ABSTRACT
In the pathogenesis of major depressive disorder, chronic stress-related neuroinflammation hinders favorable prognosis and antidepressant response. Mitochondrial DNA may be an inflammatory trigger, after its release from stress-induced dysfunctional central nervous system mitochondria into peripheral circulation. This evidence supports the potential use of peripheral mitochondrial DNA as a neuroinflammatory biomarker for the diagnosis and treatment of major depressive disorder. Herein, we critically review the neuroinflammation theory in major depressive disorder, providing compelling evidence that mitochondrial DNA release acts as a critical biological substrate, and that it constitutes the neuroinflammatory disease pathway. After its release, mitochondrial DNA can be carried in the exosomes and transported to extracellular spaces in the central nervous system and peripheral circulation. Detectable exosomes render encaged mitochondrial DNA relatively stable. This mitochondrial DNA in peripheral circulation can thus be directly detected in clinical practice. These characteristics illustrate the potential for mitochondrial DNA to serve as an innovative clinical biomarker and molecular treatment target for major depressive disorder. This review also highlights the future potential value of clinical applications combining mitochondrial DNA with a panel of other biomarkers, to improve diagnostic precision in major depressive disorder.
ABSTRACT
Fish diversity plays a critical role in maintaining the balance of water ecosystems, especially in the Chongqing section of the National Nature Reserve for Rare and Endemic Fishes in the upper Yangtze River, which serves as an important habitat for rare and endemic fish, as well as an important channel for the replenishment of fishery resources in the Three Gorges Reservoir. Under a 10-year ban on fishing in the Yangtze River basin, we investigate fish diversity and seasonal variation in the Reserve by using environmental DNA (eDNA) metabarcoding. We found fishes belonging to 85 genera, 24 families, and 8 orders in the Reserve. A comparison of eDNA metabarcoding results with the diversity of a recent fish catch revealed that eDNA metabarcoding not only enables rapid and efficient fish monitoring but also has a high sensitivity. Furthermore, the study demonstrates that eDNA metabarcoding can be used as a tool for monitoring seasonal variations of fish composition in freshwater ecosystems. The alpha and beta diversity analysis both showed compositional differences in the fish community in accordance with seasonal variations. In addition, changes in eDNA relative sequence abundance and the detection of fish species at different sampling sites may reflect shifts in habitat use and distribution. Thus, we provide detailed seasonal data on fish diversity in the Chongqing section of the Reserve. This will contribute to conservation and to the understanding of fish diversity and community dynamics in the Chongqing section of the Reserve.
ABSTRACT
Nardosinone, a predominant bioactive product from Nardostachys jatamansi DC, is well-known for its promising therapeutic applications, such as being used as a drug on anti-inflammatory, antidepressant, cardioprotective, anti-neuroinflammatory, anti-arrhythmic, anti-periodontitis, etc. However, its stability under varying environmental conditions and its degradation products remain unclear. In this study, four main degradation products, including two previously undescribed compounds [2-deoxokanshone M (64.23%) and 2-deoxokanshone L (1.10%)] and two known compounds [desoxo-narchinol A (2.17%) and isonardosinone (3.44%)], were firstly afforded from the refluxed products of nardosinone in boiling water; their structures were identified using an analysis of the extensive NMR and X-ray diffraction data and the simulation and comparison of electronic circular dichroism spectra. Compared with nardosinone, 2-deoxokanshone M exhibited potent vasodilatory activity without any of the significant anti-neuroinflammatory activity that nardosinone contains. Secondly, UPLC-PDA and UHPLC-DAD/Q-TOF MS analyses on the degradation patterns of nardosinone revealed that nardosinone degraded more easily under high temperatures and in simulated gastric fluid compared with the simulated intestinal fluid. A plausible degradation pathway of nardosinone was finally proposed using nardosinonediol as the initial intermediate and involved multiple chemical reactions, including peroxy ring-opening, keto-enol tautomerization, oxidation, isopropyl cleavage, and pinacol rearrangement. Our findings may supply certain guidance and scientific evidence for the quality control and reasonable application of nardosinone-related products.
Subject(s)
Sesquiterpenes , Sesquiterpenes/chemistry , Temperature , Polycyclic Sesquiterpenes , Anti-Inflammatory AgentsABSTRACT
A novel ternary deep eutectic solvent (DES), consisted of choline chloride, oxalic acid and ethylene glycol, was developed as a green, low-cost and recyclable pretreatment system for multi-stage utilization of Eucommia ulmoides seed shells. Under optimum conditions, 79.7 % hemicellulose and 65.6 % lignin were quickly removed while 84.0 % cellulose was retained. After DES pretreatment, the yield and purity of gutta-percha achieved 85.1 mg/g and 96.2 %, which increased 1.4 and 1.8 folds higher than that of un-treatment ones. Meanwhile, 69.1 % enzymatic digestibility of cellulose was obtained, that was 2.3 folds higher than that of raw substrates. Moreover, 53.6 % low-condensation lignin with aromatic structures and valuable aryl-ether linkages was well collected. Importantly, the DES that has been recycled five runs can still remove 73.9 % hemicellulose and 58.0 % lignin. Overall, the DES was determined to efficiently promote the separation and conversion of high-quality gutta-percha, value-added lignin and high-yield glucose from Eucommia ulmoides seed shells.
Subject(s)
Eucommiaceae , Lignin , Lignin/chemistry , Eucommiaceae/chemistry , Gutta-Percha , Deep Eutectic Solvents , Monosaccharides , Solvents/chemistry , Hydrolysis , Cellulose/chemistry , Seeds , BiomassABSTRACT
The appearance of antiferroelectrics (AFE) in the ferrimagnetism (FM) system would give birth to a new type of multiferroic candidate, which is significant to the development of novel devices for energy storage. Here we demonstrate the realization of full antiferroelectrics in a magnetic La0.5Sr0.5Fe12O19 system (AFE+FM), which also presents a strong magnetodielectric response (MD) and magnetoresistance (MR) effect. The antiferroelectric phase was achieved at room temperature by replacing 0.5 Sr2+ ions with 0.5 La2+ ions in the SrFe12O19 compound, whose phase transition temperature of ferroelectrics (FE) to antiferroelectrics was brought down from 174 °C to -141 °C, while the temperature of antiferroelectrics converting to paraelectrics (PE) shifts from 490 °C to 234 °C after the substitution. The fully separated double P-E hysteresis loops reveal the antiferroelectrics in La0.5Sr0.5Fe12O19 ceramics. The magnitude of exerting magnetic field enables us to control the generation of spin current, which induces MD and MR effects. A 1.1T magnetic field induces a large spin current of 15.6 n A in La0.5Sr0.5Fe12O19 ceramics, lifts up dielectric constants by 540%, and lowers the resistance by -89%. The magnetic performance remains as usual. The multiple functions in one single phase allow us to develop novel intelligent devices.
ABSTRACT
Recent years, China has released a series of policies to encourage drug research and development in pediatric populations, aiming to meet pediatric populations' medical needs. Because of the physical and psychological developmental characteristics, tolerance of aversive feelings when taking medications are different between pediatric population and adults. So pediatric populations are at a relatively higher risk of not taking medications as prescribed when the medication tasted unpleasant. Therefore, sound design and evaluation of oral sensory features have important clinical significance and value in developing pediatric medications. "Technical guidance for the design and evaluation of the oral sensory features of pediatric drugs (trial version) " was released in November 2022, by Centre for Drug Evaluation, National Medical Products Administration of China. Based on the guidance, this article will introduce the drafting background and review considerations, hoping to provide reference for the design and evaluation of oral sensory features, and promote drug developing in pediatric population.
ABSTRACT
We describe a novel approach for screening fragments against a protein that combines the sensitivity of DNA-encoded library technology with the ability of fragments to explore what will bind. Each of the members of the library consists of a fragment which is linked to a photoactivatable diazirine moiety. Split and pool synthesis combines each fragment with a set of linkers with the version of the library reported here containing some 70k different compounds, each with an individual DNA code. Incubation of the library with a protein sample is followed by photoactivation, washing and subsequent PCR and sequencing which allows the individual fragment hits to be identified. We illustrate how the approach allows successful hit fragment identification using only microgram quantities of material for two targets. PAK4 is a kinase for which conventional fragment screening has generated many advance leads. The as yet undrugged target, 2-epimerase, presents a more challenging active site for identification of hit compounds. In both cases, PAC-FragmentDEL identified fragments validated as hits by ligand-observed NMR measurements and crystal structure determination of off-DNA sample binding to the proteins.
ABSTRACT
Pig diarrhea is a universal problem in the process of pig breeding, which seriously affects the development of the pig industry. Porcine enteric coronaviruses (PECoVs) are common pathogens causing diarrhea in pigs, currently including transmissible gastroenteritis virus (TGEV), porcine epidemic diarrhea virus (PEDV), porcine deltacoronavirus (PDCoV) and swine acute diarrhea syndrome coronavirus (SADS-CoV). With the prosperity of world transportation and trade, the spread of viruses is becoming wider and faster, making it even more necessary to prevent PECoVs. In this paper, the host factors required for the efficient replication of these CoVs and the compounds that exhibit inhibitory effects on them were summarized to promote the development of drugs against PECoVs. This study will be also helpful in discovering general host factors that affect the replication of CoVs and provide references for the prevention and treatment of other CoVs.
Subject(s)
Coronavirus Infections , Coronavirus , Porcine epidemic diarrhea virus , Swine Diseases , Swine , Animals , Coronavirus Infections/drug therapy , Coronavirus Infections/prevention & control , Coronavirus Infections/veterinary , Diarrhea/drug therapy , Diarrhea/veterinaryABSTRACT
Introduction: The haematology external quality assessment (EQA) scheme is the most commonly used service of quality assurance. The provision of complete blood count (CBC) materials must meet the quality requirements at a reasonable cost. These requirements are the most significant challenges for EQA organisers in Vietnam. This study's objective was to evaluate the homogeneity, long-term stability, and peer-group performance of 10-parameter stabilised CBC EQA samples. Methods: The CBC EQA material was prepared using the following steps, including (1) adjusting levels of stabilised erythrocyte, leukocyte, and platelet samples, (2) mixing those cells into batches at three levels, and (3) dispensing and storing them at 2-6 °C. A set of 10 and 30 specimens were randomly chosen from each batch to study the homogeneity and long-term stability following ISO 13528:2015. In total, 166 samples at two levels were randomly distributed to 40 participants, which reported 83 automatic cell counters among six automated analyser models in the CBC EQA program. Results: The 10-parameter stabilised CBC EQA materials at three levels became homogeneous and stable in 12 weeks when preserved at 2-6 °C. Meanwhile, for five parameters (RBC, Hb, MCH, MCV, and MPV), this process was prolonged for up to 16 weeks in stock condition. In terms of peer-group performance, the CV (%) values increased at the low concentration for almost all parameters, especially in platelet counts. Conclusions: The stabilised CBC EQA samples prepared using the partial fixation method with aldehyde and gutaraldehyde in this study meet the ISO 13528:2015 requirements of homogeneity and long-term stability for the CBC EQA scheme. Analytical performance evaluation should categorise participant methods into peer groups.
ABSTRACT
INTRODUCTION: Cataracts caused by old age are one of the most frequent causes for blindness and poor vision worldwide. In this study, we aimed to clarify the possible role of rs1894720 polymorphism in the pathogenesis of age-related cataract. MATERIAL AND METHODS: Rs1894720 polymorphism genotype was detected by TaqMan. Bioinformatics analysis, luciferase assay, real-time PCR, western blot, and protein density analysis were conducted to establish the correlations between MIAT and miR-26b as well as between BCL2L2 and miR-26b. Flow cytometry and MTT assay were also performed to observe the effect of MIAT/miR-26b/BCL2L2 signalling pathway on the status of cell apoptosis and viability. RESULTS: MIAT functioned as an endogenous competing RNA to sponge miR-26b. In addition, BCL2L2 was identified as a target of miR-26b. Therefore, the expression of miR-26b was obviously suppressed by MIAT or anti-miR-26b, while the mRNA and protein expression of BCL2L2 was up-regulated in the presence of MIAT or anti-miR-26b. Moreover, the positive effect of MIAT on BCL2L2 expression was exerted via inhibition of the expression of miR-26b. In addition, the cells transfected with MIAT or anti-miR-26b showed suppressed expression of caspase-3 and reduced apoptosis index but higher cell viability, indicating that MIAT could suppress cell apoptosis via inhibition of miR-26b expression. Furthermore, the subjects carrying the GT and TT genotypes of single-nucleotide polymorphism (SNP) rs1894720 were associated with a higher risk of age-related cataracts, as indicated by their odds ratio (OR) and p-values. CONCLUSIONS: Rs1894720 SNP could down-regulate the expression of MIAT, thus leading to reduced BCL2L2 expression and enhanced epithelial cell apoptosis in the lens, eventually increasing the incidence of age-related cataract.
ABSTRACT
Porcine epidemic diarrhea virus (PEDV) is an alphacoronavirus that causes severe watery diarrhea in piglets with high morbidity and mortality, resulting in serious economic losses to the farming industry. Ergosterol peroxide (EP) is a sterol with diverse biological activities including antiviral activity. In this study, we explored whether EP extracted from the fruiting body of the mushroom Cryptoporus volvatus had the potential to inhibit PEDV infection in Vero cells. The results revealed that EP had a remarkable inhibitory effect on PEDV infection. It could significantly inhibit multiple stages of the PEDV life cycle, including internalization, replication and release, and could directly inactivate PDCoV infectivity. However, it did not affect PEDV attachment. Furthermore, EP alleviated PEDV-induced apoptosis and mitigated the decrease in mitochondrial membrane potential caused by PEDV infection. It suppressed ROS generation and p53 activation caused by PEDV infection. The ROS scavenger N-acetyl-l-cysteine (NAC) and the p53 specific inhibitor Pifithrin-α (PFT-α) suppressed PEDV-induced apoptosis and impeded viral replication, suggesting that ROS and p53 play an important role in PEDV-induced apoptosis and viral replication. Collectively, EP can prevent PEDV internalization, replication and release, possesses the ability to directly inactivate PEDV, and can inhibit PEDV-induced apoptosis by interfering with PEDV-induced ROS production and p53 activation. These findings highlight the therapeutic potential of EP against PEDV infection.
Subject(s)
Coronavirus Infections/veterinary , Ergosterol/analogs & derivatives , Porcine epidemic diarrhea virus/drug effects , Reactive Oxygen Species/metabolism , Swine Diseases/virology , Tumor Suppressor Protein p53/metabolism , Animals , Apoptosis/drug effects , Chlorocebus aethiops , Coronavirus Infections/drug therapy , Coronavirus Infections/virology , Ergosterol/pharmacology , Polyporaceae/chemistry , Swine , Swine Diseases/drug therapy , Vero Cells , Virus Replication/drug effectsABSTRACT
Norisoboldine (NOR), an isoquinoline alkaloid, has previously been shown to ameliorate collagen-induced arthritis (CIA) by modulating the function of multiple cells such as T lymphocytes and fibroblast-like synoviocytes. To further study its anti-arthritis mechanism, the effect of NOR on the systemic metabolism regulation was investigated using an NMR-based untargeted metabolomics approach. CIA model rats were orally administered with NOR (30 mg/kg) for 14 consecutive days. The alterations of endogenous metabolites in the urine samples were quantified by 1H NMR. While NOR significantly mitigated CIA in rats as evidenced by the reduced clinical scores and histopathological changes, the results indicated that the treatment restored the levels of 22 metabolites that were significantly changed by arthritis, and most of which were related to lipid metabolism. Further studies demonstrated that NOR up-regulated the expression of carnitine palmitoyltransferase 1 (CPT-1) and down-regulated the expression of fatty acid synthase (FASN) in the spleens and the synovial tissues of CIA rats. Together these results revealed a strong association between RA and the system in metabolic disorders. The differential metabolites and their related pathways may also serve as novel therapeutic targets for RA.
Subject(s)
Alkaloids/pharmacology , Arthritis, Experimental/drug therapy , Arthritis, Experimental/metabolism , Lipid Metabolism/drug effects , Magnetic Resonance Spectroscopy/methods , Alkaloids/therapeutic use , Animals , Arthritis, Experimental/pathology , Arthritis, Experimental/urine , Carnitine O-Palmitoyltransferase/genetics , Carnitine O-Palmitoyltransferase/metabolism , Collagen/toxicity , Fatty Acid Synthase, Type I/genetics , Fatty Acid Synthase, Type I/metabolism , Female , Metabolomics , Multivariate Analysis , Rats, Wistar , Spleen/drug effects , Spleen/metabolism , Synovial Membrane/drug effects , Synovial Membrane/metabolism , Urine/chemistryABSTRACT
Porcine deltacoronavirus (PDCoV) is a newly discovered swine enteropathogenic coronavirus with worldwide distribution. However, efficient strategies to prevent or treat the infection remain elusive. Our in vitro study revealed that ergosterol peroxide (EP) from the mushroom Cryptoporus volvatus has efficient anti-PDCoV properties. The aim of this study is to evaluate the potential of EP as a treatment for PDCoV in vivo and elucidate the possible mechanisms. Seven-day-old piglets were infected with PDCoV by oral administration in the presence or absence of EP. Piglets infected with PDCoV were most affected, whereas administration of EP reduced diarrhea incidence, alleviated intestinal lesion, and decreased viral load in feces and tissues. EP reduced PDCoV-induced apoptosis and enhanced tight junction protein expressions in the small intestine, maintaining the integrity of the intestinal barrier. EP showed immunomodulatory effect by suppressing PDCoV-induced pro-inflammatory cytokines and the activation of IκBα and NF-κB p65, and upregulating IFN-I expression. Knockdown of p38 inhibited PDCoV replication and alleviated PDCoV-induced apoptosis, implying that EP inhibited PDCoV replication and alleviated PDCoV-induced apoptosis via p38/MAPK signaling pathway. Collectively, ergosterol peroxide can protect piglets from PDCoV, revealing the potential of EP for development as a promising strategy for treating and controlling the infection of PDCoV.
Subject(s)
Apoptosis/drug effects , Coronavirus Infections/veterinary , Deltacoronavirus , Ergosterol/analogs & derivatives , Swine Diseases/virology , Tight Junctions/drug effects , Animals , Coronavirus Infections/drug therapy , Coronavirus Infections/virology , Deltacoronavirus/drug effects , Ergosterol/pharmacology , Ergosterol/therapeutic use , Intestine, Small/drug effects , Intestine, Small/virology , LLC-PK1 Cells , Male , Swine , Swine Diseases/drug therapyABSTRACT
Porcine deltacoronavirus (PDCoV) is a swine enteropathogenic coronavirus (CoV) that continues to spread globally, placing strain on economic and public health. Currently, the pathogenic mechanism of PDCoV remains largely unclear, and effective strategies to prevent or treat PDCoV infection are still limited. In this study, the interaction between autophagy and PDCoV replication in LLC-PK1 cells was investigated. We demonstrated that PDCoV infection induced a complete autophagy process. Pharmacologically induced autophagy with rapamycin increased the expression of PDCoV N, while pharmacologically inhibited autophagy with wortmannin decreased the expression of PDCoV N, suggesting that PDCoV-induced autophagy facilitates virus replication. Further experiments showed that PDCoV infection activated p38 signaling pathway to trigger autophagy. Besides, ergosterol peroxide (EP) alleviated PDCoV-induced activation of p38 to suppress autophagy, thus exerting its antiviral effects. Finally, we employed a piglet model of PDCoV infection to demonstrate that EP prevented PDCoV infection by suppressing PDCoV-induced autophagy via p38 signaling pathway in vivo. Collectively, these findings accelerate the understanding of the pathogenesis of PDCoV infection and provide new insights for the development of EP as an effective therapeutic strategy for PDCoV.
Subject(s)
Antiviral Agents/pharmacology , Autophagy , Coronavirus Infections/veterinary , Deltacoronavirus/drug effects , Ergosterol/analogs & derivatives , MAP Kinase Signaling System , Virus Replication/drug effects , Animals , Coronavirus Infections/prevention & control , Coronavirus Infections/virology , Deltacoronavirus/physiology , Ergosterol/pharmacology , LLC-PK1 Cells , Swine , Swine Diseases/virologyABSTRACT
A sulfa-Michael/aldol/lactonization cascade reaction has been established to construct isotetronic acid-fused thiochromanes in a highly stereoselective fashion (≥11:1 dr, 35-98% ee). The tricyclic products were obtained in 35-99% isolated yields in the presence of a bifunctional squaramide. Three reactive sites of ß,γ-unsaturated α-ketoester, including the less-explored ester carbonyl group, were sequentially utilized to construct two fused heterocycles in a one-pot operation.
Subject(s)
Esters , Catalysis , StereoisomerismABSTRACT
Porcine deltacoronavirus (PDCoV) is an emerging swine enteropathogenic coronavirus (CoV) that poses economic and public health burdens. Currently, there are no effective antiviral agents against PDCoV. Cryptoporus volvatus often serves as an antimicrobial agent in Traditional Chinese Medicines. This study aimed to evaluate the antiviral activities of ergosterol peroxide (EP) from C. volvatus against PDCoV infection. The inhibitory activity of EP against PDCoV was assessed by using virus titration and performing Quantitative Reverse transcription PCR (RT-qPCR), Western blotting and immunofluorescence assays in LLC-PK1 cells. The mechanism of EP against PDCoV was analyzed by flow cytometry, RT-qPCR and Western blotting. We found that EP treatment inhibited PDCoV infection in LLC-PK1 cells in a dose-dependent manner. Subsequently, we demonstrated that EP blocked virus attachment and entry using RT-qPCR. Time-of-addition assays indicated that EP mainly exerted its inhibitory effect at the early and middle stages in the PDCoV replication cycle. EP also inactivated PDCoV infectivity directly as well as suppressed PDCoV-induced apoptosis. Furthermore, EP treatment decreased the phosphorylation of IκBα and p38 MAPK induced by PDCoV infection as well as the mRNA levels of cytokines (IL-1ß, IL-6, IL-12, TNF-α, IFN-α, IFN-ß, Mx1 and PKR). These results imply that EP can inhibit PDCoV infection and regulate host immune responses by downregulating the activation of the NF-κB and p38/MAPK signaling pathways in vitro. EP can be used as a potential candidate for the development of a new anti-PDCoV therapy.
Subject(s)
Antiviral Agents/pharmacology , Deltacoronavirus/drug effects , Deltacoronavirus/immunology , Ergosterol/analogs & derivatives , MAP Kinase Signaling System/drug effects , NF-kappa B/metabolism , Animals , Apoptosis/drug effects , Cell Line , Coronavirus Infections/drug therapy , Coronavirus Infections/immunology , Coronavirus Infections/virology , Cytokines/metabolism , Ergosterol/chemistry , Ergosterol/pharmacology , I-kappa B Proteins/metabolism , Immunologic Factors/chemistry , Immunologic Factors/pharmacology , LLC-PK1 Cells , Polyporaceae , Swine , Swine Diseases , Transcription Factor RelA/metabolism , Virion/drug effects , Virus Replication/drug effectsABSTRACT
The recent experience with SARS-COV-2 has raised our alarm about the cross-species transmissibility of coronaviruses and the emergence of new coronaviruses. Knowledge of this family of viruses needs to be constantly updated. Porcine deltacoronavirus (PDCoV), a newly emerging member of the genus Deltacoronavirus in the family Coronaviridae, is a swine enteropathogen that causes diarrhea in pigs and may lead to death in severe cases. Since PDCoV diarrhea first broke out in the United States in early 2014, PDCoV has been detected in many countries, such as South Korea, Japan and China. More importantly, PDCoV can also infect species other than pigs, and infections have even been reported in children, highlighting its potential for cross-species transmission. A thorough and systematic knowledge of the epidemiology and pathogenesis of PDCoV will not only help us control PDCoV infection, but also enable us to discover the common cellular pathways and key factors of coronaviruses. In this review, we summarize the current knowledge on the prevalence, pathogenicity and infection dynamics, pathogenesis and immune evasion strategies of PDCoV. The existing anti-PDCoV strategies and corresponding mechanisms of PDCoV infection are also introduced, aiming to provide suggestions for the prevention and treatment of PDCoV and zoonotic diseases.
ABSTRACT
Mastitis affects cows in all regions of the world and Escherichia coli (E. coli) is by far the most common reason of mastitis. Now antibiotic therapy is still the preferred approach of treating mastitis. However, antibiotic usage is easy to lead to antibiotic resistance. There is an urgent need for developing efficacious alternative antimicrobials. Pheromonicin-NM (PMC-NM) is a new engineered bactericidal peptide consisting of colicin Ia and an anti-porin A antibody mimetic. It can lead to the dissipation of cellular energy and therefore kill the bacteria rapidly. The aim of the present study was to investigate the comparative effects of PMC-NM and antibiotic ceftiofur on antibacterial and innate immune responses of bovine mammary epithelial cells (BMEC) to E. coli infection. We found that E. coli growth was inhibited by PMC-NM from 0.5 h after treatment and was completely inhibited at 3 h, indicating a rapid antibacterial activity for PMC-NM. The mRNA expression of TLR2, IL-1ß, IL-8, lactoferrin, LAP, TAP and DEFB1 was increased by PMC-NM treatment at 2 h after E. coli infection, suggesting the enhanced inflammatory responses induced by PMC-NM contribute to pathogens clearance at early phase. By contrast, in E. coli-infected BMECs, ceftiofur treatment upregulated TLR2 and NOD2 levels at 12 h, and extremely elevated transcription levels of TNF-α, IL-1ß, IL-8, lactoferrin, LAP, TAP, BNBD5, DEFB1 at 6 h. The excessive expression of these genes at later phase can induce uncontrolled inflammatory responses and finally cause damage. Taken together, PMC-NM might be used as an ideal antibacterial agent against E. coli mastitis.
Subject(s)
Anti-Bacterial Agents/pharmacology , Epithelial Cells/drug effects , Immunologic Factors/pharmacology , Mammary Glands, Animal/cytology , Animals , Cattle , Cells, Cultured , Cephalosporins/pharmacology , Cytokines/genetics , Cytokines/immunology , Epithelial Cells/immunology , Epithelial Cells/microbiology , Escherichia coli/drug effects , Escherichia coli Infections/drug therapy , Escherichia coli Infections/immunology , Female , Mastitis, Bovine/drug therapy , Mastitis, Bovine/immunology , Mucin-1/genetics , Mucin-1/immunologyABSTRACT
The on-DNA synthesis of highly substituted cyclobutanes was achieved through a photocatalytic [2 + 2] cycloaddition reaction in aqueous solution. Readily available DNA-tagged styrene derivatives were reacted with structurally diverse cinnamates in the presence of an iridium-based photocatalyst, Ir(ppy)2(dtbbpy)PF6, to forge two new C(sp3)-C(sp3) bonds. This transformation was demonstrated to have excellent functional group tolerance and allowed for the facile installation of a variety of heteroaromatic substituents on a densely functionalized cyclobutane scaffold.
Subject(s)
Cyclobutanes/chemistry , DNA/chemical synthesis , Catalysis , Cycloaddition Reaction , DNA/chemistry , Molecular Structure , Photochemical ProcessesABSTRACT
The aim of this study was to develop and test an optimal vaccination strategy against bovine viral diarrhea virus (BVDV) based on the E2 glycoprotein of the BJ1305 strain. To achieve higher E2-specific antibody titers and to broaden the cellular immune response, a plasmid encoding the E2 protein (pcDNA3.1-E2) was constructed and a purified recombinant E2 protein was generated. The E2 protein was emulsified in the adjuvant ISA 61 VG prior to administration. We immunized mice three times with pcDNA3.1-E2 or the recombinant E2 protein or primed twice with pcDNA3.1-E2 and boosted once with the E2 protein. To evaluate the protection against BVDV conferred by the vaccines, the mice were challenged with BVDV strain Oregon C24V after the third immunization. Although all immunized mice developed humoral and cellular immune responses, the E2-specific antibody titers in the DNA prime-protein boost group were significantly higher than those elicited by either the DNA or the protein vaccine. In addition, vaccination with the E2 DNA vaccine induced higher percentages of CD4+IFN-γ+ T cells and CD8+IFN-γ+ T cells among total CD3+ T cells than the other regimens. The predominant antibody subclass in the vaccinated mice was IgG1. Serum tumor necrosis factor alpha (TNF-α) levels in the DNA prime-protein boost group were significantly higher after the third immunization than in the other groups. Moreover, the mice treated with the DNA prime-protein boost vaccination regimen acquired protection against BVDV challenge, as shown by a significant reduction of viremia, only minor pathological changes, and a lower viral antigen burden than in the control and solo vaccinated mice. These results demonstrate the potential advantage of a DNA prime-protein boost vaccination approach over a solo vaccination for the prevention of BVDV. The ability of this vaccine strategy to control and eradicate BVD in herds warrants further investigation.