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

The Influence of a Prolonged Meteorological Drought on the Catchment Water Storage Capacity: A Hydrological Model Perspective

Abstract. Understanding the propagation process of prolonged meteorological droughts (i.e., decade) helps solve the problem of increasing water scarcity around the world. Historical literature studied the propagation between different drought types (e.g., from meteorological to hydrological drought) with mainly statistical approaches, however, little attention has been paid to the causality between the meteorological drought with potential changes in the Catchment Water Storage Capacity (CWSC) where the latter plays a critical role in catchment response behavior to former. This study used the temporal variation in the estimated value of a model parameter that denotes the CWSC in its model structure to reflect the potential changes in real CWSC. The most likely Change points of the CWSC were determined based on the Bayesian Change point analysis. Also, the possible association and linkage between the shift in the CWSC and the time-lag of the catchment (i.e., time-lag between the occurrence of the drought with the Change point) with multiple catchment properties and climate characteristics have been studied. Catchments from southeastern Australia were used as a study area to verify the effectiveness of the proposed approach. Results indicated that (1) in 62.7 % of the catchments, the sustained drought causes significant shifts in the CWSC. The shift led to the opposite response in two subsets of catchments, i.e., 48.2 % of catchments had lower runoff generation rates for a given rainfall while 14.5 % of catchments had higher runoff generation rate. (2) Catchments with larger elevation and slope, lower forest coverage of Evergreen Broadleaf Forest are more likely to have increase in the CWSC during a chronic drought while smaller catchments with lower elevation, lower coverage of the Evergreen Broadleaf Forest are more likely to have a decrease in the CWSC. (3) The changed catchments were not equally susceptible to the pressure due to persistent meteorological drought. Catchments with a lower proportion of Evergreen Broadleaf Forest usually have longer time-lag and are more resilient. This study improves our understanding of possible changes in CWSC induced by a prolonged meteorological drought, which will help improve our ability to simulate the hydrological system. </jats:p

H₂O₂/Glucose Sensor Based on a Pyrroloquinoline Skeleton-Containing Molecule Modified Gold Cavity Array Electrode

H2O2-related metabolites are essential indicators in clinical diagnosis because the accumulation of such reactive oxygen species could cause the risk of cardiovascular disease. Herein, we reported an electrochemical sensor to determine H2O2 and glucose. The pyrroloquinoline skeleton containing molecules (PQT) were used as the electrocatalyst and the gold cavity array (GCA) electrodes as the supporting electrode. The GCA electrode was fabricated by electrodeposition using high-ordered two-dimensional polystyrene spheres as the template. The strong absorbability of iodide ions (I−) displaced adventitious materials from the metal surface and the I− monolayer was subsequently removed by electrochemical oxidation to get a clean electrode surface. PQT molecules were firmly immobilized on the GCA electrode and performed an excellent electrocatalytic effect on H2O2/glucose detection, manifested by a small overpotential and a significantly increased reduction current. A good linear correlation was observed over a wide range of 0.2 μmol/L–1.0 mmol/L with the limit of detection of 0.05 μmol/L. Moreover, the sensor can realize sensitive, accurate, and the highly selective detection of actual samples, proving its application prospect in clinical diagnosis

H2O2/Glucose Sensor Based on a Pyrroloquinoline Skeleton-Containing Molecule Modified Gold Cavity Array Electrode

H2O2-related metabolites are essential indicators in clinical diagnosis because the accumulation of such reactive oxygen species could cause the risk of cardiovascular disease. Herein, we reported an electrochemical sensor to determine H2O2 and glucose. The pyrroloquinoline skeleton containing molecules (PQT) were used as the electrocatalyst and the gold cavity array (GCA) electrodes as the supporting electrode. The GCA electrode was fabricated by electrodeposition using high-ordered two-dimensional polystyrene spheres as the template. The strong absorbability of iodide ions (I−) displaced adventitious materials from the metal surface and the I− monolayer was subsequently removed by electrochemical oxidation to get a clean electrode surface. PQT molecules were firmly immobilized on the GCA electrode and performed an excellent electrocatalytic effect on H2O2/glucose detection, manifested by a small overpotential and a significantly increased reduction current. A good linear correlation was observed over a wide range of 0.2 μmol/L–1.0 mmol/L with the limit of detection of 0.05 μmol/L. Moreover, the sensor can realize sensitive, accurate, and the highly selective detection of actual samples, proving its application prospect in clinical diagnosis.</jats:p

miR-10a rejuvenates aged human mesenchymal stem cells and improves heart function after myocardial infarction through KLF4

Abstract Background Aging is one of the key factors that regulate the function of human bone marrow mesenchymal stem cells (hBM-MSCs) and related changes in microRNA (miRNA) expression. However, data reported on aging-related miRNA changes in hBM-MSCs are limited. Methods We demonstrated previously that miR-10a is significantly decreased in aged hBM-MSCs and restoration of the miR-10a level attenuated cell senescence and increased the differentiation capacity of aged hBM-MSCs by repressing Krüpple-like factor 4 (KLF4). In the present study, miR-10a was overexpressed or KLF4 was downregulated in old hBM-MSCs by lentiviral transduction. The hypoxia-induced apoptosis, cell survival, and cell paracrine function of aged hBM-MSCs were investigated in vitro. In vivo, miR-10a-overexpressed or KLF4-downregulated old hBM-MSCs were implanted into infarcted mouse hearts after myocardial infarction (MI). The mouse cardiac function of cardiac angiogenesis was measured and cell survival of aged hBM-MSCs was investigated. Results Through lentivirus-mediated upregulation of miR-10a and downregulation of KLF4 in aged hBM-MSCs in vitro, we revealed that miR-10a decreased hypoxia-induced cell apoptosis and increased cell survival of aged hBM-MSCs by repressing the KLF4–BAX/BCL2 pathway. In vivo, transplantation of miR-10a-overexpressed aged hBM-MSCs promoted implanted stem cell survival and improved cardiac function after MI. Mechanistic studies revealed that overexpression of miR-10a in aged hBM-MSCs activated Akt and stimulated the expression of angiogenic factors, thus increasing angiogenesis in ischemic mouse hearts. Conclusions miR-10a rejuvenated aged hBM-MSCs which improved angiogenesis and cardiac function in injured mouse hearts