BUSCADOR RECOLECTA

Experimental data and TEM images from the nanoparticle synthesis, and data from catalytic characterization., Peer reviewed

[Introduction]: Secreted mucins are highly O-glycosylated glycoproteins produced by goblet cells in mucosal epithelia. They constitute the protective viscous gel layer overlying the epithelia and are involved in pathogen recognition, adhesion and expulsion. The gill polyopisthocotylidan ectoparasite Sparicotyle chrysophrii, feeds on gilthead seabream (Sparus aurata) blood eliciting severe anemia., [Methods]: Control unexposed and recipient (R) gill samples of gilthead seabream experimentally infected with S. chrysophrii were obtained at six consecutive times (0, 11, 20, 32, 41, and 61 days post-exposure (dpe)). In histological samples, goblet cell numbers and their intensity of lectin labelling was registered. Expression of nine mucin genes (muc2, muc2a, muc2b, muc5a/c, muc4, muc13, muc18, muc19, imuc) and three regulatory factors involved in goblet cell differentiation (hes1, elf3, agr2) was studied by qPCR. In addition, differential expression of glycosyltransferases and glycosidases was analyzed in silico from previously obtained RNAseq datasets of S. chrysophrii-infected gilthead seabream gills with two different infection intensities., [Results and Discussion]: Increased goblet cell differentiation (up-regulated elf3 and agr2) leading to neutral goblet cell hyperplasia on gill lamellae of R fish gills was found from 32 dpe on, when adult parasite stages were first detected. At this time point, acute increased expression of both secreted (muc2a, muc2b, muc5a/c) and membrane-bound mucins (imuc, muc4, muc18) occurred in R gills. Mucins did not acidify during the course of infection, but their glycosylation pattern varied towards more complex glycoconjugates with sialylated, fucosylated and branched structures, according to lectin labelling and the shift of glycosyltransferase expression patterns. Gilthead seabream gill mucosal response against S. chrysophrii involved neutral mucus hypersecretion, which could contribute to worm expulsion and facilitate gas exchange to counterbalance parasite-induced hypoxia. Stress induced by the sparicotylosis condition seems to lead to changes in glycosylation characteristic of more structurally complex mucins., Peer reviewed

Optimization of heating time in batch synthesis (Figure S1); physical and geometrical parameters adopted for Comsol Multiphysics simulation (Table S1); N2 adsorption analysis (Table S2); reproducibility analysis (Figure S2); TEM and HRSTEM characterization of SBA-15, unsupported Ag NCs, and Ag NCs/SBA-15 with 0.16, 0.62, and 0.96 wt % (Figure S3); leaching test for propargylguanidine cyclization (Figure S4, Table S3, and Figure S5); catalytic reusability and stability study by reaction of 2-(phenylethynyl)phenol (Figure S6); continuous flow reaction and activation energy for 2-(phenylethynyl)phenol cyclization (figure S7); reaction mechanism for hydroamination of propargylguanidines (Scheme S1); general procedure for the synthesis of starting material with corresponding characterization data., Peer reviewed

[Introduction]: Secreted mucins are highly O-glycosylated glycoproteins produced by goblet cells in mucosal epithelia. They constitute the protective viscous gel layer overlying the epithelia and are involved in pathogen recognition, adhesion and expulsion. The gill polyopisthocotylidan ectoparasite Sparicotyle chrysophrii, feeds on gilthead seabream (Sparus aurata) blood eliciting severe anemia., [Methods]: Control unexposed and recipient (R) gill samples of gilthead seabream experimentally infected with S. chrysophrii were obtained at six consecutive times (0, 11, 20, 32, 41, and 61 days post-exposure (dpe)). In histological samples, goblet cell numbers and their intensity of lectin labelling was registered. Expression of nine mucin genes (muc2, muc2a, muc2b, muc5a/c, muc4, muc13, muc18, muc19, imuc) and three regulatory factors involved in goblet cell differentiation (hes1, elf3, agr2) was studied by qPCR. In addition, differential expression of glycosyltransferases and glycosidases was analyzed in silico from previously obtained RNAseq datasets of S. chrysophrii-infected gilthead seabream gills with two different infection intensities., [Results and Discussion]: Increased goblet cell differentiation (up-regulated elf3 and agr2) leading to neutral goblet cell hyperplasia on gill lamellae of R fish gills was found from 32 dpe on, when adult parasite stages were first detected. At this time point, acute increased expression of both secreted (muc2a, muc2b, muc5a/c) and membrane-bound mucins (imuc, muc4, muc18) occurred in R gills. Mucins did not acidify during the course of infection, but their glycosylation pattern varied towards more complex glycoconjugates with sialylated, fucosylated and branched structures, according to lectin labelling and the shift of glycosyltransferase expression patterns. Gilthead seabream gill mucosal response against S. chrysophrii involved neutral mucus hypersecretion, which could contribute to worm expulsion and facilitate gas exchange to counterbalance parasite-induced hypoxia. Stress induced by the sparicotylosis condition seems to lead to changes in glycosylation characteristic of more structurally complex mucins., Peer reviewed

Crystallographic structure of [Au25(Cys)18], intensity distribution and emission wavelength of white light, time-resolved 1O2 phosphorescence decay of treated silicones, stability test of treated silicones, and comparison of photobactericidal enhancement by [Au25(Cys)18] nanoclusters synthesized through two different synthetic methods., Peer reviewed

[Introduction]: Secreted mucins are highly O-glycosylated glycoproteins produced by goblet cells in mucosal epithelia. They constitute the protective viscous gel layer overlying the epithelia and are involved in pathogen recognition, adhesion and expulsion. The gill polyopisthocotylidan ectoparasite Sparicotyle chrysophrii, feeds on gilthead seabream (Sparus aurata) blood eliciting severe anemia., [Methods]: Control unexposed and recipient (R) gill samples of gilthead seabream experimentally infected with S. chrysophrii were obtained at six consecutive times (0, 11, 20, 32, 41, and 61 days post-exposure (dpe)). In histological samples, goblet cell numbers and their intensity of lectin labelling was registered. Expression of nine mucin genes (muc2, muc2a, muc2b, muc5a/c, muc4, muc13, muc18, muc19, imuc) and three regulatory factors involved in goblet cell differentiation (hes1, elf3, agr2) was studied by qPCR. In addition, differential expression of glycosyltransferases and glycosidases was analyzed in silico from previously obtained RNAseq datasets of S. chrysophrii-infected gilthead seabream gills with two different infection intensities., [Results and Discussion]: Increased goblet cell differentiation (up-regulated elf3 and agr2) leading to neutral goblet cell hyperplasia on gill lamellae of R fish gills was found from 32 dpe on, when adult parasite stages were first detected. At this time point, acute increased expression of both secreted (muc2a, muc2b, muc5a/c) and membrane-bound mucins (imuc, muc4, muc18) occurred in R gills. Mucins did not acidify during the course of infection, but their glycosylation pattern varied towards more complex glycoconjugates with sialylated, fucosylated and branched structures, according to lectin labelling and the shift of glycosyltransferase expression patterns. Gilthead seabream gill mucosal response against S. chrysophrii involved neutral mucus hypersecretion, which could contribute to worm expulsion and facilitate gas exchange to counterbalance parasite-induced hypoxia. Stress induced by the sparicotylosis condition seems to lead to changes in glycosylation characteristic of more structurally complex mucins., Peer reviewed

Peer reviewed

Figure S1. Flow chart illustrating the workflow for characterizing the probability that a given Sus scrofa high density single nucleotide polymorphism (SNP) genotype could be described under the null hypothesis – descending strictly from contributions from domestic pigs – relative to the alternative hypothesis of allowing for European wild boar ancestry. Table S1. A complete list of the high-density single nucleotide polymorphism (SNP) Sus scrofa genotypes included in the study, with 1,421 reference samples organized into five groups (mixed-commercial breeds, Durocs, heritage breeds, primitive breeds, and European wild boar) for the development of the Delta Likelihood test static, and, for evaluation of the Delta Likelihood approach, a test set of 435 genotypes from 29 additional domestic breeds and 6,566 wild pigs sampled across the invaded range within the contiguous United States. The original publishing source for genotypes is presented in the table with complete references below. Figure S2. Illustration of the genetic structure, as revealed with principal component analysis (plotting principal component 1 versus 2), among the five reference groups of Sus scrofa genotyped with a high-density single nucleotide polymorphism (SNP) array for the development of the Delta Likelihood statistic – an approach to probabilistically describe that a given genotype possesses wild boar ancestry in full or in part (i.e., a wild boar-domestic pig hybrid). Figure S3. Barplots illustrating genetic structure among the 1,421 genotypes that comprise five reference groups of Sus scrofa – organized as mixed-commercial breeds of domestic pig, Durocs, heritage breeds, primitive breeds, and European wild boar – as revealed with high-density single nucleotide polymorphism (SNP) genotypes and leave-one-out analyses conducted in ADMIXTURE (v1.3.0; Alexander et al., 2009) in a supervised framework as described in Smyser et al. (2020). Figure S4. Delta Likelihood statistics calculated from high-density single nucleotide polymorphism (SNP) genotypes for Sus scrofa reference genotypes, organized into reference groups representing mixed-commercial breeds (n = 310), Durocs (n = 159), heritage breeds (n =381), primitive breeds (n = 189), and European wild boar (n = 382); plots represent kernel density estimates of Delta Likelihood statistics for the respective reference groups in which the loci retained for analysis were pruned for linkage disequilbrium (LD) base on the specific reference group listed in the plot title. Pruning for LD based on the heritage reference group resulted in the greatest difference between the upper tail of the kernel density estimate observed among the domestic pig reference groups versus the lower tail of the wild boar reference group (difference of 36.07 when LD pruning was based on the heritage reference group versus 32.72 for the mixed-commercial reference group, 28.79 for Duroc, 35.17 for primitive, and 30.79 for wild boar); thus, the set of 18,790 loci based on the heritage reference set LD prune was retained for subsequent analyses. Table S2. The statistical distribution and associated parameters for respective distributions fitted to Delta Likelihood statistics calculated for domestic pig reference groups (i.e., mixedcommercial breeds, Durocs, heritage breeds, primitive breeds) based on bootstrapped iterations in which the composition of the specific genotypes included in the reference groups were resampled with replacement 1,000 times. Figure S5. Kernel density estimates of Delta Likelihood statistics calculated with 18,790 single nucleotide polymorphic (SNP) loci for Sus scrofa reference genotypes, organized into reference groups representing mixed-commercial breeds (n = 310), Durocs (n = 159), heritage breeds (n = 381), primitive breeds (n = 189), European wild boar (n = 382) and an empirical test set of an additional 29 domestic breeds, represented by 435 genotypes, that were excluded from the reference set. Figure S6. Delta Likelihood statistics calculated from simulated high-density single nucleotide polymorphism (SNP) genotypes (18,790 loci) for Sus scrofa in which genotypes were simulated with R package gscramble (Anderson 2023, R Development Core Team 2023) from fourgeneration pedigrees established by randomly sampling without replacement a total of 16 genotypes from one of four domestic pig reference groups (16 – 1 genotypes drawn from either mixed-commercial, Duroc, heritage, or primitive reference groups) and a complementary number of genotypes (0 – 15) drawn from the wild boar reference group. Following four generations of one-to-one pairing with no inbreeding conducted within the simulation, while allowing for biologically realistic rates of recombination (1 centimorgan per megabase) to reflect the linkage structure among proximate loci and tracking the proportion of each genotype attributable to wild boar ancestry versus domestic pig ancestry, we randomly selected a single simulated genotype for analysis from the 64,000 iterations conducted., Peer reviewed

(A) CBK1 cbk1Δ cdc15-2 (YMF3869), (B) cbk1-6E cbk1Δ cdc15-2 (YMF3763), (C) cbk1-5E-E409S cbk1Δ cdc15-2 (YMF4279), (D) cbk1-5E-E574T cbk1Δ cdc15-2 (YMF4280), and (E) cbk1-9A cbk1Δ cdc15-2 (YMF3764) cells were grown in YPD and arrested in late anaphase by raising the temperature to 37 °C before rapamycin was added to half of the culture for 20 min. Subsequently, to allow progression through the cell cycle, cells were released in the absence (i) or presence (ii) of rapamycin. Samples were taken at the indicated times to study cell-cycle progression by flow cytometry. Schematic illustration of cbk1-9A mutant in which phosphosites containing serines or threonines followed by prolines were changed to alanine to block phosphorylations (iii). FACS graphs can be found in the supplementary FACS file (S1 File)., pbio.3002263.s006.pdf, Peer reviewed

(A) DSE4-6HA cdc15-2 (YMF4029) cells were grown in YPD and arrested in late anaphase by shifting the temperature to 37 °C before the addition of DMSO (−) or rapamycin (+) for 20 min. Subsequently, cells were released from the anaphase arrest in the absence (−) or presence (+) of rapamycin before protein extracts were prepared from shown time points and analysed by immunoblotting. Raw data for blot can be found in Supporting information (S6A Raw Images). (B) iHA-CTS1 cdc14-1 cells (YMF4088) were grown and processed as in A. Protein extracts were prepared from indicated time points and analysed by immunoblotting. Raw data for blot can be found in Supporting information (S6B Raw Images). (C) CTS1-GFPEnvy cdc14-1 cells (YMF4231) were grown as in C. Samples were taken at the indicated times to determine the proportion of cells with Cts1 at the division site in the presence (i) of rapamycin. Examples of cells are shown for the 60 min time point after the release at 24 °C in the presence (ii) of rapamycin. Scale bar indicates 5 μm. Underlying data for all the graphs can be found in S1 Data file., pbio.3002263.s007.pdf, Peer reviewed