Degree of Master of Science in Geospatial Technologies.

Paracoccidioidomycosis: no genetic damage in human peripheral blood cells of patients assessed by single-cell gel (comet) assay

Paracoccidioidomycosis is a systemic fungal infection caused by Paracoccidioides brasiliensis. As infectious diseases can cause DNA damage, the authors aimed at analyzing DNA breakage in peripheral blood cells of patients with paracoccidioidomycosis by using the comet assay. The results suggested that paracoccidioidomycosis does not cause genotoxicity.

Characterization of molecular damage induced by UV photons and carbon ions on biomimetic heterostructures

The study of the effect of radiation on living tissues is a rather complex task to address mainly because they are made of a set of complex functional biological structures and interfaces. Particularly if one is looking for where damage is taking place in a first stage and what are the underlying reaction mechanisms. In this work a new approach is addressed to study the effect of radiation by making use of well identified molecular hetero-structures samples which mimic the biological environment. These were obtained by assembling onto a solid support deoxyribonucleic acid (DNA) and phospholipids together with a soft water-containing polyelectrolyte precursor in layered structures and by producing lipid layers at liquid/air interface with DNA as subphase. The effects of both ultraviolet (UV) radiation and carbon ions beams were systematically investigated in these heterostructures, namely damage on DNA by means vacuum ultraviolet (VUV), infrared (IR), X-Ray Photoelectron (XPS) and impedance spectroscopy. Experimental results revealed that UV affects furanose, PO2-, thymines, cytosines and adenines groups. The XPS spectrometry carried out on the samples allowed validate the VUV and IR results and to conclude that ionized phosphate groups, surrounded by the sodium counterions, congregate hydration water molecules which play a role of UV protection. The ac electrical conductivity measurements revealed that the DNA electrical conduction is arising from DNA chain electron hopping between base-pairs and phosphate groups, with the hopping distance equal to the distance between DNA base-pairs and is strongly dependent on UV radiation exposure, due loss of phosphate groups. Characterization of DNA samples exposed to a 4 keV C3+ ions beam revealed also carbon-oxygen bonds break, phosphate groups damage and formation of new species. Results from radiation induced damage carried out on biomimetic heterostructures having different compositions revealed that damage is dependent on sample composition, with respect to functional targeted groups and extent of damage. Conversely, LbL films of 1,2-dipalmitoyl-sn-Glycero-3-[Phospho-rac-(1-glycerol)] (Sodium Salt) (DPPG) liposomes, alternated with poly(allylamine hydrochloride) (PAH) revealed to be unaffected, even by prolonged UV irradiation exposure, in the absence of water molecules. However, DPPG molecules were damaged by the UV radiation in presence of water with cleavage of C-O, C=O and –PO2- bonds. Finally, the study of DNA interaction with the ionic lipids at liquid/air interfaces revealed that electrical charge of the lipid influences the interaction of phospholipid with DNA. In the presence of DNA in the subphase, the effects from UV irrladiation were seen to be smaller, which means that ionic products from biomolecules degradation stabilize the intact DPPG molecules. This mechanism may explain why UV irradiation does not cause immediate cell collapse, thus providing time for the cellular machinery to repair elements damaged by UV.

Prevalence of cholelithiasis in patients with chagasic megaesophagus

INTRODUCTION: The prevalence of cholelithiasis in the general population ranges from 9 to 18%. This prevalence is known to be higher in the presence of parasympathetic nerve damage of the biliary tract either due to surgery (vagotomy) or neuronal destruction (Chagas disease). The objective of this study was to evaluate the association of cholelithiasis and chagasic or idiopathic megaesophagus. METHODS: The ultrasound scans of 152 patients with megaesophagus submitted to cardiomyotomy and subtotal esophagectomy surgery were evaluated. The presence of cholelithiasis was compared between chagasic and idiopathic esophagopathy and ultrasound and clinical findings were correlated with age, sex and race. RESULTS: A total of 152 cases of megaesophagus, including 137 with chagasic megaesophagus and 15 with idiopathic megaesophagus, were analyzed. The mean age was 56.7 years (45-67) in the 137 patients with chagasic megaesophagus and 35.6 years (27-44) in the 15 cases of idiopathic megaesophagus, with a significant difference between the two groups (p < 0.0001). The group with chagasic megaesophagus consisted of 59 (43%) women and 78 (56.9%) men, while the group with idiopathic megaesophagus consisted of 8 (53.3%) women and 7 (46.6%) men, showing no significant difference between the groups. Of the 137 patients with confirmed chagasic megaesophagus, 39 (28.4%) presented cholelithiasis versus one case (6.6%) in the 15 patients with idiopathic megaesophagus. CONCLUSIONS: The prevalence of cholelithiasis is high in patients with chagasic megaesophagus and preoperative ultrasound should be performed routinely in these patients in order to treat both conditions during the same surgical procedure.

Evaluation of neuroprotective effects of natural extracts obtained from portuguese agro-food residues

Countries are currently faced with problems derived from changes in lifespan and an increase in lifestyle-related diseases. Neurodegenerative disorders such Parkinson’s (PD) and Alzheimer’s (AD) diseases are an increasing problem in aged societies. Data from World Alzheimer Report 2011 indicate that 36 million people worldwide are living with dementia. Oxidative stress has been associated with the development of AD and PD. Therefore there is interest to search for effective compounds or therapies to combat the oxidative damage in these diseases. Current evidence strongly supports a contribution of phenolic compounds present in fruits and vegetables to the prevention of neurodegenerative diseases such AD and PD. The industrial processing of a wide variety of fruits results in the accumulation of by-products without commercial value. Opuntia ficus-indica (cactus pear) is consumed fresh and processed like in juice. Prunnus avium (sweet cherry) is consumed fresh but the organoleptics characteristics of the fruits leads to the smaller and ragged fruits have no commercial value. Fruit extracts of both species has described to be rich in phenolic compounds and to have high antioxidant activities due to its composition. The aim of this work was assessing the efficacy of O. ficus-indica and P. avium by-products extracts obtained with conventional solvent extraction and pressurized liquid extraction in a neurodegeneration cell model. All extracts have protected neuroblastoma cells from H2O2-induced death at low, non-toxic levels, which approach to physiologically-relevant serum concentration. However, cherry extract has a slighter neuroprotective activity. The protective effect of Opuntia extracts are not conducted by a direct antioxidant activity since there are not decreases in intracellular ROS levels in cell treated with extracts and challenged with H2O2, while cherry extract neuroprotection seems to be due to a direct scavenging activity. Extracts from different biological matrixes seems to protect neuronal cells trough different cellular mechanisms.

Degree of disability, pain levels, muscle strength, and electromyographic function in patients with Hansen's disease with common peroneal nerve damage

INTRODUCTION: This study evaluated the degree of disability, pain levels, muscle strength, and electromyographic function (RMS) in individuals with leprosy. METHODS: We assessed 29 individuals with leprosy showing common peroneal nerve damage and grade 1 or 2 disability who were referred for physiotherapeutic treatment, as well as a control group of 19 healthy participants without leprosy. All subjects underwent analyses of degree of disability, electromyographic tests, voluntary muscle force, and the Visual Analog Pain Scale. RESULTS: McNemar's test found higher levels of grade 2 of disability (Δ = 75.9%; p = 0.0001) among individuals with leprosy. The Mann-Whitney test showed greater pain levels (Δ = 5.0; p = 0.0001) in patients with leprosy who had less extension strength in the right and left extensor hallucis longus muscles (Δ = 1.28, p = 0.0001; Δ = 1.55, p = 0.0001, respectively) and dorsiflexion of the right and left feet (Δ = 1.24, p = 0.0001; Δ = 1.45, p = 0.0001, respectively) than control subjects. The Kruskal-Wallis test showed that the RMS score for dorsiflexion of the right (Δ = 181.66 m·s-2, p = 0.001) and left (Δ = 102.57m·s-2, p = 0.002) feet was lower in patients with leprosy than in control subjects, but intragroup comparisons showed no difference. CONCLUSIONS: Leprosy had a negative influence on all of the study variables, indicating the need for immediate physiotherapeutic intervention in individuals with leprosy. This investigation opens perspectives for future studies that analyze leprosy treatment with physical therapeutic intervention.

Microvascularização arterial da cóclea

RESUMO: Na descrição deste estudo foi utilizada a terminologia anatómica da Sociedade Brasileira de Anatomia adaptada ao português por J. A. Esperança-Pina de acordo com o tratado Anatomia Humana da Relação. Os actuais estudos sobre hipoacusia sensorioneural implicam um grupo crescente de situações, em que a lesão se situa ao nível da microvascularização coclear, daí que o conhecimento exacto da angiomorfologia normal se torne essencial na fase actual do conhecimento. A autora tem vindo a estudar, desde 1986, a angiomorfologia do ouvido Interno no modelo experimental, o Cobaio, utilizando várias técnicas microvasculares. sendo dado enfâse particular neste estudo à técnica de microscopia electrónica de varrimento em moldes vasculares. Os animais usados no presente estudo pertencem à espécie cavia porcellus, cobaio, por serem considerados na comunidade cientifica internacional como o melhor modelo experimental para estudo do ouvido interno, pelo facto de a morfologia coclear ser muito semelhante à do Homem e por isso ser um modelo fiável para cirurgia experimental e microdissecção. Este estudo foi realizado em 100 cobaios, cavia porcellus, de ambos os sexos com peso médio de 450g. A vascularização do ouvido interno, no cobaio como no homem, faz-se através dos ramos de divisão da artéria auditiva interna ou labiríntica. A artéria labiríntica origina-se como ramo colateral da artéria cerebelosa ântero-inferior a qual tem origem na artéria basilar ou na artéria vertebral. Embora no homem a artéria auditiva interna possa também destacar-se da artéria basilar e até da artéria vertebral, no cobaio em todos os casos estudados a sua origem verificou-se sempre na artéria cerebelosa ântero-inferior. A artéria labiríntica, ao passar abaixo do meato auditivo interno, divide-se na artéria vestibular anterior e na artéria coclear comum.A artéria vestibular anterior dirige-se para o nervo vestibular, emite vasa nervorum para este nervo e vasculariza o utrículo e os canais semicirculares. A artéria coclear comum origina dois ramos principais, a artéria vestíbulo‑coclear ou vestibular posterior no cobaio, a qual se destaca junto à espira basal da cóclea e a artéria coclear, como ramo terminal, que passa a denominar-se de artéria modiolar ou espiralada, após entrar no modíolo. A artéria modiolar ascende no modíolo promovendo através dos seus ramos colaterais e dos seus ramos terminais a microvascularização coclear, numa vascularização de órgão de tipo terminal. Ao longo do seu trajecto verificou‑se de modo constante uma redução gradual de calibre em cada uma das espiras, por emissão de ramos colaterais, sendo que o calibre da artéria na base da cóclea apresenta um valor que diminui gradualmente até ao ápice. A artéria modiolar origina em todo o seu trajecto ramos colaterais, cujo número diminui em valor absoluto da base para o ápice: Arteríolas radiárias internas, arteríolas de trajecto flexuoso que caminham junto às estruturas sensorioneurais da parede interna da cóclea, junto ao lábio timpânico da lâmina espiral óssea e na parede do próprio modíolo, que se relacionam intimamente com este. As arteríolas radiárias internas originam‑se no flanco da artéria modiolar espiralada. Contam‑se dez a doze em cada espira, extraordinariamente flexuosas desde a sua origem. As arteríolas radiárias internas originam como ramos colaterais, vários grupos de arteríolas de menor calibre, que vascularizam distintas regiões da parede interna da cóclea, as arteríolas do gânglio espiral, a rede espiral interna, as arteríolas de origem dos glomérulos de Schwalbe e a arteríola da lâmina basilar. As arteríolas radiárias externas importantes ramos colaterais da artéria modiolar espiralada promovem a vascularização de importantes estruturas da parede externa. Ao atingir o limite externo do ligamento espiral, as arteríolas radiárias externas dividem‑se em vários ramos arteriolares de menor calibre, ao longo da convexidade do limite externo do ligamento espiral, originando a rede capilar pós-estriada que ocupa a porção lateral do ligamento espiral e a rede capilar ad‑ -estriada, na sua porção mais medial em íntima relação com a estria vascular. A espira basal da cóclea apresenta grande riqueza de vascularização, com características particulares apenas a esta espira, a qual é metabolicamente a mais exigente. A arteríola da janela da cóclea aborda a janela da cóclea pela sua convexidade e divide-se numa rica rede vascular da qual emergem arteríolas pré-capilares que se ramificam em capilares, os quais se dirigem em profundidade penetrando a rampa timpânica da cóclea ao nível da espira basal. Importou neste estudo verificar quais as semelhanças em termos de calibre de estruturas análogas, na parede interna e na parede externa da cóclea, com particular incidência na rede capilar. Do estudo estatístico realizado com testes paramétricos de Tamahane e não paramétricos de Mann-Whitney, verifica-se que comparando todas as estruturas consideradas estas têm calibres diferentes, com excepção dos capilares da estria vascular e do ligamento espiral, pertencentes à parede externa da cóclea que têm calibres iguais aos capilares da rede espiral interna e aos capilares da parede interna da cóclea, dependentes das arteríolas da rede espiral interna. As redes capilares dependentes das arteríolas radiárias internas que vascularizam as estruturas sensorioneurais junto á parede interna do modiolo são em tudo semelhantes em termos de calibre às redes capilares da parede externa da cóclea, incluindo os capilares da estria vascular. Esta particularidade traduz num órgão com vascularização de tipo terminal,um mecanismo de controlo do fluxo sanguíneo coclear tão importante na parede interna como na parede externa da cóclea. ------------ ABSTRACT:Current studies on sensorineural hearing loss, imply a growing group of situations in which the lesion is located at the level of the cochlear microvasculature, hence the exact knowledge of normal angiomorfology becomes essential in current state of knowledge. The author has been studying since 1986, the angiomorfology of inner on the experimental model, the guinea pig, using various microvascular techniques being given particular emphasis in this study to the results of the technique of scanning electron microscopy on corrosion casts. The animals used in this study belong to the species cavia porcellus, guinea pig, to be considered in the international scientific community as the best experimental model for the study of the inner ear, the cochlear morphology is very similar to human and therefore a reliable model for experimental surgery and microdissection. This study was performed in 100 guinea pigs of both sexes with average weight of 450g. There shall be a brief description of embryology, anatomy and cochlear physiology in the light of developmental biology, regarding also the spatial location of the cochlea and the determinism of morphogenetic fields in their development and function. The cochlear transduction mechanism converts the sound wave in stimuli sound and so afferent auditory nerve fibres and deafness are closely related to the cochlear microvasculature. Cochlear ischemia is accompanied by immediate hearing loss. The different type of cochlear injury that leads to sensorineural deafness is well studied in presbycusis where an objective link with the audiometric pattern as been established. The sensory type of deafness, is closely related to the degeneracy of the organ of Corti and damage to the outer hair cells at the basal turn of the cochlea. Keeping in mind cochlear tonotopy with location of high frequency sounds at the level of the base of the cochlea, it explains the audiometric pattern with loss in high frequencies. The neural type of deafness, is characterized by neuronal loss with loss of descendant important neuronal afferents, with audiometric translation on a gradually curve with important loss of auditory discrimination. The metabolic type of deafness results in atrophy of the vascular stria, with consequent change in the potential of the endolymph by decreasing the vascular stria cells and changes in K + recycling mechanism. There is also a change in the morphology of the spiral ligament and the audiometric patern as a flattened curve with loss at all frequencies. Bearing in mind cochlear tonotopy and being characterized all types of sensorineural deafness, we may inquire to what extent the cochlear microvasculature, considering not only the cochlea as a whole but different regions of the inner wall and the outer wall of the cochlea, contributes to deafness. We analysed the entire cochlear morphology on scanning electron microscopy with particular emphasis on bone and membranous cochlea. The inner wall of the cochlea and intramodiolar structures such as the spiral ganglion, the morphology of its cell bodies and their axons are analyzed. The morphology of Corti’s organ is described in detail, with description and large detail of the inner and outer hair cells. Is then presented the study of the microvasculature itself. The spiral modiolar artery is observed with the diaphanization technique and the technique of scanning electron microscopy on corrosion vascular casts. After emergence of collateral branches of the greatest importance, the radiating internal and external arterioles, the modiolar artery gives rise to its terminal branches, the arterioles of the cochear apex. Arterial vasa vasorum and vasa nervorum are displayed with a great detail, which was not yet described in such detail in previous microvascular studies. The arterial radiating arterioles originate in the flank of the spiral modiolar artery in number of ten to twelve in each loop, and they vascularize through their branches the inner wall cochlear sensorineural structures located in the modiolus as the spiral ganglion and structures near the organ of Corti. Their caliber is above 20 μm on the basal turn and in the second loop it decreases to values between 12 and 20 μm, decreasing progressively to the apex of the cochlea.They arise near the modiolus or on their way in the spiral lamina forming vascular loops, and divide without presenting vascular constrictions in their divisions, originating new vascular loops of lower caliber. Internal ratiating arterioles originate as collateral branches several groups of smaller caliber arterioles, which vascularize distinct regions of the inner wall of the cochlea namely, the arterioles of the spiral ganglion, the internal spiral network, the arterioles of origin of the glomeruli of Schwalbe and the arterioles of the basilar membrane. The glomeruli of Schwalbe play an important functional role as relay-stations, in hemodynamic terms, to control the cochlear microvasculature. External radiating arterioles have their origin in the spiral modiolar artery, they are directed towards the outer wall of the cochlea and run through the roof of the scala vestibuli. Above the insertion of Reissner’s membrane on the external wall the external radiating arterioles originate the spiral ligament arterioles, which vascularize the spiral ligament, they divide into several arteriolar branches of smaller caliber, along the convexity of the outer edge of the spiral ligament. The connective tissue of the spiral ligament forms a mesh with supporting function of the highly specialized epithelium, where pericytes were identifiable. Next to its base there is the microvascular network of stria vascularis. The adstriated vascular network which is divided into a capillary network, the capillary network of stria vascularis. The stria vascularis, the only vascularized epithelium of the human body, plays an important role, forming an haemato-labyrintine barrier to assure labyrinthine endocochlear potential and transport of ions, essential for the mechanism of transduction of external hair cells. The cochlear basal turn has a special feature on its external wall, the region of the windows, the round windows giving access to scala tympani and the oval window thatleads into scala vestibuli, and so it is metabolic demanding. For their role in cochlear tonotopy the sensorineural structures and those of the external wall of the cochlea, are particularly vulnerable to hypoxia. Although the complementarity of all the techniques was important for three- -dimensional reconstruction of the microvasculature of the cochlea, the scanning electron microscopy technique, especially when we used the system Semafore was fundamental to perform precise morphometric mesures regarding all vascular structures.Regarding the capillaries of the inner and outer wall of the cochlea networks this technique allowed their characterization in morphometric terms. To conclude the capillaries of the inner wall and of the external wall of the cochlea have similar size. So although located at different cochlear regions, with a different functional role, in cochlear physiology these networks consist of capillaries of similar caliber. It seems to translate a cochlear blood flow control mechanism that is so important in the inner wall as in and the external wall of the cochlea to provide for in inner ear homeosthasia.