Effect of polyunsaturated fatty acids (PUFAs) on the proliferation and extracellular matrix mineralization and gene expression of gilthead seabream skeletal cell lines

The aquaculture industry aims at replacing significant amounts of marine fish oil by vegetable oils in fish diet. Dietary lipids have been shown to alter the fatty acid composition of bone compartments, which would impact the local production of factors controlling bone formation. Knowledge on the mechanisms underlying the nutritional regulation of bone metabolism is however scarce in fish. Two in vitro bone-derived cell systems developed from seabream (an important species for aquaculture in the Mediterranean region) vertebra, capable of in vitro mineralization and exhibiting prechondrocyte (VSa13) and pre-osteoblast (VSa16) phenotype, were used to assess the effect of certain polyunsaturated fatty acids (PUFAs; arachidonic (AA), eicosapentaenoic (EPA) and docosahexaenoic (DHA) acids) on cell proliferation, extracellular matrix (ECM) mineralization and gene expression. While all PUFAs promoted morphological changes in both cell lines, VSa16 cell proliferation appeared to be stimulated by PUFAs in a dose dependent manner until 100M, whereas proliferation of VSa13 cells was impaired at concentrations above 10M. AA, EPA and DHA inhibited VSa13 ECM mineralization, alone and in combination, while VSa16 ECM mineralization was only inhibited by AA and EPA. DHA had the opposite effect, increasing mineralization almost by 2 fold. When EFAs were combined, DHA apparently compensated for the inhibitory effect of AA and EPA. Expression of marker genes for bone and lipid metabolisms has been investigated by qPCR and shown to be regulated in pre-osteoblasts exposed to individual PUFAs. Our results show that PUFAs are effectors of fish bone cell lines, altering cell morphology, proliferation and mineralization when added to culture medium. This work also demonstrates the suitability of our in vitro cell systems to get insights into mineralization-related effects of PUFAs in vivo and to evaluate the replacement of fish oils by vegetable oil sources in fish feeds.Um dos objetivos da indústria de aquacultura é a substituição, na dieta dos peixes, de óleos de peixe por óleos vegetais. Sendo que os lípidos, presentes na dieta, alteram a composição de ácidos gordos nos compartimentos de osso, este facto afectaria a produção local de factores que controlam a formação óssea. O conhecimento de mecanismos subjacentes à regulação nutricional do metabolismo ósseo é, no entanto, escasso em peixes. Neste estudo, foram utilizadas duas linhas celulares, derivadas de vértebra da dourada, com capacidade de mineralização in vitro, quando submetidas a determinadas condições. Estas exibem um fenótipo de pré-condrócitos (VSa13) e préosteoblástos (VSa16) e foram utilizadas para avaliar o efeito de ácidos gordos polinsaturados, selecionados (PUFAs; araquidónico (AA), eicosapentaenóico (EPA) e docosahexaenóico (DHA)) sobre a proliferação celular, mineralização da matriz extra celular (ECM) e expressão genética. Embora se tenham observado alterações morfológicas em ambas as linhas celulares, devido ao tratamento com PUFAs, houve, também, a estimulação da proliferação nas células VSa16, dependendo da dose utilizada e até uma concentração de 100M. Por outro lado, a proliferação das células VSa13 é inibida em concentrações acima de 10M. Os tratamentos com AA, EPA e DHA mostraram uma inibição da mineralização nas células VSa13, isoladamente e em combinação. No caso das células VSa16 a mineralização foi afectada, negativamente pelo uso de AA e EPA e, positivamente pelos tratamentos com DHA. Aquando da combinação dos PUFAs, o DHA aparentemente compensou o efeito inibitório dos AA e EPA. A expressão de genes envolvidos no metabolismo do osso e de lípidos foi investigada por qPCR e mostra a existência de regulação em pré-osteoblastos, expostos a PUFAs individuais. Os nossos resultados mostram que os PUFAs alteram a morfologia celular, proliferação e mineralização em linhas de células de peixes derivadas de osso. Este trabalho demonstra, também, a aptidão dos nossos sistemas celulares in vitro na obtenção de conhecimento relacionado com o efeito que a utilização de PUFAs tem na mineralização in vivo, bem como permite avaliar os efeitos da substituição de óleos de peixe por fontes de óleos vegetais na alimentação de peixes

Tegaserod for Female Patients Suffering From IBS With Mixed Bowel Habits or Constipation: A Randomized Controlled Trial

Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/73726/1/j.1572-0241.2008.01808.x.pd

Evidence for a Physically Compact Narrow-Line Region in the Seyfert 1 Galaxy NGC 5548

We have combined HST/FOS and ground-based spectra of the Seyfert 1 galaxy NGC 5548 to study the narrow emission lines over the 1200 -- 10,000 angstrom region. All of the spectra were obtained when the broad emission line and continuum fluxes were at an historic low level, allowing us to accurately determine the contribution of the narrow-line region (NLR) to the emission lines. We have generated multicomponent photoionization models to investigate the relative strength of the high ionization lines compared to those in Seyfert 2 galaxies, and the weakness of the narrow Mg II 2800 line. We present evidence for a high ionization component of NLR gas that is very close to the nucleus (~1 pc). This component must be optically thin to ionizing radiation at the Lyman edge (tau = 2.5) to avoid producing [O I] and Mg II in a partially ionized zone. The very high ionization lines (N V, [Ne V], [Fe VII], [Fe X]) are stronger than the predictions of our standard model, and we show that this may be due to supersolar abundances and/or a ``blue bump'' in the extreme ultraviolet (although recent observations do not support the latter). An outer component of NLR gas (at only ~70 pc from the continuum source) is needed to produce the low ionization lines. We show that the outer component may contain dust, which further reduces the Mg II flux by depletion and by absorption of the resonance photons after multiple scatterings. We show that the majority of the emission in the NLR of NGC 5548 must arise within about ~70 pc from the nucleus. Thus, the NLR in this Seyfert 1 galaxy is very physically compact, compared to the typical NLR in Seyfert 2 galaxies.Comment: 38 pages, Latex, includes 2 figures (postscript), to appear in Ap

Comparative analysis of zebrafish bone morphogenetic proteins 2, 4 and 16: molecular and evolutionary perspectives

BMP2, BMP4 and BMP16 form a subfamily of bone morphogenetic proteins acting as pleiotropic growth factors during development and as bone inducers during osteogenesis. BMP16 is the most recent member of this subfamily and basic data regarding protein structure and function, and spatio-temporal gene expression is still scarce. In this work, insights on BMP16 were provided through the comparative analysis of structural and functional data for zebrafish BMP2a, BMP2b, BMP4 and BMP16 genes and proteins, determined from three-dimensional models, patterns of gene expression during development and in adult tissues, regulation by retinoic acid and capacity to activate BMP-signaling pathway. Structures of Bmp2a, Bmp2b, Bmp4 and Bmp16 were found to be remarkably similar; with residues involved in receptor binding being highly conserved. All proteins could activate the BMP-signaling pathway, suggesting that they share a common function. On the contrary, stage-and tissue-specific expression of bmp2, bmp4 and bmp16 suggested the genes might be differentially regulated (e.g. different transcription factors, enhancers and/or regulatory modules) but also that they are involved in distinct physiological processes, although with the same function. Retinoic acid, a morphogen known to interact with BMP-signaling during bone formation, was shown to downregulate the expression of bmp2, bmp4 and bmp16, although to different extents. Taxonomic and phylogenetic analyses indicated that bmp16 diverged before bmp2 and bmp4, is not restricted to teleost fish lineage as previously reported, and that it probably arose from a whole genomic duplication event that occurred early in vertebrate evolution and disappeared in various tetrapod lineages through independent events

Managing the Reproductive Development of Grasses by Grazing Practices

Grazing natural grassland communities is necessary for both productive (feeding herbivores) and environmental (maintenance of open landscapes) objectives. Management guidelines should take into account the functional diversity of plant species between and within grassland communities. The management of the heading stage of grasses by grazing is an important tool to maintain acceptable forage quality and to avoid the seeding of low-valued species. The heading stage should be managed even in extensive systems and this needs a good knowledge of the phenological development of dominant species. In this work we illustrate the approach through the study of the development of four contrasting grasses and analysing the consequences for grazing management. This work concerned only diversity of the plant components of the grassland ecosystem

Suspended sediment dynamics on a seasonal scale in the Mandovi and Zuari estuaries, Central West coast of India

Suspended particulate matter (SPM) collected at regular stations from the Mandovi and Zuari estuaries indicates that the peaks of high SPM coincide with peaks of high rainfall and low salinity and also with peaks of moderate/low rainfall coupled with high salinity during the monsoon. The estuarine turbidity maximum (ETM) is a characteristic feature, it occurs in the channel accompanying spring tide during the monsoon and pre-monsoon, and shifts to the bay on neap tide during post-monsoon. ETM remains at the same position in the Mandovi River, both during the monsoon and pre-monsoon, whereas in Zuari it stretched upstream during monsoon and migrates seaward of the channel during pre-monsoon. The ETM coincides with the freshwater-seawater interface during the monsoon and is formed by the interaction between tidal currents and river flows. The ETM during pre-monsoon is associated with high salinities and is generated by tidal and wind-induced currents. The turbidity maximum on neap tide during post-monsoon may be due to the erosion and resuspension of sediments from the emergent tidal flats and transport of these turbid waters into the bay. Funneling effect of the narrowing bay in the Zuari estuary and associated physical processes effectively enhance the magnitude of the currents and transports sediments to the channel. SPM retention percentage indicates that the estuarine channel is prone to siltation

Effect of variable levels of dietary cholesterol and plant sterols on the growth performance and bone metabolism in gilthead seabream (Sparus aurata) juveniles

Cholesterol is found in all animal tissues and is an important component of biological cell membranes with functions such as precursor to bile acids, hormones and vitamins. Fish meal and fish oil are cholesterol-rich ingredients. Replacement of these marine-derived ingredients by plant proteins and vegetable oils tends to reduce dietary cholesterol levels

Anti-osteogenic activity of cadmium in zebrafish

Among the many anthropogenic chemicals that end up in the aquatic ecosystem, heavy metals, in particular cadmium, are hazardous compounds that have been shown to affect developmental, reproductive, hepatic, hematological, and immunological functions in teleost fish. There is also evidence that cadmium disturbs bone formation and skeletal development, but data is scarce. In this work, zebrafish was used to further characterize the anti-osteogenic/osteotoxic effects of cadmium and gain insights into underlying mechanisms. Upon exposure to cadmium, a reduction of the opercular bone growth was observed in 6-days post-fertilization (dpf) larvae and an increase in the incidence of skeletal deformities was evidenced in 20-dpf post-larvae. The extent and stiffness of newly formed bone was also affected in adult zebrafish exposed to cadmium while regenerating their caudal fin. A pathway reporter assay revealed a possible role of the MTF-1 and cAMP/PKA signaling pathways in mechanisms of cadmium osteotoxicity, while the expression of genes involved in osteoblast differentiation and matrix production was strongly reduced in cadmium-exposed post-larvae. This work not only confirmed cadmium anti-osteogenic activity and identified targeted pathways and genes, but it also suggested that cadmium may affect biomechanical properties of bone.Portuguese Science and Technology Foundation (FCT) - UID/Multi/04326/2019 (CCMAR), PEst-C/MAR/LA0015/2011 (CCMAR); SFRH/BD/128634/2017 and SFRH/BD/52425/2013info:eu-repo/semantics/publishedVersio

Reduction of blood C-reactive protein concentration complements the resolution of sputum bacillary load in patients on anti-tuberculosis therapy

Funding: This study was conducted under the PanACEA Biomarkers Expansion (PanBIOME) programme and the establishment of Maputo Tuberculosis Trial Unit (MaTuTU Project) which was funded in parts through the European and Developing Countries Clinical Trials Partnership (EDCTP), PZA-study and Federal Ministry of Education and Research (BMBF), Germany.Background: Tuberculosis (TB) is a difficult-to-treat disease requiring the combination of four antibiotics for a minimum of 6 months. Rapid and quantitative biomarkers to monitor treatment response are urgently needed for individual patient management and clinical trials. C-reactive protein (CRP) is often used clinically as a rapid marker of inflammation caused by infection. We assessed the relationship of TB bacillary load and CRP as biomarkers of treatment response. Methods: Xpert MTB/RIF-confirmed pulmonary TB cases were enrolled for treatment response assessment in Mozambique. Treatment response was measured using the Tuberculosis Molecular Bacterial Load Assay (TB-MBLA) in comparison with standard-of-care Mycobacterium Growth Indicator Tube (MGIT) culture at baseline and at weeks 1, 2, 4, 8, 12, 17, and 26 of treatment. Blood CRP concentration was measured at baseline, week 8, and week 26. Treatment response was defined as increase in MGIT culture time to positivity (TTP), and reduction in TB-MBLA-measured bacillary load and blood CRP concentration. Results: Out of the 81 screened presumptive TB cases, 69 were enrolled for 6-month treatment follow-up resulting in 94% treatment completion rate. Four participants did not complete TB treatment and 22 participants had missing CRP or TB-MBLA results and were excluded from TB-MBLA-CRP analysis. The remaining 43 participants—median age, 31 years old [interquartile range (IQR): 18–56]; 70% (30/43) male; and 70% (30/43) infected with HIV—were considered for analysis. Culture TTP and bacillary load were inversely correlated, Spearman’s r = −0.67, p < 0.0001. Resolution of sputum bacillary load concurred with reduction of blood CRP, r = 0.70, p < 0.0001. At baseline, bacillary load had a median (IQR) of 6.4 (5.5–7.2), which reduced to 2.4 (0.0–2.9) and 0.0 (0.0–0.0) log10 CFU/ml at months 2 and 6 of treatment, respectively. Correspondingly, blood CRP reduced from 1.9 (1.6–2.1) at baseline to 1.3 (0.9–1.7) and 0.4 (0.1–0.8) log10 mg/dl at months 2 and 6 of treatment, respectively. CRP reduction trialed bacteriological resolution at a rate of −0.06 log10 mg/dl compared to a bacillary load of 0.23 log10 CFU/ml per week. Consequently, 14 (33%) and 37 (88%) patients had reduced CRP to normal concentration and bacillary load to zero by the end of treatment, respectively. Pre-treatment CRP concentration and bacillary load, and resolution during treatment were slightly lower in HIV co-infected patients but not significantly different from HIV-uninfected TB patients. Conclusion: TB-MBLA-measured bacillary load and blood CRP complement each other in response to anti-TB therapy. Slow CRP reduction probably reflects residual TB bacilli in the lung not expectorated in sputum. Combining both measures can improve the accuracy of these biomarkers for monitoring TB treatment response and shorten turnaround time since the results of both assays could be available in 24 h.Publisher PDFPeer reviewe