Key exploited species as surrogates for coastal conservation in an oceanic archipelago: insights from topshells and limpets from Madeira (NE Atlantic Ocean)

As lapas e os caramujos estão entre os herbívoros mais bem adaptados ao intertidal do Atlântico Nordeste. Estas espécies-chave fornecem serviços ecossistémicos valiosos, desempenhando um papel fundamental no equilíbrio ecológico do intertidal e têm um elevado valor económico, estando sujeitas a altos níveis de exploração e representando uma das atividades económicas mais rentáveis na pesca de pequena escala no arquipélago da Madeira. Esta dissertação visa preencher as lacunas existentes na história de vida e dinâmica populacional destas espécies, e aferir os efeitos da regulamentação da apanha nos mananciais explorados. A abordagem conservacionista implícita ao longo desta tese pretende promover: (i) a regulamentação adequada da apanha de caramujos (Phorcus sauciatus) e (ii) a avaliação dos efeitos da regulamentação da apanha de lapas nas populações exploradas (Patella aspera, Patella candei). Atualmente, os mananciais de lapas e caramujos são explorados perto do rendimento máximo sustentável, e a monitorização e fiscalização são fundamentais para evitar a futura sobre-exploração. A regulamentação da apanha de lapas produziu um efeito positivo nas espécies de lapas exploradas, com um aumento no tamanho, na proporção de indivíduos reprodutores, no tamanho de maturação e num maior equilíbrio na proporção de sexos. A apanha de caramujos não está regulamentada e com o atual nível de exploração ocorrem alterações na estrutura de tamanhos, abundância e potencial reprodutivo das populações exploradas, pelo que urge implementar a regulamentação da apanha desta espécie, por forma a mitigar os efeitos negativos desta atividade. O efeito da proximidade das populações humanas e acessibilidade costeira na estrutura de tamanhos e abundância de gastrópodes explorados mostrou que a proporção de reprodutores e a abundância eram geralmente menores em áreas mais próximas das populações humanas e em áreas mais acessíveis. Os efeitos das Áreas Marinhas Protegidas na proteção das populações de lapas resultaram num aumento diferencial do tamanho, da maturidade sexual e da captura por unidade de esforço de acordo com o grau de proteção. O esclarecimento e envolvimento das comunidades locais, reguladores, decisores políticos e partes interessadas, baseados em informação e educação, são cruciais para uma gestão eficaz e sustentável destes gastrópodes marinhos e ecossistemas a médio e longo prazo.Limpets and topshells are among the most successful intertidal grazers in the North-eastern Atlantic. These keystone species play a pivotal role in structuring rocky shores communities, and provoding valuable ecosystem services. Than have an important economic value, being subject to high levels of exploitation and representing one of the most profitable economic activities in small-scale fisheries in the archipelago of Madeira. This thesis aims to fill the gaps on the life-traits and population dynamics of these species, and assess the effects of harvesting regulations on the exploited stocks. A focus on conservation is implicit throughout this thesis since it addresses the promotion of: (i) proper regulation of the unregulated harvesting of topshells (Phorcus sauciatus) and (ii) provide additional information on the effects of harvesting regulations on limpets (Patella aspera, Patella candei). Currently, limpets and topshells stocks are being exploited near the maximum sustainable yield and monitoring and enforcement must be accomplished to avoid future overexploitation. Conservation measures prompted a positive effect on both exploited limpet species with an increase in length, reproductive individuals, size-at-first maturity and a more balanced sex-ratio after harvesting regulations. The harvesting of topshells is not regulated and with the current level of exploitation there are changes in the size structure, abundance and reproductive potential of the exploited populations, so it is imperative to implement the harvesting regulations for this species, in order to mitigate the negative effects of harvesting. The effect of proximity to human settlements and coastal accessibility on the size-structure and abundance of exploited gastropods showed that the mean-size, proportion of reproductive individuals and abundance were generally smaller in areas closer to human settlements and in more accessible areas. The effects of protection from the Marine Protected Areas on limpet populations resulted in a differential increase on size, size-at-first maturity and catch-per-unit-effort according to the degree of protection. The understanding and commitment of local communities, regulators, policymakers and stakeholders, based on information and education are crucial to the effective management and to ensure the sustainability of these marine gastropods and ecosystems at medium and long term

Activated Leukocyte Cell Adhesion Molecule Expression and Shedding in Thyroid Tumors

Activated leukocyte cell adhesion molecule (ALCAM, CD166) is expressed in various tissues, cancers, and cancer-initiating cells. Alterations in expression of ALCAM have been reported in several human tumors, and cell adhesion functions have been proposed to explain its association with cancer. Here we documented high levels of ALCAM expression in human thyroid tumors and cell lines. Through proteomic characterization of ALCAM expression in the human papillary thyroid carcinoma cell line TPC-1, we identified the presence of a full-length membrane-associated isoform in cell lysate and of soluble ALCAM isoforms in conditioned medium. This finding is consistent with proteolytically shed ALCAM ectodomains. Nonspecific agents, such as phorbol myristate acetate (PMA) or ionomycin, provoked increased ectodomain shedding. Epidermal growth factor receptor stimulation also enhanced ALCAM secretion through an ADAM17/TACE-dependent pathway. ADAM17/TACE was expressed in the TPC-1 cell line, and ADAM17/TACE silencing by specific small interfering RNAs reduced ALCAM shedding. In addition, the CGS27023A inhibitor of ADAM17/TACE function reduced ALCAM release in a dose-dependent manner and inhibited cell migration in a wound-healing assay. We also provide evidence for the existence of novel O-glycosylated forms and of a novel 60-kDa soluble form of ALCAM, which is particularly abundant following cell stimulation by PMA. ALCAM expression in papillary and medullary thyroid cancer specimens and in the surrounding non-tumoral component was studied by western blot and immunohistochemistry, with results demonstrating that tumor cells overexpress ALCAM. These findings strongly suggest the possibility that ALCAM may have an important role in thyroid tumor biology

In vitro culturing of porcine tracheal mucosa as an ideal model for investigating the influence of drugs on human respiratory mucosa

It has been previously shown that fresh mucosa from different mammals could serve as raw material for in vitro culturing with the differentiation of cilia, which are the most important morphological structures for the function of the mucociliary system. Increasing legal restrictions on the removal of human tissue and changing surgical techniques have led to a lack of fresh human mucosa for culturing. Most of the animals that have been used as donors up to now are genetically not very close to human beings and must all be sacrificed for such studies. We, therefore, established a modified system of culturing mucosa cells from the trachea of pigs, which is available as a regular by-product after slaughtering. With respect to the possibility of developing “beating” cilia, it could be shown that the speed of cell proliferation until adhesion to the coated culture dishes, the formation of conjunctions of cell clusters and the proliferation of cilia were comparable for porcine and human mucosa. Moreover, it could be demonstrated that the porcine cilia beat frequency of 7.57 ± 1.39 Hz was comparable to the human mucosa cells beat frequency of 7.3 ± 1.4 Hz and that this beat frequency was absolutely constant over the investigation time of 360 min. In order to prove whether the reaction to different drugs is comparable between the porcine and human cilia, we initially tested benzalkonium chloride, which is known to be toxic for human cells, followed by naphazoline, which we found in previous studies on human mucosa to be non-toxic. The results clearly showed that the functional and morphological reactions of the porcine ciliated cells to these substances were similar to the reaction we found in the in vitro cultured human mucosa

Two Bee-Pollinated Plant Species Show Higher Seed Production when Grown in Gardens Compared to Arable Farmland

Background Insect pollinator abundance, in particular that of bees, has been shown to be high where there is a super-abundance of floral resources; for example in association with mass-flowering crops and also in gardens where flowering plants are often densely planted. Since land management affects pollinator numbers, it is also likely to affect the resultant pollination of plants growing in these habitats. We hypothesised that the seed or fruit set of two plant species, typically pollinated by bumblebees and/or honeybees might respond in one of two ways: 1) pollination success could be reduced when growing in a floriferous environment, via competition for pollinators, or 2) pollination success could be enhanced because of increased pollinator abundance in the vicinity. Methodology/Principal Findings We compared the pollination success of experimental plants of Glechoma hederacea L. and Lotus corniculatus L. growing in gardens and arable farmland. On the farms, the plants were placed either next to a mass-flowering crop (oilseed rape, Brassica napus L. or field beans, Vicia faba L.) or next to a cereal crop (wheat, Triticum spp.). Seed set of G. hederacea and fruit set of L. corniculatus were significantly higher in gardens compared to arable farmland. There was no significant difference in pollination success of G. hederacea when grown next to different crops, but for L. corniculatus, fruit set was higher in the plants growing next to oilseed rape when the crop was in flower. Conclusions/Significance The results show that pollination services can limit fruit set of wild plants in arable farmland, but there is some evidence that the presence of a flowering crop can facilitate their pollination (depending on species and season). We have also demonstrated that gardens are not only beneficial to pollinators, but also to the process of pollination

Energy Metabolism in H460 Lung Cancer Cells: Effects of Histone Deacetylase Inhibitors

BACKGROUND: Tumor cells are characterized by accelerated growth usually accompanied by up-regulated pathways that ultimately increase the rate of ATP production. These cells can suffer metabolic reprogramming, resulting in distinct bioenergetic phenotypes, generally enhancing glycolysis channeled to lactate production. In the present work we showed metabolic reprogramming by means of inhibitors of histone deacetylase (HDACis), sodium butyrate and trichostatin. This treatment was able to shift energy metabolism by activating mitochondrial systems such as the respiratory chain and oxidative phosphorylation that were largely repressed in the untreated controls. METHODOLOGY/PRINCIPAL FINDINGS: Various cellular and biochemical parameters were evaluated in lung cancer H460 cells treated with the histone deacetylase inhibitors (HDACis), sodium butyrate (NaB) and trichostatin A (TSA). NaB and TSA reduced glycolytic flux, assayed by lactate release by H460 cells in a concentration dependent manner. NaB inhibited the expression of glucose transporter type 1 (GLUT 1), but substantially increased mitochondria bound hexokinase (HK) activity. NaB induced increase in HK activity was associated to isoform HK I and was accompanied by 1.5 fold increase in HK I mRNA expression and cognate protein biosynthesis. Lactate dehydrogenase (LDH) and pyruvate kinase (PYK) activities were unchanged by HDACis suggesting that the increase in the HK activity was not coupled to glycolytic flux. High resolution respirometry of H460 cells revealed NaB-dependent increased rates of oxygen consumption coupled to ATP synthesis. Metabolomic analysis showed that NaB altered the glycolytic metabolite profile of intact H460 cells. Concomitantly we detected an activation of the pentose phosphate pathway (PPP). The high O(2) consumption in NaB-treated cells was shown to be unrelated to mitochondrial biogenesis since citrate synthase (CS) activity and the amount of mitochondrial DNA remained unchanged. CONCLUSION: NaB and TSA induced an increase in mitochondrial function and oxidative metabolism in H460 lung tumor cells concomitant with a less proliferative cellular phenotype

A Meta-Analysis of Probiotic Efficacy for Gastrointestinal Diseases

Background: Meta-analyses on the effects of probiotics on specific gastrointestinal diseases have generally shown positive effects on disease prevention and treatment; however, the relative efficacy of probiotic use for treatment and prevention across different gastrointestinal diseases, with differing etiology and mechanisms of action, has not been addressed. Methods/Principal Findings: We included randomized controlled trials in humans that used a specified probiotic in the treatment or prevention of Pouchitis, Infectious diarrhea, Irritable Bowel Syndrome, Helicobacter pylori, Clostridium difficile Disease, Antibiotic Associated Diarrhea, Traveler’s Diarrhea, or Necrotizing Enterocolitis. Random effects models were used to evaluate efficacy as pooled relative risks across the eight diseases as well as across probiotic species, single vs. multiple species, patient ages, dosages, and length of treatment. Probiotics had a positive significant effect across all eight gastrointestinal diseases with a relative risk of 0.58 (95 % (CI) 0.51–0.65). Six of the eight diseases: Pouchitis, Infectious diarrhea, Irritable Bowel Syndrome, Helicobacter pylori, Clostridium difficile Disease, and Antibiotic Associated Diarrhea, showed positive significant effects. Traveler’s Diarrhea and Necrotizing Enterocolitis did not show significant effects of probiotcs. Of the 11 species and species mixtures, all showed positive significant effects except for Lactobacillus acidophilus, Lactobacillus plantarum, and Bifidobacterium infantis. Across all diseases and probiotic species, positive significant effects of probiotics were observed for all age groups, single vs. multiple species, and treatment lengths

Activation of store-operated calcium entry in airway smooth muscle cells: insight from a mathematical model

Intracellular dynamics of airway smooth muscle cells (ASMC) mediate ASMC contraction and proliferation, and thus play a key role in airway hyper-responsiveness (AHR) and remodelling in asthma. We evaluate the importance of store-operated entry (SOCE) in these dynamics by constructing a mathematical model of ASMC signaling based on experimental data from lung slices. The model confirms that SOCE is elicited upon sufficient depletion of the sarcoplasmic reticulum (SR), while receptor-operated entry (ROCE) is inhibited in such conditions. It also shows that SOCE can sustain agonist-induced oscillations in the absence of other influx. SOCE up-regulation may thus contribute to AHR by increasing the oscillation frequency that in turn regulates ASMC contraction. The model also provides an explanation for the failure of the SERCA pump blocker CPA to clamp the cytosolic of ASMC in lung slices, by showing that CPA is unable to maintain the SR empty of . This prediction is confirmed by experimental data from mouse lung slices, and strongly suggests that CPA only partially inhibits SERCA in ASMC