Oligopotent stem cells are distributed throughout the mammalian ocular surface.

The integrity of the cornea, the most anterior part of the eye, is indispensable for vision. Forty-five million individuals worldwide are bilaterally blind and another 135 million have severely impaired vision in both eyes because of loss of corneal transparency; treatments range from local medications to corneal transplants, and more recently to stem cell therapy. The corneal epithelium is a squamous epithelium that is constantly renewing, with a vertical turnover of 7 to 14 days in many mammals. Identification of slow cycling cells (label-retaining cells) in the limbus of the mouse has led to the notion that the limbus is the niche for the stem cells responsible for the long-term renewal of the cornea; hence, the corneal epithelium is supposedly renewed by cells generated at and migrating from the limbus, in marked opposition to other squamous epithelia in which each resident stem cell has in charge a limited area of epithelium. Here we show that the corneal epithelium of the mouse can be serially transplanted, is self-maintained and contains oligopotent stem cells with the capacity to generate goblet cells if provided with a conjunctival environment. Furthermore, the entire ocular surface of the pig, including the cornea, contains oligopotent stem cells (holoclones) with the capacity to generate individual colonies of corneal and conjunctival cells. Therefore, the limbus is not the only niche for corneal stem cells and corneal renewal is not different from other squamous epithelia. We propose a model that unifies our observations with the literature and explains why the limbal region is enriched in stem cells.

High-resolution 3D whole-heart coronary MRA: a study on the combination of data acquisition in multiple breath-holds and 1D residual respiratory motion compensation.

OBJECT: To study a scan protocol for coronary magnetic resonance angiography based on multiple breath-holds featuring 1D motion compensation and to compare the resulting image quality to a navigator-gated free-breathing acquisition. Image reconstruction was performed using L1 regularized iterative SENSE. MATERIALS AND METHODS: The effects of respiratory motion on the Cartesian sampling scheme were minimized by performing data acquisition in multiple breath-holds. During the scan, repetitive readouts through a k-space center were used to detect and correct the respiratory displacement of the heart by exploiting the self-navigation principle in image reconstruction. In vivo experiments were performed in nine healthy volunteers and the resulting image quality was compared to a navigator-gated reference in terms of vessel length and sharpness. RESULTS: Acquisition in breath-hold is an effective method to reduce the scan time by more than 30 % compared to the navigator-gated reference. Although an equivalent mean image quality with respect to the reference was achieved with the proposed method, the 1D motion compensation did not work equally well in all cases. CONCLUSION: In general, the image quality scaled with the robustness of the motion compensation. Nevertheless, the featured setup provides a positive basis for future extension with more advanced motion compensation methods.

Contribuição para o Estudo de um Sistema Integrado de Abastecimento de Água e Tratamento de Água Residual Doméstica. Caso de Estudo: Cidade da Praia

O problema de escassez de água em muitas regiões do globo acompanhado do acelerado crescimento da população e das decorrentes alterações climáticas, constituem um conjunto de preocupações ligadas à gestão dos recursos hídricos, principalmente no que diz respeito à satisfação das necessidades de consumo quer em termos quantitativos, quer em termos qualitativos. Tanto o abastecimento de água para o consumo humano, como a descarga e reutilização de águas residuais urbanas, requerem, à priori, um conjunto de processos e operações de tratamento tanto mais exigentes quanto maior o nível de qualidade final pretendido, tendo sempre em consideração aspectos como a saúde e a exposição humana como também o tipo de actividade a que se destina a água. Com o presente trabalho pretende-se dar um contributo para a avaliação da viabilidade de implementação de um sistema integrado de abastecimento de água e de tratamento de águas residuais domésticas na cidade da Praia, em Cabo Verde, tendo por base a situação actual, e um ano horizonte de 20 anos (2015 a 2035). Cabo Verde é um país com sérios problemas no que se refere aos recursos hídricos uma vez que carece de chuva e de reservas de águas subterrâneas. Circundado pelo mar, quase a totalidade dos municípios é abastecida pela água proveniente de processos de dessalinização da água do mar, através de tecnologias de membranas - Osmose Inversa (OI). Neste estudo, através de inquérito directo (260 inquéritos) e visitas presenciais a instituições, empresas e directamente à população, fez-se a caracterização dos sistemas de gestão da água e de águas residuais da referida cidade, por forma a perceber o funcionamento dos mesmos. Em 2014, cerca de 90% da água distribuída à população da Praia (10.000 m3.dia-1) proveio da água dessalinizada na central da ELECTRA. No mesmo ano, o caudal médio diário de água residual afluente à ETAR do Palmarejo foi de 2245 m3.dia-1. A deficiente cobertura da rede de drenagem de águas residuais da cidade (26,5%, segundo resultados do inquérito) é a principal razão pela baixa afluência à ETAR. Neste trabalho, procedeu-se ainda, à avaliação comparativa de custos para produção de água para a rega de áreas verdes da cidade, quer a partir de técnicas de dessalinização quer a partir da reutilização de água residual tratada. Com a reutilização de água residual tratada na rega das áreas verdes da cidade da Praia (92,4 ha), concluiu-se que, ao fim dos próximos 21 anos, poupar- se-á, na dessalinização, cerca de 32 milhões de euros (3.520 milhões de ECV). Da análise efectuada, concluiu-se que o volume necessário para o sistema de armazenamento da água residual tratada (estimado com base no mês de maior necessidade de rega do ano horizonte, Junho de 2035, assumindo um tempo de retenção hidráulico de um dia) é de 5.000 m3.

Nutrientes y renovación del agua en aiguamolls de l’Empordà. (NE de la península Ibérica). Uso potencial de agua residual para la inundación de zonas húmedas

The monitoring of the ecological water quality of 'Aiguamolls de l’Empordà' Natural Park was carried out between September 1996 to August 1997. The aim was to evaluate the impact of effluent from a nearby wastewater treatment plant, on the flooding of marshes in the Natural Park. In this paper we present physical and chemical data together nutrient concentrations and its changes in the lentic system of the 'Aiguamolls de l’Empordà'. The ultimate aim was to evaluate the effect of reclaimed wastewater on the salt marsh system. Seasonal variation in nutrients concentrations and their relationship to turnover and source of water were analysed. Significant variation in the N/P ratio was related to water turnover. Confined systems presented lower values for this ratio. Taking into account the nutrient concentrations of the reclaimed water, which is rich in nitrogen, it is recommended flooding applied in zones with high water turnover and higher N/P ratios

Electrically assisted ocular gene therapy.

Electrotransfer and iontophoresis are being developed as innovative non-viral gene delivery systems for the treatment of eye diseases. These two techniques rely on the use of electric current to allow for higher transfection yield of various ocular cell types in vivo. Short pulses of relatively high-intensity electric fields are used for electrotransfer delivery, whereas the iontophoresis technique is based on the application of low voltage electric current. The basic principles of these techniques and their potential therapeutic application for diseases of the anterior and posterior segments of the eye are reviewed. Iontophoresis has been found most efficient for the delivery of small nucleic acid fragments such as antisense oligonucleotides, siRNA, or ribozymes. Electrotransfer, on the other hand, is being developed for the delivery of oligonucleotides or custom designed plasmids. The wide range of strategies already validated and the potential for targeting specific types of cells confirm the promising early observations made using electrotransfer and iontophoresis. These two nonviral delivery systems are safe and can be used efficiently for targeted gene delivery to ocular tissues in vivo. At the present, their application for the treatment of ocular human diseases is nearing its final stages of adaptation and practical implementation at the bedside.

Fluorescein and indocyanine-green angiography in ocular syphilis: an exploratory study.

BACKGROUND: Fluorescein (FA) and indocyanine-green angiography (ICGA) may offer valuable information concerning disease severity and prognosis in ocular syphilis. The aim of the present study is to describe angiographic patterns encountered in the context of ocular syphilis, and to explore the associations between specific angiographic manifestations and severity of disease presentation, as well as disease evolution after treatment. METHODS: We performed a retrospective institutional study with the inclusion of 23 patients with ocular syphilis presenting to the uveitis clinic of the Jules-Gonin Eye Hospital in a 10-year period. FA and ICGA were performed following a standard protocol for posterior uveitis. Patterns of fluorescence were noted, and statistical associations between each angiographic pattern and any demographic, clinical, or laboratory parameter at baseline and after treatment were sought. RESULTS: The presence of any dark dots in ICGA was significantly associated with anterior uveitis (p = 0.031). The presence of hot spots in ICGA was significantly associated with longer duration of symptoms prior to initial visit (p = 0.032) and with male gender (p = 0.012). Weak non-significant trends were found associating vascular staining in FA with anterior uveitis (p = 0.066), vitritis (p = 0.069), and younger age (p = 0.061), as well as disc hyperfluorescence in FA with seropositivity for HIV (p = 0.089) and macular edema in FA with longer disease duration (p = 0.061). The presence of any dark dots in ICGA exhibited a weak trend of association with anterior uveitis and/or vitritis (p = 0.079). CONCLUSIONS: Out of the several associations identified implicating specific angiographic features, we underline the possible role of the presence of dark dots in ICGA for identifying active inflammation, and the role of hot spots in ICGA as markers of long-standing disease. Vascular staining in FA appears to be more common in patients with severe ocular inflammation with presence of anterior uveitis and/or vitritis.

Efeito residual do calcário na produção de milho e soja em solo Glei Pouco Húmico

O cultivo de milho e soja em solos ácidos requer a aplicação de calcário, para elevar o pH, reduzir a toxidez de alumínio e fornecer os nutrientes cálcio e magnésio. O calcário pode ter efeito residual prolongado, sendo necessário quantificar a durabilidade e a eficiência da calagem no solo por um período longo de cultivos sucessivos. Neste trabalho, deu-se continuidade a um experimento instalado na EMBRAPA Cerrados, em 1986, em solo Glei Pouco Húmico argiloso, com doses iniciais de 0, 2, 4, 6 e 8 t ha-1 de calcário dolomítico a lanço. A partir de 1990, foi cultivado o milho em rotação com a soja, em um total de sete cultivos sucessivos. Os dados de produtividade mostraram que o calcário aplicado em 1986 tem apresentado efeito residual. A resposta das duas culturas até o quarto cultivo e da soja, no sexto cultivo, foi significativamente maior a partir da primeira dose de 2 t ha-1 de calcário. No quinto e no sétimo ano de cultivo, a produtividade do milho foi significativamente maior, respectivamente, a partir das doses de 4 e 6 t ha-1 de calcário. A produtividade relativa máxima, para a média de quatro cultivos de milho e três cultivos de soja em sucessão, foi obtida com a saturação por bases do solo de 42% para o milho e 38% para a soja. A partir do quarto ano de aplicação do calcário, a perda anual média de cálcio + magnésio trocáveis do solo nos tratamentos com calcário foi de 2 mmol c dm-3.

Management of patients with uveal metastases at the Liverpool Ocular Oncology Centre.

BACKGROUND: Uveal metastasis is the most common intraocular malignancy. METHODS: This was a retrospective study of all patients with uveal metastases referred to the Liverpool Ocular Oncology Centre between January 2007 and December 2012. Biopsy was performed as a primary investigation if the clinical examination suggested metastasis with no evidence of any extraocular metastases. RESULTS: Ninety-six patients (109 eyes) were included. Breast and lung carcinomas were the most common primary malignancies, affecting 41 and 27 patients, respectively. The median time interval between detection of primary cancer and uveal metastasis was 24 months (range 1-288 months). Thirty-nine patients underwent ocular biopsy, confirming the diagnosis in all patients. The biopsy indicated the site of origin in 24 out of the 27 without a known primary tumour. In 7 of these 27 cases, previous systemic investigations had failed to identify the primary tumour. Seventy-three patients received external beam irradiation; two patients received photodynamic therapy; and two patients had Ru-106 plaque radiotherapy. The visual acuity was stable or improved in 75.5% of the cases. CONCLUSIONS: Immediate biopsy provides a quick diagnosis that may expedite treatment and improve any opportunities for conserving vision while facilitating the general oncologic management on these patients.

Controle da volatilização de amônia em compostagem, mediante adição de gesso agrícola e superfosfatos com diferentes níveis de acidez residual

Tem sido recomendada a adição de superfosfato simples ou gesso agrícola nas pilhas de estercos e de outros resíduos orgânicos, com a finalidade de reduzir as perdas de amônia por volatilização durante a compostagem. Experimentos, contudo, têm mostrado resultados divergentes, quando avaliam a eficiência desses produtos e as doses em que devem ser aplicados. É provável que variações na acidez residual do gesso e do superfosfato simples possam explicar essas divergências. Para verificar essa hipótese, realizou-se um experimento, utilizando, como aditivos, o gesso agrícola, o superfosfato simples e o superfosfato triplo, cada um com dois valores de acidez residual (respectivamente, 0,13 e 0,20%; 7,02 e 2,36%; 2,38 e 1,86%). Em frascos de vidro de 1,6 litro de capacidade, foram colocados 10 g de substrato orgânico obtido por mistura de quantidades iguais, em massa, de estercos secos e frescos de galinha e de gado. Os aditivos foram misturados ao substrato em doses equivalentes a 0, 50, 100, 150 e 200 kg t-1. A quantidade de amônia perdida por volatilização foi determinada aos 7, 14, 21, 28, 35 e 42 dias, coletando-se o gás em solução de ácido sulfúrico encerrada em pequeno recipiente colocado no interior do frasco e titulando-se o ácido remanescente após o período de exposição à amônia, com solução de NaOH 0,01 mol L-1. As amostras de gesso agrícola estudadas não se mostraram eficientes em reduzir a volatilização de amônia da mistura de estercos. O superfosfato simples diminuiu a volatilização de NH3 em até 4,8 vezes, tendo sido a amostra com maior acidez residual a mais eficiente. O superfosfato triplo foi o aditivo mais eficiente no controle da volatilização de NH3, quando aplicado em doses baixas, igualando-se ao superfosfato simples na maior dose. Aparentemente, a ação dos aditivos deveu-se à formação de compostos amoniacais mais estáveis por meio de reações do gás NH3 com ácidos e, ou, fosfato monocálcico presentes naqueles produtos.

Ultrasound assessment of ocular vascular effects of repeated intravitreal injections of ranibizumab for wet age-related macular degeneration.

PURPOSE: Determine the effect of repeated intravitreal injections of ranibizumab (0.5 mg; 0.05 ml) on retrobulbar blood flow velocities (BFVs) using ultrasound imaging quantification in twenty patients with exudative age-related macular degeneration treated for 6 months. METHODS: Visual acuity (ETDRS), central macular thickness (OCT), peak-systolic, end-diastolic and mean-BFVs in central retinal (CRA), temporal posterior ciliary (TPCA) and ophthalmic (OA) arteries were measured before, 2 days, 3 weeks and 6 months after the first injection. Patients were examined monthly and received 1-5 additional injections depending on ophthalmologic examination results. RESULTS: Six months after the first injection, a significant increase in visual acuity 50.9 ± 25.9 versus 44.4 ± 21.7 (p < 0.01) and decrease in mean central macular thickness 267 ± 74 versus 377 ± 115 μm (p < 0.001) were observed compared to baseline. Although mean-BFVs decreased by 16%±3% in CRA and 20%±5% in TPCA (p < 0.001) 2 days after the first injection, no significant change was seen thereafter. Mean-BFVs in OA decreased by 19%±5% at week 3 (p < 0.001). However, the smallest number of injections (two injections) was associated with the longest time interval between the last injection and month 6 (20 weeks) and with the best return to baseline levels for mean-BFVs in CRA, suggesting that ranibizumab had reversible effects on native retinal vascular supply after its discontinuation. Moreover, a significant correlation between the number of injections and percentage of changes in mean-BFVs in CRA was observed at month 6 (R = 0.74, p < 0.001) unlike TPCA or OA. CONCLUSION: Ranibizumab could impair the native choroidal and retinal vascular networks, but its effect seems reversible after its discontinuation.

Ocular drug delivery targeting the retina and retinal pigment epithelium using polylactide nanoparticles.

PURPOSE: To study the kinetics of polylactide (PLA) nanoparticle (NP) localization within the intraocular tissues and to evaluate their potential to release encapsulated material. METHODS: A single intravitreous injection (5 micro L) of an NP suspension (2.2 mg/mL) encapsulating either Rh-6G (Rh) or Nile red (Nr) was performed. Animals were killed at various times, and the NPs localization within the intraocular tissues was studied by environmental scanning electron microscopy (ESEM), confocal microscopy, light microscopy histology, fluorescence microscopy, and immunohistochemistry. Eyes injected with blank NPs, free Rh, or PBS solution were used as the control. RESULTS: ESEM showed the flow of the NPs from the site of injection into the vitreous cavity and their rapid settling on the internal limiting membrane. Histology demonstrated the anatomic integrity of the injected eyes and showed no toxic effects. A mild inflammatory cell infiltrate was observed in the ciliary body 6 hours after the injection and in the posterior vitreous and retina at 18 to 24 hours. The intensity of inflammation decreased markedly by 48 hours. Confocal and fluorescence microscopy and immunohistochemistry showed that a transretinal movement of the NPs was gradually taking place with a later localization in the RPE cells. Rh encapsulated within the injected NPs diffused and stained the retina and RPE cells. PLA NPs were still present within the RPE cells 4 months after a single intravitreous injection. CONCLUSIONS: Intravitreous injection of PLA NPs appears to result in transretinal movement, with a preferential localization in the RPE cells. Encapsulated Rh diffuses from the NPs and stains the neuroretina and the RPE cells. The findings support the idea that specific targeting of these tissues is feasible. Furthermore, the presence of the NPs within the RPE cells 4 months after a single injection shows that a steady and continuous delivery of drugs can be achieved.