cDNA and Gene Analyses Imply a Novel Structure for a Rat Carcinoembryonic Antigen-related Protein

The gene encoding the human tumor marker carcinoembryonic antigen (CEA) belongs to a gene family which can be subdivided into the CEA and the pregnancy-specific glycoprotein subgroups. The corresponding proteins are members of the immunoglobulin superfamily, characterized through the presence of one IgV-like domain and a varying number of IgC-like domains. Since the function of the CEA family is not well understood, we decided to establish an animal model in the rat to study its tissue- specific and developmental stage-dependent expression. To this end, we have screened an 18-day rat placenta cDNA library with a recently isolated fragment of a rat CEA-related gene. Two overlapping clones containing the complete coding region for a putative 709 amino acid protein (rnCGM1; Mr = 78,310) have been characterized. In contrast to all members of the human CEA family, this rat CEA-related protein consists of five IgV-like domains and only one IgC-like domain. This novel structure, which has been confirmed at the genomic level might have important functional implications. Due to the rapid evolutionary divergence of the rat and human CEA gene families it is not possible to assign rnCGM1 to its human counterpart. However, the predominant expression of the rnCGM1 gene in the placenta suggests that it could be analogous to one of the human pregnancy-specific glycoprotein genes

Spatiotemporal Expression of Pregnancy-Specific Glycoprotein Gene rnCGMl in Rat Placenta

As a basis towards a better understanding of the role of the pregnancy-specific glycoprotein (PSG) family in the maintenance of pregnancy, detailed investigations are described on the expression of a recently identified rat PSG gene (rnCGM1) at the mRNA and protein levels. Using specific oligonucleotide primers, rnCGM1 transcripts were identified after reverse transcription, polymerase chain reaction, and hybridization with a radiolabelled, internal oligonucleotide. Transcripts were only found in significant amounts in placenta. In situ hybridization visualized rnCGM1 transcripts at day 14 post coitum (p.c.), in secondary trophoblast giant cells and in the spongiotrophoblast. Only those secondary giant cells lining the maternal decidua were positive. In contrast, primary giant cells did not contain rnCGM1 mRNA. At day 18 p.c., rnCGM1. transcripts were almost exclusively detectable in the spongiotrophoblast. No rnCGM1 transcripts were found in rat embryos of these two developmental stages. Rabbit antisera were generated against the amino-terminal immunoglobulin variable-like domain and against a synthetic peptide containing the last 13 carboxy-terminal amino acids of rnCGM1. Bothe antisera recognized a 124 kDa protein in day 18 rat placental extracts as identified by Western blot analysis. The anti-peptide antiserum recognized a 116 kDa protein in the serum of a 14 day p.c. pregnant rat that is absent from the sera of non-pregnant females. Taken together, these results confirm exclusive expression of rnCGM1 in the rat trophoblast, but unlike human PSG, negligible or no expression is found in other organs, such as fetal liver or salivary glands, indicating a more specialized function of rnCGM1. Its spatiotemporal expression pattern is conducive with a potential role of PSG in protecting the fetus against the maternal immune system and/or in regulating the invasive growth of trophoblast cells

Table of Contents and Prologue

Editorial board, Table of contents, and Prologue, an introduction to volume 2

Identification of a Carcinoembryonic Antigen Gene Family in the Rat

The existence of a carcinoembryonic antigen (CEA)-like gene family in rat has been demonstrated through isolation and sequencing of the N- terminal domain exons of presumably five discrete genes (rnCGM1-5). This finding will allow for the first time the study of functional and clinical aspects of the tumor marker CEA and related antigens in an animal model. Sequence comparison with the corresponding regions of members of the human CEA gene family revealed a relatively low similarity at the amino acid level, which indicates rapid divergence of the CEA gene family during evolution and explains the lack of cross- reactivity of rat CEA-like antigens with antibodies directed against human CEA. The N-terminal domains of the rat CEA-like proteins show structural similarity to immunoglobulin variable domains, including the presence of hypervariable regions, which points to a possible receptor function of the CEA family members. Although so far only one of the five rat CEA-like genes could be shown to be transcriptionally active, multiple mRNA species derived from other members of the rat CEA-like gene family have been found to be differentially expressed in rat placenta and liver

Comparison of Statistical and Model-Based Hindcasts of Subtidal Water Levels in Chesapeake Bay

Subtidal water levels in Chesapeake Bay, which can have amplitudes as large as 1 m at Baltimore, are an important component of total water levels. The most importance forcing mechanisms for these variations are surface winds over the Bay and coastal subtidal water levels. Two methods for hindcasting subtidal water levels in the Bay were developed: statistical prediction (based on multiple linear regression) and a barotropic numerical circulation model-based prediction. The hindcast water levels were compared with the observed values at three key locations (Chesapeake Bay Bridge Tunnel (CBBT) in the lower bay near the mouth, Solomons Island at midbay, and Baltimore in the upper bay) by a. variety of statistical measures. The hindcast results show that in both annually averaged differences and in the incidence of outliers the numerical model-based hindcasts are slightly more accurate than the statistical hindcasts, although on a monthly basis the statistical hindcast was often equal to or better than the model hindcast. Errors in both methods follow a seasonal pattern, being smallest in the summer months and largest in winter. Both hindcast methods explain significant portions of the observed variance. In addition, several techniques were used to estimate the relative importance of coastal water level forcing and wind forcing in the subtidal variability. In general, the two forcings were about of equal importance at Baltimore, while coastal forcing was dominant at Solomons Island and CBBT

The Proper Candidate: An Examination of the 1525 Debate Between Ulrich Zwingli and Balthasar Hubmaier Concerning Baptism

This thesis examines the written debate that occurred in 1525-1526 between Balthasar Hubmaier and Ulrich Zwingli on the proper candidate of baptism. Both pastors held to the reformation principle of sola scriptura yet came to different conclusions as to what the Bible teaches about baptism. Hubmaier argued that baptism should be administered to believers only. Only those who have mindfully repented and have chosen to follow Christ are eligible for Christian baptism. Hubmaier\u27s arguments for believers\u27 baptism are presented in the second chapter. Zwingli, on the other hand, argued that the children of believers are entitled to the Christian baptism. Just as circumcision was a sign of the Old Testament covenant with Israel, baptism is the sign of the New Covenant with the church. The arguments for infant baptism by Zwingli are set forth in the third chapter. Next, Hubmaier\u27s rebuttal of Zwingli is examined. Hubmaier pointed out inconsistencies in Zwingli\u27s arguments. His strongest argument is the lack of example or instruction of infant baptism in Scripture. The fifth chapter outlines Zwingli\u27s refutation of Hubmaier. Zwingli suggests that Hubmaier is causing division in the church and ignoring the Old Testament. His best argument is the example of entire households being baptized into the church. These households, conceivably, would have included children. Finally, the final chapter is an analysis of the arguments presented by Zwingli and Hubmaier, specifically in light of sola scriptura. While Zwingli made compelling arguments, Hubmaier\u27s argument that baptism is for believers was founded more upon Scripture

Differential Uptake of Gold Nanoparticles by 2 Species of Tadpole, the Wood Frog (Lithobates Sylvaticus) and the Bullfrog (Lithobates Catesbeianus)

Engineered nanoparticles are aquatic contaminants of emerging concern that exert ecotoxicological effects on a wide variety of organisms. We exposed cetyltrimethylammonium bromide–capped spherical gold nanoparticles to wood frog and bullfrog tadpoles with conspecifics and in combination with the other species continuously for 21 d, then measured uptake and localization of gold. Wood frog tadpoles alone and in combination with bullfrog tadpoles took up significantly more gold than bullfrogs. Bullfrog tadpoles in combination with wood frogs took up significantly more gold than controls. The rank order of weight-normalized gold uptake was wood frogs in combination \u3e wood frogs alone \u3e bullfrogs in combination \u3e bullfrogs alone \u3e controls. In all gold-exposed groups of tadpoles, gold was concentrated in the anterior region compared with the posterior region of the body. The concentration of gold nanoparticles in the anterior region of wood frogs both alone and in combination with bullfrogs was significantly higher than the corresponding posterior regions. We also measured depuration time of gold in wood frogs. After 21 d in a solution of gold nanoparticles, tadpoles lost \u3e83% of internalized gold when placed in gold-free water for 5 d. After 10 d in gold-free water, tadpoles lost 94% of their gold. After 15 d, gold concentrations were below the level of detection. Our finding of differential uptake between closely related species living in similar habitats with overlapping geographical distributions argues against generalizing toxicological effects of nanoparticles for a large group of organisms based on measurements in only one species

Pattern Recognition Receptors and the Innate Immune Response to Viral Infection

The innate immune response to viral pathogens is critical in order to mobilize protective immunity. Cells of the innate immune system detect viral infection largely through germline-encoded pattern recognition receptors (PRRs) present either on the cell surface or within distinct intracellular compartments. These include the Toll-like receptors (TLRs), the retinoic acid-inducble gene I-like receptors (RLRs), the nucleotide oligomerization domain-like receptors (NLRs, also called NACHT, LRR and PYD domain proteins) and cytosolic DNA sensors. While in certain cases viral proteins are the trigger of these receptors, the predominant viral activators are nucleic acids. The presence of viral sensing PRRs in multiple cellular compartments allows innate cells to recognize and quickly respond to a broad range of viruses, which replicate in different cellular compartments. Here, we review the role of PRRs and associated signaling pathways in detecting viral pathogens in order to evoke production of interferons and cytokines. By highlighting recent progress in these areas, we hope to convey a greater understanding of how viruses activate PRR signaling and how this interaction shapes the anti-viral immune response

Subaru optical observations of the old pulsar PSR B0950+08

We report the B band optical observations of an old (17.5 Myr) radiopulsar PSR B0950+08 obtained with the Suprime-Cam at the Subaru telescope. We detected a faint object, B=27.07(16). Within our astrometrical accuracy it coincides with the radio position of the pulsar and with the object detected earlier by Pavlov et al. (1996) in UV with the HST/FOC/F130LP. The positional coincidence and spectral properties of the object suggest that it is the optical counterpart of PSR B0950+08. Its flux in the B band is two times higher than one would expect from the suggested earlier Rayleigh-Jeans interpretation of the only available HST observations in the adjacent F130LP band. Based on the B and F130LP photometry of the suggested counterpart and on the available X-ray data we argue in favour of nonthermal origin of the broad-band optical spectrum of PSR B0950+08, as it is observed for the optical emission of the younger, middle-aged pulsars PSR B0656+14 and Geminga. At the same time, the optical efficiency of PSR B0950+08, estimated from its spin-down power and the detected optical flux, is by several orders of magnitude higher than for these pulsars, and comparable with that for the much younger and more energetic Crab pulsar. We cannot exclude the presence of a compact, about 1'', faint pulsar nebula around PSR B0950+08, elongated perpendicular to the vector of its proper motion, unless it is not a projection of a faint extended object on the pulsar position.Comment: 8 pages, LaTeX, aa.cls style, 5 PS figures, submitted to A&A. Image is available in FITS format at http://www.ioffe.rssi.ru/astro/NSG/obs/0950-subar

Crystal Structure of SARS-CoV-2 Main Protease in Complex with the Non-Covalent Inhibitor ML188

Viral proteases are critical enzymes for the maturation of many human pathogenic viruses and thus are key targets for direct acting antivirals (DAAs). The current viral pandemic caused by SARS-CoV-2 is in dire need of DAAs. The Main protease (M(pro)) is the focus of extensive structure-based drug design efforts which are mostly covalent inhibitors targeting the catalytic cysteine. ML188 is a non-covalent inhibitor designed to target SARS-CoV-1 M(pro), and provides an initial scaffold for the creation of effective pan-coronavirus inhibitors. In the current study, we found that ML188 inhibits SARS-CoV-2 M(pro) at 2.5 microM, which is more potent than against SAR-CoV-1 M(pro). We determined the crystal structure of ML188 in complex with SARS-CoV-2 M(pro) to 2.39 A resolution. Sharing 96% sequence identity, structural comparison of the two complexes only shows subtle differences. Non-covalent protease inhibitors complement the design of covalent inhibitors against SARS-CoV-2 main protease and are critical initial steps in the design of DAAs to treat CoVID 19