Purified Immunoglobulin F(ab′) 2 Protects Mice and Rhesus Monkeys against Lethal Ricin Intoxication

Ricin is a highly toxic ribosome-inactivating lectin derived from castor beans. To date, no antidote is available to treat ricin-poisoned patients, and the development of a safe and effective antidote is urgently needed. First, ricin was prepared and used to construct a mouse model and a rhesus monkey model of ricin intoxication. Second, pepsin-digested F(ab′) 2 fragments of serum IgG from horses injected with Freund’s-adjuvanted purified ricin were prepared. Third, the protective efficacy was evaluated in mouse and rhesus monkey models of lethal ricin intoxication. The purity quotient of the prepared ricin and F(ab′) 2 fragments exceeded 90% and 85% in the mouse and monkey models, respectively. The LD 50 of ricin in mice and rhesus monkeys was 2.7 and 9 μg/kg, respectively. A quantity of 6.25 and 1.85 mg/kg F(ab′) 2 was sufficient to treat lethal ricin intoxication in the mice and rhesus monkeys, respectively. Finally, the effect of this therapeutic antibody on peripheral blood immune cells was examined by analysis of peripheral blood immune cells through single cell sequencing. The underlying mechanism was found to involve restraining neutrophil activation, proliferation, and differentiation. Purified F(ab′) 2 fragments administered with needle-free devices fully protect mice and rhesus monkeys against lethal doses of ricin intoxication

Construction and Evaluation of the Brucella Double Gene Knock-out Vaccine Strain MB6 Δbp26ΔwboA (RM6)

Brucellosis is a serious zoonotic infection worldwide. To date, vaccination is the most effective measure against brucellosis. This study was aimed at obtaining a vaccine strain that has high protective efficacy and low toxicity, and allows vaccination to be differentiated from infection. Using homologous recombination, we constructed a double gene-deletion Brucella strain MB6 Δbp26ΔwboA (RM6) and evaluated its characteristics, safety and efficacy. The RM6 strain had good proliferative ability and stable biological characteristics in vivo and in vitro. Moreover, it had a favorable safety profile and elicited specific immune responses in mice and sheep. The RM6 strain may have substantial practical application value

Development and Efficacy Evaluation of an SP01-adjuvanted Inactivated Escherichia Coli Mutant Vaccine Against Bovine Coliform Mastitis

Escherichia coli ( E. coli ) is one of the most common pathogens causing clinical mastitis in cattle, but no vaccine is available to prevent this disease in China. Therefore, development of an E. coli vaccine against bovine clinical mastitis is urgently needed. The candidate vaccine (Ch-O111-1) and challenge (LZ06) strains were screened from milk samples of cows with clinical mastitis. To extend the cross-protection of the Ch-O111-1 strain, we deleted the galE gene fragment of the Ch-O111-1 strain through homologous recombination between the Ch-O111-1 strain and pCVD442/ΔgalE plasmid, which was identified through conventional methods, including PCR, SDS-PAGE and sequencing. The Ch-O111-1/ΔgalE (Z9) strain was characterized by extensive cross-reactivity and attenuated virulence. We prepared inactivated Z9 vaccines with different adjuvants. Immunization of inactivated Z9 antigen induced adjuvant-, dosage- and inoculation time-dependent antibody titers in cows and mice. Furthermore, immunization with SP01-adjuvanted inactivated Z9 vaccine protected cows against severe clinical mastitis caused by LZ06 and protected mice against death due to LZ06. An SP01-adjuvanted inactivated Z9 vaccine was successfully developed and found to protect cows against severe mastitis caused by Escherichia coli

Preparation of Equine Immunoglobulin F(ab′) 2 against Smallpox and Evaluation of its Immunoprotective Effect

Smallpox, a severe infectious disease caused by the smallpox virus, causes a death rate as high as 30% within 15-20 days after infection. Therefore, development of anti-Smallpox product as a strategic reserve is urgently needed. We prepared and tested pepsin-digested F(ab′) 2 fragments of serum IgG from horses. Transmission electron microscopy indicated that the purified virus showed morphology consistent with VVTT. The titer was above 1.0 × 10 7 PFU/mL. The purity of the antigen exceeded 90%, according to HPLC. After purification and cleavage, the yield of the purified product F(ab′) 2 was approximately 1.3%, its purity exceeded 90%, and the neutralizing antibody titer exceeded 1:3200. F(ab′) 2 fragments had good preventive and therapeutic effects in mice at antibody doses of 5.2 mg/mL and 2.6 mg/mL. The viral loads of the drug-treated mice were suppressed to varying degrees, and the higher dose groups (5.2 and 2.6 mg/mL) showed a 2-3 fold lower viral load than that in the control group. A process for producing equine immunoglobulin F(ab′) 2 against VVTT was established. The prepared horse anti-smallpox immunoglobulin product had good neutralizing antibody effects on VVTT. The highly purified preparation may serve as a potential candidate for smallpox treatment

Nucleotide diversity maps reveal variation in diversity among wheat genomes and chromosomes

<p>Abstract</p> <p>Background</p> <p>A genome-wide assessment of nucleotide diversity in a polyploid species must minimize the inclusion of homoeologous sequences into diversity estimates and reliably allocate individual haplotypes into their respective genomes. The same requirements complicate the development and deployment of single nucleotide polymorphism (SNP) markers in polyploid species. We report here a strategy that satisfies these requirements and deploy it in the sequencing of genes in cultivated hexaploid wheat (<it>Triticum aestivum</it>, genomes AABBDD) and wild tetraploid wheat (<it>Triticum turgidum </it>ssp. <it>dicoccoides</it>, genomes AABB) from the putative site of wheat domestication in Turkey. Data are used to assess the distribution of diversity among and within wheat genomes and to develop a panel of SNP markers for polyploid wheat.</p> <p>Results</p> <p>Nucleotide diversity was estimated in 2114 wheat genes and was similar between the A and B genomes and reduced in the D genome. Within a genome, diversity was diminished on some chromosomes. Low diversity was always accompanied by an excess of rare alleles. A total of 5,471 SNPs was discovered in 1791 wheat genes. Totals of 1,271, 1,218, and 2,203 SNPs were discovered in 488, 463, and 641 genes of wheat putative diploid ancestors, <it>T. urartu</it>, <it>Aegilops speltoides</it>, and <it>Ae. tauschii</it>, respectively. A public database containing genome-specific primers, SNPs, and other information was constructed. A total of 987 genes with nucleotide diversity estimated in one or more of the wheat genomes was placed on an <it>Ae. tauschii </it>genetic map, and the map was superimposed on wheat deletion-bin maps. The agreement between the maps was assessed.</p> <p>Conclusions</p> <p>In a young polyploid, exemplified by <it>T. aestivum</it>, ancestral species are the primary source of genetic diversity. Low effective recombination due to self-pollination and a genetic mechanism precluding homoeologous chromosome pairing during polyploid meiosis can lead to the loss of diversity from large chromosomal regions. The net effect of these factors in <it>T. aestivum </it>is large variation in diversity among genomes and chromosomes, which impacts the development of SNP markers and their practical utility. Accumulation of new mutations in older polyploid species, such as wild emmer, results in increased diversity and its more uniform distribution across the genome.</p

Structural characteristics and deep-water hydrocarbon accumulation model of the Scotian Basin, Eastern Canada

Commercial hydrocarbon reservoirs have been discovered in shallow-water areas of the Scotian Basin, Eastern Canada. However, knowledge about the structure and hydrocarbon accumulation characteristics of the basin is still insufficient, which constrains the oil and gas exploration in deep-water areas. Based on comprehensive data of magnetic anomalies, seismic survey, and drilling, this study determines the structure characteristics of the Scotian Basin and its hydrocarbon accumulation conditions in deep waters and evaluates the deep-water hydrocarbon exploration potential. The transform faults and basement structures in the northern basin control the sedimentary framework showing thick strata in east and thin strata in west of the basin. The bowl-shaped depression formed by thermal subsidence during the transitional phase and the confined environment (micro basins) caused by salt tectonics provide favorable conditions for the development of source rocks during the depression stage (also referred to as the depression period sequence) of the basin. The progradation of large shelf-margin deltas during the drift phase and steep continental slope provide favorable conditions for the deposition of slope-floor fans on continental margins of the basin. Moreover, the source-reservoir assemblage comprising the source rocks within the depression stage and the turbidite sandstones on the continental margin in the deep waters may form large deep-water turbidite sandstone reservoirs. This study will provide a valuable reference for the deep-water hydrocarbon exploration in the Scotian Basin

Architecture and exploration target areas of the Senegal Basin, West Africa

The recent discovery of large oil and gas fields in the deep-water of the Senegal Basin has drawn global attention. Despite this, several exploration wells in this area fail, which can be primarily contributed to a lack of understanding of the basin's structures and hydrocarbon accumulation conditions. This study examines these characteristics utilizing gravity, seismic and drilling data, and finally makes a comparison with the Cote d’Ivoire Basin, a typical transform margin basin in the South Atlantic. The results suggest that the Senegal Basin, influenced by multiple transform faults and a weak Paleozoic basement, experienced three evolutionary stages: rifting, transitional, and drifting. Each stage contributed to the development of distinct depositional sequences - syn-rift sequences, sag sequences, and continental margin sequences, respectively. The Triassic - Early Jurassic rifting stage predominantly formed continental deposits, like fluvial, lacustrine, and deltaic deposits, in the syn-rift sequences. The Middle-Late Jurassic transitional stage, influenced by transform faults, witnessed the formation of marginal ridges or submarine uplift zones. These zones, in conjunction with landward high terrains, formed a restricted environment promoting the development of source rocks in the sag sequences. During the drifting stage, three types of reservoirs, namely platform carbonate rocks, deltas, and slope-floor fans were formed. Notably, large-scale hydrocarbon reservoirs have been found in the deltas and the slope-floor fans both in the Senegal Basin and the Cote d’Ivoire Basin. The Upper Jurassic - Aptian platforms exhibit thick carbonate rocks and organic reefs on their edges, suggesting substantial potential for hydrocarbon exploration in the Senegal Basin

Structural characteristics and exploration fields in passive continental margin basins of Central Atlantic

The petroleum resources of the passive continental margin basins in the Central Atlantic are rich, but the insufficient knowledge of basin structural characteristics and accumulation conditions in the area restricts the exploration of deepwater oil and gas. Based on the integral analysis of seismic, drilling, gravity anomalies and magnetic anomalies data, the basin structural characteristics and hydrocarbon accumulation conditions in deepwater area of the passive continental margin basins of the Central Atlantic are studied and the exploration fields are predicted with the Senegal and Scotia basins as key anatomical objects. It can be concluded that the passive continental margin basins of the Central Atlantic have experienced three evolution stages: the rift period, the transition period and the drift period, with corresponding development of three tectonic layers: the rift layer, the depression layer and the continental margin layer, and the basin structures are controlled by transform faults and basement properties. From Triassic to Early Jurassic rift period, a series of horst-graben structures were formed, and the sediments were mainly continental ones such as rivers, lakes and deltas. From Middle to Late Jurassic transition period, the marginal ridge or submarine uplift zone were developed due to the activity of transform faults, and a landward high terrain limited environment was formed due to the stretch, thinning and subsidence of the Paleozoic weak basement, providing a favorable condition for the development of source rocks in the depression layer. During Cretaceous drift period, platform margin reef and delta-slope floor fan reservoirs were developed. Both the deepwater slope floor fan in the Scotia Basin and the platform margin reef in the Senegal Basin have great exploration potential

Purification, Preliminary Structure and Antitumor Activity of Exopolysaccharide Produced by <i>Streptococcus thermophilus</i> CH9

In the present study, the preliminary structure and in vitro antitumor activity of three exopolysaccharides (EPSs) from Streptococcus thermophilus CH9 were investigated. Then, three purified fractions of EPS-1a, EPS-2a, and EPS-3a were obtained by chromatography using DEAE-52 cellulose and Sephadex G-100, respectively. The average molecular weight of EPS-1a, EPS-2a, and EPS-3a, were 1.80 &#215; 106, 1.06 &#215; 106 and 1.05 &#215; 106. The monosaccharide composition of EPS-3a was dramatically different from the others. The EPS-1a and EPS-2a were mainly composed of mannose, in a ratio of 69.82% and 57.09%, respectively, while EPS-3a was mainly composed of glucose (63.93%), without mannose. In addition, the surface morphology observed suggested that there were protein particles on the sugar chain of EPS-3a and EPS-3a was a protein-containing polysaccharide. Furthermore, EPS-3a exhibited higher antitumor activity against human liver cancer HepG2 cells in vitro. The antitumor activity of EPS-3a in HepG2 cells was associated with cell apoptosis. HE staining and Hoechst 33342 staining showed that with the treatment of EPS-3a, HepG2 cells had typical morphological changes. Flow cytometry analysis showed that the cell cycle was arrested at G0/G1 phase