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  • Determination of the biochemical methane potential of swine hydrolyzate
    Publication . Domingues, Paulo Simão; Sousa, Helena Pala; Oliveira, Nelson Simões; Ribeiro, Ana M.; Ferreira, Alexandre
    Swine farming generates remains of births and animals that die during the process. Implementing a plan to eliminate these materials by hydrolysis process results in an animal by-product called swine hydrolyzate. The treatment of this by-product through its application in the anaerobic digestion process can also represent its conversion into a resource for energy production. Thus, the present study focuses on this by-product as a potential substrate for anaerobic digestion, evaluating the methane production potential. The results show biogas production with an average methane content of about 70 %. Theoretical biochemical methane potential was 967.95 ± 0.11 mL g VS−1, which represented a bioconversion efficiency of 59.97 ± 3.54 % and 62.26 ± 0.43 %, considering the experimental results of 580.47 ± 34.29 mL g VS−1 and 602.67 ± 4.16 mL g VS−1, respectively. By comparing the experimental biochemical methane potential with the predicted value using the modified Gompertz model, it was possible to conclude that the maximum methane production rate was 73.46 ± 0.36 mL g VS−1 day−1, with a digestion time of 18 days to obtain 90 % of the methane production potential.
  • Removal of Hydrophobic Organic Pollutants and Copper by Alginate-Based and Polycaprolactone Materials
    Publication . Marques-da-Silva, Dorinda; Lopes, João M.; Correia, Iris; Silva, João S.; Lagoa, Ricardo
    Organic pollutants (OPs) and heavy metals are environmental toxicants associated with great concerns. Decontamination processes are urgent for both, and the possibility to achieve their simultaneous removal from polluted waters is highly interesting. Additionally, in many cases, the effect of organic matter in the removal process is overlooked and must be considered. This work aimed to study the potential of alginate-based and polycaprolactone (PCL) materials to remove OPs and copper ions in the absence and presence of organic matter. The OPs investigated were the polycyclic aromatic hydrocarbons anthracene and benzo[a]pyrene, and the pesticide chlorpyrifos, both hydrophobic compounds. Copper (II) ions were used as a model of heavy metals. Alginate-based spheres were prepared by gelation, and PCL microparticles were obtained by oil/water emulsion solvent evaporation. The materials with the highest efficiencies for OP removal from aqueous solutions were those with activated carbon and PCL. Furthermore, the spheres with activated carbon could remove anthracene and copper simultaneously, even in the presence of humic acid. This work points to activated carbon–alginate spheres as a multifunctional adsorbent able to remove different pollutants and to PCL for potential applications in OP decontamination processes.
  • Complexation and DFT studies of lower rim hexahomotrioxacalix[3]arene derivatives bearing pyridyl groups with transition and heavy metal cations. Cone versus partial cone conformation
    Publication . Marcos, Paula M.; Teixeira, Filipa A.; Segurado, Manuel A. P.; Ascenso, José R.; Bernardino, Raul; Peter J. Cragg; Sylvia Michel; Véronique Hubscher-Bruder; Françoise Arnaud-Neu
    The binding of representative alkali, alkaline earth, transition and heavymetal cations by 2-pyridylmethoxy derivatives (1b, in cone and partial cone conformations) of p-tert-butylhexahomotrioxacalix[3]arene was studied. Binding was assessed by extraction studies of the metal picrates from water into dichloromethane and by stability constant measurements in acetonitrile and methanol, using spectrophotometric and potentiometric techniques. Microcalorimetric studies of some selected complexes in acetonitrile were performed, as well as proton NMR titrations. Computational methods (density functional theory calculations) were also employed to complement the NMR data. The results are compared with those obtained with the dihomooxacalix[4]arene 2b and the calix[4]arene 3b derivative analogues. Partial cone-1b is the best extractant for transition and heavy metal cations. Both conformers of 1b exhibit very high stability constants for soft and intermediate cations Pb2+, Cd2+, Hg2+, Zn2+ and Ni2+, with cone-1b the strongest binder (ML, log b ≥7) and partial cone-1b the most selective. Both derivatives show a slight preference for Na+. Besides the formation of ML complexes, ML2 and M2L species were also observed. The former complexes were, in general, formed with the transition and heavy metal cations, whereas the latter were obtained with Ag+ and Hg2+ and partial cone-1b. In most cases, these species were corroborated by the proton NMR and density functional theory studies.
  • Comparison of Laccases and Hemeproteins Systems in Bioremediation of Organic Pollutants
    Publication . Lopes, João M.; Marques-da-Silva, Dorinda; Videira, Paula Q.; Lagoa, Ricardo L.
    Aim: Laccases and peroxidases have attracted great interest for industrial and environmental applications. These enzymes have a broad substrate range and a robust oxidizing ability. Moreover, using mediators or co-oxidants makes it possible to increase their catalytic activity and extend their substrate scope to more resistant chemical structures. Background: Fungal laccases and ligninolytic peroxidases, mainly lignin and manganese peroxidases, are the privileged oxidoreductases for bioremediation processes. Nonetheless, an increasing diversity of laccases and peroxidase-type enzymes has been proposed for environmental technologies. Objective: This article aims to provide an overview of these enzymes and compare their applicability in the degradation of organic pollutants. Methods: Fundamental properties of the proteins are covered and applications towards polycyclic aromatic hydrocarbons (PAHs) and pesticides are specially focused. Results: Laccases are multicopper oxidases initially studied for applications in the pulp and paper in-dustry but able to oxidize a variety of environmentally concerning compounds. Relying on O2, laccases do not require peroxides nor auxiliary agents, like Mn2+, although suitable redox mediators are needed to attack the more recalcitrant pollutants (e.g., PAHs). True and pseudo-peroxidases use a stronger oxi-dant (H2O2) and the redox chemistry at the heme site generates high potential species that allow the oxidation of dyes and some pesticides. Conclusion: Lately, research efforts have been directed to enzyme discovery, testing with micropollu-tants, and improving biocatalysts’ stability by immobilization and protein engineering. Further under-standing of the effects of natural media components and solvents on the enzymes might lead to compet-itive enzymatic treatments of highly toxic media.
  • Complexation and DFT studies of lanthanide ions by (2-pyridylmethoxy)homooxacalixarene derivatives
    Publication . Marcos, Paula M.; Teixeira, Filipa A.; Segurado, Manuel A.P.; Ascenso, José R.; Bernardino, Raul; Cragg, Peter J.; Michel, Sylvia; Hubscher-Bruder, Véronique; Arnaud-Neu, Françoise
    The binding of lanthanide cations by 2-pyridylmethoxy derivatives of p-tert-butyldihomooxacalix[4]arene (1b), in the cone conformation, and p-tert-butylhexahomotrioxacalix[3]arene (2b), in both cone and partial cone conformations, was studied. These properties were assessed by extraction studies of the metal picrates from water into dichloromethane and stability constant measurements in methanol and acetonitrile, using spectrophotometric and microcalorimetric techniques. Proton NMR titrations with La3+ and Yb3+ cations were done in order to get information on the binding sites. Computational methods (density functional theory (DFT) calculations) were also used to complement the NMR data. The p-tert-butylcalix[4]arene analogue (3b) was also studied, and the results of the four ligands were compared. Partial cone-2b is the best extractant for lanthanide ions, showing some preference for the heavy lanthanides. In complexation, all four ligands show the same trend and a high selectivity for Yb3+ (ML, log β ≥ 7). Besides the formation of ML complexes, ML2 species were also obtained. In most cases, these species were corroborated by the proton NMR studies. For partial cone-2b with Pr3+ the complexation process is enthalpically driven, whereas for 3b the formation of the ML2 species with this cation is due to a favourable entropy term. DFT studies indicate that ligand 3b forms the most stable complex with La3+, followed by partial cone-2b.
  • Autosomal recessive hypotrichosis with loose anagen hairs associated with TKFC mutations*
    Publication . Onoufriadis, A.; Cabezas, A.; Ng, J. C. F.; Canales, J.; Costas, M. J.; Ribeiro, J. M.; Rodrigues, J. R.; McAleer, M. A.; Castelo‐Soccio, L.; Simpson, M. A.; Fraternali, F.; Irvine, A. D.; Cameselle, J. C.; McGrath, J. A.
    Background: Loose anagen hair is a rare form of impaired hair anchorage in which anagen hairs that lack inner and outer root sheaths can be gently and painlessly plucked from the scalp. This condition usually occurs in children and is often self-limiting. A genetic basis for the disorder has been suggested but not proven. A better understanding the aetiology of loose anagen hair may improve prevention and treatment strategies. Objectives: To identify a possible genetic basis of loose anagen hair using next-generation DNA sequencing and functional analysis of variants identified. Methods: In this case study, whole-exome sequencing analysis of a pedigree with one affected individual with features of loose anagen hair was performed. Results: The patient was found to be compound heterozygous for two single-nucleotide substitutions in TKFC resulting in the following missense mutations: c.574G> C (p.Gly192Arg) and c.682C> T (p.Arg228Trp). Structural analysis of human TKFC showed that both mutations are located near the active site cavity. Kinetic assays of recombinant proteins bearing either of these amino acid substitutions showed almost no dihydroxyacetone kinase or D-glyceraldehyde kinase activity, and FMN cyclase activity reduced to just 10% of wildtype catalytic activity. Conclusions: TKFC missense mutations may predispose to the development of loose anagen hairs. Identification of this new biochemical pathobiology expands the metabolic and genetic basis of hypotrichosis.
  • A Combined Vermifiltration-Hydroponic System for Swine Wastewater Treatment
    Publication . Ispolnov, Kirill; Aires, Luis M. I.; Lourenço, Nídia D.; Vieira, Judite S.
    Intensive swine farming causes strong local environmental impacts by generating effluents rich in solids, organic matter, nitrogen, phosphorus, and pathogenic bacteria. Insufficient treatment of hog farm effluents has been reported for common technologies, and vermifiltration is considered a promising treatment alternative that, however, requires additional processes to remove nitrate and phosphorus. This work aimed to study the use of vermifiltration with a downstream hydroponic culture to treat hog farm effluents. A treatment system comprising a vermifilter and a downstream deep-water culture hydroponic unit was built. The treated effluent was reused to dilute raw wastewater. Electrical conductivity, pH, and changes in BOD5, ammonia, nitrite, nitrate, phosphorus, and coliform bacteria were assessed. Plants were monitored throughout the experiment. Electrical conductivity increased due to vermifiltration; pH stayed within a neutral to mild alkaline range. Vermifiltration removed 83% of BOD5, 99% of ammonia and nitrite, and increased nitrate by 11%. Hydroponic treatment removed BOD5 (63%), ammonia (100%), nitrite (66%), nitrate (27%), and phosphorus (47% total and 44% dissolved) from vermifiltered water. Coliforms were reduced by vermifiltration but recovered in the hydroponic unit. Plants showed the ability to grow on vermi-filtered wastewater, although requiring nutrient supplementation. Vermifiltration combined with hydroponics is a promising treatment for swine wastewater, although optimization will be needed for a sustainable real-scale implementation.
  • Anaerobic Mesophilic Co-Digestion of Swine Slurry and Hidrolyzate in Batch Reactors: A Case Study
    Publication . Domingues, Paulo S.; Pala, Helena; Oliveira, Nelson S.
    The management of residual flows from swine farms, such as slurry and hydrolyzate, is a serious environmental problem in Portugal, so the search for new solutions is important, especially if they can be implemented in production sites, avoiding the transport of waste. Anaerobic co-digestion can configure a sustainable method of management and valorization of these flows in swine farms, resulting in biogas to produce energy and a digestate with agronomic value. The swine hydrolyzate is the product from the elimination of swine carcasses on the farms, through a hydrolysis process. Its use in the anaerobic process wasn’t found in the literature, identifying the need to evaluate its potential. Thus, anaerobic co-digestion of swine slurry and hydrolyzate was carried out and the main purpose of this research was to find the best feeding ratio between the two substrates in batch test, focusing on biogas production. The study was developed in two phases, both under mesophilic conditions. In the first one, using 100 ml syringes, the effect of the relative proportion of substrates on the anaerobic co-digestion potential was investigated and the effect of pressure sterilization of the substrates on biogas production was verified. In the second phase, a larger scale study was carried out in a 4,500 ml digestor, ran under the conditions identified as the most favorable in the preliminary tests. The results obtained allowed us to conclude that the largest volume of accumulated biogas was obtained with the proportion of 90.9% swine slurry: 9.1% swine hydrolyzate (v/v); and that the sterilization of the substrates doesn’t constitute an effective thermal pre-treatment. The larger scale test revealed an inefficient anaerobic process due to the inhibitory effect caused by the accumulation of ammonia and volatile organic acids. However, the production yield was 606.8 LBiogas/kgVS and 431.6 LMethane/kgVS, indicating that process allowed a production of biogas and methane higher than the values cited in literature for anaerobic digestion of swine slurry. This result showed that the use of swine hydrolyzate as a co-substrate results in a better balance of nutrients, promoting a better development of microorganisms.
  • Low-Protein Diets, Malnutrition, and Bone Metabolism in Chronic Kidney Disease
    Publication . Pereira, Cidália D.; Guimarães, Carla; Ribeiro, Vânia S.; Vaz, Daniela C.; Martins, Maria João
    Chronic kidney disease (CKD) has a high prevalence worldwide, with increasing incidence in low- and middle-income countries, and is associated with high morbidity and mortality, particularly from cardiovascular disease. Protein-restricted diets are one of the most widely used non-pharmacological approaches to slow the progression of CKD and prevent associated metabolic abnormalities. However, some concerns have been raised about the long-term safety of these diets, particularly with regard to patients’ nutritional status and bone and mineral disorders. Therefore, the aim of this article is to review the most recent scientific evidence on the relevance of using protein-restricted diets (with or without keto-analogue supplementation) and, in particular, their relationships with malnutrition and mineral and bone disorders in people with CKD without kidney replacement therapies. Although protein-restricted diets, especially when supplemented with keto-analogues and highly personalized and monitored, do not appear to be associated with malnutrition, research on their effects on bone and mineral disorders is scarce, deserving further investigation.
  • Should Children Drink Water with Very Low Mineral Content? Implications of the Global Expansion of Water Filtration Systems and Relevance of Consumption of Water with Higher Mineralization Levels
    Publication . Pereira, Cidália D.; Martins, Maria João
    The consumption of water with very low mineral content (W-VLMC; water with total dissolved solids below 50 mg/L), despite limited and inconsistent evidence and the resulting knowledge gaps, has not been associated with health risks for the general population. However, certain population subgroups (those eating very unbalanced diets or avoiding certain foods, engaged in prolonged periods of fasting, and/or doing prolonged or strenuous exercise as well as pregnant or breastfeeding women) should be mindful of maintaining sufficient intake of all essential minerals through their food if regularly using this type of water as their main beverage. The rapid expansion of water filtration systems—often producing W-VLMC—creates a timely and valuable opportunity to advance research on the health implications of W-VLMC intake. As these systems become increasingly common in educational settings and homes, children represent a subgroup experiencing rising exposure to W-VLMC. Additional studies are needed to assess the health effects of such exposure from early childhood. A complementary yet contrasting perspective is that the use of water intended for human consumption—with stringent quality control standards—and natural mineral waters—inherently pure, thus eliminating the need for filtration—with higher mineralization in both types of water, may provide an additional dietary source of essential minerals, especially for all the population subgroups mentioned above.