Attenuated replication and pathogenicity of SARS-CoV-2 B.1.1.529 Omicron.

The Omicron (B.1.1.529) variant of SARS-CoV-2 emerged in November 2021 and is rapidly spreading among the human population1. Although recent reports reveal that the Omicron variant robustly escapes vaccine-associated and therapeutic neutralization antibodies2-10, the pathogenicity of the virus remains unknown. Here we show that the replication of Omicron is substantially attenuated in human Calu3 and Caco2 cells. Further mechanistic investigations reveal that Omicron is inefficient in its use of transmembrane serine protease 2 (TMPRSS2) compared with wild-type SARS-CoV-2 (HKU-001a) and previous variants, which may explain its reduced replication in Calu3 and Caco2 cells. The replication of Omicron is markedly attenuated in both the upper and lower respiratory tracts of infected K18-hACE2 mice compared with that of the wild-type strain and Delta (B.1.617.2) variant, resulting in its substantially ameliorated lung pathology. Compared with wild-type SARS-CoV-2 and the Alpha (B.1.1.7), Beta (1.351) and Delta variants, infection by Omicron causes the lowest reduction in body weight and the lowest mortality rate. Overall, our study demonstrates that the replication and pathogenicity of the Omicron variant of SARS-CoV-2 in mice is attenuated compared with the wild-type strain and other variants.

Type-II IFN inhibits SARS-CoV-2 replication in human lung epithelial cells and ex vivo human lung tissues through indoleamine 2,3-dioxygenase-mediated pathways.

Interferons (IFNs) are critical for immune defense against pathogens. While type-I and -III IFNs have been reported to inhibit SARS-CoV-2 replication, the antiviral effect and mechanism of type-II IFN against SARS-CoV-2 remain largely unknown. Here, we evaluate the antiviral activity of type-II IFN (IFNγ) using human lung epithelial cells (Calu3) and ex vivo human lung tissues. In this study, we found that IFNγ suppresses SARS-CoV-2 replication in both Calu3 cells and ex vivo human lung tissues. Moreover, IFNγ treatment does not significantly modulate the expression of SARS-CoV-2 entry-related factors and induces a similar level of pro-inflammatory response in human lung tissues when compared with IFNß treatment. Mechanistically, we show that overexpression of indoleamine 2,3-dioxygenase 1 (IDO1), which is most profoundly induced by IFNγ, substantially restricts the replication of ancestral SARS-CoV-2 and the Alpha and Delta variants. Meanwhile, loss-of-function study reveals that IDO1 knockdown restores SARS-CoV-2 replication restricted by IFNγ in Calu3 cells. We further found that the treatment of l-tryptophan, a substrate of IDO1, partially rescues the IFNγ-mediated inhibitory effect on SARS-CoV-2 replication in both Calu3 cells and ex vivo human lung tissues. Collectively, these results suggest that type-II IFN potently inhibits SARS-CoV-2 replication through IDO1-mediated antiviral response.

Enhancing immune responses of ESC-based TAA cancer vaccines with a novel OMV delivery system.

Embryonic stem cell (ESC)-derived epitopes can act as therapeutic tumor vaccines against different types of tumors Jin (Adv Healthc Mater 2023). However, these epitopes have poor immunogenicity and stimulate insufficient CD8+ T cell responses, which motivated us to develop a new method to deliver and enhance their effectiveness. Bacterial outer membrane vesicles (OMVs) can serve as immunoadjuvants and act as a delivery vector for tumor antigens. In the current study, we engineered a new OMV platform for the co-delivery of ESC-derived tumor antigens and immune checkpoint inhibitors (PD-L1 antibody). An engineered Staphylococcal Protein A (SpA) was created to non-specifically bind to anti-PD-L1 antibody. SpyCatcher (SpC) and SpA were fused into the cell outer membrane protein OmpA to capture SpyTag-attached peptides and PD-L1 antibody, respectively. The modified OMV was able to efficiently conjugate with ESC-derived TAAs and PD-L1 antibody (SpC-OMVs + SpT-peptides + anti-PD-L1), increasing the residence time of TAAs in the body. The results showed that the combination therapy of ESC-based TAAs and PD-L1 antibody delivered by OMV had significant inhibitory effects in mouse tumor model. Specifically, it was effective in reducing tumor growth by enhancing IFN-γ-CD8+ T cell responses and increasing the number of CD8+ memory cells and antigen-specific T cells. Overall, the new OMV delivery system is a versatile platform that can enhance the immune responses of ESC-based TAA cancer vaccines.

Biodegradation of di(2-ethylhexyl) phthalate by a new bacterial consortium.

Di(2-ethylhexyl) phthalate (DEHP) with continuous high concentration was used as the sole carbon and energy source to isolate a new bacterial consortium (K1) from agricultural soil covered with plastic film for a long time. Unclassified Comamonadaceae, Achromobacter, and Pseudomonas in K1 were identified as major genera of the consortium by high-throughput sequencing, and unclassified Commanadaceae was first reported to be related to DEHP degradation. Response surface method (RSM) showed that the optimum conditions for K1 to degrade DEHP were 31.4 °C, pH 7.3, and a concentration of 420 mg L-1. K1 maintains normal cell viability and stable DEHP degradation efficiency in the range of 10-3000 mg L-1 DEHP concentration, which is superior to existing research. The biodegradation of DEHP followed first-order kinetics when the initial concentration of DEHP was between 100 and 3,000 mg L-1. GC-MS analysis of different treatment groups showed that DEHP was degraded by the consortium group through the de-esterification pathway, and treatment effect was significantly better than that of the single bacteria treatment group. The subsequent substrate utilization experiment further confirmed that K1 could quickly mineralize DEHP. In addition, K1 has high degradation capacity for the most common phthalate acid esters in the environment.

Heavy metals pollution of soil in central plains urban agglomeration (CPUA), China: human health risk assessment based on Monte Carlo simulation.

The present study conducted the concentration evaluation, pollution assessment, source analysis, and risk assessment of heavy metals in the soil of the CPUA, China, to contribute to the smooth construction of urban agglomeration. Elevated levels of mean concentrations of cadmium (Cd), chromium (Cr), and copper (Cu) in the soils were shown compared to background values. Cu and zinc (Zn) and also lead (Pb) and Cd exhibited spatial similarity. Manganese (Mn) and Cr exhibited point source characteristics such as the concentrations at a point much higher than the surrounding area. The potential ecological risk in the northern region belonged to the moderate risk level category. Cd contributed over 90% to the potential ecological risk. The health risk among children was higher than that among adults. The major exposure pathways were different for adults and children. Exposure, as shown using Hazard Index (HI), to adults was mainly through the skin contact route, while to children was through both the skin contact and ingestion route. The primary CR (carcinogenic risk) to adults was through the inhalation route, while that to children was through the ingestion route. In both children and adults, Cr was the main contributor to HI and CR. According to the Monte Carlo simulation results, the cumulative probability of exceeding the critical value of HI for children was approximately 2.8-3.0 times that for adults. According to the sensitivity analysis results, non-carcinogenic risk prevention should begin mainly by reducing exposure duration and skin contact. The cancer risk may be reduced primarily by decreasing the exposure duration and controlling ingestion. The PMF (Positive Matrix Factorization) source analysis revealed that Pb mainly came from transportation sources. In addition, Cu, Pb, and Mn were derived mainly from agricultural sources. Cr was derived mostly from a natural source, and Cd originated mainly from an industrial source.

Hyperbranched Polyesters Based on Indole- and Lignin-Derived Monomeric Aromatic Aldehydes as Effective Nonionic Antimicrobial Coatings with Excellent Biocompatibility.

This research aims to investigate nonionic hyperbranched polyesters (HBPs) derived from indole and lignin resources as new nontoxic antimicrobial coatings. Three nonionic HBPs with zero to two methoxy ether substituents on each benzene ring in the polymer backbones were synthesized by melt-polycondensation of three corresponding AB2 monomers. The molecular structures and thermal properties of the obtained HBPs were characterized by gel permeation chromatography, nuclear magnetic resonance spectroscopy, Fourier transform infrared spectroscopy, thermogravimetric analysis, and differential scanning calorimetry analyses. These HBPs were conveniently spin-coated on a silicon substrate, which exhibited significant antibacterial effect against Gram-negative (Escherichia coli and Pseudomonas aeruginosa) and Gram-positive bacteria (Staphylococcus aureus and Enterococcus faecalis). The presence of methoxy substituents enhanced the antimicrobial effect, and the resulting polymers showed negligible leakage in water. Finally, the polymers with the methoxy functionality exhibited excellent biocompatibility according to the results of hemolysis and MTT assay, which may facilitate their biomedical applications.

Polycyclic aromatic hydrocarbons in soils of Central Plains Urban Agglomeration, China: The bidirectional effects of urbanization and anthropogenic activities.

To investigate the variations in environmental behavior (levels, distribution, sources, and soil toxicity) of polycyclic aromatic hydrocarbons (PAHs) under the impact of anthropogenic activities during the urbanization process, we collected soil samples from 195 sites in the Central Plains Urban Agglomeration (CPUA), North China, and analyzed 16 U.S. Environmental Protection Agency (EPA) PAH priority pollutants. We divided the sampling sites into three groups (urban area, industrial area, and farmland) and collected soil samples (0-20 cm surface layer). ∑16PAHs concentrations in the soils of the urban area, industrial area, and farmland ranged from 24.2 to 4400 ng/g, 12.3-8780 ng/g, and 20.9-852 ng/g (the average value of 349, 634, and 186 ng/g), respectively. The 4 to 5 ring PAHs were dominant compounds in three soil types, accounting for 65-80% of the ∑16PAHs. The results of the source analysis showed that the PAHs in the soils of CPUA were mainly from energy consumption. PAH levels in urban and industrial soils had a potential low cancer risk. The impact of urbanization on PAHs in the soil was bidirectional. On the one hand, the level of PAHs in the farmland soil might increase due to burning coal and agricultural machinery, which releases diesel or petrol fumes. On the other hand, in the urbanization process, the PAH content in urban soil and industrial soil showed a downward trend due to the implementation of environmental protection policies in China, which have reduced the atmospheric input of PAHs into the soil.

Fatty infiltration of hip muscles and trochanteric soft tissue thickness are associated with hip fractures in the elderly.

PURPOSE: Apart from bone conditions, muscle and soft tissue parameters might also influence hip fractures. We aimed to evaluate the association between hip muscle and trochanteric soft tissue parameters and hip fractures. METHODS: We retrospectively reviewed 60 patients with hip fractures and 114 controls without hip fractures. Cases and controls were matched for age, sex, and body mass index using propensity score matching. Muscle cross-sectional area (CSA), mean attenuation, and fatty infiltration rate (FIR) (proportion of intramuscular fat content) were measured on CT images for the gluteus maximus, the gluteus medius/minimus, and the anterior and medial compartments of the upper thigh. Trochanteric soft tissue thickness (TSTT) and femoral neck attenuation were also measured. Univariate and multivariate analyses were conducted to identify potential risk factors of hip fractures. RESULTS: Patients with hip fractures had significantly lower femoral neck attenuation, TSTT, and CSA of the gluteus maximus and anterior compartment than controls. FIR of all hip muscle groups were significantly higher in hip fracture patients than controls. Multivariate analysis revealed that every 1% increase in FIR of medial compartment independently increased the odds of hip fractures by 23.7% (OR = 1.237, 95% CI = 1.093-1.401) and every 1 cm longer TSTT independently decreased the odds by 32.8% (OR = 0.672, 95% CI = 0.477-0.946). CONCLUSION: Fatty infiltration of hip muscles can better discriminate hip fractures than muscle area. Increased TSTT is independently associated with low fracture risk.

Degradation of tetracycline wastewater by suspended biochar as carriers in moving bed biofilm reactor.

To improve the removal efficiency of antibiotics in moving bed biofilm reactor, suspended biochar block was prepared by the one-pot process and was used as carriers to construct a reaction device to study the treatment effect of antibiotic wastewater. The characteristics of the hanging biofilm in wastewater were investigated. And the mechanism of biochar as a biological carrier has been studied. The results showed that in the 45-day experiment, the maximum number of biofilms for suspended biochar carriers was twice 3.4 times that of the high-density polyethylene carriers. When 10 mg/L tetracycline was added to the reactor, the removal efficiency of the tetracycline removal rate was 71.85% and the chemical oxygen demand (COD), total nitrogen (TN), and NH4+-N removal efficiency reached to 89.95, 61.91 and 85.47% respectively. Suspension biochar carriers can reduce fluctuations in redox potentials, thereby improving the cellular efficiency of microorganisms. Meanwhile, it inhibits the production of soluble microbial products and extracellular polymers, reduces toxic effects, and enhances the adhesion between microorganisms and carriers. The microbial communities of the two carriers were investigated by high-throughput sequencing techniques. Suspended biochar significantly increased the relative abundance of Hydrogenophaga and Comamonas, and improved the ability of nitrification and denitrification. Comamonas could be responsible for tetracycline degradation.

Emerging roles of circular RNAs in osteoporosis.

Osteoporosis is one bone disease characterized with skeletal impairment, bone strength reduced and fracture risk enhanced. The regulation processes of bone metabolism are associated with several factors such as mechanical stimulation, epigenetic regulation and hormones. However, the mechanism of osteoporosis remains unsatisfactory. Increasing high-throughput RNA sequencing and circular RNAs (circRNAs) microarray studies indicated that circRNAs are differentially expressed in osteoporosis. Growing functional studies further pinpointed specific deregulated expressed circRNAs (e.g., circ_28313, circ_0016624, circ_0006393, circ_0076906 and circ_0048211) for their functions involved in bone metabolism, including bone marrow stromal cells (BMSCs) differentiation, proliferation and apoptosis. Moreover, CircRNAs (circ_0002060, Circ_0001275 and Circ_0001445) may be acted as diagnostic biomarkers for osteoporosis. This review discussed recent progresses in the circRNAs expression profiling analyses and their potential functions in regulating BMSCs differentiation, proliferation and apoptosis.

Safety and Feasibility of Radiotherapy Plus Camrelizumab for Locally Advanced Esophageal Squamous Cell Carcinoma.

LESSONS LEARNED: Radiotherapy plus anti-PD-1 antibody as first-line therapy is safe and feasible in locally advanced esophageal squamous cell carcinoma (ESCC). Tumor-infiltrating and peripheral lymphocytes were associated with patient survival. Further studies combining chemoradiotherapy with immunotherapy in locally advanced ESCC and exploration of predictive biomarkers are warranted. BACKGROUND: We conducted a phase Ib study of radiotherapy plus programmed cell death protein 1 (PD-1) monoclonal antibody camrelizumab as first-line treatment for locally advanced esophageal squamous cell carcinoma (ESCC). METHODS: We planned to enroll 20 patients with newly diagnosed locally advanced ESCC. Patients received 60 Gy radiation (2.0 Gy/fraction, 5 fractions/week), with camrelizumab (200 mg every 2 weeks) starting with radiotherapy and continuing for 32 weeks (i.e., for 16 cycles). The primary endpoints were safety and feasibility. Secondary endpoints were rates of radiologic and pathologic response, overall survival (OS), and progression-free survival (PFS). Study data were collected by the week during radiotherapy (RT), every month during the maintenance camrelizumab treatment, and every 3 months after treatment. Tumor microenvironment and peripheral blood were monitored at baseline and after 40 Gy radiation for association with efficacy. RESULTS: Twenty patients were enrolled and received treatment. One patient (patient 10) was excluded upon discovery of a second tumor in the bladder during treatment, leaving 19 patients for analysis. Toxicity was deemed tolerable. Fourteen (74%) patients had assessed objective response. At a median follow-up time of 31.0 months (95% confidence interval [CI], 27.0-35.1), median OS and PFS times were 16.7 months (95% CI, 5.9-27.9) and 11.7 months (95% CI, 0-30.3), respectively. OS and PFS rates at 24 months were 31.6% and 35.5%, respectively. Kaplan-Meier analysis revealed associations between the following factors and OS/PFS: tumor programmed cell death ligand 1 (PD-L1) expression, PD-1+ CD8+ , PD-1+ CD4+ T cells, and PD-L1+ CD4+ T cells; peripheral blood CD4+ , CD8+ , CD4+ regulatory T cells, and their subsets. CONCLUSION: Radiotherapy plus camrelizumab had manageable toxicity and antitumor efficacy for locally advanced ESCC. Several biomarkers were associated with clinical benefit and deserve further study.

Synthesis, Enzymatic Degradation, and Polymer-Miscibility Evaluation of Nonionic Antimicrobial Hyperbranched Polyesters with Indole or Isatin Functionalities.

Most macromolecular antimicrobials are ionic and thus lack miscibility/compatibility with nonionic substrate materials. In this context, nonionic hyperbranched polyesters (HBPs) with indole or isatin functionality were rationally designed, synthesized, and characterized. Antimicrobial disk diffusion assay indicated that these HBPs showed significant antibacterial activity against 8 human pathogenic bacteria compared to small molecules with indole or isatin groups. According to DSC measurements, up to 20% indole-based HBP is miscible with biodegradable polyesters (polyhydroxybutyrate or polycaprolactone), which can be attributed to the favorable hydrogen bonding between the N-H moiety of indole and the C═O of polyesters. HBPs with isatin or methylindole were completely immiscible with the same matrices. None of the HBPs leaked out from plastic matrix after being immersed in water for 5 days. The incorporation of indole into HBPs as well as small molecules facilitated their enzymatic degradation with PETase from Ideonella sakaiensis, while isatin had a complex impact. Molecular docking simulations of monomeric molecules with PETase revealed different orientations of the molecules at the active site due to the presence of indole or isatin groups, which could be related to the observed different enzymatic degradation behavior. Finally, biocompatibility analysis with a mammalian cell line showed the negligible cytotoxic effect of the fabricated HBPs.

The health risk assessment of Heavy Metals (HMs) in road dust based on Monte Carlo simulation and bio-toxicity: a case study in Zhengzhou, China.

Heavy metals (HMs) in road dust pose a significant threat to human health. The analysis of human health risks of HMs is an important theoretical basis for risk screening and management. The chemical forms and characteristics of HMs in road dust were analyzed. Based on the bio-toxicity of the different fractions of the HMs and Monte Carlo, three assessment models, including the health risk assessment based on bio-toxicity, the health risk assessment based on the Monte Carlo simulation, and the health risk assessment based on the Monte Carlo simulation and bio-toxicity, were established. Under the Traditional Model, the non-carcinogenic risks were only harmful to children, while the carcinogenic risks were not harmful to adults and children. Under the M-Traditional Model, the probability of non-carcinogenic risks being harmful to children's health was 83.17%. The probability that carcinogenic risks pose a threat to children's health was 28.61%. Considering the bio-toxicity of HMs in different chemical forms, non-carcinogenic risks and carcinogenic risks under the B-Traditional Model were all less than the corresponding critical values, indicating that the HMs in the road dust did no harm to both the adults and children. Based on the MB-Traditional Model, the chance of non-carcinogenic risks being harmful to children's health is 15.43%. Among different HMs, the non-carcinogenic risks of As are highest and the carcinogenic risks of Cr were the highest, so As and Cr should be listed as priority control contamination. MB-Traditional Model established in this study simultaneously considered bio-toxicity and random simulation and obtained more accurate results, which could provide a theoretical basis for risk analysis and management.

Synthesis, Molecular Docking Simulation, and Enzymatic Degradation of AB-Type Indole-Based Polyesters with Improved Thermal Properties.

We report the facile synthesis of a series of indole-based hydroxyl-carboxylate (AB-type) monomers by a one-step procedure. These monomers were successfully polymerized by melt polycondensation to yield AB-type polyesters with a varied number of flexible methylene units in the backbones. These indole-based AB-polyesters showed decent thermal stability according to the TGA results (onset thermal degradation temperature of >330 °C), and their glass transition temperatures are dependent on the length of the methylene bridge (Tg ≈ 62-102 °C) according to the DSC results. Furthermore, DSC and WAXD measurements revealed that these polymers did not crystallize from melt, but the ones with flexible structures could crystallize from solution. Molecular docking simulations showed favorable interactions between indole-based polyesters and polyethylene terephthalate hydrolase (PETase) from Ideonella sakaiensis. This was corroborated by the experimental results, which indicated that the PETase enzyme has degrading activity on the indole-based AB polyesters except for the one with the highest degree of crystallinity.

TBX6-associated congenital scoliosis (TACS) as a clinically distinguishable subtype of congenital scoliosis: further evidence supporting the compound inheritance and TBX6 gene dosage model.

PURPOSE: To characterize clinically measurable endophenotypes, implicating the TBX6 compound inheritance model. METHODS: Patients with congenital scoliosis (CS) from China(N = 345, cohort 1), Japan (N = 142, cohort 2), and the United States (N = 10, cohort 3) were studied. Clinically measurable endophenotypes were compared according to the TBX6 genotypes. A mouse model for Tbx6 compound inheritance (N = 52) was investigated by micro computed tomography (micro-CT). A clinical diagnostic algorithm (TACScore) was developed to assist in clinical recognition of TBX6-associated CS (TACS). RESULTS: In cohort 1, TACS patients (N = 33) were significantly younger at onset than the remaining CS patients (P = 0.02), presented with one or more hemivertebrae/butterfly vertebrae (P = 4.9 × 10‒8), and exhibited vertebral malformations involving the lower part of the spine (T8-S5, P = 4.4 × 10‒3); observations were confirmed in two replication cohorts. Simple rib anomalies were prevalent in TACS patients (P = 3.1 × 10‒7), while intraspinal anomalies were uncommon (P = 7.0 × 10‒7). A clinically usable TACScore was developed with an area under the curve (AUC) of 0.9 (P = 1.6 × 10‒15). A Tbx6-/mh (mild-hypomorphic) mouse model supported that a gene dosage effect underlies the TACS phenotype. CONCLUSION: TACS is a clinically distinguishable entity with consistent clinically measurable endophenotypes. The type and distribution of vertebral column abnormalities in TBX6/Tbx6 compound inheritance implicate subtle perturbations in gene dosage as a cause of spine developmental birth defects responsible for about 10% of CS.

Canopy Nitrogen Concentration Monitoring Techniques of Summer Corn Based on Canopy Spectral Information.

Crop nitrogen monitoring techniques, particularly choosing sensitive monitoring bands and suitable monitoring models, have great significance both in theory and in practice for achieving non-destructive monitoring of nitrogen concentration and accurate management of water and fertilizer in large-scale areas. In this study, a lysimeter experiment was carried out to examine the characteristics of canopy spectral reflectance variation of summer corn under different fertilization levels. The relationship between canopy spectral reflectance and nitrogen concentration was investigated, based on which sensitive bands for the corn canopy nitrogen monitoring were selected and a suitable spectral index model was determined. The results suggest that under different fertilization levels, the canopy spectral reflectance of summer corn decreases with the increase of the canopy nitrogen concentration in the visible light band, but varies in the opposite direction in the near-infrared band, with a premium put on a higher correlation between the spectral reflectance of the characteristic bands and their first derivatives and the canopy nitrogen concentration. The most sensitive bands for monitoring the canopy nitrogen concentration using spectral reflectance and its first derivative are found to be 762 nm and 726 nm and the correlation coefficients are 0.550 and 0.795, respectively. The optimal band combination, generated by multivariate stepwise regression analysis, is composed of 762 nm, 944 nm and 957 nm bands. From the 55 reported spectral index models of crop nitrogen concentration monitoring, the most suitable index model, NDRE, is chosen such that this index model has the highest correlation with the canopy nitrogen concentration in summer corn. This model has a significant positive correlation with the canopy nitrogen concentration at each growth period, and the correlation coefficient is up to 0.738 during the whole growth period. Spectral monitoring models of canopy nitrogen concentration are constructed using sensitive bands, and a combination of bands and the spectral index, suggesting that these models perform well in monitoring. The models arranged in descending order of simulation accuracy are as follows: the suitable spectral index model, the optimal band combination model, the sensitive band reflectance first derivative model, the sensitive band reflectance model. The determination coefficients are 0.754, 0.711, 0.639 and 0.306, respectively.

Live attenuated Salmonella typhimurium vaccines delivering SaEsxA and SaEsxB via type III secretion system confer protection against Staphylococcus aureus infection.

BACKGROUND: Staphylococcus aureus (S. aureus) causes a wide range of infectious diseases in human and animals. The emergence of antibiotic-resistant strains demands novel strategies for prophylactic vaccine development. In this study, live attenuated S. enterica subsp. enterica serotype Typhimurium (S. Typhimurium) vaccine against S. aureus infection was developed, in which Salmonella Pathogenesis Island-1 Type 3 Secretion System (SPI-1 T3SS) was employed to deliver SaEsxA and SaEsxB, two of ESAT-6-like (Early Secreted Antigenic Target-6) virulence factors of S. aureus. METHODS: Antigens SaEsxA and SaEsxB were fused with the N-terminal secretion and translocation domain of SPI-1 effector SipA. And cytosolic delivery of Staphylococcal antigens into macrophages was examined by western blot. BALB/c mice were orally immunized with S. Typhimurium-SaEsxA and S. Typhimurium-SaEsxB vaccines. Antigen-specific humoral and Th1/Th17 immune responses were examined by ELISA and ELISPOT assays 7-9 days after the 2nd booster. For ELISPOT assays, the statistical significance was determined by Student's t test. The vaccine efficacy was evaluated by lethal challenge with two S. aureus clinical isolates Newman strain and USA 300 strain. Statistical significance was determined by Log rank (Mantel-Cox) analysis. And a P value of < 0.05 was considered statistically significant. RESULTS: Oral administration of S. Typhimurium-SaEsxA and S. Typhimurium-SaEsxB vaccines induced antigen-specific humoral and Th1/Th17 immune responses, which increased the survival rate for vaccinated mice when challenged with S. aureus strains. CONCLUSIONS: The newly developed S. Typhimurium-based vaccines delivering SaEsxA and SaEsxB by SPI-1 T3SS could confer protection against S. aureus infection. This study provides evidence that translocation of foreign antigens via Salmonella SPI-1 T3SS into the cytosol of antigen presenting cells (APCs) could induce potent immune responses against pathogens.

Immunotherapy Targeting Adenosine Synthase A Decreases Severity of Staphylococcus aureus Infection in Mouse Model.

Staphylococcusaureus is a severe pathogen found in the community and in hospitals. Most notably, methicillin-resistant S. aureus (MRSA) is resistant to almost all antibiotics, which is a growing public health concern. The emergence of drug-resistant strains has prompted the search for alternative treatments such as immunotherapeutic approaches. Previous research showed that S. aureus exploit the immunomodulatory attributes of adenosine to escape host immunity. In this study, we investigated adenosine synthase A (AdsA), an S. aureus cell wall-anchored enzyme as possible targets for immunotherapy. Mice vaccinated with aluminum hydroxide-formulated recombinant AdsA (rAdsA) induced high-titer anti-AdsA antibodies, thereby providing consistent protection in 3 mouse infection models when challenged with 2 S. aureus strains. The importance of anti-AdsA antibody in protection was demonstrated by passive transfer experiments. Moreover, AdsA-specific antisera promote killing S. aureus by immune cells. Altogether, our data demonstrate that the AdsA is a promising target for vaccines and therapeutics development to alleviate severe S. aureus diseases.

Cellulose-Organic Montmorillonite Nanocomposites as Biomacromolecular Quorum-Sensing Inhibitor.

The aim of this study was to develop simple cellulose nanocomposites that can interfere with the quorum-sensing (QS)-regulated physiological process of bacteria, which will provide a sustainable and inexpensive solution to the serious challenges caused by bacterial infections in various products like food packaging or biomedical materials. Three cellulose nanocomposites with 1-5 w% octadecylamine-modified montmorillonite (ODA-MMT) were prepared by regeneration of cellulose from ionic liquid solutions in the presence of ODA-MMT suspension. Structural characterization of the nanocomposites showed that the ODA-MMT can be exfoliated or intercalated, depending on the load level of the nanofiller. Thermal gravimetric analysis showed that the incorporation of ODA-MMT nanofiller can improve the thermal stability of the nanocomposites compared with regenerated cellulose. Evaluation of the anti-QS effect against a pigment-producing bacteria C. violaceum CV026 by disc diffusion assay and flask incubation assay revealed that the QS-regulated violacein pigment production was significantly inhibited by the cellulose nanocomposites without interfering the bacterial vitality. Interestingly, the nanocomposite with the lowest load of ODA-MMT exhibited the most significant anti-QS effect, which may be correlated to the exfoliation of nanofillers. To our knowledge, this is the first report on the anti-QS effect of cellulose nanocomposites without the addition of any small molecular agents. Such inexpensive and nontoxic biomaterials will thus have great potential in the development of new cellulosic materials that can effectively prevent the formation of harmful biofilms.

Exploring the Loading Capacity of Generation Six to Eight Dendronized Polymers in Aqueous Solution.

Aspects of size, structural (im)perfection, inner density, and guest molecule loading capacity of dendronized polymers (DPs) of high generation (6≤g≤8) in aqueous solution are studied using electron paramagnetic resonance spectroscopy on amphiphilic, spin-labeled guest molecules. The results show that the interior of the charged DPs is strongly polar, especially in comparison to their lower generation (1-4) analogues. This is a direct sign that large amounts of water penetrate the DP surface, reflecting the structural (im)perfections of these high-generation DPs and much lower segmental densities than theoretically achievable. Images obtained with atomic force microscopy reveal that the high-generation DPs do not aggregate and give further insights into the structural imperfections. Electron paramagnetic resonance spectroscopic data further show that despite their structural imperfections, these DPs can bind and release large numbers of amphiphilic molecules. It is concluded that attention should be paid to their synthesis, for which a protocol needs to be developed that avoids the relatively large amount of defects generated in the direct conversion of a generation g=4 DP to a generation g=6 DP, which had to be used here.