Revalorization of spent coffee residues by a direct agronomic approach

Spent coffee grounds (SCG) represent a high-volume food waste worldwide, and several reuse approaches have been attempted. Herein, a greenhouse field experiment was carried out by cultivating Batavia lettuce with 5%, 10%, 15%, 20% and 30% (v/v) espresso SCG directly composted in the soil. Healthy vegetables were obtained for all treatments, without yield loss for up to 10% SCG. A progressive increment of green color intensity with increasing SCG content was observed, corroborated by the increase of their photosynthetic pigments (chlorophylls and carotenoids). Furthermore, total ascorbic acid and tocopherols showed statistical significant increases (p < 0.001) between control and all tested groups. Marked variations of nutritionally relevant minerals, particularly potassium, phosphorous and sodium were also revealed at higher percentage treatments (20% and 30%). This approach constitutes a clean, direct, simple and cost-effective measure to produce value-added vegetables, while reducing food waste disposal.

Implementação de um sistema para simulação por Monte Carlo da passagem de fotões através do olho humano mediante a utilização da plataforma GEANT4 - I

Dissertação para obtenção do Grau de Mestre em Engenharia Biomédica

Implementação de um sistema para simulação por Monte Carlo da passagem de fotões através do olho humano mediante a utilização da plataforma GEANT4 – II

Dissertação para obtenção do Grau de Mestre em Engenharia Biomédica

Cultivo e avaliação do potencial da microalga Chlorella zofingiensis para a produção de biodiesel e produtos de valor acrescentado

O presente trabalho tem como objetivo o cultivo da microalga Chlorella zofingiensis, e a avaliação da sua potencial aplicação na produção de biodiesel e de produtos de valor acrescentado, de entre os quais se destacam os antioxidantes. Com o intuito da produção de biocombustível é necessário efetuar o cultivo da microalga num volume que permita a obtenção de elevada quantidade de biomassa para a concretização do trabalho. Além deste biocombustível, existe ainda a possibilidade de valorização de alguns produtos com valor comercial, como é o caso da astaxantina, a saber na área farmacêutica, alimentar ou até mesmo cosmética. O cultivo da microalga foi feito em meio Bold’s Basal Medium (BBM), inicialmente em matrazes de 5 L e, quando se obteve uma cultura suficientemente densa, inocularam-se fotobiorreatores de 50 L. Conseguiu-se atingir uma concentração máxima de 0,76 g/L, no reator de 5 L, após cerca de 6 semanas de ensaio. Por sua vez, em fotobiorreatores de 50 L, a concentração máxima obtida foi de 0,4 g/L, após 4 semanas de ensaio. Nestas culturas foi possível obter-se uma percentagem lipídica de 7 %, apresentado concentração de pigmentos por litro de cultura na ordem dos 10 mg/L, 4 mg/L e 2 mg/L de clorofila a, clorofila b e carotenoides totais, respetivamente. Com esta percentagem lipídica recuperaram-se 400 mg de óleo, obtendo-se posteriormente 280 mg de biodiesel. Pela análise à amostra de biodiesel obtida foi possível obter o perfil lipídico desta microalga, quando cultivada em meio BBM, sendo 41% de ácido palmítico (C16:0), 9% de ácido esteárico (C18:0), 27% de ácido oleico (C18:1) e 23% de ácido linoleico (C18:2). Os resultados obtidos mostram que a Chlorella zofingiensis é uma microalga com interesse potencial para a produção de clorofila e carotenóides, mas não para o óleo para a produção de biodiesel.

Torpedo Maculopathy with an Anisometropic Amblyopia in a 5-Year-Old Caucasian Girl: Case Report

The aim of this study is to report a clinical case of asymptomatic female Caucasian children with torpedo maculopathy. A 5-year-old girl was referred to our clinic for routine evaluation. The ophthalmic examination revealed best-corrected visual acuity of 20/20 in both eyes, without any changes in the biomicroscopy. Fundus examination showed normal findings in one eye, whereas in the contralateral eye it disclosed, in the temporal sector of the macular region, a whitish, atrophic, oval chorioretinal lesion with clearly defined margins. Posterior evaluations documented the stability of the lesion. Torpedo maculopathy diagnosis is based on its characteristic shape and peculiar location. The differential diagnosis has to be established versus choroidal lesions (melanoma and nevus), congenital or iatrogenic hyperplasia of the retinal pigment epithelium (RPE) and particularly versus the congenital pigmented lesions associated with Gardner's syndrome.

The Impact of Multifocal Intraocular Lens in Retinal Imaging with Optical Coherence Tomography

Multifocal intraocular lenses (MF IOLs) have concentric optical zones with different dioptric power, enabling patients to have good visual acuity at multiple focal points. However, several optical limitations have been attributed to this particular design. The purpose of this study is to access the effect of MF IOLs design on the accuracy of retinal optical coherence tomography (OCT). Cross-sectional study conducted at the Refractive Surgery Department of Central Lisbon Hospital Center. Twenty-three eyes of 15 patients with a diffractive MF IOL and 27 eyes of 15 patients with an aspheric monofocal IOL were included in this study. All patients underwent OCT macular scans using Heidelberg Spectralis®. Macular thickness and volume values and image quality (Q factor) were compared between the two groups. There were no statistically significant differences between both groups regarding macular thickness or volume measurements. Retinal OCT image quality was significantly lower in the MF IOL group (p < 0.01). MF IOLs are associated with a significant decrease in OCT image quality. However, this fact does not seem to compromise the accuracy of spectral domain OCT retinal measurements.

Espessura da Coróide na Oclusão de Ramo Venoso da Retina

Objectivo: Avaliar a espessura da coróide na área macular em doentes com oclusão de ramo venoso da retina (ORVR) unilateral através de tomografia de coerência óptica de domínio espectral (SD-OCT) em modo enhanced depth imaging (EDI). Material e Métodos: Estudo retrospectivo não randomizado que incluiu 34 olhos de 17 doentes com ORVR unilateral (média de idade 68,6 ± 11,2 anos). Foi realizada análise estatística para comparar a espessura da coróide através de 3 medições (subfoveal e 750 µm temporal e nasal à fóvea) em cada uma de 7 linhas nos 15⁰x5⁰ centrais à fóvea para cada um dos olhos afectados e adelfos (21 medições em cada). Foi ainda realizada comparação entre a espessura macular central da retina e a espessura da coróide, para além da relação entre esta última e o tempo de evolução. Relacionou-se ainda a idade com a espessura da coróide no grupo controlo. Resultados: A média da espessura da coróide nos 17 olhos com ORVR foi de 211,8 ±55,97µm, o que foi superior à média verificada nos 17 olhos adelfos (185,7±46,1µm), sendo a diferença estatisticamente significativa (p⁼0,019). A coróide foi mais espessa a nível subfoveal (197,5±40,3µm) e mais delgada a nível nasal (176,9±54,9µm) no grupo controlo. Não se demonstrou haver relação entre o tempo de evolução e a espessura da coróide nos olhos com ORVR. Por outro lado houve relação entre a espessura da coróide e a espessura macular central (r⁼0,6; r²⁼0,36). Verificou-se uma correlação negativa, embora fraca, entre a idade e a espessura da coróide no grupo controlo (r⁼⁻0,022). Conclusões: A espessura da coróide pode ser avaliada através do EDI SD-OCT. Segundo alguns relatos, a mesma parece diminuir com a idade, tendência essa que se revelou também neste estudo. Tal como verificado na única publicação sobre a espessura da coróide na OVCR, demonstrou-se haver alteração na espessura da coróide na área macular em olhos com ORVR. Contudo são necessários mais estudos, com amostras maiores, que confirmem a alteração desta camada em olhos com ORVR e investiguem a sua influência na fisiopatologia da doença, no prognóstico visual e na resposta ao tratamento.

Intraocular Pressure in Eyes Receiving Intravitreal Antivascular Endothelial Growth Factor Injections

PURPOSE: The aim of the this study was to determine the effect of intravitreal antivascular endothelial growth factor injections on intraocular pressure (IOP) and identify possible risk factors for the development of increased IOP. MATERIALS AND METHODS: This prospective study included a total of 106 eyes receiving intravitreal injection of bevacizumab as treatment for macular edema or active choroidal neovascularization. IOP was measured by Goldmann applanation tonometry immediately before the intravitreal injection and 5 min, 1 h and 15 days after the procedure. The records of the study patients were reviewed for age, gender, history of glaucoma, diabetes mellitus, phakic status, systemic and topical medication and number of previous injections. Subconjunctival reflux was registered. IOP elevation was defined as IOP ≥21 mm Hg and/or a change from baseline of ≥5 mm Hg recorded at least on two or more measurements on the same visit. RESULTS: Mean preoperative IOP was 15.31 ± 3.90 mm Hg and postoperative IOP values were 27.27 ± 11.87 mm Hg (after 5 min), 17.59 ± 6.24 mm Hg (after 1 h) and 16.86 ± 3.62 mm Hg (after 15 days). The IOP variation was statistically significant between pre- and postoperative measurements (p < 0.05). Subconjunctival reflux was recorded in 11.3%, and in this subgroup the IOP at 5 min and at 1 h was lower than preoperative IOP (p < 0.05). CONCLUSIONS: More than one third of the eyes achieved IOPs >30 mm Hg 5 min after injection. Subconjunctival reflux contributed to a lower mean postoperative IOP (p < 0.05). Considerations for the management include prophylactic IOP lowering with medical therapy and/or preinjection ocular decompression for patients with a history of glaucoma or ocular hypertension and switching to an as-needed injection protocol in patients suffering a marked IOP rise in previous injections. © 2015 S. Karger AG, Basel.

Bull's Eye Maculopathy in a Patient Treated with Efavirenz

Objective: To report a case of retinal toxicity associated with efavirenz in an adult. Methods: We describe a case of gradual-onset blurry vision in both eyes in a 37-year-old HIV Caucasian woman, on antiretroviral therapy (ART), including efavirenz. Results: The patient presented with a best corrected visual acuity of 20/100 for the right eye (RE) and 20/125 for the left eye (LE). Fundoscopy revealed mottled atrophic changes of the macular retinal pigment epithelium (RPE) in both eyes. Fluorescein angiography revealed an annular pattern of RPE atrophy in both eyes. Full-field electretinography (ERG) was normal. Conclusions: Based on our patient’s history and on previous reports, efavirenz seems to be the culprit in this case. Our report provides evidence in support of routine ophthalmological evaluation of patients on efavirenz.

American cutaneous leishmaniasis: presentation and problems of patient management

We report our experience with the diagnosis and treatment of 60 patients with American cutaneous leishmaniasis. They were infected in Panama (55), Brazil (4) or Colombia (I). Among 35 patients with a 3 week exposure in Panama, the mean maximum incubation period was 33 days (range 4-81 days). Diagnosis was delayed an average of 93 days after onset of skin lesions, due to the patient's delay in seeking medical attention (31 days), medical personnel's delay in considering the diagnosis (45 days), and the laboratory's delay in confirming the diagnosis (17 days). Forty-four patients (73%) developed ulcers typical of cutaneous leishmaniasis. Sixteen additional patients (27%) had atypical macular, papular, squamous, verrucous or acneiform skin lesions that were diagnosed only because leishmanial cultures were obtained. Of the 59 patients treated with pentavalent antimonial drugs, only 34 (58%) were cured after the first course of treatment. Lesions which were at least 2 cm in diameter, ulcerated, or caused by Leishmania braziliensis were less likely to be cured after a single course of treatment than were lesions smaller than 2 cm, nonulcerated or caused by Leishmania mexicana or Leishmania donovani.

New roles of Rab GTPases on eye diseases: targeting VEGF secretion

RESUMO: A retina é composta, entre outras estruturas, pelo epitélio pigmentar da retina (EPR)e pela coróide. A região central da retina denomina-se mácula, e é a zona mais afetada na degenerescência macular relacionada com a idade, a forma mais comum de degenerescência da retina. Nesta doença, a secreção de fatores de crescimento pelo EPR é afetada, nomeadamente a do fator de crescimento vascular endotelial (VEGF), e pouco se sabe ainda sobre os mecanismos moleculares conducentes a esta condição. A família de proteínas Rab GTPases está envolvida nas vias intracelulares de sinalização e tráfego membranares, essenciais na transdução de sinais extracelulares em respostas biológicas. A sua crucial importância nestes mecanismos levou-nos a considerar o seu potencial envolvimento nas vias de secreção do VEGF, e a questionar-nos se teriam algum papel regulador sobre as mesmas. O principal objetivo deste trabalho é identificar Rab GTPases importantes para as vias de secreção e endocitose do VEGF no EPR. Essa identificação ajudará a esclarecer a patogénese da degenerescência macular da retina, e poderá servir para uma procura mais direcionada de novos agentes terapêuticos. A caracterização de dois modelos in vitro do EPR, células primárias isoladas de murganho e a linha celular B6-RPE07,levou-nos a concluir que são ambos semelhantes. Contudo, a linha celular foi escolhida como protótipo do EPR por permitir o acesso a um número ilimitado de células. No decurso deste trabalho, desenvolvemos e caracterizámos uma biblioteca de ferramentas moleculares que nos permitiram reduzir os níveis proteicos das proteínas Rab GTPases, com base na tecnologia de ácido ribonucleico (ARN) de interferência. O papel das proteínas Rab GTPases na secreção do VEGF no EPR foi estudado com base no silenciamento de apenas uma proteína, ou combinando várias, segundo a sua localização e funções intracelulares descritas. Este trabalho permitiu-nos concluir que as proteínas Rab GTPases são importantes intervenientes no processo de secreção de VEGF pelo EPR, e confirmar dados anteriores que relatam o envolvimento de algumas Rab GTPases endocíticas no processo. Propomos ainda um novo modelo para a interação destas proteínas no EPR, e sugerimos que a Rab10 e a Rab14 atuam negativamente sobre a Rab8, controlando o seu funcionamento. Os nossos resultados evidenciam a importância das proteínas Rab GTPases na secreção do VEGF pelas células do EPR, e servem de base a futuros estudos que melhor procurem compreender este mecanismo e de que modo a sua alteração se relaciona com a degenerescência da retina.--------ABSTRACT: Retinal pigment epithelium (RPE) and choroid are components of the mammalian retina, of which the central region is called macula. The most common form of retinaldegeneration, age-related macular degeneration (AMD), involves primarily deregulation of growth factors secretion by the RPE. Very little is known about the molecular mechanisms that lead to impairment of RPE’s homeostatic intracellular processes, namely the secretion of vascular endothelial growth factor (VEGF). Rab GTPases’ family regulates membrane targeting and traffic, being essential in the transduction of signal pathways. Given Rab proteins’ role in intracellular trafficking, we propose to identify key regulatory Rab proteins involved in either the secretory or the recycling pathways of VEGF in RPE. Understanding how Rab proteins’ function disruption could lead to retinal and choroidal pathology would ultimately contribute to find new therapeutic agents. Here, we characterized two mouse RPE in vitro cell models, primary cells and B6-RPE07 cell line, and concluded that both display important epithelial features as the RPE presents in vivo. Considering unlimited cell number and results reproducibility, we chose B6-RPE07 cells to further study Rab proteins’ function. To scrutinize the consequences of Rab proteins’ absence or diminished levels, we have developed novel molecular tools to achieve silencing of these key proteins using miRNA technology. We further addressed the effect of Rab proteins’ absence on VEGF secretion by performing an extensive screening where different Rab proteins were silenced, both individually and in multiple combinations considering their cellular/ compartment location. We conclude that Rab GTPases are important intervenients in VEGF secretion by RPE cells, confirming endocytic Rab proteins’ role in regulation of VEGF biology. We also propose a novel model for Rab proteins’ interaction in RPE. Our results suggest that Rab10 and Rab14 might influence Rab8 in a negative feedback mechanism, important for controlling VEGF secretion. Our achievements’ unravel Rab proteins’ role in VEGF secretion by RPE cells and are the basis for future studies to better understand RPE molecular secretory machinery.

Spectroscopic characterization of natural dyes by their non-invasive identification on pre-Columbian codices: the Maya yellow

Mesoamerican cultures had a strong tradition of written and pictorial manuscripts, called the codices. In studies already performed it was found the use of Maya Blue, made from a mixture of indigo and a clay called palygorskite, forming an incredibly stable material where the dye is trapped inside the nanotubes of the clay, after heating. However, a bigger challenge lies in the study of the yellows used, for these civilizations might have used this clay-dye mixture to produce their yellow colorants. As a first step, it was possible to provide identification, by non-invasive methods, of two colorants (a flavonoid and a carotenoid). While the flavonoid absorbed between 368-379 nm, the carotenoid would absorb around 455 nm. A temperature study also conducted allowed to set 140ºC as the desirable temperature to heat the samples without degrading them. FT-IR, conventional Raman and SERS allowed us to understand the existence of a reaction between the dyes and the clays (palygorskite and kaolinite), however it is difficult to understand it in a molecular point of view. As a second step, five species of Mexican dyes were selected on the basis of historical sources. The Maya yellow samples were produced adapting the recipe proposed by Reyes-Valerio, supporting the yellow dyes extracted from the dried plants on the clays, with addition of water, and then heated at 140ºC. It was found that the addition of water in palygorskite would increase the pH, hence deprotonating the molecules having a clear negative effect in the color. A second recipe was developed, without the addition of water; however, it was found that the use of water based binders would still alter the color of the samples with palygorskite. In this case, kaolinite without heating yield better results as a Maya yellow hybrid. It was found that the Maya chemistry might not have been the same for all the colors. The Mesoamericans might have found that different dyes could work better to their desires if matched with different clays. It was noticeable that for a clear distinction between flavonoids and carotenoids the reflectance and emission studies suffice, but when clay is added, Raman techniques will perform better. For this reason, conventional Raman and SERS were employed in order to create a database for the Mesoamerican dyestuffs for a future identification.

Evaluating the potential of yeast strains to produce added value products for the food and/or pharmaceutical industries

This study focus in the valorization of the apple pomace with the main goal of obtaining added value products. For that, hot compressed water technology was used for the extraction of phenolic compounds and hydrolysis of polysaccharides presents in the lignocellulosic structure of apple pomace to obtain simple sugars. The sugars have been utilized as alternative carbon source for growth, lipid accumulation and carotenoids production by five different yeast Yarrowia lipolytica, Rhodotorula mucilaginosa, Rhodotorula glutinis, Rhodosporidium babjevae and Rhodosporidium toruloides. Hydrolysis experiments were carried out with constant pressure of 100 bar, flow rate of 2mL/min and temperatures between 50°C and 250°C. The amount of total sugars present in apple pomace hydrolysates showed maximum values for the hydrolysis temperatures of 110°C and 190°C. In fact, these temperatures revealed the best results regarding the monosaccharides quantities. The amount of 5-HMF and furfural in each hydrolysate varied through the different temperatures. Maximum values for 5-HMF were obtained with 170°C, while furfural showed to be maximum at 210°C. Extraction of phenolic compounds were performed in simultaneously with hydrolysis reactions. Total phenolic compounds (TPC) increased along the temperature, however with small variations between 170°C and 250°C. Hydrolysates were then used as alternative carbon source to yeast growth. R. mucilaginosa shows the highest optical density, with the hydrolysate obtained at 130°C. Carotenoids produced by these yeast scored a total of 7.02μg carotenoids/g cell dry weight, while for the control assay, the same yeast scored 9.31μg caratonoides/g cell dry weight. β-carotene was quantified by HPLC, were 33% of the carotenoid production by R. mucilaginosa with hydrolysate as carbon source, corresponded to β-caroteno.

Valorization of agroindustrial waste

The main objective of this work is the valorization of residues from agro-industry giving them an added value. The valorization was performed by using a "green" and sustainable solvent - supercritical fluid, in this case carbon dioxide. Two residues and one biomass were used to produce two different final products, thereby emphasizing the versatility of the waste recovery - spent coffee grounds and microalgae Chlorella protothecoides to produce biodiesel, and tomato pomace to extract carotenoids. In the first part of this work it was demonstrated the possibility to obtain a conversion of coffee spent grounds oil into biodiesel, through an enzymatic transesterification reaction, of 98.01% with the following operating conditions: molar ratio oil:methanol 1:24, residence time 0.8 min, pressure 25 MPa, temperature 313,15K. In this first phase, it was also used the microalgae Chlorella protothecoides, a biomass, to produce biodiesel and favorable results were obtained with this green process compared with a traditional process - basic catalysis / acid. In the second part of this work, by an extraction with supercritical CO2 it was obtained 3.38% oil from tomato pomace under the following conditions: pressure 35.1 MPa, temperature 313,15K. It was found that this oil contains various carotenoids: β-carotene, lutein and lycopene. The latter is present in larger amount.

Causes of low vision and use of optical aids in the elderly

PURPOSE: To determine the causes of low vision in an elderly population attended by a university visual rehabilitation service and to check for the use of prescribed optical aids. METHOD: A cross-sectional study was carried out on patients aged 60 years or over attending for the first time a university low vision service in 2001. Ophthalmic reevaluation and interview were performed by means of a structured questionnaire in 2002. RESULTS: The sample comprised 50 subjects aged between 60 and 90 years. Severe low vision (<20/200) was present in 68.0% of patients. The main cause of low vision was age-related macular degeneration (44.0%). Regarding literacy, 16.0% were illiterate and 72.0% had completed fundamental schooling. Thirty-one patients (62.0%) had been prescribed optical aids; 54.8% of these patients stated that they use them. A majority (70.6%) held a favorable opinion of these aids. CONCLUSIONS: The main cause of low vision was age-related macular degeneration. Approximately half of those receiving prescriptions reported actually using the aids in their daily activities. Making best use of residual vision in the elderly population with visual impairment is a priority, given the social context, if the independence necessary for enhanced quality of life is to be achieved.