An Improved Protection-Free One-Pot Chemical Synthesis of 2′-Deoxynucleoside-5′-Triphosphates

A facile, straightforward, reliable, and an efficient method for the gram-scale chemical synthesis of both purine deoxynucleotides such as 2 ′-deoxyguanosine-5 ′-triphosphate (dGTP) and 2 ′-deoxyadenosine-5 ′-triphosphate (dATP) and pyrimidine deoxynucleotides such as 2 ′-deoxycytidine-5 ′-triphosphate (dCTP), thymidine-5 ′-triphosphate (TTP), and 2 ′-deoxyuridine-5 ′-triphosphate (dUTP) starting from the corresponding nucleoside is described. This improved “one-pot, three step” Ludwig synthetic strategy involves the monophosphorylation of nucleoside followed by reaction with tributylammonium pyrophosphate and hydrolysis of the resulting cyclic intermediate to provide the corresponding dNTP in good yields (65%–70%).     

院前明胶海绵栓塞治疗肾血管损伤的动物研究

目的：探讨在新型综合手术救治方舱中明胶海绵栓塞治疗肾血管损伤的可行性和疗效。方法：在新型综合手术救治方舱中构建3例动物肾腹（背）干动脉损伤模型、3例肾段动脉损伤模型,先用1mm×1mm×1mm明胶海绵颗粒对损伤血管进行栓塞治疗,若未能止血,改用2mm×2mm×2mm颗粒栓塞,观察栓塞后15分钟、1月后的肾动脉造影,栓塞后1月的组织病理表现。结果：3例肾段动脉和1例肾腹干动脉使用1mm×1mm×1mm明胶海绵颗粒一次栓塞成功;1例肾腹干动脉和1例肾背干动脉用1mm×1mm×1mm颗粒栓塞未能完全止血,改用2mm×2mm×2mm颗粒栓塞成功。6例动物术后生命体征平稳,全部存活,一个月后造影复查见栓塞动脉来再通,但其余分支动脉血供得到保留。大体标本观栓塞侧肾明显缩小,来见脓腔和脓性分泌物,病理切片示栓塞后局部肾组织有不同范围缺血性梗死。结论：野战方舱中明胶海绵栓塞治疗出血效果明确,对于一般肾血管损伤定位准确,止血迅速、彻底,有良好的可行性和疗效。     

Multi-scale mineralogical characterization of the hypercalcified sponge Petrobiona massiliana (Calcarea, Calcaronea)

The massive basal skeleton of a few remnant living hypercalcified sponges rediscovered since the 1960s are valuable representatives of ancient calcium carbonate biomineralization mechanisms in basal Metazoa. A multi-scale mineralogical characterization of the easily accessible Mediterranean living hypercalcified sponge belonging to Calcarea, Petrobiona massiliana (Vacelet and Lévi, 1958), was conducted. Oriented observations in light and electron microscopy of mature and growing areas of the Mg-calcite basal skeleton were combined in order to describe all structural levels from the submicronic to the macroscopic scale. The smallest units produced are ca. 50–100 nm grains that are in a mushy amorphous state before their crystallization. Selected area electron diffraction (SAED) further demonstrated that submicronic grains are assembled into crystallographically coherent clusters or fibers, the latter are even laterally associated into single-crystal bundles. A model of crystallization propagation through amorphous submicronic granular units is proposed to explain the formation of coherent micron-scale structural units. Finally, XRD and EELS analyses highlighted, respectively, inter-individual variation of skeletal Mg contents and heterogeneous spatial distribution of Ca ions in skeletal fibers. All mineralogical features presented here cannot be explained by classical inorganic crystallization principles in super-saturated solutions, but rather underlined a highly biologically regulated formation of the basal skeleton. This study extending recent observations on corals, mollusk and echinoderms confirms that occurrence of submicronic granular units and a possible transient amorphous precursor phase in calcium carbonate skeletons is a common biomineralization strategy already selected by basal metazoans.     

Pyrosequencing reveals highly diverse and species-specific microbial communities in sponges from the Red Sea

Marine sponges are associated with a remarkable array of microorganisms. Using a tag pyrosequencing technology, this study was the first to investigate in depth the microbial communities associated with three Red Sea sponges, Hyrtios erectus, Stylissa carteri and Xestospongia testudinaria. We revealed highly diverse sponge-associated bacterial communities with up to 1000 microbial operational taxonomic units (OTUs) and richness estimates of up to 2000 species. Altogether, 26 bacterial phyla were detected from the Red Sea sponges, 11 of which were absent from the surrounding sea water and 4 were recorded in sponges for the first time. Up to 100 OTUs with richness estimates of up to 300 archaeal species were revealed from a single sponge species. This is by far the highest archaeal diversity ever recorded for sponges. A non-negligible proportion of unclassified reads was observed in sponges. Our results demonstrated that the sponge-associated microbial communities remained highly consistent in the same sponge species from different locations, although they varied at different degrees among different sponge species. A significant proportion of the tag sequences from the sponges could be assigned to one of the sponge-specific clusters previously defined. In addition, the sponge-associated microbial communities were consistently divergent from those present in the surrounding sea water. Our results suggest that the Red Sea sponges possess highly sponge-specific or even sponge-species-specific microbial communities that are resistant to environmental disturbance, and much of their microbial diversity remains to be explored.     

Questioning the evidence of organic compounds called sponge biomarkers

Elevated concentrations of an organic compound, 24‐isopropylcholestane, found in the Precambrian Huqf Supergroup of Oman may provide the oldest known sponge ‘fossil’. This evidence is of critical importance for a properly balanced understanding of the origin of animals. Several different pelagophyte (Class Pelagophyceae part of the Stramenopiles within the Chromaveolata) algae are also capable of producing these exact compounds, and may similarly have done so in deep time. Modern marine algae are also reported to produce structural isomers that are compositionally identical to the sponge marker; they do this in copious quantities. Further, 24‐isopropylcholestane can be produced by diagenetic alteration of compounds produced in large quantities by algae. It is also possible that contamination by petroleum derived lubricating oil used when coring while extracting these compounds from subsurface layers, has affected the data. All extinct organisms that may have produced this compounds are unavailable for analysis by the modern organic chemist and cannot be eliminated from the list of possible producers of the sponge marker. There are also significant uncertainties regarding the dating of the strata from which these ancient compounds are found. Although the compounds are widely reported as c. 751 Ma, they are younger than 645 Ma. It seems more likely that these compounds represent algal biochemical evolution at a time when algal burial occurred in great quantity with well known coeval algal fossils but no sponge fossils. The macroalgal biomass may have declined during the agronomic revolution at the base of the Cambrian Period owing to processing by metazoans, accounting for the comparative scarcity of these sponge markers in Phanerozoic sediments, after which time sponge spicules and body fossils become evident.     

Reproductive processes in sponges: a critical evaluation of current data and views

The nature of a number of fundamental processes occurring during reproduction in sponges still remains in doubt. Among the more significant of these are: the true status of sponges described as dioecious, namely whether some are actually successive hermaphrodites; the origin of oogonia, which have recently been claimed to be derived from choanocytes; the origin and mechanism of formation of large spermatogenic masses; the specific pathway leading to fertilization taken by sperm cells within the sponge tissue of viviparous species; the role played during larval metamorphosis by somatic cells which are incorporated into embryos; the cell lineage of choanocytes which form flagellated chambers during larval metamorphosis; the specific relationship of somatic growth and dormancy to gametogenesis; the role of budding and fragmentation in population maintenance; the role, if any, of gemmules in dispersion. It is considered mandatory that new techniques be developed in order to further elucidate these and other reproductive processes and to gather definitive data concerning them. The employment of only microscopic techniques is ultimately insufficient for investigating the dynamic relationships of reproductive processes.     

Single-cell genomics reveals complex carbohydrate degradation patterns in poribacterial symbionts of marine sponges

Many marine sponges are hosts to dense and phylogenetically diverse microbial communities that are located in the extracellular matrix of the animal. The candidate phylum Poribacteria is a predominant member of the sponge microbiome and its representatives are nearly exclusively found in sponges. Here we used single-cell genomics to obtain comprehensive insights into the metabolic potential of individual poribacterial cells representing three distinct phylogenetic groups within Poribacteria. Genome sizes were up to 5.4 Mbp and genome coverage was as high as 98.5%. Common features of the poribacterial genomes indicated that heterotrophy is likely to be of importance for this bacterial candidate phylum. Carbohydrate-active enzyme database screening and further detailed analysis of carbohydrate metabolism suggested the ability to degrade diverse carbohydrate sources likely originating from seawater and from the host itself. The presence of uronic acid degradation pathways as well as several specific sulfatases provides strong support that Poribacteria degrade glycosaminoglycan chains of proteoglycans, which are important components of the sponge host matrix. Dominant glycoside hydrolase families further suggest degradation of other glycoproteins in the host matrix. We therefore propose that Poribacteria are well adapted to an existence in the sponge extracellular matrix. Poribacteria may be viewed as efficient scavengers and recyclers of a particular suite of carbon compounds that are unique to sponges as microbial ecosystems.     

Amperometric Detection of Reduced Nicotinamide Adenine Dinucleotide Using a Film-covered Organic Salt Electrode

A new N-acylated serinol, inconspicamide (1), was isolated from the marine sponge, Stelletta inconspicua, together with a glyceryl ether (2). Their structures were determined on the basis of spectroscopic data and the modified Mosher analysis. They exhibited moderate cytotoxic activity against HeLa human cervical cancer cells.     

Metachromins X and Y from a marine sponge Spongia sp. and their effects on cell cycle progression

New sesquiterpene quinones, metachromins X (1) and Y (2), together with the known metachromins C (3), J (4), and T (5), were isolated as inhibitors of cell cycle progression in the HeLa/Fucci2 cells. The structure of 1 was assigned by spectroscopic data and confirmed by a total synthesis. The planar structure of 2 was determined by interpretation of spectroscopic data, whereas its absolute configuration was analyzed by a combination of chiral HPLC and CD spectroscopy. Metachromins X (1) and C (3) arrested the cell cycle progression of HeLa/Fucci2 cells at S/G2/M phase.