Given its complex metamorphosis and digestive system ontogeny, Senegalese sole larvae capacity to digest and utilize dietary protein is likely to change throughout development. In the present study, ...we hypothesized that the manipulation of dietary protein complexity may affect Senegalese sole larvae capacity to digest, absorb and retain protein during metamorphosis, as well as the mRNA expression of genes encoding for the precursors of proteolytic enzymes of the digestive tract and the enterocyte peptide transporter PepT1, which may have further impact on somatic growth. Three diets were formulated using approximately the same practical ingredients, except for the main protein source. The Intact diet protein content was mostly based on intact plant protein where the target peptide molecular weight (MW) would be >70kDa. The PartH diet protein fraction was mostly based on a protein hydrolysate with a high content of 5–70kDa peptides. The HighH diet protein fraction was mostly based on a protein hydrolysate with a high content of 5kDa peptides. A growth trial was performed with larvae reared at 19°C under a co-feeding regime from mouth opening. The transcription of pga, tryp1c, ialp, ampn and pepT1 (encoding respectively for PepsinogenA, Trypsinogen1C, Intestinal alkaline phosphatase, Aminopeptidase N and for the enterocyte peptide transporter 1) was quantified by qPCR, during the metamorphosis climax (16DAH) and after the metamorphosis was completed (28DAH). An in vivo method of controlled tube-feeding was used to assess the effect on the larvae capacity to utilize polypeptides with different MW (1.0 and 7.2kDa) representing a typical peptide MW of each of the hydrolysates included in the diets. The PartH diet stimulated growth in metamorphosing larvae (16DAH), whereas the Intact diet stimulated growth after 36DAH. The Intact diet stimulated the larvae absorption capacity for 1.0kDa peptides at 16DAH, which may have contributed for enhanced growth in later stages. The PartH diet stimulated the transcription of tryp1c and pept1 at 28DAH, which seemed to reflect on increased post-larvae capacity to retain dietary 7.2kDa polypeptides. That may indicate a possible strategy to optimize the digestion and utilisation of the PartH dietary protein, though it did not reflect into increased growth. The Intact diet promoted the transcription of pepsinogenA, which may reflect a reduced gastrointestinal transit time, which could have enhanced the dietary nutrients assimilation, ultimately improving growth. The present results suggest that, whereas pre-metamorphic sole larvae utilize better dietary protein with a moderate degree of hydrolysis, post-metamorphic sole make a greater use of intact protein.
•Proteolytic capacity is a limiting factor for protein digestion in sole early stage larvae.•Sole larvae adjust the way they utilize protein in response to dietary formulation.•Pre-metamorphic sole larvae grow better upon moderately hydrolysed dietary protein.•Dietary intact protein seems to be suitable to sole post-larvae and young juveniles.•Microdiets protein for Senegalese sole should be adapted to each developmental stage.
Due to its high protein synthesis and deposition rates, skeletal muscle protein deposition is a major determinant of fish growth. Dietary protein complexity is likely to influence protein utilization ...and deposition in skeletal muscle, possibly affecting fish myogenesis. In this study, three microdiets were formulated with different degree of hydrolysis of dietary protein as the changing factor: one diet contained a mix of intact protein sources targeting a peptide with molecular weight >20 kDa (Intact); a second diet contained a hydrolysate with polypeptides ranging from 5 to 70 kDa (PartH); and a third diet contained a high level of a protein hydrolysate mostly composed of small peptides (<5 kDa) (HighH). A possible effect on the regulation of muscle growth in Senegalese sole larvae was evaluated through white muscle cellularity and the expression of muscle growth-related genes at 16 and 36 DAH. The PartH diet promoted white muscle growth during the metamorphosis climax (16 DAH), which was reflected on increased body weight. At 36 DAH, different diets induced different expression patterns of genes encoding for the myogenic regulatory factors, which affected muscle growth dynamics, ultimately promoting growth potential in the Intact group. A lower recruitment of small-sized fibres in the PartH and HighH groups led to reduced potential for muscle growth, which resulted on further reduced somatic growth. Accordingly, fish fed the Intact diet grew better up to a late juvenile stage (60 DAH) and were still heavier than others even after 30 days of feeding all groups on the same commercial diet, at 90 DAH. The up-regulation in the transcript levels of genes encoding for de novo DNA methyltransferases in the HighH group suggest a potential for nutritional programming in this species.
•Moderately hydrolysed protein promotes muscle growth of pre-metamorphic sole larvae.•Dietary intact protein increased growth potential in sole post-larvae and juveniles.•Changes in dietary protein complexity affected myogenesis in sole larvae.•Changes in dietary protein complexity affected the expression of de novo DNMTs.
Conventional wisdom is that previous infection with omicron subvariant BA.1 or BA.2 does not protect against BA.5, but data from Portugal show considerable protection against BA.5 infection from ...previous BA.1 or BA.2 infection.
A new photonic structure is produced from cellulose nanocrystal iridescent films reflecting both right and left circularly polarized light. Micrometer‐scale planar gaps perpendicular to the films' ...cross‐section between two different left‐handed films' cholesteric domains are impregnated with a nematic liquid crystal. This photonic feature is reversibly tuned by the application of an electric field or a temperature variation.
•Evaluate environmental land use conflicts (LUC) in rural watersheds.•Investigate the ecological quality of surface water in LUC areas.•Characterize the structure of macroinvertebrate communities ...within LUC areas.•Assess the overall impact of LUC on the riverine ecosystem.•Propose measures of soil conservation and water protection to mitigate impacts of LUC.
Starting from a diagnosis of areas with different environmental land use conflicts located in various rural sub-basins of the River Sordo basin (northern Portugal), the present study analyzed the ecological quality of surface water in small mountain streams to establish a relationship between land use, water and aquatic biota. Environmental land use conflicts were set up on the basis of land use and land capability maps, coded as follows: 1 – agriculture, 2 – pasture, 3 – pasture/forest, and 4 – forest. Land capability was assessed by the ruggedness number methodology (RN). The difference between the codes of capability and use defines a conflict class, where a negative or null value means no conflict and a positive value means class i conflict. Within and without the conflict areas, ecological quality of surface water was evaluated by the metrics EPT taxa, IPtIN index, diversity of Shannon–Wiener and Evenness index. Macroinvertebrate communities are strongly correlated to conflict classes, because sites without (reference sites) or with minor physicochemical and hydromorphological degradation (Class 1) presented high diversity, evenness, EPT taxa and IPtIN index, while more impacted sites (Class 2) presented an ecological status not fulfilling the demands of the European Union Water Framework Directive (2000/60). The present study indicates a significant impact of land use on water quality which has straight influence on the distribution of biota, emphasizing the key role of riparian vegetation in the conservation of aquatic ecosystems. The highest impacts on macroinvertebrate assemblages were associated with changes in water quality parameters such as temperature, oxygen saturation (%), turbidity, total suspended solids (TSS), nitrates, phosphates and sulphates, conductivity and dissolved oxygen, as well as hydromorphological alterations driven by the total absence of riparian vegetation as a consequence of terrace building, agriculture and the resectioning/reinforcement of the banks associated with the culture of vine. Macroinvertebrates proved reliable to distinguish conflict classes and separate seasons. The main conservation measures required to improve the conflict areas in the short and medium term mainly include the adoption of agroforestry practices as these not only improve the quality of water and soil, maintaining land resources over a long period of use, but also brings many benefits to the landowner.
Wheat blast, caused by
(PoT), is an emerging threat to global wheat production. The current understanding of the population biology of the pathogen and epidemiology of the disease has been based on ...phylogenomic studies that compared the wheat blast pathogen with isolates collected from grasses that were invasive to Brazilian wheat fields. In this study, we performed a comprehensive sampling of blast lesions in wheat crops and endemic grasses found in and away from wheat fields in Minas Gerais. A total of 1,368 diseased samples were collected (976 leaves of wheat and grasses and 392 wheat heads), which yielded a working collection of 564
isolates. We show that, contrary to earlier implications, PoT was rarely found on endemic grasses, and, conversely, members of grass-adapted lineages were rarely found on wheat. Instead, most lineages were host-specialized, with constituent isolates usually grouping according to their host of origin. With regard to the dominant role proposed for signalgrass in wheat blast epidemiology, we found only one PoT member in 67 isolates collected from signalgrass grown away from wheat fields and only three members of
-adapted lineages among hundreds of isolates from wheat. Cross-inoculation assays on wheat and a signalgrass used in pastures (
) suggested that the limited cross-infection observed in the field may be due to innate compatibility differences. Whether or not the observed level of cross-infection would be sufficient to provide an inoculum reservoir, or serve as a bridge between wheat growing regions, is questionable and, therefore, deserves further investigation.
STUDY QUESTION
What is the contribution of the provision, at no cost for users, of long acting reversible contraceptive methods (LARC; copper intrauterine device IUD, the levonorgestrel-releasing ...intrauterine system LNG-IUS, contraceptive implants and depot-medroxyprogesterone DMPA injection) towards the disability-adjusted life years (DALY) averted through a Brazilian university-based clinic established over 30 years ago.
SUMMARY ANSWER
Over the last 10 years of evaluation, provision of LARC methods and DMPA by the clinic are estimated to have contributed to DALY averted by between 37 and 60 maternal deaths, 315–424 child mortalities, 634–853 combined maternal morbidity and mortality and child mortality, and 1056–1412 unsafe abortions averted.
WHAT IS KNOWN ALREADY
LARC methods are associated with a high contraceptive effectiveness when compared with contraceptive methods which need frequent attention; perhaps because LARC methods are independent of individual or couple compliance. However, in general previous studies have evaluated contraceptive methods during clinical studies over a short period of time, or not more than 10 years. Furthermore, information regarding the estimation of the DALY averted is scarce.
STUDY DESIGN, SIZE AND DURATION
We reviewed 50 004 medical charts from women who consulted for the first time looking for a contraceptive method over the period from 2 January 1980 through 31 December 2012.
PARTICIPANTS/MATERIALS, SETTING, METHODS
Women who consulted at the Department of Obstetrics and Gynaecology, University of Campinas, Brazil were new users and users switching contraceptive, including the copper IUD (n = 13 826), the LNG-IUS (n = 1525), implants (n = 277) and DMPA (n = 9387). Estimation of the DALY averted included maternal morbidity and mortality, child mortality and unsafe abortions averted.
MAIN RESULTS AND THE ROLE OF CHANCE
We obtained 29 416 contraceptive segments of use including 25 009 contraceptive segments of use from 20 821 new users or switchers to any LARC method or DMPA with at least 1 year of follow-up. The mean (±SD) age of the women at first consultation ranged from 25.3 ± 5.7 (range 12–47) years in the 1980s, to 31.9 ± 7.4 (range 16–50) years in 2010–2011. The most common contraceptive chosen at the first consultation was copper IUD (48.3, 74.5 and 64.7% in the 1980s, 1990s and 2000s, respectively). For an evaluation over 20 years, the cumulative pregnancy rates (SEM) were 0.4 (0.2), 2.8 (2.1), 4.0 (0.4) and 1.3 (0.4) for the LNG-IUS, the implants, copper IUD and DMPA, respectively and cumulative continuation rates (SEM) were 15.1 (3.7), 3.9 (1.4), 14.1 (0.6) and 7.3 (1.7) for the LNG-IUS, implants, copper IUD and DMPA, respectively (P < 0.001). Over the last 10 years of evaluation, the estimation of the contribution of the clinic through the provision of LARC methods and DMPA to DALY averted was 37–60 maternal deaths; between 315 and 424 child mortalities; combined maternal morbidity and mortality and child mortality of between 634 and 853, and 1056–1412 unsafe abortions averted.
LIMITATIONS, REASONS FOR CAUTION
The main limitations are the number of women who never returned to the clinic (overall 14% among the four methods under evaluation); consequently the pregnancy rate could be different. Other limitations include the analysis of two kinds of copper IUD and two kinds of contraceptive implants as the same IUD or implant, and the low number of users of implants. In addition, the DALY calculation relies on a number of estimates, which may vary in different parts of the world.
WIDER IMPLICATIONS OF THE FINDINGS
LARC methods and DMPA are highly effective and women who were well-counselled used these methods for a long time. The benefit of averting maternal morbidity and mortality, child mortality, and unsafe abortions is an example to health policy makers to implement more family planning programmes and to offer contraceptive methods, mainly LARC and DMPA, at no cost or at affordable cost for the underprivileged population.
STUDY FUNDING/COMPETING INTEREST(S)
This study received partial financial support from the Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP), grant # 2012/12810-4 and from the National Research Council (CNPq), grant #573747/2008-3. B.F.B., M.P.G., and V.M.C. were fellows from the scientific initiation programme from FAPESP. Since the year 2001, all the TCu380A IUD were donated by Injeflex, São Paulo, Brazil, and from the year 2006 all the LNG-IUS were donated by the International Contraceptive Access Foundation (ICA), Turku, Finland. Both donations are as unrestricted grants. The authors declare that there are no conflicts of interest associated with this study.
Aromatic plants are a remarkable source of natural products.
Paláu (Verbenaceae), commonly known as lemon verbena, is a relevant source of essential oils with potential applications due to its lemony ...scent and bioactive properties. Studies carried out on this species have focused on the volatile composition of the essential oil obtained by Clevenger hydrodistillation (CHD), with little information available on alternative extraction methodologies or the biological properties of the oil. Therefore, this work aimed to compare the volatile composition, antioxidant activity, cytotoxicity, anti-inflammatory and antibacterial activities of the essential oil extracted by conventional hydrodistillation by Clevenger (CHD) and Microwave-Assisted Hydrodistillation (MAHD). Significant differences (
< 0.05) were observed for some compounds, including the two major ones, geranial (18.7-21.1%) and neral (15.3-16.2%). Better antioxidant activity was exhibited by the MAHD essential oil in DPPH radical scavenging and reducing power assays, while no differences were observed in the cellular antioxidant assay. The MADH essential oil also presented higher inhibition against four tumoral cell lines and exhibited lower cytotoxicity in non-tumoral cells as compared with Clevenger-extracted essential oil. In contrast, the latter showed higher anti-inflammatory activity. Both essential oils were able to inhibit the growth of eleven out of the fifteen bacterial strains tested.
Animal venoms are large, complex libraries of bioactive, disulphide-rich peptides. These peptides, and their novel biological activities, are of increasing pharmacological and therapeutic importance. ...However, recombinant expression of venom peptides in Escherichia coli remains difficult due to the significant number of cysteine residues requiring effective post-translational processing. There is also an urgent need to develop high-throughput recombinant protocols applicable to the production of reticulated peptides to enable efficient screening of their drug potential. Here, a comprehensive study was developed to investigate how synthetic gene design, choice of fusion tag, compartment of expression, tag removal conditions and protease recognition site affect levels of solubility of oxidized venom peptides produced in E. coli.
The data revealed that expression of venom peptides imposes significant pressure on cysteine codon selection. DsbC was the best fusion tag for venom peptide expression, in particular when the fusion was directed to the bacterial periplasm. While the redox activity of DsbC was not essential to maximize expression of recombinant fusion proteins, redox activity did lead to higher levels of correctly folded target peptides. With the exception of proline, the canonical TEV protease recognition site tolerated all other residues at its C-terminus, confirming that no non-native residues, which might affect activity, need to be incorporated at the N-terminus of recombinant peptides for tag removal.
This study reveals that E. coli is a convenient heterologous host for the expression of soluble and functional venom peptides. Using the optimal construct design, a large and diverse range of animal venom peptides were produced in the µM scale. These results open up new possibilities for the high-throughput production of recombinant disulphide-rich peptides in E. coli.
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