ANKRD22 promotes resolution of psoriasiform skin inflammation by antagonizing NIK-mediated IL-23 production.

Inflammation resolution is an essential process for preventing the development of chronic inflammatory diseases. However, the mechanisms that regulate inflammation resolution in psoriasis are not well understood. Here, we report that ANKRD22 is an endogenous negative orchestrator of psoriasiform inflammation because ANKRD22-deficient mice are more susceptible to IMQ-induced psoriasiform inflammation. Mechanistically, ANKRD22 deficiency leads to excessive activation of the TNFRII-NIK-mediated noncanonical NF-κB signaling pathway, resulting in the hyperproduction of IL-23 in DCs. This is due to ANKRD22 being a negative feedback regulator for NIK because it physically binds to and assists in the degradation of accumulated NIK. Clinically, ANKRD22 is negatively associated with IL-23A expression and psoriasis severity. Of greater significance, subcutaneous administration of an AAV carrying ANKRD22-overexpression vector effectively hastens the resolution of psoriasiform skin inflammation. Our findings suggest ANKRD22, an endogenous supervisor of NIK, is responsible for inflammation resolution in psoriasis, and may be explored in the context of psoriasis therapy.

A completely precious metal-free alkaline fuel cell with enhanced performance using a carbon-coated nickel anode.

SignificanceWe present a groundbreaking advance in completely nonprecious hydrogen fuel cell technologies achieving a record power density of 200 mW/cm2 with Ni@CNx anode and Co-Mn cathode. The 2-nm CNx coating weakens the O-binding energy, which effectively mitigates the undesirable surface oxidation during hydrogen oxidation reaction (HOR) polarization, leading to a stable fuel cell operation for Ni@CNx over 100 h at 200 mA/cm2, superior to a Ni nanoparticle counterpart. Ni@CNx exhibited a dramatically enhanced tolerance to CO relative to Pt/C, enabling the use of hydrogen gas with trace amounts of CO, critical for practical applications. The complete removal of precious metals in fuel cells lowers the catalyst cost to virtually negligible levels and marks a milestone for practical alkaline fuel cells.

Esketamine induces apoptosis of nasopharyngeal carcinoma cells through the PERK/CHOP pathway.

Nasopharyngeal carcinoma, a malignant tumor prevalent in southeast Asia and north Africa, still lacks effective treatment. Esketamine, an N-methyl-D-aspartatic acid (NMDA) receptor (NMDAR) antagonist, is widely used in clinical anesthesia. Emerging evidence suggests that esketamine plays an important role in inhibiting tumor cell activity. However, the underlying mechanisms of esketamine on nasopharyngeal carcinoma remain unknown. In this study, we found that esketamine inhibited the proliferation and migration of nasopharyngeal carcinoma cells. Mechanically, transcriptome sequencing and subsequent verification experiments revealed that esketamine promoted the apoptosis of nasopharyngeal carcinoma cells through endoplasmic reticulum stress PERK/ATF4/CHOP signaling pathway mediated by NMDAR. Additionally, when combined with esketamine, the inhibitory effect of cisplatin on the proliferation of nasopharyngeal carcinoma cells was significantly enhanced. These findings provide new insights into future anti-nasopharyngeal carcinoma clinical strategies via targeting the NMDAR/PERK/CHOP axis alone or in combination with cisplatin.

Rational design and synthesis of triazene-amonafide derivatives as novel potential antitumor agents causing oxidative damage towards DNA through intercalation mode.

In this work, we rationally designed and synthesized two novel triazene-amonafide derivatives 2-(2-(diisopropylamino)ethyl)-5-(3,3-dimethyltriaz-1-en-1-yl)-1H-benzo[de]isoquinoline-1,3(2H)-dione (D-11) and 5-(3,3-diethyltriaz-1-en-1-yl)-2-(2-(diisopropylamino)ethyl)-1H-benzo[de]isoquinoline-1,3(2H)-dione (D-12) as potential antitumor agents. The DNA damage induced by the intercalation mode of D-11 (D-12) towards DNA was electrochemically detected through the construction of efficient biosensors. The consecutive processes of reversible redox of naphthylimide ring and irreversible oxidation of triazene moiety were elucidated on the surface of glassy carbon electrode (GCE) by CV, SWV, and DPV methods. Electrochemical biosensors were obtained through the immobilization of ctDNA, G-quadruplexes, poly(dG), and poly(dA), respectively, on the clean surface of GCE. After the incubation of biosensors with D-11 or D-12, the peaks of dGuo and dAdo decreased prominently, and the peak of 8-oxoGua appeared at +0.50 V, suggesting that the interaction between D-11 (D-12) and DNA could result in the oxidative damage of guanine. Unexpected, the as-prepared DNA biosensor possessed satisfactory anti-interference property and good practicability in real samples. UV-vis and fluorescence spectra, and gel electrophoresis assays were employed to further confirm the intercalation mode of D-11 (D-12) towards DNA base pairs. Moreover, D-11 was proved to exhibit stronger anti-proliferation activity than mitionafide and amonafide against both A549 and HeLa cell lines.

Impact of relative and absolute values on orienting attention in time.

Reward has been known to render the reward-associated stimulus more salient to block effective attentional orienting in space. However, whether and how reward influences goal-directed attention in time remains unclear. Here, we used a modified attentional cueing paradigm to explore the effect of reward on temporal attention, in which the valid targets were given a low monetary reward and invalid targets were given a high monetary reward. The results showed that the temporal cue validity effect was significantly smaller when the competitive reward structure was employed (Experiment 1), and we ruled out the possibility that the results were due to the practice effect (Experiment 2a) or a reward-promoting effect (Experiment 2b). When further strengthening the intensity of the reward from 1:10 to 1:100 (Experiment 3), we found a similar pattern of results to those in Experiment 1. These results suggest that reward information which was based on relative instead of absolute values can weaken, but not reverse, the orienting attention in time.

Effect of prophylactic antiviral intervention on T cell immunity in hepatitis B virus-infected pregnant women.

BACKGROUND: Antiviral intervention in hepatitis B virus (HBV)-infected pregnant women can effectively reduce mother-to-child transmission. However, the immunological characteristics of pregnant women with chronic HBV infection and the effects of antiviral intervention during pregnancy on maternal immune response remain unknown. We aimed to investigate these effects by comparing mothers who received antiviral intervention during pregnancy with those who did not. METHODS: Pregnant women positive for hepatitis B surface antigen and hepatitis B e-antigen (HBsAg+ HBeAg+) were enrolled at delivery, including 34 received prophylactic antiviral intervention during pregnancy (AVI mothers) and 15 did not (NAVI mothers). T lymphocyte phenotypes and functions were analysed using flow cytometry. RESULTS: At delivery, maternal regulatory T cell (Treg) frequency in AVI mothers was significantly higher than that in NAVI mothers (P < 0.002), and CD4+ T cells in AVI mothers displayed a decreased ability to secrete IFN-γ (P = 0.005) and IL-21 (P = 0.043), but an increased ability to secrete IL-10 and IL-4 (P = 0.040 and P = 0.036), which represented a higher Treg frequency, enhanced Th2 response and suppressed Th1 response. Treg frequency among AVI mothers was correlated negatively with serum HBsAg and HBeAg levels. After delivery, the ability of CD4+ T cells or CD8+ T cells to secrete IFN-γ or IL-10 was similar and no significant difference in Treg frequency was found between the two groups. CONCLUSIONS: Prophylactic antiviral intervention during pregnancy has an effect on T cell immunity in pregnant women, which was characterised by increased maternal Treg frequency, enhanced Th2 response and suppressed Th1 response at delivery.

Focused attention: its key role in gaze and arrow cues for determining where attention is directed.

Others' gaze direction and traffic arrow signal lights play significant roles in guiding observers' attention in daily life. Previous studies have shown that gaze and arrow cues can direct attention to the cued location. However, it is ambiguous where gaze and arrow cues guide attention: the cued location or a broader cued region. Therefore, the present study adopted a primary cue-target task and manipulated possible target locations to explore this issue. The results revealed that due to the different physical characteristics of non-predictive gaze and arrow cues, physically unfocused-pointing gaze cues guided attention to a broader cued region, whereas focused-pointing arrow cues guided attention to the exact cued location. Furthermore, gaze cues could also direct attention to the exact cued location when observers' attention was focused in a top-down manner (with highly predictive probability). These findings suggest that where gaze and arrow cues direct attention depends on whether observers' attention is focused by the cues, either in a bottom-up or top-down manner. Accordingly, a preliminary framework called the "Focused-Diffused Attentional Orienting Model" is proposed to explain how gaze and arrow cues direct humans' attention. The present study enhances our understanding of human attentional orienting systems from a behavioral perspective.

Metabolomic analyses reveal new stage-specific features of COVID-19.

The current pandemic of coronavirus disease 2019 (COVID-19) has affected >160 million individuals to date, and has caused millions of deaths worldwide, at least in part due to the unclarified pathophysiology of this disease. Identifying the underlying molecular mechanisms of COVID-19 is critical to overcome this pandemic. Metabolites mirror the disease progression of an individual and can provide extensive insights into their pathophysiological significance at each stage of disease. We provide a comprehensive view of metabolic characterisation of sera from COVID-19 patients at all stages using untargeted and targeted metabolomic analysis. As compared with the healthy controls, we observed different alteration patterns of circulating metabolites from the mild, severe and recovery stages, in both the discovery cohort and the validation cohort, which suggests that metabolic reprogramming of glucose metabolism and the urea cycle are potential pathological mechanisms for COVID-19 progression. Our findings suggest that targeting glucose metabolism and the urea cycle may be a viable approach to fight COVID-19 at various stages along the disease course.

Exposure to neonicotinoid insecticides and their characteristic metabolites: Association with human liver cancer.

Neonicotinoid insecticides (NEOs) are commonly applied for pest control in China and around the world. Previous studies reported that NEOs are hepatotoxic to mammals. However, limited studies have explored the associations between NEOs exposure and liver disease. In the present study, we detected six parent NEOs (p-NEOs), including acetamiprid, thiacloprid, dinotefuran, clothianidin, imidacloprid, and thiamethoxam, and five characteristic metabolites (m-NEOs), including 5-hydroxy-imidacloprid, olefin-imidacloprid, N-desmethyl-acetamiprid, 1-methyl-3-(tetrahydro-3-furylmethyl) guanidine and 1-methyl-3-(tetrahydro-3-furyl methyl) urea, in blood samples collected from healthy donors (n = 100; females vs. males: 45 vs. 55; age: 22-91 years) and liver cancer patients (n = 274; females vs. males: 118 vs. 156; age: 11-88 years) in one hospital from Guangzhou city, South China. NEOs were frequently detected (61%-94%) in blood samples, with median concentrations ranging from 0.19 ng/mL to 1.28 ng/mL and 0.20 ng/mL to 2.03 ng/mL for healthy and liver cancer populations, respectively. olefin-imidacloprid was the most abundant NEOs in healthy and liver cancer populations, accounting for 23.4% and 20.7%, respectively. Significant positive correlations among most m-NEOs concentrations were found, and associations between m-NEOs and their corresponding p-NEOs were positively correlated. These findings indicated that the sources of m-NEOs were both endogenous and exogeneous. Females had higher median concentrations of NEOs and their metabolites than males. Moreover, the α-fetoprotein values and blood concentrations of target analytes (r = 0.428-0.601, p < 0.05) were positively correlated. Meanwhile, associations between the concentrations of p-NEOs and m-NEOs and liver cancer were found (odds ratio = 2.33-9.02, 95% confidence interval = 0.31-22.7, p < 0.05), indicating that human exposure to NEOs and their metabolites might increase the odds of liver cancer prevalence. Our work provided a new insight into the hepatotoxicity of NEOs and their metabolites, and human health risks of exposure to these pollutants warranted further studies.

Nano-silica embedded polydimethylsiloxane on interdigitated sensor as adhesive polymer for detecting lung cancer mutation.

Non-small cell lung cancer (NSCLC) incited by epidermal growth factor receptor (EGFR) mutation makes up ∼85% of lung cancer diagnosed and death cases worldwide. The presented study introduced an alternative approach in detecting EGFR mutation using nano-silica integrated with polydimethylsiloxane (PDMS) polymer on interdigitated electrode (IDE) sensor. A 400 µm gap-sized aluminum IDE was modified with nano-polymer layer, which was made up of silica nanoparticles and PDMS polymer. IDE and PDMS-coated IDE (PDMS/IDE) were imaged using electron microscopes that reveals its smooth and ideal sensor morphology. The nano-silica-integrated PDMS/IDE surface was immobilized with EGFR probe and target to specify the lung cancer detection. The sensor specificity was justified through the insignificant current readouts with one-base mismatch and noncomplementary targets. The sensitivity of nano-silica-integrated PDMS/IDE was examined with mutant target spiked in human serum, where the resulting current affirms the detection of EGFR mutation. Based on the slope of the calibration curve, the sensitivity of nano-silica-integrated PDMS/IDE was 2.24E-9 A M-1 . The sensor recognizes EGFR mutation lowest at 1 aM complementary mutant target; however, the detection limit obtained based on 3σ calculation is 10 aM with regression value of 0.97.

Characterization of T cell immunity in chronic hepatitis B virus-infected mothers with postpartum alanine transaminase flare.

BACKGROUND: Postpartum alanine transaminase (ALT) flares occur frequently in chronic hepatitis B virus (HBV)-infected mothers with antepartum antiviral therapy (AVT). We aimed to characterize the T cell immunity in HBV-infected mothers experiencing postpartum ALT flares. METHODS: Twenty HBV-infected pregnant women who received AVT at 26-28 weeks of gestation were enrolled and followed up until 15-18 weeks postpartum. Among the 20 HBV-infected pregnant women, 6 experienced postpartum ALT flare (AF mothers), while 14 did not (NAF mothers). T lymphocyte phenotypes and functions were analyzed using flow cytometry. RESULTS: Compared to NAF mothers, the quantitative HBsAg levels in AF mothers decreased significantly at 6-8 or 15-18 weeks postpartum. Significant differences in HBeAg levels between these groups were only found at delivery. Regulatory T cell (Treg) numbers in AF mothers were lower than those of NAF mothers before AVT; however, there were no significant differences in Treg numbers at other follow-up points. Expression of other T cell phenotypes were similar between the two groups. T cells in AF mothers produced more pro-inflammatory cytokines (IFN-γ, IL-21, TNF-α, IL-2) or less anti-inflammatory cytokine (IL-10) than those in NAF mothers before, during, or after antiviral treatment. The ratio of IFN-γ to IL-10 producing by CD4+ T cells or CD8+ T cells was higher in AF mothers than that in NAF mothers during pregnancy or after delivery. CONCLUSIONS: The characteristics of T cell immunity was distinct between mothers with postpartum ALT flare and those without ALT flare from pregnancy to postpartum, which indicated that T cell immunity might get involved in postpartum ALT flare.

Occurrence and distribution of neonicotinoids and characteristic metabolites in paired urine and indoor dust from young adults: Implications for human exposure.

Neonicotinoid insecticides (NEOs) are widely used for pest control worldwide. The profile of NEOs in paired urine and indoor dust has not yet been reported in China. In this study, 40 paired samples (i.e., 160 urine and 40 indoor dust) were collected from university students and dormitories from Guangzhou City of China to measure the concentrations of six NEOs and their three metabolites. Target analytes were frequently detected in paired urine (81%-98%) and indoor dust (75%-95%) samples, with median concentrations ranging from 0.02 [specific gravity (SG) adjusted: 0.02] to 2.08 (SG-adjusted: 2.38) ng/mL in urine and from 0.05 to 2.74 ng/g in indoor dust. 5-Hydroxy-imidacloprid was predominant in urine, while N-desmethyl acetamiprid was predominant in indoor dust samples, accounting for 56% and 37%, respectively. 1-Methyl-3-(tetrahydro-3-furylmethyl) urea, a dinotefuran degradate, was measured for the first time in indoor dust, with the median level of 1.02 ng/g. Significant gender-related differences (p < 0.05) in the urinary concentrations of most NEOs were found. We calculated the estimated daily intake (EDI) of target compounds from urine and indoor measurements. The EDIs of target analytes varied among all urine and indoor dust samples, with median values ranging from 0.51 (SG-adjusted: 0.56) to 51.6 (SG-adjusted: 52.8) ng/kg bw/day and from 0.04 to 2.10 pg/kg bw/day, respectively. Moreover, the median EDIsurine of most target analytes in females were significantly higher than (p < 0.05) those in males. The median EDIsdust of target compounds in dust from female dormitories were slightly higher than that in dust from male dormitories. These findings indicated that females were more exposed to NEO than males. Thus, the potential health risks of exposure to NEOs and their metabolites in female adults should be addressed in future studies. To our knowledge, this study is the first to report the profiles of NEOs and their metabolites in paired urine and indoor dust samples from young adults in China.

Profiles of neonicotinoid insecticides and their metabolites in paired saliva and periodontal blood samples in human from South China: Association with oxidative stress markers.

Neonicotinoid insecticides (NEOs) are widely used around the world. The distribution of NEOs in paired saliva and periodontal blood samples was not previously documented in China. In this study, the concentrations of six NEOs and three corresponding metabolites were measured in 188 paired saliva and periodontal blood samples collected from South China. NEOs and their metabolites were frequently detected (68-94%) in paired saliva and periodontal blood, with median levels of 0.01-0.99 ng/mL. 1-Methyl-3-(tetrahydro-3-furylmethyl) urea was the most predominant NEO in paired saliva (39%) and periodontal blood (42%). Gender-related differences in NEOs and their metabolite concentrations were found: males showed lower levels than females. We calculated the concentration ratios between saliva and periodontal blood (S/PB ratios), and found that the median S/PB ratios of NEO and their metabolites were higher than 1, indicating that NEOs and their metabolites were easily excreted via saliva. 8-Hydroxy-2'-deoxyguanosine (8-OHdG) was measured in paired saliva and periodontal blood as a marker of oxidative stress. 8-OHdG concentrations in saliva and periodontal blood were significantly and positively correlated (p < 0.05) with the concentrations of most NEOs and their metabolites in saliva and periodontal blood samples. These findings indicated that exposure to NEOs and their metabolites is associated with oxidative stress. This study is the first to report NEOs and their metabolites in paired saliva and periodontal blood samples collected from South China.

Effects of intrauterine exposure to maternal-derived HBeAg on T cell immunity in cord blood.

Immature immune system and immune tolerance induced by exposure to HBeAg in utero and/or shortly after infection in newborns were reportedly the causes of chronic HBV infection. To investigate the effect of maternal-derived HBeAg on neonatal T cell immunity, we analysed and compared T cell phenotypes and functions among neonates born to HBsAg+ /HBeAg+ mothers (HBeAg+ neonates), HBsAg+ /HBeAg- mothers (HBeAg- neonates) and healthy control mothers (HC neonates), using flow cytometry. The results showed that neonatal T cell phenotypes were similar regardless of HBeAg exposure. Upon anti-CD3 and anti-CD28 stimulation in HBeAg+ neonates, CD4+ T cell production of IFN-γ (P < .05) was significantly enhanced, while CD8+ T cells secreted significantly more IL-2 compared with those in HBeAg- and HC groups (P < .05). Moreover, similar levels of IFN-γ and IL-10 were observed in the culture supernatant after stimulation with rHBsAg, rHBcAg or rHBeAg among HBeAg+ , HBeAg- and HC neonates, whereas HBeAg+ neonates produced more TNF-α than HBeAg- neonates upon stimulation with rHBcAg. In conclusion, the results indicated that the HBsAg+ /HBeAg+ maternal environment did not influence the phenotypes of cord blood T cells but boosted neonatal non-specific Th1-type cytokine production.

Occurrence of parabens and their metabolites in the paired urine and blood samples from Chinese university students: Implications on human exposure.

Parabens, a group of p-hydroxybenzoic acid esters, are extensively used in cosmetics, personal care products, pharmaceuticals, and foodstuff. In the present study, the total forms (free plus conjugated) of four parent parabens, such as methylparaben (MeP), ethylparaben (EtP), propylparaben (PrP), and butylparaben (BuP), and four metabolites, namely methyl protocatechuate (OH-MeP), ethyl protocatechuate (OH-EtP), p-hydroxy benzoic acid (4-HB), and 3,4-dihydroxy benzoic acid (3,4-DHB), were detected in paired urine and blood samples collected from 196 Chinese university students. The median urinary and blood parabens and their metabolites concentrations ranged from 0.24 to 167 ng/mL and from <0.02 to 2.88 ng/mL, respectively. MeP was the predominant parent parabens, accounting for 68% and 52% of urine and blood samples, respectively. Furthermore, 4-HB predominantly contributed to the parabens and their metabolites in urine (54%) and blood (41%). Significant positive correlations were observed between the urinary levels and blood levels. Moreover, relatively high levels of parabens and their metabolites were detected in urine samples. Our results imply that urinary concentrations are good predictors of human exposure to parabens and metabolites. Gender-related difference in urinary concentrations of parabens and their metabolites were found. The median urinary levels of the tested compounds in females were significantly higher than those in males (Mann Whitney U test, p < 0.05 or 0.01). The estimated daily intakes (EDIs) of MeP, EtP, and PrP were also evaluated. The median values of EDIMeP, EDIEtP, and EDIPrP for all of the university students were estimated to be 25.9, 1.61, and 3.82 µg/kg bw/day, respectively. The median values (µg/kg bw/day) of EDIMeP, EDIEtP, and EDIPrP were higher in females (53.7, 8.65, and 5.22) than in males (8.41, 0.85, and 2.57). This study is the first study to report the occurrence of parabens and their metabolites in paired urine and blood samples in China.

Development and application of a triplex real-time PCR assay for simultaneous detection of avian influenza virus, Newcastle disease virus, and duck Tembusu virus.

BACKGROUND: Pathogens including duck-origin avian influenza virus (AIV), duck-origin Newcastle disease virus (NDV) and duck Tembusu virus (DTMUV) posed great harm to ducks and caused great economic losses to the duck industry. In this study, we aim to develop a triplex real-time polymerase chain reaction (PCR) assay to detect these three viruses as early as possible in the suspicious duck flocks. RESULTS: The detection limit of the triplex real-time PCR for AIV, NDV, and DTMUV was 1 × 101 copies/µL, which was at least 10 times higher than the conventional PCR. In addition, the triplex assay was highly specific, and won't cross-react with other duck pathogens. Besides, the intra-day relative standard deviation and inter-day relative standard deviation were lower than 4.44% for these viruses at three different concentrations. Finally, a total of 120 clinical samples were evaluated by the triplex real-time PCR, the conventional PCR and virus isolation, and the positive rates for these three methods were 20.83, 21.67, 19.17%, respectively. Taking virus isolation as the gold standard, the diagnostic specificity and positive predictive value of the three viruses were all above 85%, while the diagnostic sensitivity and negative predictive value of the three viruses were all 100%. CONCLUSION: The developed triplex real-time PCR is fast, specific and sensitive, and is feasible and effective for the simultaneous detection of AIV, NDV, and DTMUV in ducks.

Different temporal dynamics of object-based attentional allocation for reward and non-reward objects.

Previous studies have confirmed that both non-reward objects (such as rectangles) and reward objects (such as banknotes) can guide the allocation of our attention; however, it is unclear whether the allocation mode of attention for reward objects is the same as for non-reward objects. This study aims to evaluate different modes of object-based attentional selection elicited by two types of objects: reward objects and non-reward objects. In our analysis, we used a two-rectangle paradigm in which two objects were presented visually. In a series of four experiments, we found a constant object-based effect with non-reward objects, such as rectangles and umbrellas, as stimuli in all of the stimulus onset asynchrony (SOA) conditions (Experiments 1 and 4), but the object-based effect disappeared only at longer SOA with reward objects such as monetary and food objects as stimuli (Experiments 2 and 3). Moreover, we found that monetary and food objects induced similar object-based effects. These results suggest that the temporal dynamics of object-based attentional allocation are different with respect to reward and non-reward objects, and different types of reward objects can guide attentional allocation in a similar way.

Development and application of multiplex PCR method for simultaneous detection of seven viruses in ducks.

BACKGROUND: Major viruses, including duck-origin avian influenza virus, duck-origin Newcastle disease virus, novel duck parvovirus, duck hepatitis A virus, duck Tembusu virus, fowl adenovirus, and duck enteritis virus, pose great harm to ducks and cause enormous economic losses to duck industry. This study aims to establish a multiplex polymerase chain reaction (m-PCR) method for simultaneous detection of these seven viruses. RESULTS: Specific primers were designed and synthesized according to the conserved region of seven viral gene sequences. Then, seven recombinant plasmids, as the positive controls, were reconstructed in this study. Within the study, D-optimal design was adopted to optimize PCR parameters. The optimum parameters for m-PCR were annealing temperature at 57 °C, Mg2+ concentration at 4 mM, Taq DNA polymerase concentration at 0.05 U/µL, and dNTP concentration at 0.32 mM. With these optimal parameters, the m-PCR method produced neither cross-reactions among these seven viruses nor nonspecific reactions with other common waterfowl pathogens. The detection limit of m-PCR for each virus was 1 × 104 viral DNA copies/µL. In addition, the m-PCR method could detect a combination of several random viruses in co-infection analysis. Finally, the m-PCR method was successfully applied to clinical samples, and the detection results were consistent with uniplex PCR. CONCLUSION: Given its rapidity, specificity, sensitivity, and convenience, the established m-PCR method is feasible for simultaneous detection of seven duck-infecting viruses and can be applied to clinical diagnosis of viral infection in ducks.

Impact of mycolic acid deficiency on cells of Corynebacterium glutamicum ATCC13869.

Mycolic acid (MA) plays important role in Corynebacterium glutamicum, but the key enzymes in the biosynthetic pathway of MA in C. glutamicum ATCC13869 have not been characterized. Since the locus BBD29_RS14045 in C. glutamicum ATCC13869 shows high similarity to the gene Cgl2871, which encodes Pks13, the key enzyme for synthesizing MA in C. glutamicum ATCC13032, it was deleted, resulting in the mutant WG001. Compared with the wild-type ATCC13869, MA was not synthesized in WG001, but more phosphatidylglycerol and phosphatidylinositol containing longer unsaturated fatty acids were produced. WG001 cells also show hindered cell growth and defective cell separation when compared with ATCC13869 cells. Transcriptomic analysis shows that many genes relevant to the pathways of fatty acids, inositol, phospholipids, cell wall, and cell division were significantly regulated in WG001 cells when compared with ATCC13869 cells. This study demonstrates that the locus BBD29_RS14045 encodes a key enzyme that plays important role for synthesizing MA in C. glutamicum ATCC13869.

Autophagy-independent functions of UVRAG are essential for peripheral naive T-cell homeostasis.

UV radiation resistance-associated gene (UVRAG) encodes a tumor suppressor with putative roles in autophagy, endocytic trafficking, and DNA damage repair but its in vivo role in T cells is unknown. Because conditional homozygous deletion of Uvrag in mice results in early embryonic lethality, we generated T-cell-specific UVRAG-deficient mice that lacked UVRAG expression specifically in T cells. This loss of UVRAG led to defects in peripheral homeostasis that could not be explained by the increased sensitivity to cell death and impaired proliferation observed for other autophagy-related gene knockout mice. Instead, UVRAG-deficient T-cells exhibited normal mitochondrial clearance and activation-induced autophagy, suggesting that UVRAG has an autophagy-independent role that is critical for peripheral naive T-cell homeostatic proliferation. In vivo, T-cell-specific loss of UVRAG dampened CD8(+) T-cell responses to LCMV infection in mice, delayed viral clearance, and impaired memory T-cell generation. Our data provide novel insights into the control of autophagy in T cells and identify UVRAG as a new regulator of naïve peripheral T-cell homeostasis.