Microbiome changes through the ontogeny of the marine sponge Crambe crambe.

BACKGROUND: Poriferans (sponges) are highly adaptable organisms that can thrive in diverse marine and freshwater environments due, in part, to their close associations with internal microbial communities. This sponge microbiome can be acquired from the surrounding environment (horizontal acquisition) or obtained from the parents during the reproductive process through a variety of mechanisms (vertical transfer), typically resulting in the presence of symbiotic microbes throughout all stages of sponge development. How and to what extent the different components of the microbiome are transferred to the developmental stages remain poorly understood. Here, we investigated the microbiome composition of a common, low-microbial-abundance, Atlantic-Mediterranean sponge, Crambe crambe, throughout its ontogeny, including adult individuals, brooded larvae, lecithotrophic free-swimming larvae, newly settled juveniles still lacking osculum, and juveniles with a functional osculum for filter feeding. RESULTS: Using 16S rRNA gene analysis, we detected distinct microbiome compositions in each ontogenetic stage, with variations in composition, relative abundance, and diversity of microbial species. However, a particular dominant symbiont, Candidatus Beroebacter blanensis, previously described as the main symbiont of C. crambe, consistently occurred throughout all stages, an omnipresence that suggests vertical transmission from parents to offspring. This symbiont fluctuated in relative abundance across developmental stages, with pronounced prevalence in lecithotrophic stages. A major shift in microbial composition occurred as new settlers completed osculum formation and acquired filter-feeding capacity. Candidatus Beroebacter blanensis decreased significatively at this point. Microbial diversity peaked in filter-feeding stages, contrasting with the lower diversity of lecithotrophic stages. Furthermore, individual specific transmission patterns were detected, with greater microbial similarity between larvae and their respective parents compared to non-parental conspecifics. CONCLUSIONS: These findings suggest a putative vertical transmission of the dominant symbiont, which could provide some metabolic advantage to non-filtering developmental stages of C. crambe. The increase in microbiome diversity with the onset of filter-feeding stages likely reflects enhanced interaction with environmental microbes, facilitating horizontal transmission. Conversely, lower microbiome diversity in lecithotrophic stages, prior to filter feeding, suggests incomplete symbiont transfer or potential symbiont digestion. This research provides novel information on the dynamics of the microbiome through sponge ontogeny, on the strategies for symbiont acquisition at each ontogenetic stage, and on the potential importance of symbionts during larval development.

Silica-associated proteins from hexactinellid sponges support an alternative evolutionary scenario for biomineralization in Porifera.

Metazoans use silicon traces but rarely develop extensive silica skeletons, except for the early-diverging lineage of sponges. The mechanisms underlying metazoan silicification remain incompletely understood, despite significant biotechnological and evolutionary implications. Here, the characterization of two proteins identified from hexactinellid sponge silica, hexaxilin and perisilin, supports that the three classes of siliceous sponges (Hexactinellida, Demospongiae, and Homoscleromorpha) use independent protein machineries to build their skeletons, which become non-homologous structures. Hexaxilin forms the axial filament to intracellularly pattern the main symmetry of the skeletal parts, while perisilin appears to operate in their thickening, guiding extracellular deposition of peripheral silica, as does glassin, a previously characterized hexactinellid silicifying protein. Distant hexaxilin homologs occur in some bilaterians with siliceous parts, suggesting putative conserved silicifying activity along metazoan evolution. The findings also support that ancestral Porifera were non-skeletonized, acquiring silica skeletons only after diverging into major classes, what reconciles molecular-clock dating and the fossil record.

Highly efficient grafting of hetero-complementary amidinium and carboxylate hydrogen-bonding/ionic pairs onto polymer surfaces.

We describe a grafting methodology, based on thiol-fluoroarene chemistry, to efficiently incorporate complementary hydrogen-bonding carboxylate and amidinium groups into polymer backbones. The process was optimized both in solution and on the surface of processed films, with the aim to produce materials showing hetero-complementary adhesion.

Phylomitogenomics bolsters the high-level classification of Demospongiae (phylum Porifera).

Class Demospongiae is the largest in the phylum Porifera (Sponges) and encompasses nearly 8,000 accepted species in three subclasses: Keratosa, Verongimorpha, and Heteroscleromorpha. Subclass Heteroscleromorpha contains ∼90% of demosponge species and is subdivided into 17 orders. The higher level classification of demosponges underwent major revision as the result of nearly three decades of molecular studies. However, because most of the previous molecular work only utilized partial data from a small number of nuclear and mitochondrial (mt) genes, this classification scheme needs to be tested by larger datasets. Here we compiled a mt dataset for 136 demosponge species-including 64 complete or nearly complete and six partial mt-genome sequences determined or assembled for this study-and used it to test phylogenetic relationships among Demospongiae in general and Heteroscleromorpha in particular. We also investigated the phylogenetic position of Myceliospongia araneosa, a highly unusual demosponge without spicules and spongin fibers, currently classified as Demospongiae incertae sedis, for which molecular data were not available. Our results support the previously inferred sister-group relationship between Heteroscleromorpha and Keratosa + Verongimorpha and suggest five main clades within Heteroscleromorpha: Clade C0 composed of order Haplosclerida; Clade C1 composed of Scopalinida, Sphaerocladina, and Spongillida; Clade C2 composed of Axinellida, Biemnida, Bubarida; Clade C3 composed of Tetractinellida; and Clade C4 composed of Agelasida, Clionaida, Desmacellida, Merliida, Suberitida, Poecilosclerida, Polymastiida, and Tethyida. The inferred relationships among these clades were (C0(C1(C2(C3+C4)))). Analysis of molecular data from M. araneosa placed it in the C3 clade as a sister taxon to the highly skeletonized tetractinellids Microscleroderma sp. and Leiodermatium sp. Molecular clock analysis dated divergences among the major clades in Heteroscleromorpha from the Cambrian to the Early Silurian, the origins of most heteroscleromorph orders in the middle Paleozoic, and the most basal splits within these orders around the Paleozoic to Mesozoic transition. Overall, the results of this study are mostly congruent with the accepted classification of Heteroscleromorpha, but add temporal perspective and new resolution to phylogenetic relationships within this subclass.

Mass mortality of the keratose sponge Sarcotragus foetidus in the Aegean Sea (Eastern Mediterranean) correlates with proliferation of Vibrio bacteria in the tissues.

In the last two decades, episodes of mass mortality in benthic communities have often been associated with climatic anomalies, but the ultimate mechanisms through which they lead to death have rarely been identified. This study reports a mass mortality of wild sponges in the Aegean Sea (Turkey, Eastern Mediterranean), which affected the keratose demosponge Sarcotragus foetidus in September 2021. We examined the occurrence of thermo-dependent bacteria of the genus Vibrio in the sponges, identified through 16S rRNA of colonies isolated from sponge tissue in specific culturing media. Six Vibrio sequences were identified from the sponges, three of them being putatively pathogenic (V. fortis, V. owensii, V. gigantis). Importantly, those Vibrios were isolated from only tissues of diseased sponges. In contrast, healthy individuals sampled in both summer and winter led to no Vibrio growth in laboratory cultures. A 50 years record of sea surface temperature (SST) data for the study area reveals a progressive increase in temperature from 1970 to 2021, with values above 24°C from May to September 2021, reaching an absolute historical maximum of 28.9°C in August 2021. We hypothesize that such elevated SST values maintained for several months in 2021 promoted proliferation of pathogenic Vibrio species (thermo-dependent bacteria) in S. foetidus, triggering or aggravating the course of sponge disease. Thus, vibrioisis emerges as one of the putative mechanisms through which global water warming in the Mediterranean Sea translates into sponge mortality. The historical time course of temperature data for the studied area in the Aegean Sea predicts that recurrent waves of elevated SST are likely to occur in the coming summers. If so, recurrent disease may eventually eliminate this abundant sponge from the sublittoral in the midterm, altering the original bathymetric distribution of the species and compromising its ecological role.

Use of vinasse and coffee waste as chelating agent of photo-Fenton landfill leachate treatment.

This research studies the use of vinasse (VS) coming from Pisco and caffeic acid (Caa) from solid coffee waste as chelating agents of this process, to carry out a photo-Fenton process using UVc lamps of 254-nm wavelength for 60 min, at the natural pH of the landfill leachate (8.9). Without the chelating agent, there was a removal of UV 254 and COD of 54.2% and 54.7%, respectively, when the photo-Fenton reaction was carried out at pH 3; at pH 6, the removal of UV 254 and COD was 13.1% and 39.2%, respectively, and at pH 8.9, the elimination of UV 254 and COD was 10.8% and 16.1%, respectively. When Caa was used in the landfill leachate (LL) for the photo-catalytic processes carried out at pH 8.9, a removal of 24.1%, 43.0%, and 47.4% of UV 254 was obtained using 5 mg/L, 50 mg/L, and 100 mg/L of Caa. The removal of UV 254 was 27.3%, 30.7%, and 36.3% using 5 mg/L, 50 mg/L, and 100 mg/L of VS, respectively, and the removal of COD was 32.2%, 35.4%, and 39.2% using 5 mg/L, 50 mg/L, and 100 mg/L of VS, respectively. When Caa was used in the LL at pH 8.9, the concentration of total Fe went from 37.5 to 33.2, from 40.2 to 36.8, and from 45.2 to 42.1, using 5 mg/L, 50 mg/L, and 100 mg/L of caffeic acid, respectively. Using VS in the LL at pH 8.9, the concentration of total Fe along the run went from 35.1 to 32.2, from 39.4 to 34.8, and from 42.1 to 40.2, using 5 mg/L, 50 mg/L, and 100 mg/L of VS, respectively. As a result of these processes, it was noted that the use of Caa and VS increases the solubility of Fe at a higher pH.

Antimicrobial Resistance in the Environment: Towards Elucidating the Roles of Bioaerosols in Transmission and Detection of Antibacterial Resistance Genes.

Antimicrobial resistance (AMR) is continuing to grow across the world. Though often thought of as a mostly public health issue, AMR is also a major agricultural and environmental problem. As such, many researchers refer to it as the preeminent One Health issue. Aerial transport of antimicrobial-resistant bacteria via bioaerosols is still poorly understood. Recent work has highlighted the presence of antibiotic resistance genes in bioaerosols. Emissions of AMR bacteria and genes have been detected from various sources, including wastewater treatment plants, hospitals, and agricultural practices; however, their impacts on the broader environment are poorly understood. Contextualizing the roles of bioaerosols in the dissemination of AMR necessitates a multidisciplinary approach. Environmental factors, industrial and medical practices, as well as ecological principles influence the aerial dissemination of resistant bacteria. This article introduces an ongoing project assessing the presence and fate of AMR in bioaerosols across Canada. Its various sub-studies include the assessment of the emissions of antibiotic resistance genes from many agricultural practices, their long-distance transport, new integrative methods of assessment, and the creation of dissemination models over short and long distances. Results from sub-studies are beginning to be published. Consequently, this paper explains the background behind the development of the various sub-studies and highlight their shared aspects.

Symbiont transmission in marine sponges: reproduction, development, and metamorphosis.

Marine sponges (phylum Porifera) form symbioses with diverse microbial communities that can be transmitted between generations through their developmental stages. Here, we integrate embryology and microbiology to review how symbiotic microorganisms are transmitted in this early-diverging lineage. We describe that vertical transmission is widespread but not universal, that microbes are vertically transmitted during a select developmental window, and that properties of the developmental microbiome depends on whether a species is a high or low microbial abundance sponge. Reproduction, development, and symbiosis are thus deeply rooted, but why these partnerships form remains the central and elusive tenet of these developmental symbioses.

Microbes from Mum: symbiont transmission in the tropical reef sponge Ianthella basta.

Most marine sponge species harbour distinct communities of microorganisms which contribute to various aspects of their host's health and physiology. In addition to their key roles in nutrient transformations and chemical defence, these symbiotic microbes can shape sponge phenotype by mediating important developmental stages and influencing the environmental tolerance of the host. However, the characterisation of each microbial taxon throughout a sponge's life cycle remains challenging, with several sponge species hosting up to 3000 distinct microbial species. Ianthella basta, an abundant broadcast spawning species in the Indo-Pacific, is an emerging model for sponge symbiosis research as it harbours only three dominant symbionts: a Thaumarchaeotum, a Gammaproteobacterium, and an Alphaproteobacterium. Here, we successfully spawned Ianthella basta, characterised its mode of reproduction, and used 16S rRNA gene amplicon sequencing, fluorescence in situ hybridisation, and transmission electron microscopy to characterise the microbial community throughout its life cycle. We confirmed I. basta as being gonochoric and showed that the three dominant symbionts, which together make up >90% of the microbiome according to 16S rRNA gene abundance, are vertically transmitted from mother to offspring by a unique method involving encapsulation in the peri-oocytic space, suggesting an obligate relationship between these microbes and their host.

Seasonally Driven Sexual and Asexual Reproduction in Temperate Tethya Species.

Marine organisms that rely on environmental cues for reproduction are likely to experience shifts in reproductive phenology and output due to global climate change. To assess the role that the environment may play in the reproductive timing for temperate sponges, this study examined sexual and asexual reproduction in New Zealand sponge species (Tethya bergquistae and the Tethya burtoni complex) and correlated reproductive output with temperature, chlorophyll-a concentration, and rainfall. Histological analyses of sponges collected monthly (from February 2015 to February 2017) revealed that these sponges are oviparous and gonochoristic and that they sexually reproduce annually during the austral summer. Both monthly collections and in situ monitoring revealed that Tethya spp. asexually bud continuously, but with greater intensity in the austral spring and summer. Temperature was positively associated with both sexual reproduction and budding, with seasonal cues appearing important. Future shifts in the environment that alter such cues are expected to affect population dynamics of these sponges.

Multivalent Lactose-Ferrocene Conjugates Based on Poly (Amido Amine) Dendrimers and Gold Nanoparticles as Electrochemical Probes for Sensing Galectin-3.

Galectin-3 is considered a cancer biomarker and bioindicator of fibrosis and cardiac remodeling and, therefore, it is desirable to develop convenient methods for its detection. Herein, an approach based on the development of multivalent electrochemical probes with high galectin-3 sensing abilities is reported. The probes consist of multivalent presentations of lactose-ferrocene conjugates scaffolded on poly (amido amine) (PAMAM) dendrimers and gold nanoparticles. Such multivalent lactose-ferrocene conjugates are synthesized by coupling of azidomethyl ferrocene-lactose building blocks on alkyne-functionalized PAMAM, for the case of the glycodendrimers, and to disulfide-functionalized linkers that are then used for the surface modification of citrate-stabilized gold nanoparticles. The binding and sensing abilities toward galectin-3 of both ferrocene-containing lactose dendrimers and gold nanoparticles have been evaluated by means of isothermal titration calorimetry, UV-vis spectroscopy, and differential pulse voltammetry. The highest sensitivity by electrochemical methods to galectin-3 was shown by lactosylferrocenylated gold nanoparticles, which are able to detect the lectin in nanomolar concentrations.

Synthesis of Phosphoramidite Monomers Equipped with Complementary Bases for Solid-Phase DNA Oligomerization.

We describe the preparation of two monomers that bear complementary nucleobases at the edges (guanine-2'-deoxycytidine and 2-aminoadenine-2'-deoxyuridine) and that are conveniently protected and activated for solid-phase automated DNA synthesis. We report the optimized synthetic routes leading to the four nucleobase derivatives involved, their cross-coupling reactions into dinucleobase-containing monomers, and their oligomerization in the DNA synthesizer.

Hiperparatiroidismo primario: reporte de caso clínico / Primary Hyperparathyroidism: Case report

Antecedentes: El hiperparatiroidismo primario, tercer trastorno metabólico más común a nivel mundial, provoca un aumento del calcio sérico y de la hormona paratiroidea, causado por una hiperactividad de las glándulas paratiroideas. En el 85% de los casos, se debe a un adenoma paratiroideo. Es predominantemente asintomático (>80%), pero sus manifestaciones pueden ser musculo-esqueléticas, neurológicas, psiquiátricas, renales, cardiovasculares, y gastrointestinales. Descripción del Caso Clínico: Paciente femenina de 53 años de edad, con antecedente de hipercolesterolemia familiar heterocigoto e hipertensión arterial; presenta insomnio, náuseas, pérdida de peso, mialgias, astenia, relujo gastroesofágico e irritabilidad. Ante indicación de exámenes de labo-ratorio muestra hipercalcemia leve. Se comprueba un adenoma paratiroideo derecho y nódulos tiroideos coloides benignos en lóbulo izquierdo mediante gammagrafía Tc99m MIBI, siendo sometida a su resección con posterior evolución satisfactoria. Conclusiones: Considerando la predilección asintomática del hiperparatiroidismo primario y su preferencia de presentación como adenoma, resulta indispensable su sospecha ante toda hipercalcemia. Requiere entonces una concientización amplia no solo en el ámbito médico sino también en la comunidad, ampliando esfuerzos entre el clínico, cirujano, y patólogo. Se recomienda medir el calcio y fósforo sérico de forma rutinaria en la evaluación clínica cotidiana de cada paciente para su detección...(AU)

Effects of sediment resuspension on the larval stage of the model sponge Carteriospongia foliascens.

Sponges are important components of many marine communities and perform key functional roles. Little is known on the processes that drive larval dispersal and habitat selection in sponges, and in particular under stress scenarios. The increase in sediment in the marine environment is a growing concern for the health of ecosystems, but scarce information exists on the effects of sediment on sponge larvae. This study assessed the effects of suspended and deposited sediment on the larva of Carteriospongia foliascens. A suspended sediment concentration (SSC) of 100 mg L-1 caused homogenisation of the natural pattern of phototactic responses, leading to 100% of photonegative behaviours and a reduction of swim speeds by 27%. After 24 h exposure to suspended sediments, fine particles were found attached to larval cilia, causing abnormal swimming behaviours. Larvae did not have the ability to remove the attached sediment that led to a transformation of the larval body into a cocoon-like morphology and death. Mortality tripled from 3 mg L-1 (9%) to 300 mg L-1 (30%) and the relative SSC EC10 and EC50 values corresponded to 2.6 mg L-1 and 17.6 mg L-1 respectively. Survival, as determined by live swimming larvae, exceeded 50% even in the highest SSC of 300 mg L-1, however settlement success decreased by ~20%. Larvae were able to settle onto substrate having deposited sediment levels (DSLs) up to 3 mg cm-2 (~24%), but recorded a 25 × chance of dislodgement compared to settlers on substrate with DSL of 0.3 mg cm-2. Larvae avoided settling onto substrates with DSLs >10 mg cm-2 and preferentially settled onto alternative vertical substrate that were free of sediment. While C. foliascens larvae have some ability to survive and settle through conditions of elevated sediment, detrimental effects are also clear.

In situ determination of Si, N, and P utilization by the demosponge Tethya citrina: A benthic-chamber approach.

Sponges consume dissolved silicon (DSi) to build their skeletons. Few studies have attempted to quantify DSi utilization by these organisms and all available determinations come from laboratory measurements. Here we measured DSi consumption rates of the sponge Tethya citrina in its natural habitat, conducting 24h incubations in benthic chambers. Sponges consumed DSi at an average rate of 0.046 ± 0.018 µmol h-1 mL-1 when DSi availability in its habitat was 8.3 ± 1.8 µM. Such DSi consumption rates significantly matched the values predicted by a kinetic model elsewhere developed previously for this species through laboratory incubations. These results support the use of laboratory incubations as a suitable approach to learn about DSi consumption. During the field incubations, utilization of other dissolved inorganic nutrients by this low-microbial-abundance (LMA) sponge was also measured. The sponges were net sources of ammonium (-0.043 ± 0.031 µmol h-1 mL-1), nitrate (-0.063 ± 0.031 µmol h-1 mL-1), nitrite (-0.007 ± 0.003 µmol h-1 mL-1), and phosphate (-0.004 ± 0.005 µmol h-1 mL-1), in agreement with the general pattern in other LMA species. The detected effluxes were among the lowest reported for sponges, which agreed with the low respiration rates characterizing this species (0.35 ± 0.11 µmol-O2 h-1 mL-1). Despite relatively low flux, the dense population of T. citrina modifies the availability of dissolved inorganic nutrients in the demersal water of its habitat, contributing up to 14% of nitrate and nitrite stocks. Through these effects, the bottom layer contacting the benthic communities where siliceous LMA sponges abound can be partially depleted in DSi, but can benefit from inputs of N and P dissolved inorganic nutrients that are critical to primary producers.

Improved landfill leachate quality using ozone, UV solar radiation, hydrogen peroxide, persulfate and adsorption processes.

Advanced oxidation processes based on ozonation, UV solar radiation, hydrogen peroxide, and persulfate were tested for the complete treatment of a specific landfill leachate. As a post-treatment of the advanced oxidation process, an additional adsorption process was carried out using a natural zeolite. Application of the UVsolar/O3/H2O2 process required 140 min of irradiation and the consumption of 0.67 g/L H2O2 to eliminate 56% and 17% of colour and chemical oxygen demand (COD), respectively. When adding persulfate to the system (UVsolar/O3/H2O2/S2O8-2), colour and COD were reduced by 29% and 77%, respectively, with a 0.2 g/L concentration of S2O8-2 requiring 250 min of irradiation time. In an experimental run with landfill leachate, adsorption post-treatment with a natural zeolite resulted in 36%, 99%, and 18% total reductions in COD, ammonium, and chloride, respectively. In another experimental run, adsorption with a zeolite was used as a pre-treatment for the advanced oxidation process (UVsolar/O3/H2O2 and UVsolar/O3/H2O2/S2O8-2). The results were similar to those for adsorption post-treatment, with 30%, 90%, and 20% total reductions in COD, ammonium, and chloride, respectively. Despite improvements, some measured parameters of treated landfill leachate still exceeded Chilean regulations for water quality. Furthermore, Lactuca sativa seeds did not germinate after irrigation with 100% treated landfill leachate or dilutions above 3%. Finally, EC50 values were enhanced during treatment, going from 0.002% for raw landfill leachate to 1.179% after the adsorption process.

Selective N-terminal acylation of peptides and proteins with a Gly-His tag sequence.

Methods for site-selective chemistry on proteins are in high demand for the synthesis of chemically modified biopharmaceuticals, as well as for applications in chemical biology, biosensors and more. Inadvertent N-terminal gluconoylation has been reported during expression of proteins with an N-terminal His tag. Here we report the development of this side-reaction into a general method for highly selective N-terminal acylation of proteins to introduce functional groups. We identify an optimized N-terminal sequence, GHHHn- for the reaction with gluconolactone and 4-methoxyphenyl esters as acylating agents, facilitating the introduction of functionalities in a highly selective and efficient manner. Azides, biotin or a fluorophore are introduced at the N-termini of four unrelated proteins by effective and selective acylation with the 4-methoxyphenyl esters. This Gly-Hisn tag adds the unique capability for highly selective N-terminal chemical acylation of expressed proteins. We anticipate that it can find wide application in chemical biology and for biopharmaceuticals.

Peptide Half-Life Extension: Divalent, Small-Molecule Albumin Interactions Direct the Systemic Properties of Glucagon-Like Peptide 1 (GLP-1) Analogues.

Noncovalent binding of biopharmaceuticals to human serum albumin protects against enzymatic degradation and renal clearance. Herein, we investigated the effect of mono- or divalent small-molecule albumin binders for half-life extension of peptides. For proof-of-principle, the clinically relevant glucagon-like peptide 1 (GLP-1) was functionalized with diflunisal, indomethacin, or both. In vitro, all GLP-1 analogues had subnanomolar GLP-1 receptor potency. Surface plasmon resonance revealed that both small molecules were able to confer albumin affinity to GLP-1 and indicated that affinity is increased for divalent analogues. In lean mice, the divalent GLP-1 analogues were superior to monovalent analogues with respect to control of glucose homeostasis and suppression of food intake. Importantly, divalent GLP-1 analogues showed efficacy comparable to liraglutide, an antidiabetic GLP-1 analogue that carries a long-chain fatty acid. Finally, pharmacokinetic investigations of a divalent GLP-1 analogue demonstrated a promising gain in circulatory half-life and absorption time compared to its monovalent equivalent.

Folding Topology of a Short Coiled-Coil Peptide Structure Templated by an Oligonucleotide Triplex.

The rational design of a well-defined protein-like tertiary structure formed by small peptide building blocks is still a formidable challenge. By using peptide-oligonucleotide conjugates (POC) as building blocks, we present the self-assembly of miniature coiled-coil α-helical peptides guided by oligonucleotide duplex and triplex formation. POC synthesis was achieved by copper-free alkyne-azide cycloaddition between three oligonucleotides and a 23-mer peptide, which by itself exhibited multiple oligomeric states in solution. The oligonucleotide domain was designed to furnish a stable parallel triplex under physiological pH, and to be capable of templating the three peptide sequences to constitute a small coiled-coil motif displaying remarkable α-helicity. The formed trimeric complex was characterized by ultraviolet thermal denaturation, gel electrophoresis, circular dichroism (CD) spectroscopy, small-angle X-ray scattering (SAXS), and molecular modeling. Stabilizing cooperativity was observed between the trimeric peptide and the oligonucleotide triplex domains, and the overall molecular size (ca. 12 nm) in solution was revealed to be independent of concentration. The topological folding of the peptide moiety differed strongly from those of the individual POC strands and the unconjugated peptide, exclusively adopting the designed triple helical structure.

Chemoselective Reactions for the Synthesis of Glycoconjugates from Unprotected Carbohydrates.

Glycobiology is the comprehensive biological investigation of carbohydrates. The study of the role and function of complex carbohydrates often requires the attachment of carbohydrates to surfaces, their tagging with fluorophores, or their conversion into natural or non-natural glycoconjugates, such as glycopeptides or glycolipids. Glycobiology and its "omics", glycomics, require easy and robust chemical methods for the construction of these glycoconjugates. This review gives an overview of the rapidly expanding field of chemical reactions that selectively convert unprotected carbohydrates into glycoconjugates through the anomeric position. The discussion is divided in terms of the anomeric bond type of the newly formed glycoconjugates, including O-, N-, S-, and C-glycosides.