On the way from Asia to America: eutriconodontan mammals from the Early Cretaceous of Yakutia, Russia.

Eutriconodonta are an important group of early crown mammals with a wide distribution in the Jurassic-Cretaceous of the Northern Hemisphere and few occurrences in the Southern Hemisphere. Three taxa of eutriconodontans are known from the Early Cretaceous high-latitude Teete vertebrate assemblage in Yakutia, Russia: Sangarotherium aquilonium (Eutriconodonta incertae sedis), Gobiconodon sp. A (large), and Gobiconodon sp. B (small) (Gobiconodontidae). These three taxa are based on four specimens and indicate a remarkable taxonomic diversity of eutriconodontans at this locality. The coexistence of two Gobiconodon species, large and small, is characteristic for several Early Cretaceous vertebrate assemblages in Asia. Gobiconodon sp. A from the Teete locality is the largest species of this genus known from Asia, but is smaller than the North American G. ostromi. The spreading of Gobiconodon from Asia to North America likely occurred during the Aptian-Albian faunal dispersal event. The discovery of Gobiconodon in the Teete locality is further evidence for a dispersal route via Beringia from Asia to North America which previously has been postulated based on the occurrence of Asian dinosaur taxa in western North America at this time. The questionable record of Gobiconodon from Europe and its lack from eastern North America make a dispersal from Asia to North America via Europe less probable.

Wear patterns and dental functioning in an Early Cretaceous stegosaur from Yakutia, Eastern Russia.

Isolated stegosaurian teeth from the Early Cretaceous high-latitude (palaeolatitude estimate of N 62°- 66.5°) Teete locality in Yakutia (Eastern Siberia, Russia) are characterized by a labiolingually compressed, slightly asymmetrical and mesiodistally denticulated (9-14 denticles) crown, a pronounced ring-like cingulum, as well as a "complex network of secondary ridges". The 63 teeth (found during on-site excavation in 2012, 2017-2019 and screen-washing in 2017-2019) most likely belong to one species of a derived (stegosaurine) stegosaur. Most of the teeth exhibit a high degree of wear and up to three wear facets has been observed on a single tooth. The prevalence of worn teeth with up to three wear facets and the presence of different types of facets (including steeply inclined and groove-like) indicate the tooth-tooth contact and precise dental occlusion in the Teete stegosaur. The microwear pattern (mesiodistally or slightly obliquely oriented scratches; differently oriented straight and curved scratches on some wear facets) suggest a complex jaw mechanism with palinal jaw motion. Histological analysis revealed that the Teete stegosaur is characterized by relatively short tooth formation time (95 days) and the presence of a "wavy enamel pattern". Discoveries of a "wavy enamel pattern" in the Teete stegosaur, in a Middle Jurassic stegosaur from Western Siberia, and in the basal ceratopsian Psittacosaurus, suggest that this histological feature is common for different ornithischian clades, including ornithopods, marginocephalians, and thyreophorans. A juvenile tooth in the Teete sample indicates that stegosaurs were year-round residents and reproduced in high latitudes. The combination of high degree of tooth wear with formation of multiple wear facets, complex jaw motions, relatively short tooth formation time and possibly high tooth replacement rates is interpreted as a special adaptation for a life in high-latitude conditions or, alternatively, as a common stegosaurian adaptation making stegosaurs a successful group of herbivorous dinosaurs in the Middle Jurassic-Early Cretaceous and enabeling them to live in both low- and high-latitude ecosystems.

Impact of catalyst load, chemical oxygen demand and nitrite on disinfection and removal of contaminants during catalytic ozonation of wastewater.

Calcium-silicate mineral Polonite® and aluminum-based catalyst (AL-1010S), previously identified as promising materials for catalytic ozonation, were used as catalysts to investigate the impact of some operating conditions (ratio ozone feed concentration to catalyst load) and wastewater characteristics (chemical oxygen demand - COD and nitrite - NO2 concentration) on the disinfection and removal of contaminants of emerging concern (CECs) during catalytic ozonation of wastewater. Tests conducted in synthetic wastewater using two different ozone gas concentration (4 and 8 g (nm3)) and 6 different catalyst loads provided ratios of 0.08, 0.11, 0.16, and 0.32. Results from the experiments indicated that the ratio of 0.11 was optimal and reached residual disinfection below 2 MPN mL-1 from the initial concentration of 5 ±â€¯2 × 105 MPN mL-1 and removal of atrazine (ATZ) above 80% from the initial concentration of 100 ±â€¯10 µg L-1 for an ozone dose of 41-45 mg L-1. Catalytic ozonation with the selected materials enhanced disinfection and ATZ removal from synthetic wastewater (SWW) in comparison to non-catalytic ozonation by making the treatment performance less sensitive to increased chemical oxygen demand (COD) and nitrite (NO2) in the matrix. Validation of the results in real wastewater effluents confirmed that catalytic ozonation enhanced disinfection. Catalytic ozonation using Polonite® and AL-1010S provided residual bacteria level of 0.6 ±â€¯0.42 MPN mL-1 and 0.29 ±â€¯0.41 MPN mL-1, while non-catalytic ozonation lead to an average residual bacteria level of 1.26 ±â€¯0.09 MPN mL-1 for the same range of transferred ozone dose. However, under the conditions tested, a limited number of CECs were extracted at levels above the limits of quantification and further validation work required to evaluate the performance of catalytic ozonation for the removal of CECs.

A high-latitude fauna of mid-Mesozoic mammals from Yakutia, Russia.

The Early Cretaceous (?Berriasian-Barremian) Teete vertebrate locality in Western Yakutia, East Siberia, Russia, has produced mammal remains that are attributed to three taxa: Eleutherodontidae indet. cf. Sineleutherus sp. (Haramiyida; an upper molariform tooth), Khorotherium yakutensis gen. et sp. nov. (Tegotheriidae, Docodonta; maxillary fragment with three molariform teeth and dentary fragment with one molariform tooth), and Sangarotherium aquilonium gen. et sp. nov. (Eutriconodonta incertae sedis; dentary fragment with one erupted molariform tooth and one tooth in crypt). This is the second occurrence of Mesozoic mammals in high latitudes (paleolatitude estimate N 63-70°) of the Northern Hemisphere. In spite of the presumed Early Cretaceous age based on freshwater mollusks, the Teete mammal assemblage has a distinctive Jurassic appearance, being most similar to the Middle-Late Jurassic mammal assemblages known from Siberia, Russia and Xinjiang, China. The smooth transition from Jurassic to Cretaceous biota in Northern Asia is best explained by stable environmental conditions.

Novel materials for catalytic ozonation of wastewater for disinfection and removal of micropollutants.

The purpose of this work was to identify novel materials that could be used for the catalytic ozonation of wastewater for improved removal of micropollutants and disinfection. The materials chosen for investigation were selected based on their commercial availability and composition characteristics that suggested that they could act as catalysts. Synthetic wastewater (SWW) was used to mimic municipal secondary effluent wastewater while obtaining constant and reproducible matrix characteristics. Polonite®, wollastonite, zeolite, TiO2-Al2O3 (8%/92%), and AL-1010S (AlO2-based) were tested for their potential impact on efficiency of disinfection, based on the removal of E. coli bacteria and removal of contaminants of emerging concern (CECs), relative to conventional ozonation. Atrazine (ATZ), ibuprofen (IBP), naproxen (NPX), and gemfibrozil (GBZ) were used as indicator compounds. Zeolite and wollastonite did not promote disinfection and CECs removal; TiO2-Al2O3 and AL-1010S provided improvement for both criteria, but to a lesser extent in SWW than in Milli-Q water; and Polonite® did not enhance the removal of CECs but led to the higher E. coli inactivation. These results suggest that Polonite®, AL-1010S, and TiO2-Al2O3 can act as catalysts and provide mechanisms to lower the ozone dose required to reach disinfection. The apparent kinetic constant of the reaction for the catalytic ozonation of ATZ, in the ultrapure water, was determined for Polonite®, TiO2-Al2O3 and AL-1010S. The reusability of the catalysts was demonstrated over four consecutive cycles of 6 h of treatment in a continuous flow ozonation system.

Intracerebroventricular Administration of 192IgG-Saporin Alters Expression of Microglia-Associated Genes in the Dorsal But Not Ventral Hippocampus.

One of important aspects of development of Alzheimer's disease is degeneration of septal cholinergic neurons that innervate the hippocampus. We took advantage of widely used model of cholinergic deficit in the hippocampus, intracerebroventricular administration of 192IgG-saporin (Ig-saporin), to analyze the postponed consequences of cholinergic deficit in different parts of the hippocampus. We studied effects of the immunotoxin on the behavior of rats and gene expression in the dorsal and ventral hippocampus using RNA-seq approach. We found that under normal conditions dorsal and ventral parts of the hippocampus differ in the expression of 1129 protein-coding genes and 49 non-coding RNAs (ncRNAs) and do not differ in the expression of 10 microRNAs, which were detected in both parts of the hippocampus. Ig-saporin-induced degeneration of cholinergic septal neurons did not affect rat behavior in open field, T-maze, and passive avoidance task but impaired memory retention in Morris water maze. To analyze 192Ig-saporin-induced changes in the gene expression, we formed the following groups of genes: genes expressed exclusively in certain cell types (neurons, astrocytes, microglia, oligodendrocytes, and vascular cells) and, among universally expressed genes, a group of genes that encode ribosome-forming proteins. For all groups of genes, the alterations in the gene expression produced by the immunotoxin were stronger in the dorsal as compared to the ventral hippocampus. We found that, among groups of universally expressed genes, Ig-saporin increased the expression of ribosome-forming proteins in both dorsal and ventral hippocampus. Ig-saporin also strongly upregulated expression of microglia-specific genes only in the dorsal hippocampus. A subset of affected microglial genes comprised genes associated with inflammation, however, did not include genes related to acute inflammation such as interleukins-1b, -6, -15, and -18 as well as TNF. The expression of other cell-specific genes (genes specific for neurons, astrocytes, oligodendrocytes, and vascular cells) was unaffected. The data obtained suggest that disturbance of memory-associated behavior after administration of Ig-saporin is associated with upregulation of microglia-associated genes in the dorsal but not ventral hippocampus.

Adenine and guanine recognition of stop codon is mediated by different N domain conformations of translation termination factor eRF1.

Positioning of release factor eRF1 toward adenines and the ribose-phosphate backbone of the UAAA stop signal in the ribosomal decoding site was studied using messenger RNA (mRNA) analogs containing stop signal UAA/UAAA and a photoactivatable cross-linker at definite locations. The human eRF1 peptides cross-linked to these analogs were identified. Cross-linkers on the adenines at the 2nd, 3rd or 4th position modified eRF1 near the conserved YxCxxxF loop (positions 125-131 in the N domain), but cross-linker at the 4th position mainly modified the tripeptide 26-AAR-28. This tripeptide cross-linked also with derivatized 3'-phosphate of UAA, while the same cross-linker at the 3'-phosphate of UAAA modified both the 26-28 and 67-73 fragments. A comparison of the results with those obtained earlier with mRNA analogs bearing a similar cross-linker at the guanines indicates that positioning of eRF1 toward adenines and guanines of stop signals in the 80S termination complex is different. Molecular modeling of eRF1 in the 80S termination complex showed that eRF1 fragments neighboring guanines and adenines of stop signals are compatible with different N domain conformations of eRF1. These conformations vary by positioning of stop signal purines toward the universally conserved dipeptide 31-GT-32, which neighbors guanines but is oriented more distantly from adenines.

Three distinct peptides from the N domain of translation termination factor eRF1 surround stop codon in the ribosome.

To study positioning of the polypeptide release factor eRF1 toward a stop signal in the ribosomal decoding site, we applied photoactivatable mRNA analogs, derivatives of oligoribonucleotides. The human eRF1 peptides cross-linked to these short mRNAs were identified. Cross-linkers on the guanines at the second, third, and fourth stop signal positions modified fragment 31-33, and to lesser extent amino acids within region 121-131 (the "YxCxxxF loop") in the N domain. Hence, both regions are involved in the recognition of the purines. A cross-linker at the first uridine of the stop codon modifies Val66 near the NIKS loop (positions 61-64), and this region is important for recognition of the first uridine of stop codons. Since the N domain distinct regions of eRF1 are involved in a stop-codon decoding, the eRF1 decoding site is discontinuous and is not of "protein anticodon" type. By molecular modeling, the eRF1 molecule can be fitted to the A site proximal to the P-site-bound tRNA and to a stop codon in mRNA via a large conformational change to one of its three domains. In the simulated eRF1 conformation, the YxCxxxF motif and positions 31-33 are very close to a stop codon, which becomes also proximal to several parts of the C domain. Thus, in the A-site-bound state, the eRF1 conformation significantly differs from those in crystals and solution. The model suggested for eRF1 conformation in the ribosomal A site and cross-linking data are compatible.

Different modes of stop codon restriction by the Stylonychia and Paramecium eRF1 translation termination factors.

In universal-code eukaryotes, a single-translation termination factor, eukaryote class-1 polypeptide release factor (eRF1), decodes the three stop codons: UAA, UAG, and UGA. In some ciliates, like Stylonychia and Paramecium, eRF1s exhibit UGA-only decoding specificity, whereas UAG and UAA are reassigned as sense codons. Because variant-code ciliates may have evolved from universal-code ancestor(s), structural features should exist in ciliate eRF1s that restrict their stop codon recognition. In omnipotent eRF1s, stop codon recognition is associated with the N-terminal domain of the protein. Using both in vitro and in vivo assays, we show here that chimeric molecules composed of the N-terminal domain of Stylonychia eRF1 fused to the core domain (MC domain) of human eRF1 retained specificity toward UGA; this unambiguously associates eRF1 stop codon specificity to the nature of its N-terminal domain. Functional analysis of eRF1 chimeras constructed by swapping ciliate N-terminal domain sequences with the matching ones from the human protein highlighted the crucial role of the tripeptide QFM in restricting Stylonychia eRF1 specificity toward UGA. Using the site-directed mutagenesis, we show that Paramecium eRF1 specificity toward UGA resides within the NIKS (amino acids 61-64) and YxCxxxF (amino acids 124-131) motifs. Thus, we establish that eRF1 from two different ciliates relies on different molecular mechanisms to achieve specificity toward the UGA stop codon. This finding suggests that eRF1 restriction of specificity to only UGA might have been an early event occurring in independent instances in ciliate evolutionary history, possibly facilitating the reassignment of UAG and UAA to sense codons.

NMR assignments of the C-terminal domain of human polypeptide release factor eRF1.

We report NMR assignments of the protein backbone of the C-terminal domain (163 a.a.) of human class 1 translation termination factor eRF1. It was found that several protein loop residues exist in two slowly interconverting conformational states.

Invariant amino acids essential for decoding function of polypeptide release factor eRF1.

In eukaryotic ribosome, the N domain of polypeptide release factor eRF1 is involved in decoding stop signals in mRNAs. However, structure of the decoding site remains obscure. Here, we specifically altered the stop codon recognition pattern of human eRF1 by point mutagenesis of the invariant Glu55 and Tyr125 residues in the N domain. The 3D structure of generated eRF1 mutants was not destabilized as demonstrated by calorimetric measurements and calculated free energy perturbations. In mutants, the UAG response was most profoundly and selectively affected. Surprisingly, Glu55Arg mutant completely retained its release activity. Substitution of the aromatic ring in position 125 reduced response toward all stop codons. This result demonstrates the critical importance of Tyr125 for maintenance of the intact structure of the eRF1 decoding site. The results also suggest that Tyr125 is implicated in recognition of the 3d stop codon position and probably forms an H-bond with Glu55. The data point to a pivotal role played by the YxCxxxF motif (positions 125-131) in purine discrimination of the stop codons. We speculate that eRF1 decoding site is formed by a 3D network of amino acids side chains.