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We evaluated the influence of the substitution of soybean meal by detoxified castor cake on performance, digestibility of nutrients, nitrogen balance, hepatic and renal functions of pregnant goats fed with diets containing detoxified castor cake by alkaline solutions during the stages (first two-thirds and final third) of pregnancy. Three diets were provided: one based on soybean meal and the other two based on castor cake detoxified with whit calcium hydroxide or sodium hydroxide. Goats fed detoxified castor cake sodium hydroxide had lower consumption. Was no effect (P>0.05) of diets or stages on the digestibility of dry matter and nutrients. The goats that received the diets based on soybean meal and detoxified castor cake calcium hydroxide consumed larger amounts of nitrogen. The goats fed with diet the basis of SM had greater weight in the parturition day. The average levels of enzymes for hepatic and renal functions were within normal patterns. Of enzymes related to liver metabolism, only the gamma-glutamyl transferase increased in the final third of pregnancy. The present study demonstrated that detoxified castor by sodium hydroxide reduces the consumption of goats during gestation, but did not affect negatively the renal and hepatic parameters. Made available in DSpace on 2022-10-28T20:08:48Z (GMT). No. of bitstreams: 1 cnpc-2022-Art36.pdf: 295666 bytes, checksum: 609c91cabd068bbcbe282eddad5dadc3 (MD5) Previous issue date: 2022

In collaboration with the American College of Veterinary Pathologists

Objectives This study aimed to evaluate the repair of critical-sized bone defects grafted with autogenous bone and mercerized bacterial cellulose membranes (BCm) salified with alendronate (ALN). Methods Forty-eight male Wistar rats underwent surgery to create a 5 mm-diameter bone defect in the calvarium. The removed bone was particularized, regrafted into the defect, and covered by a BCm according to the group: control group (CG), simply mercerized BCm; group 1 (G1), negatively charged BCm (BCm-CM-) salified with ALN; and group 2 (G2), positively charged BCm (BCm-DEAE+) salified with ALN. Serum samples were collected preoperatively and before euthanasia to analyze osteoprotegerin (OPG), parathyroid hormone (PTH), sclerostin (SOST), and fibroblast growth factor 23 (FGF23) levels. The animals were euthanized after 15 or 60 d. Calvaria were analyzed using quantitative microtomography (μCT). Results There was an increased level of PTH in the CG compared to the G2 group, at day 60 (p = 0.019). When analyzing the same group over time, G1 presented an increased FGF23 level on days 15 and 60 (p Conclusions Salification of ionized BCm with ALN had no direct effect on bone repair; however, BCm-CM- increased the levels of FGF23 over time. BCm-DEAE+ decreased PTH levels compared to mercerized BCm. BCm-CM-salified with ALN-induced superior bone quality, with respect to trabecular separation, compared to BCm-DEAE+.

The bioactivity of natural by-products in food and pharmaceutical applications is the subject of numerous studies. Cherry production and processing generates large amounts of biowaste, most of which is not used. The recovery of these by-products is essential for promoting the circular economy and to improving sustainability in the food industry. In this work, we explored the anti-inflammatory and antimicrobial potential of two different extracts from stems, leaves, and flowers of Portuguese cherries. The anti-inflammatory potential was studied on lipopolysaccharide (LPS)-stimulated mouse macrophages (RAW 264.7) by evaluating the effect of by-products on cellular viability and nitric oxide (NO) production. Disc diffusion and minimum inhibitory concentration (MIC) were used to determine antimicrobial activity. The cherry by-products had no cytotoxic effect on RAW 264.7 cells, and were able to inhibit nitrite production in a dose-dependent manner. Moreover, all aqueous infusions showed good antioxidant activity against NO radicals. Moreover, leaf extracts showed the best activity against most of the strains studied. The results revealed, for the first time, interesting anti-inflammatory and antimicrobial properties of cherry by-products. This could potentially be of interest for their therapeutic use in the treatment of inflammation-related diseases or in controlling the growth of microorganisms.

Regulation of inflammation is a critical process for maintaining physiological homeostasis. The λ-carrageenan (λ-CGN) is a mucopolysaccharide extracted from the cell wall of red algae (Chondrus crispus) capable of inducing acute intestinal inflammation, which is translated into the production of acute phase reactants secreted into the blood circulation. However, the associated mechanisms in vertebrates are not well understood. Here, we investigated the crucial factors behind the inflammatory milieu of λ-CGN-mediated inflammation administered at 0, 1.75, and 3.5% (v/w) by i.p. injection into the peritoneal cavity of adult zebrafish (ZF) (Danio rerio). We found that polymorphonuclear leukocytes (neutrophils) and lymphocytes infiltrating the ZF peritoneal cavity had short-term persistence. Nevertheless, they generate a strong pattern of inflammation that affects systemically and is enough to produce edema in the cavity. Consistent with these findings, cell infiltration, which causes notable tissue changes, resulted in the overexpression of several acute inflammatory markers at the protein level. Using reversed-phase high-performance liquid chromatography followed by a hybrid linear ion-trap mass spectrometry shotgun proteomic approach, we identified 2938 plasma proteins among the animals injected with PBS and 3.5% λ-CGN. First, the bioinformatic analysis revealed the composition of the plasma proteome. Interestingly, 72 commonly expressed proteins were recorded among the treated and control groups, but, surprisingly, 2830 novel proteins were differentially expressed exclusively in the λ-CGN-induced group. Furthermore, from the commonly expressed proteins, compared to the control group 62 proteins got a significant (p < 0.05) upregulation in the λ-CGN-treated group, while the remaining ten proteins were downregulated. Next, we obtained the major protein-protein interaction networks between hub protein clusters in the blood plasma of the λ-CGN induced group. Moreover, to understand the molecular underpinnings of these effects based on the unveiled protein sets, we performed a bioinformatic structural similarity analysis and generated overlapping 3D reconstructions between ZF and humans during acute inflammation. Biological pathway analysis pointed to the activation and abundance of diverse classical immune and acute phase reactants, several catalytic enzymes, and varied proteins supporting the immune response. Together, this information can be used for testing and finding novel pharmacological targets to treat human intestinal inflammatory diseases.

Elevational gradients along mountain slopes offer opportunities to study key factors shaping species diversity patterns. Several environmental factors change over short distances along the elevational gradient in predictable ways. However, different taxa respond to these factors differently resulting in various proposed models for biodiversity patterns along elevational transects. Using a multi-taxa approach, we investigated the effects of elevation, area, habitat and soil characteristics on species richness, individual abundance and species composition of six groups of ground-dwelling arthropods along four transect lines in the Swiss National Park and its surroundings (Eastern Alps). Spiders, millipedes, centipedes, ants, ground beetles and rove beetles were sampled using standardized methods (pitfall traps, cardboard traps, visual search) in 65 sites spanning an elevational range from 1800 to 2750 m a.s.l.. A total of 14,782 individuals comprising 248 species were collected (86 spider, 74 rove beetle, 34 ground beetle, 21 millipede, 19 centipede and 14 ant species). Linear mixed model-analysis revealed that rarefied species richness in five out of the six arthropod groups was affected by elevation (the quadratic term of elevation provided the best fit in most cases). We found three different patterns (linear decrease in centipedes, low elevation plateau followed by a decrease in ants and rove beetles, and midpoint peak in spiders and millipedes). These patterns were only partially mirrored when considering individual abundance. Elevation influenced species composition in all groups examined. Overall, elevation was the most important factor explaining the diversity patterns, while most local habitat and soil characteristics have little influence on these patterns. Our study supports the importance of using multi-taxa approaches when examining effects of elevational gradients. Considering only a single group may result in misleading findings for overall biodiversity.

Canine visceral leishmaniasis (CVL) is caused by Leishmania infantum and is endemic in many areas of southeastern Brazil. We have hypothesized that the prevalence of infection by L. infantum in dogs housed in open-admission animal shelters is beyond the range of 3.4 - 9.6% reported among dogs domiciled in similar CVL-endemic areas. Hence, this study aimed to determine the rate of L. infantum infection among dogs maintained in shelters and to investigate the epidemiology of CVL in such environments by analyzing hematological and biochemical parameters. A total of 627 dogs from 17 different shelters across the State of Minas Gerais were screened using the Dual-Path Platform test and enzyme-linked immunosorbent assay and 211 (33.6%) were found to be seropositive in both tests. Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) was performed on skin, bone marrow and lymphoid tissues of 118 seropositive dogs with inconclusive CVL clinical diagnosis and, of these, 78 (66.1%) were PCR+ for L. infantum and 7 (5.9%) were PCR+ for L. amazonensis. One dog presented a PCR-RFLP profile that was consistent with co-infection by both parasites. Leishmania amazonensis DNA was detected in skin samples of six single-infected dogs and this constitutes a novel finding. Dogs infected only with L. amazonensis were less debilitated than those infected by L. infantum, which showed typical clinical manifestations of CVL. The co-infected dog showed only mild clinical signs. The results presented herein not only support our original hypothesis but also suggest that dogs are potential reservoirs of L. amazonensis. Public health authorities should acknowledge their responsibility towards animals in collective shelters, recognize that they are potential foci of zoonotic diseases, and establish proper functioning directives to minimize transmission to humans and to other dogs.

Background The colonization of mosquitoes susceptible to Plasmodium vivax via direct membrane feeding assay (DMFA) has the potential to significantly advance our knowledge of P. vivax biology, vector-parasite interaction and transmission-blocking vaccine research. Anopheles darlingi and Anopheles deaneorum are important vectors of malaria in the Western Brazilian Amazon. Since 2018, well-established colonies of these species have been maintained in order to mass produce mosquitoes destined for P. vivax infection. Plasmodium susceptibility was confirmed when the colonies were established, but susceptibility needs to be maintained for these colonies to remain good models for pathogen transmission. Thus, the susceptibility was assessed of colonized mosquitoes to P. vivax isolates circulating in the Western Amazon. Methods Laboratory-reared mosquitoes from F10-F25 generations were fed on P. vivax blood isolates via DMFA. Susceptibility was determined by prevalence and intensity of infection as represented by oocyst load seven days after blood feeding, and sporozoite load 14 days after blood feeding. The effect of infection on mosquito survival was evaluated from initial blood feeding until sporogonic development and survival rates were compared between mosquitoes fed on infected and uninfected blood. Correlation was calculated between gametocytaemia and prevalence/intensity of infection, and between oocyst and sporozoite load. Results Significant differences were found in prevalence and intensity of infection between species. Anopheles darlingi showed a higher proportion of infected mosquitoes and higher oocyst and sporozoite intensity than An. deaneorum. Survival analysis showed that An. deaneorum survival decreased drastically until 14 days post infection (dpi). Plasmodium vivax infection decreased survival in both species relative to uninfected mosquitoes. No correlation was observed between gametocytaemia and prevalence/intensity of infection, but oocyst and sporozoite load had a moderate to strong correlation. Conclusions Colonized An. darlingi make excellent subjects for modelling pathogen transmission. On the other hand, An. deaneorum could serve as a model for immunity studies due the low susceptibility under current colonized conditions. In the application of DMFA, gametocyte density is not a reliable parameter for predicting mosquito infection by P. vivax, but oocyst intensity should be used to schedule sporozoite experiments.

Cherries have largely been investigated due to their high content in phenolics in order to fully explore their health-promoting properties. Therefore, this work aimed to assess, for the first time, the anti-inflammatory potential of phenolic-targeted fractions of the Saco cherry, using RAW 264.7 macrophages stimulated with lipopolysaccharide. Additionally, the cytotoxic effects on gastric adenocarcinoma (AGS), neuroblastoma (SH-SY5Y) and normal human dermal fibroblast (NHDF) cells were evaluated, as well as the ability to protect these cellular models against induced oxidative stress. The obtained data revealed that cherry fractions can interfere with cellular nitric oxide (NO) levels by capturing NO radicals and decreasing inducible nitric oxide synthase and cyclooxygenase-2 expression. Furthermore, it was observed that all cherry fractions exhibited dose-dependent cytotoxicity against AGS cells, presenting cytotoxic selectivity for these cancer cells when compared to SH-SY5Y and NHDF cells. Regarding their capacity to protect cancer cells against oxidative injury, in most assays, the total cherry extract was the most effective. Overall, this study reinforces the idea that sweet cherries can be incorporated into new pharmaceutical products, smart foods and nutraceuticals.

The males of many species of New World Phlebotomines produce volatile terpenoid chemicals, shown in Lutzomyia longipalpis s.l. to be sex/aggregation pheromones. Pheromone is produced by secretory cells which surround a cuticular reservoir which collects the pheromone and passes it through a cuticular duct to the surface of the insect. The pheromone then passes through specialised cuticular structures on the abdominal surface prior to evaporation. The shape and distribution of the specialised structures are highly diverse and differ according to species. In this study we used SEM to examine the interior cuticular pheromone collection and transport structures of 3 members of the Lu. longipalpis s.l. species complex and Migonemyia migonei. We found a new structure which we have called the manifold which appears to be a substantial extension of the interior tergal cuticle connected in-line with the cuticular duct and reservoir. The manifold of the Campo Grande member of the complex is longer and wider than the Jacobina member whereas the manifold of the Sobral member was shorter than both other members of the complex. Overall, the secretory apparatus of the Sobral member was smaller than the other two. The manifold of M. migonei was very different to those found in Lu. longipalpis s.l. and was positioned in a pit-like structure within the tergal cuticle. The secretory reservoir was connected by a short duct to the manifold. Differences in the size and shape of the manifold may be related to the chemical structure of the pheromone and may have taxonomic value. Examination of the interior cuticle by SEM may help to locate the secretory apparatus of vector species where pheromonal activity has been inferred from behavioural studies but the external secretory structures or pheromones have not yet been found., Author summary The males of many species of New World sand flies produce volatile chemicals shown in Lutzomyia longipalpis s.l. to be attractive to females and other males. These sex/aggregation pheromones are produced by secretory cells which surround a cuticular reservoir that collects the pheromone and from which it passes through a cuticular duct and a specialised structure on the surface of the abdomen, from where it evaporates. We examined by SEM the structure of the interior cuticular secretory apparatus of 3 members of the Lutzomyia longipalpis s.l. species complex and Migonemyia migonei and found a new structure associated with pheromone release that we have called the manifold. The manifold appears to be a substantial extension of the interior tergal cuticle that is connected in-line with the duct and reservoir. Significant differences in size and shape of the manifold between members of the Lu. longipalpis complex were observed. The M. migonei secretory apparatus were substantially different in appearance to those of Lu. longipalpis with the manifold positioned in a pit-like structure within the tergal cuticle. These differences which may have taxonomic value and may be related to the chemical structure of the pheromone or phylogenetic differences.

In the present study, we have modified the traditional May-Holling-Tanner predator-prey model used to represent the interaction between least-weasel and field-vole population by adding an Allee effect (strong and weak) on the field-vole population and alternative food source for the weasel population. It is shown that the dynamic is different from the original May-Holling-Tanner predator-prey interaction since new equilibrium points have appeared in the first quadrant. Moreover, the modified model allows the extinction of both species when the Allee effect (strong and weak) on the prey is included, while the inclusion of the alternative food source for the predator shows that the system can support the coexistence of the populations, extinction of the prey and coexistence and oscillation of the populations at the same time. Furthermore, we use numerical simulations to illustrate the impact that changing the predation rate and the predator intrinsic growth rate have on the basin of attraction of the stable equilibrium point or stable limit cycle in the first quadrant. These simulations show the stabilisation of predator and prey populations and/or the oscillation of these two species over time.

Summary Background Cholesterol microbial transformation has been widely studied using in vitro fermentation assays, but less information is available on the biotransformation of plant sterols (PS). The excretion percentage of animal sterols (AS) (67–73%) is considerably greater than that of PS (27–33%) in feces from healthy humans following a Western diet. However, a lower content of AS in feces from subjects following a vegetarian, vegan or low-fat animal diet has been seen when compared to omnivorous subjects. Although only one human study has reported fecal sterol excretion after the consumption of PS-enriched food (8.6 g PS/day), it was found that the target group showed an increase in the excretion of cholesterol and a 57% decrease in its metabolites compared to the control group. Objective Evaluation of the impact of a PS-enriched milk based fruit beverage intake on fecal sterol excretion and the microbial conversion of sterols in postmenopausal women with mild hypercholesterolemia. Methods Forty postmenopausal women participated in a randomized, double-blind, crossover study with two beverages, with a PS-enriched (2 g PS/day) or without. The women were divided in two groups: 20 women consumed the PS-enriched beverage and the other 20 women consumed a placebo (without PS) beverage for 6 weeks. After a four-week washout period, the type of beverage was exchanged and consumed for another 6 weeks. Feces were collected at the start (0 and 10 weeks) and end of each intervention period (6 and 16 weeks), and fecal sterols were determined by capillary gas chromatography with mass spectrometry. Results The intake of the PS-enriched beverage modified the fecal sterol excretion profile. A significant increase mainly in PS and their metabolites versus the placebo intervention period was observed. Although the same effect was not observed in the case of AS, a tendency towards increased cholesterol and decreased coprostanol (the main metabolite of cholesterol) was recorded after PS-enriched beverage intake versus placebo. Furthermore, the PS-enriched beverage also modified the microbial conversion of sterols. In this context, an important decrease in the conversion percentage of cholesterol in 16 women (between 11% and 50%) and of sitosterol in 24 women (between 15% and 61%) was observed. Conclusions The results obtained suggest that the microbiota could preferably use PS as a substrate, when present in a greater proportion compared with cholesterol. Besides, a lower sitosterol and cholesterol conversion trend would mean that intake of the PS-enriched beverage could modulate the metabolic activity of the gut microbiota. Therefore, further studies on the impact of PS-enriched foods upon gut microbiota modulation are needed. Clinical Trial Registry Number: NCT 02065024 listed on the NIH website: ClinicalTrials.gov . Clinical Trial Registry Name: Food Matrix and Genetic Variability as Determinants of Bioavailability and Biological Effects of Beta-cryptoxanthin and Phytosterols (foodmagenpol). The full trial protocol is available upon request to the corresponding author.

15 pags, 7 figs, 2 tabs, Parkinson's disease (PD) is a neurodegenerative disorder characterized by fibrillar neuronal inclusions composed of aggregated α-synuclein (α-syn). These inclusions are associated with behavioral and pathological PD phenotypes. One strategy for therapeutic interventions is to prevent the formation of these inclusions to halt disease progression. α-Synuclein exists in multiple structural forms, including disordered, nonamyloid oligomers, ordered amyloid oligomers, and fibrils. It is critical to understand which conformers contribute to specific PD phenotypes. Here, we utilized a mouse model to explore the pathological effects of stable β-amyloid-sheet oligomers compared with those of fibrillar α-synuclein. We biophysically characterized these species with transmission EM, atomic-force microscopy, CD spectroscopy, FTIR spectroscopy, analytical ultracentrifugation, and thioflavin T assays. We then injected these different α-synuclein forms into the mouse striatum to determine their ability to induce PD-related phenotypes. Wefound that β-sheet oligomers produce a small but significant loss of dopamine neurons in the substantia nigra pars compacta (SNc). Injection of small β-sheet fibril fragments, however, produced the most robust phenotypes, including reduction of striatal dopamine terminals, SNc loss of dopamine neurons, and motor-behavior defects. Weconclude that although the β-sheet oligomers cause some toxicity, the potent effects of the short fibrillar fragments can be attributed to their ability to recruit monomeric α-synuclein and spread in vivo and hence contribute to the development of PD-like phenotypes. These results suggest that strategies to reduce the formation and propagation of β-sheet fibrillar species could be an important route for therapeutic intervention in PD and related disorders., This work was supported in part by the Michael J. Fox Foundation (to L.V.-D. and N.C.) and Grant P50NS108675 (Alabama Udall Center). The authors declare that they have no conflicts of interest with the contents of this article.

Background PCV3 is a member of the Circovirus family, associated with disease and mortality in pigs. It is not clear whether PCV3 putatively causes clinical symptoms and disease. In the present case, we reported a gilt infected with PCV3 associated with reproductive failures, vertical transmission, tissue lesions, viral replication by in situ hybridization, and the hypothesis that some strains of PCV3 clade one are associated with reproductive failures at the field level. Case presentation In May 2019, a pig farm in Colombia reported increased reproductive failures, and the presence of PCV3 in gilts and sows was established in a single form or coinfections, mainly with PCV2 and PPV7. Ten sows with a single infection with PCV3 were found, and one gilt with a pre-farrowing serum viral load above 103 was studied. This gilt was followed up during the pre-farrowing, farrowing period and on her litter for 6 weeks. During dystocic farrowing, a mummy and ten piglets were released, including two weak-born piglets. The highest viral loads for PCV3 were found in the mummy and the placenta. In the weak-born piglets, there were viral loads both in serum and in tissues, mainly in the mesenteric ganglia and lung. Replication of PCV3 in these tissues was demonstrated by in situ hybridizations. PCV3 was also found in the precolostrum sera of piglets and colostrum, showing vertical transmission. The viral load in piglets decreased gradually until week six of life. The viral genome’s complete sequencing was made from the mummy, and its analysis classified it as PCV3 clade one. Conclusions This report confirms that PCV3 can cause disease at the field level, and putatively, in this case, we find the generation of reproductive failures. The ability of PCV3 to cause disease as a putative pathogen may be associated with the viral load present in the pig and the strain that is affecting the farm. For this case, we found that viral loads above 103 (4.93 log genomic copies / mL) in the gilt were associated with clinical manifestation and that some PCV3 strains belonging to clade one are more associated with the reproductive presentation.

This paper is primarily a compilation of published data on the staphylinid beetle species reported for El Salvador. It is based on taxonomic and ecological literature, with inclusion of some new records from different entomological collections. Thirty eight genera and 96 species are reported in the list, including a first country record: Eulissus chalybaeus. Country-level distribution outside of El Salvador, locality references and topographic zones are included for each species. In the list, three endemic species are recorded: Seeversiella badia, S. minima and Stenus salvadorensis. It is remarkable that 14 genera (37%) and 52 species (54%) have only been reported at a country level in the literature, without a specific locality of collection, indicating the need for local inventories of this coleopteran family in the salvadoran territory.

The hemolymph and insect gut together have an essential role in the immune defense against microorganisms, including the production of antimicrobial peptides (AMP). AMPs are mainly induced by two specific signaling pathways, Toll and immune deficiency (IMD). Here, we characterize the expression profile of four genes from both pathways and describe the importance of AgraRelish in the immune defense of Anthonomus grandis against the entomopathogenic fungus Metarhizium anisopliae by RNA interference (RNAi).To characterize the pathway that is activated early during the A. grandis-M. anisopliae interaction, we assessed the expression profiles of AgraMyD88 and AgraDorsal (Toll pathway), AgraIMD and AgraRelish (IMD pathway), and several AMP genes. Interestingly, we found that IMD pathway genes are upregulated early, and Toll pathway genes are upregulated just 3 days after inoculation (DAI). Furthermore, nine AMPs were upregulated 24 h after fungus inoculation, including attacins, cecropins, coleoptericins, and defensins. AgraRelish knockdown resulted in a reduction in median lethal time (LTOur data indicate the influence of the IMD pathway on the antifungal response in A. grandis. Combining biocontrol and RNAi could significantly improve cotton boll weevil management. Hence, AgraRelish is a potential target for the development of biotechnological tools aimed at improving the efficacy of M. anisopliae against A. grandis.

Species assemblages often have a non-random nested organization, which in vertebrate scavenger (carrion-consuming) assemblages is thought to be driven by facilitation in competitive environments. However, not all scavenger species play the same role in maintaining assemblage structure, as some species are obligate scavengers (i.e., vultures) and others are facultative, scavenging opportunistically. We used a database with 177 vertebrate scavenger species from 53 assemblages in 22 countries across five continents to identify which functional traits of scavenger species are key to maintaining the scavenging network structure. We used network analyses to relate ten traits hypothesized to affect assemblage structure with the “role” of each species in the scavenging assemblage in which it appeared. We characterized the role of a species in terms of both the proportion of monitored carcasses on which that species scavenged, or scavenging breadth (i.e., the species “normalized degree”), and the role of that species in the nested structure of the assemblage (i.e., the species “paired nested degree”), therefore identifying possible facilitative interactions among species. We found that species with high olfactory acuity, social foragers, and obligate scavengers had the widest scavenging breadth. We also found that social foragers had a large paired nested degree in scavenger assemblages, probably because their presence is easier to detect by other species to signal carcass occurrence. Our study highlights differences in the functional roles of scavenger species and can be used to identify key species for targeted conservation to maintain the ecological function of scavenger assemblages., ESG, ZMR, JMB and LNA were supported by Generalitat Valenciana (SEJI/2018/024, APOSTD/2019/016, CIDEGENT/2020/030 and ACIF/2019/056, respectively), JMB, JMPG and CGC by Juan de la Cierva contracts (MEC; IJCI-2017-32149, FJCI-2015-25632 and IJC2018-036642-I, respectively), ACA by the Govern de les Illes Balears (PD/039/2017) and ESG and MM by Ramón y Cajal contracts (MEC; RYC-2019-027216-I, RYC-2015-19231). EA was supported by La Caixa-Severo Ochoa International PhD Program 2015. NS was partly supported by the National Science Centre in Poland (2013/08/M/NZ9/00469 and 2016/22/Z/NZ8/00). MK and KJ were supported by the Slovenian Research Agency (P4-0059). Contributions of HG, KLT, EFA, OER, TLD, and JCB were partially supported through funding from U.S. Department of Agriculture and the U.S. Department of Energy under (DE-EM0004391) to the University of Georgia Research Foundation. HG was also supported by the Institute of Environmental Radioactivity at Fukushima University. ALB and JDA were partially supported by Queens College and the Graduate Center at the City University of New York. JDA is currently supported by a Ramón y Cajal contract (RYC-2017-22783). ERB and EF were supported by the USA National Science Foundation Graduate Research Fellowship (1256065). CK completed study with support from Hawk Mountain Sanctuary, The Peregrine Fund, and via Pompeo M. Maresi Memorial Fund via Princeton University. JAS and CCW were supported by the USA National Science Foundation #1255913, the American Association for University Women, and the Gordon and Betty Moore Foundation. HUW acknowledges funding from the California Department of Fish & Wildlife (P0880013). PLP was supported by the Rufford Foundation and University of Wisconsin-Madison. JB and JdT thank the Percy Sladen Memorial Fund and Mr Rodney Fuhr. Several authors were funded by funds from the MEC (CGL2012-40013-C02-01/02, CGL2015-66966-C2-1-R, CGL2015-66966-C2-1-R2, CGL2017-89905-R, RTI2018-099609-B-C21 and RTI2018-099609-B-C22) and from the Junta de Andalucía (RNM-1925). POV was supported by a research contract by the University of Lleida. EES and TMN were funded and supported by Australian Geographic, Bush Heritage Australia, Australian Academy of Sciences, Ecological Society of Australia, NSW Office of Environment and Heritage, NESP Threatened Species Recovery Hub, Emirates Wolgan Valley One and Only Resort. EP, YA, MS and UY completed study under research permit by The Republic of Turkey Ministry of Agriculture and Forestry. SAL thanks PICT (BID) 0725/2014, and IAATE. ALB and JDA would like to thank the Edmund Niles Huyck Preserve, Lyme Adirondack Forest Company, and LandVest Timberland.

Graphical abstract, Highlights • 320 Argentinian Fasciola hepatica were genotyped using a panel of microsatellites. • Overall there was high genotypic richness: 263 distinct genotypes were identified. • Population structuring of F. hepatica was evident across Argentina. • Within these sub-populations there is largely random mating. • Transmission of clonemates occurs: clonal parasites accounted for 26.6% of all parasites., Fasciola hepatica, the liver fluke, is a trematode parasite that causes disease of economic importance in livestock. As a zoonosis this parasite also poses a risk to human health in areas where it is endemic. Population genetic studies can reveal the mechanisms responsible for genetic structuring (non-panmixia) within parasite populations and provide valuable insights into population dynamics, which in turn enables theoretical predictions of evolutionary dynamics such as the evolution of drug resistance. Here we genotyped 320 F. hepatica collected from 14 definitive hosts from four provinces in Argentina. STRUCTURE analysis indicated three population clusters, and principal coordinate analysis confirmed this, showing population clustering across provinces. Similarly, pairwise FST values amongst all four provinces were significant, with standardised pairwise FST (F′ST) ranging from 0.0754 to 0.6327. Therefore, population genetic structure was evident across these four provinces in Argentina. However, there was no evidence of deviation from Hardy–Weinberg equilibrium, so it appears that within these sub-populations there is largely random mating. We identified 263 unique genotypes, which gave a clonal diversity of 82%. Parasites with identical genotypes, clones, accounted for 26.6% of the parasites studied and were found in 12 of the 14 hosts studied, suggesting some clonemate transmission.

The troglobiont millipede, Cantabrodesmus lorioli Mauriès, 1971, inhabits a few caves in a small region of Northern Spain. It is considered a relict species because of its geographical isolation compared to others in its family, and its to some extent uncertain systematic position. Cantabrodesmus lorioli has been placed in three different families, and now is considered a member of the Chelodesmidae, a family with only one other known native European representative. There are few records of this species, and almost nothing is known about its biology. The only available information on this species is the description of the male and a list of caves where it has been captured. In the present work, we provide the first description of the female morphology, illustrated by scanning electron microscopy, and a series of new records from caves in a region where it had never been captured before, including the deepest known pit in Spain, Torca del Porrón. The female morphology does not show many differences from that of the male, but several characters are discussed. All of the known information on the morphology of the species is used to discuss the implications for its biology and systematic position.

27 pags., 4 figs., α-Synuclein amyloid aggregation is a defining molecular feature of Parkinson’s disease, Lewy body dementia, and multiple system atrophy, but can also be found in other neurodegenerative disorders such as Alzheimer’s disease. The process of α-synuclein aggregation can be initiated through alternative nucleation mechanisms and dominated by different secondary processes giving rise to multiple amyloid polymorphs and intermediate species. Some aggregated species have more inherent abilities to induce cellular stress and toxicity, while others seem to be more potent in propagating neurodegeneration. The preference for particular types of polymorphs depends on the solution conditions and the cellular microenvironment that the protein encounters, which is likely related to the distinct cellular locations of α-synuclein inclusions in different synucleinopathies, and the existence of disease-specific amyloid polymorphs. In this review, we discuss our current understanding on the nature and structure of the various types of α-synuclein aggregated species and their possible roles in pathology. Precisely defining these distinct α-synuclein species will contribute to understanding the molecular origins of these disorders, developing accurate diagnoses, and designing effective therapeutic interventions for these highly debilitating neurodegenerative diseases., This research was funded by the Spanish Ministry of Economy, Industry and Competitiveness (MINECO), grant number BFU2015-64119-P (N.C.), the Spanish Ministry of Science, Innovation and Universities (MICIU) and the European Commission (FEDER), grant number PGC2018-096335-B-100 (N.C.), and the National Institute of Neurological Disorders and Stroke (NINDS) of the National Health Institute (NIH), grant number R01NS102257) (L.V.-D.).

Isoliquiritigenin (IsoLQ) is a flavonoid with antioxidant properties and inducer of endoplasmic reticulum (ER) stress. In vitro and in vivo studies show that ER stress-mediated hormesis is cytoprotective, therefore, natural antioxidants and ER stress inducers have been used to prevent renal injury. Oxidative stress and ER stress are some of the mechanisms of damage involved in cisplatin (CP)-induced nephrotoxicity. This study aims to explore whether IsoLQ pretreatment induces ER stress and produces hormesis to protect against CP-induced nephrotoxicity in Lilly Laboratories Cell-Porcine Kidney 1 (LLC-PK1) cells. During the first stage of this study, both IsoLQ protective concentration and pretreatment time against CP-induced toxicity were determined by cell viability. At the second stage, the effect of IsoLQ pretreatment on cell viability, ER stress, and oxidative stress were evaluated. IsoLQ pretreatment in CP-treated cells induces expression of glucose-related proteins 78 and 94 kDa (GRP78 and GRP94, respectively), attenuates CP-induced cell death, decreases reactive oxygen species (ROS) production, and prevents the decrease in glutathione/glutathione disulfide (GSH/GSSG) ratio, free thiols levels, and glutathione reductase (GR) activity. These data suggest that IsoLQ pretreatment has a moderately protective effect on CP-induced toxicity in LLC-PK1 cells, through ER stress-mediated hormesis, as well as by the antioxidant properties of IsoLQ.

Understanding the distribution of biodiversity across the Earth is one of the most challenging questions in biology. Much research has been directed at explaining the species latitudinal pattern showing that communities are richer in tropical areas; however, despite decades of research, a general consensus has not yet emerged. In addition, global biodiversity patterns are being rapidly altered by human activities. Here, we aim to describe large-scale patterns of species richness and diversity in terrestrial vertebrate scavenger (carrion-consuming) assemblages, which provide key ecosystem functions and services. We used a worldwide dataset comprising 43 sites, where vertebrate scavenger assemblages were identified using 2,485 carcasses monitored between 1991 and 2018. First, we evaluated how scavenger richness (number of species) and diversity (Shannon diversity index) varied among seasons (cold vs. warm, wet vs. dry). Then, we studied the potential effects of human impact and a set of macroecological variables related to climatic conditions on the scavenger assemblages. Vertebrate scavenger richness ranged from species-poor to species rich assemblages (4–30 species). Both scavenger richness and diversity also showed some seasonal variation. However, in general, climatic variables did not drive latitudinal patterns, as scavenger richness and diversity were not affected by temperature or rainfall. Rainfall seasonality slightly increased the number of species in the community, but its effect was weak. Instead, the human impact index included in our study was the main predictor of scavenger richness. Scavenger assemblages in highly human-impacted areas sustained the smallest number of scavenger species, suggesting human activity may be overriding other macroecological processes in shaping scavenger communities. Our results highlight the effect of human impact at a global scale. As species-rich assemblages tend to be more functional, we warn about possible reductions in ecosystem functions and the services provided by scavengers in human-dominated landscapes in the Anthropocene., SG, JMPG, and ACA were supported by Juan de la Cierva contracts (Ministerio de Economía y Competitividad, MEC; IJCI‐2015‐24947, FJCI‐2015‐25632, and IJCI‐2014‐20744, respectively). ESG was also supported by Generalitat Valenciana (SEJI/2018/024), ACA by the Govern de les Illes Balears (PD/039/2017), and MM by a Ramón y Cajal contract (MEC; RYC‐2015‐19231). EA was supported by La Caixa‐Severo Ochoa International PhD Program 2015. NS was partly supported by the National Science Centre in Poland (2013/08/M/NZ9/00469). SAL thanks PICT (BID) 0725/2014. MK and KJ were supported by the Slovenian Research Agency (P4‐0059). Contributions of KT and JCB were partially supported through funding from U.S. Department of Agriculture, the U.S. Department of Energy under (DE‐EM0004391) to the University of Georgia Research Foundation. EB and EF were supported by the USA National Science Foundation. CK completed study under research permit NCST/5/002/R/448 with support from Hawk Mountain Sanctuary, The Peregrine Fund, and via Pompeo M. Maresi Memorial Fund via Princeton University. JAS and CCW were supported by the USA National Science Foundation #1255913 and the Gordon and Betty Moore Foundation. Several authors were funded by funds from the MEC (CGL2012‐40013‐C02‐01/02, CGL2015‐66966‐C2‐1‐R and CGL2017‐89905‐R) and from the Junta de Andalucía (RNM‐1925).

In this work, dextran based membranes with potential to be used as implantable devices in Tissue Engineering and Regenerative Medicine (TERM) were prepared by a straightforward strategy. Briefly, two polymers approved by the Food and Drug Administration, viz. dextran and poly(e-caprolactone) (PCL) were functionalized with methacrylate moieties, and subjected to photocrosslinking. Employing different weight ratios of each polymer in the formulations allowed to obtain transparent membranes with tunable physicochemical properties and low adverse host tissue response. Independently of the material, all formulations have shown to be thermally stable up to 300 °C whilst variations in the polymer ratio resulted in membranes with different glass transition temperatures (Tg) and flexibility. The swelling capacity ranged from 50% to 200%. On the other hand, in vitro hydrolytic degradation did not show to be material-dependent and all membranes maintained their structural integrity for more than 30 days, losing only 8–12% of their initial weight. Preliminary in vitro biological tests did not show any cytotoxic effect on seeded human dental pulp stem cells (hDPSCs), suggesting that, in general, all membranes are capable of supporting cell adhesion and viability. The in vivo biocompatibility of membranes implanted subcutaneously in rats’ dorsum indicate that M100/0 (100%wt dextran) and M25/75 (25 %wt dextran) formulations can be classified as “slight-irritant” and “non-irritant”, respectively. From the histological analysis performed on the main tissue organs it was not possible to detect any signs of fibrosis or necrosis thereby excluding the presence of toxic degradation by-products deposited or accumulated in these tissues. In combination, these results suggest that the newly developed formulations hold great potential as engineered devices for biomedical applications, where the biological response of cells and tissues are greatly dependent on the physical and chemical cues provided by the substrate.

In spite of the crucial role of the inflammatory state under anemic conditions, to date, no studies have directly tested the modulation of cytokines during iron overload. The aim of this work was to contribute to a better understanding of the pathophysiology and recovery from iron deficiency, by studying how fermented goat milk consumption affects inflammatory signalling during iron repletion. Eighty male Wistar rats were used for a pre-experimental period of 40 days, by dividing them into two groups (the control group receiving a normal-Fe diet and the Fe-deficient group receiving a low-Fe diet). Later, the rats were fed with a fermented goat or cow milk-based diet, with a normal-Fe content or Fe-overload (450 mg kg-1) for 30 days. After feeding the fermented milk, the anti-inflammatory and pro-inflammatory cytokines were assessed. The anti-inflammatory cytokines (IL-13, IL-10 and IL-4) were higher in both groups of animals (control and anemic) fed fermented goat milk either with normal Fe or Fe-overload with respect to the fermented cow milk. With regard to pro-inflammatory signalling, fermented goat milk consumption decreased the pro-inflammatory cytokines (IL-2, TNF-α, IL-1β, IL-12p70 and IP-10). The Fe overload increased the anti-inflammatory cytokines together with IL-1β and IP-10. Fermented goat milk consumption improves the hematological status and promotes the beneficial metabolic responses related to the inflammatory signaling in nutritional ferropenic anemia recovery, which may be a dietary strategy to lessen the evoked inflammation during iron repletion. Additionally, the parameters of inflammation should therefore be incorporated as routine biomarkers of iron deficiency or overload severity.

Tissue engineering is constantly evolving towards novel materials that mimic the properties of the replaced injured tissue or organ. A hybrid electrospun membrane of electroactive poly(l-acid lactic) (PLLA) polymer with glass reinforced hydroxyapatite (Bonelike®) microparticles placed among the polymer fibres in a morphology like "islands in the sea" was processed. The incorporation of 60 to 80wt% Bonelike® bone grafts granules with ≤150μm into the polymer solution lead to an amorphous polymeric fibre membranes, and a decrease of the average polymer fibre diameter from 550±150nm for neat PLA down to 440±170nm for the hybrid composite. The presence of Bonelike® in the polymer mats reduced the activation energy for thermal degradation from 134kJ·mol-1, obtained for the neat PLLA membranes down to 71kJ·mol-1, calculated for the hybrid composite membranes. In vitro cell culture results suggest that the developed processing method does not induce cytotoxic effects in MG 63 osteoblastic cells, and creates an environment that enhances cell proliferation, when compared to the neat PLLA membrane. The simplicity and scalability of the processing method suggests a large application potential of this novel hybrid polymer-microparticles fibre membranes for bone regenerative medicine.

16 pags., 6 figs., The self-assembly of α-synuclein (αS) into intraneuronal inclusion bodies is a key characteristic of Parkinson’s disease. To define the nature of the species giving rise to neuronal damage, we have investigated the mechanism of action of the main αS populations that have been observed to form progressively during fibril growth. The αS fibrils release soluble prefibrillar oligomeric species with cross-β structure and solvent-exposed hydrophobic clusters. αS prefibrillar oligomers are efficient in crossing and permeabilize neuronal membranes, causing cellular insults. Short fibrils are more neurotoxic than long fibrils due to the higher proportion of fibrillar ends, resulting in a rapid release of oligomers. The kinetics of released αS oligomers match the observed kinetics of toxicity in cellular systems. In addition to previous evidence that αS fibrils can spread in different brain areas, our in vitro results reveal that αS fibrils can also release oligomeric species responsible for an immediate dysfunction of the neurons in the vicinity of these species., This research was supported by the University of Florence (Fondi Ateneo to F.C. and C.C.), the Ministry of Education, Universities and Research of Italy (Progetto Dipartimento di Eccellenza “Gender Medicine” to C.C.), Parkinson’s UK (G-1508 to S.W.C. and C.M.D.), the Center for Misfolding Diseases of the University of Cambridge (S.W.C. and C.M.D.), the UK Medical Research Council (MR/N000676/1 to C.M.D.), the Agency of Science, Technology and Research of Singapore (to S.W.C.), and the Ministry of Economy and Competitiveness of Spain (MINECO RYC-2012-12068 and MINECO/FEDER EU BFU2015-64119-P to N.C.).

Any bioassay to test new chemically synthesized larvicides or phytolarvicides against Culicidae and more harmful mosquito species, such as Aedes aegypti and Aedes albopictus, which specifically transmit dengue, yellow fever, chikungunya viral fevers as well as Zika virus, or Anopheles gambiae, a vector for malaria and philariasis, requires thousands of well-developed larvae, preferably at the fourth instar stage. The natural morphogenetic cycle of Aedes spp., in the field or in the laboratory, may extend to 19 days at room temperature (e.g., 25°C) from the first permanent contact between viable eggs and water and the last stage of larval growth or metamorphosis into flying adults. Thus, accelerated sequential molting is desirable for swifter bioassays of larvicides. We achieved this goal in Aedes aegypti with very limited strategic and low-cost additions to food, such as coconut water, milk or its casein, yeast extract, and to a lesser extent, glycerol. The naturally rich coconut water was excellent for quickly attaining the population of instar IV larvae, the most advanced one before pupation, saving about a week, for subsequent larvicidal bioassays. Diluted milk, as another food source, allowed an even faster final ecdysis and adults are useful for mosquito taxonomical purpose.

In this work, dextran-based nerve tube-guides were prepared, characterized and used in a standardized animal model of neurotmesis injury. Non-porous and porous transparent tube-guides were obtained by photocrosslinking of two co-macromonomers based on dextran and poly(e-caprolactone) (PCL). Swelling capacity of the tube-guides ranged from 40–60% with no visible constriction of their inner diameter. In vitro hydrolytic degradation tests showed that the tube-guides maintained their structural integrity up to 6 months. The in vivo performance of the tube-guides was evaluated by entubulation of the rat sciatic nerve after a neurotmesis injury, with a 10 mm-gap between the nerve stumps. The results showed that the tube-guides were able to promote the regeneration of the nerve in a similar manner to what was observed with conventional techniques (nerve graft and end-to-end suture). Stereological analysis proved that nerve regeneration occurred, and both tube-guides presented fibre diameter and g-ratio closer to healthy sciatic nerves. The histomorphometric analysis of Tibialis anterior (TA) skeletal muscle showed decreased neurogenic atrophy in the porous tube-guides treated group, presenting measurements that are similar to the uninjured control.

Disruption of the continuous cutaneous membrane in the integumentary system is considered a health problem of high cost for any nation. Several attempts have been made for developing skin substitutes in order to restore injured tissue including autologous implants and the use of scaffolds based on synthetic and natural materials. Current biomaterials used for skin tissue repair include several scaffold matrices types, synthetic or natural, absorbable, degradable or non-degradable polymers, porous or dense scaffolds, and cells capsulated in hydrogels or spheroids systems so forth. These materials have advantages and disadvantages and its use will depend on the desired application. Recently, marine organisms such as jellyfish have attracted renewed interest, because both its composition and structure resemble the architecture of human dermic tissue. In this context, the present study aims to generate scaffolds from Cassiopea andromeda (C. andromeda), with application in skin tissue engineering, using a decellularization process. The obtained scaffold was studied by infrared spectroscopy (FT-IR), thermogravimetric analysis (TGA), differential scanning calorimetry analysis (DSC), and scanning electron microscopy (SEM). Crystal violet staining and DNA quantification assessed decellularization effectiveness while the biocompatibility of scaffold was determined with human dermic fibroblasts. Results indicated that the decellularization process reduce native cell population leading to 70% reduction in DNA content. In addition, SEM showed that the macro and microstructure of the collagen I-based scaffold were preserved allowing good adhesion and proliferation of human dermic fibroblasts. The C. andromeda scaffold mimics human skin and therefore represents great potential for skin tissue engineering.

Seven cobalt(II) complexes of pyrazole derivatives and dinitrobenzoate ligands were synthesized and characterized. The single-crystal X-ray diffraction structure was determined for one of the ligands and one of the complexes. The analysis and spectral data showed that all the cobalt complexes had octahedral geometries, which was supported by DFT calculations. The complexes and their free ligands were evaluated against fungal strains of Candida albicans and emerging non-albicans species and epimastigotes of Trypanosoma cruzi. We obtained antifungal activity with a minimum inhibitory concentration (MIC) ranging from 31.3 to 250 &micro, g mL&minus, 1. The complexes were more active against C. krusei, showing MIC values between 31.25 and 62.5 &micro, 1. In addition, some ligands (L1&ndash, L6) and complexes (5 and Co(OAc)2 &middot, 4H2O) significantly reduced the yeast to hypha transition of C. albicans at 500 &micro, 1 (inhibition ranging from 30 to 54%). Finally, the complexes and ligands did not present trypanocidal activity and were not toxic to Vero cells. Our results suggest that complexes of cobalt(II) with ligands derived from pyrazoles and dinitrobenzoate may be an attractive alternative for the treatment of diseases caused by fungi, especially because they target one of the most important virulence factors of C. albicans.

The worldwide incidence of bone disorders is raising, mainly due to ageing population. The lack of effective treatments is pushing the development of synthetic bone substitutes (SBSs). Most ceramic-based SBSs commercially available display limited handling properties. Attempting to solve these issues and achieve wider acceptance by the clinicians, granular ceramics have been associated with hydrogels to produce injectable/moldable SBSs. Dextrin, a low-molecular-weight carbohydrate, was used to develop a fully resorbable and injectable hydrogel. It was firstly oxidized with sodium periodate and then cross-linked with adipic acid dihydrazide. The in vivo biocompatibility and safety of the dextrin-based hydrogel (HG) was assessed by subacute systemic toxicity and skin sensitization tests, using rodent models. The results showed that the HG did not induce any systemic toxic effect, skin reaction or genotoxicity, neither impaired the bone repair/regeneration process. Then, the HG was successfully combined with granular bone substitute, registered as Bonelike® (250-500 ?m) to obtain a mouldable/injectable SBS, which was implanted in tibial fractures in goats for 3 and 6 weeks. The obtained results showed that HG allowed the stabilization of the granules into the defect, ensuring effective handling and moulding properties of the formulation, as well as an efficient cohesion of the granules. This article is protected by copyright. All rights reserved., Isabel Pereira was supported by the grant SFRH/BD/ 90066/ 2012 from FCT, Portugal. This work was funded by the project “DEXGELERATION – Advanced solutions for bone regeneration based on dextrin hydrogels” (Norte-07-0202-FEDER-038853) and the project “iBone Therapies – innovative therapies for bone regeneration” (NORTE-01-0247-FEDER-003262). The authors acknowledge the funding from FCT under the scope of the strategic funding of UID/BIO/04469/2013 and UID/BIM/04293/2013 units and COMPETE 2020 (POCI-010145-FEDER-006684), BioTecNorte operation (NORTE-010145-FEDER-000004) and NORTE-01-0145-FEDER-000012 funded by FEDER under the scope of Norte2020—Programa Operacional Regional do Norte., info:eu-repo/semantics/publishedVersion

Marine Protected Areas (MPAs) help replenish fish assemblages, though different trophic levels may show diverse recovery patterns. Long-term protection is required to achieve total recovery but poaching events may prevent the achievement of full carrying capacity. Here, we have analysed the effect of long-term protection on the entire reef fish community and the different trophic levels in the Cabo de Palos-Islas Hormigas MPA (SE Spain; SW Mediterranean Sea) in order to assess their recovery patterns after 23 years of protection. We compared the values for carrying capacity obtained with the maximum values achieved at regional scale, and we assessed the effect of a reduction in the surveillance over a few years, during which poaching events increased, on the recovery patterns. We found that, overall, biomass of fishes increased with time while density diminished. In particular, piscivorous and macro-invertivore fish increased while the other trophic groups remained constant or declined, suggesting top-down processes. For the entire study period, those trophic groups were approaching carrying capacity; however, when accounting only for the period in which enforcement was high and constant, they grew exponentially, indicating that full carrying capacity may have not been achieved yet. When compared to other Mediterranean MPAs, the Cabo de Palos-Islas Hormigas MPA showed values for biomass that were disproportionately higher, suggesting that local factors, such as habitat structure and associated oceanographic processes, may be responsible for the dynamics found. Our results help to understand the potential trajectories of fish assemblages over a consolidated MPA and highlight empirically how the reduction of surveillance in a period may change the recovery patterns.

Experiments concerning peripheral nerve regeneration have been reported since the end of the 19th century. The need to implement an effective surgical procedure in terms of functional recovery has resulted in the appearance of several approaches to solve this problem. Nerve autograft was the first approach studied and is still considered the gold standard. Since autografts require donor harvesting, other strategies involving the use of natural materials have also been studied. Nevertheless, the results were not very encouraging and attention has moved towards the use of nerve conduits made from polymers, whose properties can be easily tailored and which allow the nerve conduit to be easily processed into a variety of shapes and forms. Some of these materials are already approved by the US Food and Drug Administration (FDA), as is presented here. Furthermore, polymers with conductive properties have very recently been subject to intensive study in this field, since it is believed that such properties have a positive influence in the regeneration of the new axons. This manuscript intends to give a global view of the mechanisms involved in peripheral nerve regeneration and the main strategies used to recover motor and sensorial function of injured nerves.

We are reporting for the first time the synthesis and application of an innovative nanometric system for the controlled topic release of melatonin in the retina. The ethylcellulose nanocapsules were characterized by diverse physicochemical techniques (scanning electron microscopy, zeta potential, hydrodynamic diameters) and an in vitro release study was done. A complete ex vivo and in vivo trans-corneal permeation and an irritation study were carried out with the new formulations in albino rabbits, to which a retinal degenerative model was induced. The results obtained demonstrate that the in vitro release of melatonin (1 mg/mL and 2 mg/mL) transported by nanocapsules is slower when compared to a solution of melatonin. Greater penetration of melatonin through the cornea was demonstrated by ex vivo and in vivo tests. This can be attributable to an enhanced neuroprotective effect of melatonin on retinal ganglion cells when it is included in ethylcellulose nanocapsules compared to a solution of melatonin. These outstanding findings add promising new perspectives to current knowledge about administrations using nano-technological tools in the treatment of neurodegenerative diseases at the ocular level.

Urbanisation is increasing worldwide and is regarded a major driver of environmental change altering local species assemblages. Private domestic gardens contribute a significant share of total green area in cities, but their biodiversity has received relatively little attention. Previous studies mainly considered plants, flying invertebrates such as bees and butterflies, and birds. By using a multi-taxa approach focused on less mobile, ground-dwelling invertebrates, we examined the influence of local garden characteristics and landscape characteristics on species richness and abundance of gastropods, spiders, millipedes, woodlice, ants, ground beetles and rove beetles. We assume that most of the species of these groups are able to complete their entire life cycle within a single garden. We conducted field surveys in thirty-five domestic gardens along a rural-urban gradient in Basel, Switzerland. Considered together, the gardens examined harboured an impressive species richness, with a mean share of species of the corresponding groups known for Switzerland of 13.9%, ranging from 4.7% in ground beetles to 23.3% in woodlice. The overall high biodiversity is a result of complementary contributions of gardens harbouring distinct species assemblages. Indeed, at the garden level, species richness of different taxonomical groups were typically not inter-correlated. The exception was ant species richness, which was correlated with those of gastropods and spiders. Generalised linear models revealed that distance to the city centre is an important driver of species richness, abundance and composition of several groups, resulting in an altered species composition in gardens in the centre of the city. Local garden characteristics were important drivers of gastropod and ant species richness, and the abundance of spiders, millipedes and rove beetles. Our study shows that domestic gardens make a valuable contribution to regional biodiversity. Thus, domestic urban gardens constitute an important part of green infrastructure, which should be considered by urban planners.

Neotropical wild rodents from Costa Rica were analyzed for the presence of herpesviruses (order Herpesvirales, family Herpesviridae). Using a broadly generic PCR, herpesvirus sequences were detected in 5% (8/160) of liver and heart samples: seven putative gammaherpesviruses in samples from Talamancan oryzomys (Nephelomys devius), sprightly colilargo (Oligoryzomys vegetus), Mexican deer mouse (Peromyscus nudipes), and Chiriqui harvest mouse (Reithrodontomys creper) and one putative betaherpesvirus in long-tailed singing mouse (Scotinomys xerampelinus). Results from this study could guide ecological investigations targeting the prevalence and host associations of herpesviruses in wild rodents from Costa Rica.

Summary Horse domestication revolutionized warfare and accelerated travel, trade, and the geographic expansion of languages. Here, we present the largest DNA time series for a non-human organism to date, including genome-scale data from 149 ancient animals and 129 ancient genomes (≥1-fold coverage), 87 of which are new. This extensive dataset allows us to assess the modern legacy of past equestrian civilizations. We find that two extinct horse lineages existed during early domestication, one at the far western (Iberia) and the other at the far eastern range (Siberia) of Eurasia. None of these contributed significantly to modern diversity. We show that the influence of Persian-related horse lineages increased following the Islamic conquests in Europe and Asia. Multiple alleles associated with elite-racing, including at the MSTN “speed gene,” only rose in popularity within the last millennium. Finally, the development of modern breeding impacted genetic diversity more dramatically than the previous millennia of human management., Graphical Abstract, Highlights • Two now-extinct horse lineages lived in Iberia and Siberia some 5,000 years ago • Iberian and Siberian horses contributed limited ancestry to modern domesticates • Oriental horses have had a strong genetic influence within the last millennium • Modern breeding practices were accompanied by a significant drop in genetic diversity, Genome-wide data from 278 ancient equids provide insights into how ancient equestrian civilizations managed, exchanged, and bred horses and indicate vast loss of genetic diversity as well as the existence of two extinct lineages of horses that failed to contribute to modern domestic animals.

Group living animals produce vocalizations denominated "contact calls" to maintain contact with out-of-sight group members. These calls have been shown to vary with caller identity and distance to potential listeners. However, it is not clear whether the acoustic variation of contact calls is related to caller social isolation (e.g., inside or outside a subgroup) and listeners' responses that can be helpful to maintain contact. Here, we addressed these questions in spider monkeys (Ateles geoffroyi), a Neotropical primate that exchanges contact calls denominated "whinnies", which show graded variation related to caller immediate behavior and distance between callers. Using 566 whinnies produced by 35 free-ranging adult spider monkeys recorded at ≤ 20 m from microphones, we first analyzed whether the acoustic variation of spontaneous whinnies (i.e., whinnies that are not responses to previous whinnies) is related to caller social isolation or whether acoustic variation is related to the likelihood of eliciting a response whinny from another individual. Secondly, we assessed whether listeners' responses (i.e., time to respond vocally, acoustic characteristics of response whinnies, orienting behaviors) were related to the acoustic variation of previous whinnies. Our study revealed that callers that were outside a subgroup produced whinnies with a lower fundamental frequency (F0), which travels longer distances, and increases the likelihood of producing a response whinny. Moreover, listeners (i.e., responders) responded faster to lower F0 whinnies. However, the acoustic variation (i.e., F0 variation) in response whinnies was better explained by the separation distance between callers, than by the acoustic variation of the previous whinny. Overall, our results suggest that whinny variation facilitates vocal contact to callers that are outside a subgroup, and that context and whinny variation affect listeners' responses. peerReviewed

Many neurotoxins inflict pain by targeting receptors expressed on nociceptors, such as the polymodal cationic channel TRPV1. The tarantula double-knot toxin (DkTx) is a peptide with an atypical bivalent structure, providing it with the unique capability to lock TRPV1 in its open state and evoke an irreversible channel activation. Here, we describe a distinct gating mechanism of DkTx-evoked TRPV1 activation. Interestingly, DkTx evokes significantly smaller TRPV1 macroscopic currents than capsaicin, with a significantly lower unitary conductance. Accordingly, while capsaicin evokes aversive behaviors in TRPV1-transgenic Caenorhabditis elegans, DkTx fails to evoke such response at physiological concentrations. To determine the structural feature(s) responsible for this phenomenon, we engineered and evaluated a series of mutated toxins and TRPV1 channels. We found that elongating the DkTx linker, which connects its two knots, increases channel conductance compared with currents elicited by the native toxin. Importantly, deletion of the TRPV1 pore turret, a stretch of amino acids protruding out of the channel’s outer pore region, is sufficient to produce both full conductance and aversive behaviors in response to DkTx. Interestingly, this deletion decreases the capsaicin-evoked channel activation. Taken together with structure modeling analysis, our results demonstrate that the TRPV1 pore turret restricts DkTx-mediated pore opening, probably through steric hindrance, limiting the current size and mitigating the evoked downstream physiological response. Overall, our findings reveal that DkTx and capsaicin elicit distinct TRPV1 gating mechanisms and subsequent pain responses. Our results also indicate that the TRPV1 pore turret regulates the mechanisms of channel gating and permeation.

Field-scale data on the relationship between pollinator activity and fruit set are scarce for rabbiteye blueberries (Vaccinium virgatum Aiton). We measured the densities of Apis mellifera L. (Hymenoptera: Apidae), Habropoda laboriosa F. (Hymenoptera: Apidae), Bombus Latreille (Hymenoptera: Apidae) spp., and Xylocopa virginica L. (Hymenoptera: Apidae) in 7–21 commercial fields during each of 3 yr in Louisiana and Mississippi. Foraging bees were counted on 10 ‘Tifblue’ bushes per field on 2 d during bloom, and the density of bees per flower was calculated based on the number of flowers open during the counts. Fruit set was measured 30 d after bloom. The impact of foraging activity on fruit set was inconsistent when densities of either all foragers or foragers of individual taxa were considered. Strong associations were observed only in 2001, with fruit set increased by H. laboriosa and Bombus and with a weaker contribution by A. mellifera. Floral robbery by X. virginica had no measurable negative effects. Populations of H. laboriosa were more consistent than those of other bees across sites. Managed colonies of A. mellifera were added at two densities (12.5 or 2.5 colonies per hectare) in seven fields each in 2001. These supplemental bees did not result in greater forager densities or fruit set in stocked fields. The observations show the challenge of field-scale pollination tests but provide an initial framework for rabbiteye blueberry growers to assess the availability of foraging bees early in bloom to help decide whether to add supplemental A. mellifera to try to enhance pollination if populations of non-Apis bees are low.

Renal cell carcinoma (RCC) is asymptomatic at early stages, and thus, initial diagnosis frequently occurs at advanced or even metastatic stages, leading to a high rate of mortality. Ferric nitrilotriacetate (FeNTA)-induced RCC model is a useful tool to analyze molecular events at different stages of the carcinogenesis process in vivo. MAPKs' alterations seem to play an important role in the development and maintenance of human RCC tumors. Based on the above, p38α/β/γ, JNK1/2, and ERK1/2 statuses were studied at early stages of FeNTA-induced renal carcinogenesis (1 and 2 months of carcinogen treatment) as well as in tumor tissue. MAPKs showed distinct response along carcinogenesis process, either as total proteins and/or as their phosphorylated forms. While the increase in total and phospho-p38α/β levels became lower as carcinogenesis progressed, p38γ overexpression grew. Instead, total JNK2 diminished, but JNK1 was elevated at all studied times, and p-JNK1 levels increased at early stages, but not in tumors. In contrast, p-JNK2 rose at 2 months of treatment and in tumor tissue. Increased levels of p-ERK1/2 were observed at all stages analyzed. Very interestingly, at 1 and 2 months of FeNTA treatment, no alterations in MAPKs were found in liver or lung, where no primary tumors are induced with the scheme of FeNTA administration followed here. In conclusion, MAPKs' behavior evolved differentially as renal carcinogenesis advanced, even among isoforms of the same family, but it did not change in other tissues. All this strongly suggests a role of these kinases in FeNTA-induced RCC tumor development and maintenance.

Mesenchymal Stromal cells (MSCs) have a potential role in cell-based therapies. Foetal bovine serum (FBS) is used to supplement the basal cell culture medium but presents several disadvantages and risks. Other alternatives have been studied, including human umbilical cord blood plasma (hUCBP), aiming at the development of xeno-free culturing protocols. A comparative characterization of multicomponent metabolic composition of hUCBP and commercial FBS based on Nuclear Magnetic Resonance (NMR) spectroscopy and multivariate statistical analysis was performed. The analysis of 1H-NMR spectra revealed both similarities and differences between the two proposed supplements. Similar metabolites (amino acids, glucose, lipids and nucleotides) were found in the hUCBP and FBS NMR spectra. The results show that the major difference between the metabolic profiles of the two proposed supplements are due to the significantly higher levels of glucose and lower levels of lactate, glutamate, alanine and branched chain amino acids in hUCBP. Similar or slightly different levels of important proteinogenic amino acids, as well as of nucleotides, lipids were found in the hUCBP and FBS. In order to validate it's suitability for cell culture, umbilical cord-MSCs (UC-MSCs) and dental pulp stem cells (DPSCs) were expanded using hUCBP. In both hMSCs, in vitro culture with hUCBP supplementation presented similar to improved metabolic performances when compared to FBS. The two cell types tested expressed different optimum hUCBP percentage content. For DPSCs, the optimum hUCBP content was 6% and for UC-MSCs, 4%. Cultured hMSCs displayed no changes in senescence indicators, as well as maintained characteristic surface marker's expression. FBS substitution was associated with an increase in early apoptosis events, in a dose dependent manner, as well as to slight up- and down-regulation of targeted gene's expression. Tri-lineage differentiation capacity was also influenced by the substitution of FBS by hUCBP.

This study aimed to verify the sedative and anaesthetic effect of the essential oils of basil ( Ocimum basilicum ) (EOOB) and lemongrass (Cymbopogum flexuosus) (EOCF) in Nile tilapia juveniles. The fish were transferred to aquaria containing different concentrations of each essential oil: 10, 25, 50, 100, 200, 400 and 600 μL L -1 . The time of sedation ranged from 7 to 31 seconds and the recommended concentration was 10 or 25 μL L -1 for both essential oils. The best times for anaesthesia and recovery were found for the concentrations of 400 μL L -1 for EOOB (135.2 and 199.1 seconds, respectively) and 600 μL L -1 for EOCF (327.1 and 374.8 seconds, respectively). In conclusion, we recommend the use of EOOB and EOCF for the sedation and anaesthesia of Nile tilapia at concentrations of 10-25 (for both), 400 and 600 μL L -1 , respectively. Key words: sedation, recovery, basil, lemongrass. Correspondence to: Carlos Eduardo Copatti E-mail: carloseduardocopatti@yahoo.com.brINTRODUCTION

Understanding factors driving successful invasions is one of the cornerstones of invasion biology. Bird invasions have been frequently used as study models, and the foundation of current knowledge largely relies on species purposefully introduced during the 19th and early 20th centuries in countries colonized by Europeans. However, the profile of exotic bird species has changed radically in the last decades, as birds are now mostly introduced into the invasion process through unplanned releases from the worldwide pet and avicultural trade. Here we assessed the role of the three main drivers of invasion success (i.e., event-, species-, and location-level factors) on the establishment and spatial spread of exotic birds using an unprecedented dataset recorded throughout the last 100 y in the Iberian Peninsula. Our multimodel inference phylogenetic approach showed that the barriers that need to be overcome by a species to successfully establish or spread are not the same. Whereas establishment is largely related to event-level factors, apparently stochastic features of the introduction (time since first introduction and propagule pressure) and to the origin of introduced species (wild-caught species show higher invasiveness than captive-bred ones), the spread across the invaded region seems to be determined by the extent to which climatic conditions in the new region resemble those of the species’ native range. Overall, these results contrast with what we learned from successful deliberate introductions and highlight that different management interventions should apply at different invasion stages, the most efficient strategies being related to event-level factors., This work was funded by the Division of Mathematics and Natural Sciences of Queens College, City University of New York, and by Excellence Projects P07RNM 02918 and P08-RNM-4014 (Junta de Andalucía), Fundación Repsol, the Project Estación Biológica de Doñana-Severo Ochoa (Grant SEV-2012-0262), and AIC-A-2011-0706.

Due to several factors such as ecological conditions, group size, and social organization, primates frequently spend time out of visual contact with individuals of their own group. Through the use of long-distance vocalizations, often termed "contact calls," primates are able to maintain contact with out-of-sight individuals. Contact calls have been shown to be individually distinct, and reverberation and attenuation provide information about caller distance. It is less clear, however, whether callers actively change the structure of contact calls depending on the distance to the presumed listeners. We studied this question in spider monkeys (Ateles geoffroyi), a species with complex spatial dynamics (fission-fusion society) that produces highly frequency modulated contact calls, denominated "whinnies." We determined the acoustic characteristics of 566 whinnies recorded from 35 free-ranging spider monkeys that belong to a community located in Mexico, and used cluster analyses, discriminant function analyses, and generalized linear mixed models to assess if they varied in relation to the presumed distance to the listener. Whinnies could be grouped into five subtypes. Since the lowest frequency subtype was mainly produced by spider monkeys that exchanged whinnies at longer distances, and lower frequency calls propagate across longer distances, our results suggest that whinnies vary in order to enhance vocal contact between individuals separated by different distances. Our results also revealed that whinnies convey potential information about caller immediate behaviors and corroborated that these calls are individually distinct. Overall, our results suggest that whinny acoustic variation facilitates the maintenance of vocal contact between individuals living in a society with complex spatial dynamics.

Background: Hernia correction is a routinely performed treatment in surgical practice. The improvement of the operative technique and available materials certainly has been a great benefit to the quality of surgical results. The insertion of prostheses for hernia correction is well-founded in the literature, and has become the standard of treatment when this type of disease is discussed. Aim: To evaluate two available prostheses: the polypropylene and polypropylene coated ones in an experimental model. Methods: Seven prostheses of each kind were inserted into Wistar rats (Ratus norvegicus albinus) in the anterior abdominal wall of the animal in direct contact with the viscera. After 90 days follow-up were analyzed the intra-abdominal adhesions, and also performed immunohistochemical evaluation and videomorphometry of the total, type I and type III collagen. Histological analysis was also performed with hematoxylin-eosin to evaluate cell types present in each mesh. Results: At 90 days the adhesions were not different among the groups (p=0.335). Total collagen likewise was not statistically different (p=0.810). Statistically there was more type III collagen in the coated polypropylene group (p=0.039) while type I was not different among the prostheses (p=0.050). The lymphocytes were statistically more present in the polypropylene group (p=0.041). Conclusion: The coated prosthesis was not different from the polypropylene one regarding the adhesion. Total and type I collagen were not different among the groups, while type III collagen was more present on the coated mesh. There was a greater number of lymphocytes on the polypropylene mesh. RESUMO Racional: A correção herniária é tratamento realizado rotineiramente na prática cirúrgica. O aprimoramento da técnica operatória e dos materiais disponíveis trouxe grande benefício na qualidade dos resultados cirúrgicos. A inserção de próteses para correção herniária é bem embasada na literatura e tornou-se o padrão de tratamento. Objetivo: Avaliar em modelo experimental dois tipos de próteses diferentes, de polipropileno e polipropileno revestido. Métodos: Foram inseridas sete próteses de cada tipo em ratos Wistar (Ratus norvegicus albinus) na parede abdominal anterior do animal em contato direto com as vísceras. Após o seguimento de 90 dias analisaram-se as aderências intra-abdominais, bem como avaliação por imunoistoquímica e videomorfometria do colágeno total, tipo I e tipo III. Também, fez-se análise histológica com hematoxylina-eosina para avaliação dos tipos celulares presentes em cada tela. Resultados: Aos 90 dias as aderências não foram diferentes entre os grupos (p=0,335). O colágeno total igualmente não foi estatisticamente diferente (p=0,810). O colágeno tipo III foi estatisticamente maior no grupo polipropileno revestido (p=0,039) enquanto o tipo I não diferiu entre as próteses (p=0,050). Os linfócitos foram estatisticamente mais presentes no grupo polipropileno (p=0,041). Conclusão: A prótese revestida não foi diferente da de polipropileno na variável aderência. O colágeno total e tipo I não foram diferentes entre os grupos enquanto que o colágeno tipo III foi mais presente na tela revestida. O número de linfócitos foi maior na tela de polipropileno.