Potential targets and therapeutics for cancer stem cell-based therapy against drug resistance in hepatocellular carcinoma.

Hepatocellular carcinoma (HCC) is the most common digestive malignancyin the world, which is frequently diagnosed at late stage with a poor prognosis. For most patients with advanced HCC, the therapeutic options arelimiteddue to cancer occurrence of drug resistance. Hepatic cancer stem cells (CSCs) account for a small subset of tumor cells with the ability of self-renewal and differentiationin HCC. It is widely recognized that the presence of CSCs contributes to primary and acquired drug resistance. Therefore, hepatic CSCs-targeted therapy is considered as a promising strategy to overcome drug resistance and improve therapeutic outcome in HCC. In this article, we review drug resistance in HCC and provide a summary of potential targets for CSCs-based therapy. In addition, the development of CSCs-targeted therapeuticsagainst drug resistance in HCC is summarized in both preclinical and clinical trials. The in-depth understanding of CSCs-related drug resistance in HCC will favor optimization of the current therapeutic strategies and gain encouraging therapeutic outcomes.

Nonreciprocal dynamics of noninteracting ultracold atoms in a momentum lattice.

Realization of nonreciprocal transport is of great importance in the development of devices and systems that require the directional manipulation of signals or particles in information processing and modern physics. For ultracold atomic systems, the approaches based on synthetic dimensions have led to rapid advances in engineering quantum transport. Here, we use laser-coupled discrete momentum states of noninteracting ultracold atoms to synthesize a momentum lattice, and construct a closed ring with controllable tunneling phase in the momentum lattice. We measure the density evolution of atoms in the synthetic lattice with the single-site resolution, and observe the nonreciprocal dynamics by controlling the tunneling phase. We show the effect of both the applied phase and the coupling strength between two distinct population regions on the population distribution of atoms in the momentum lattice, and provide the optimal parameters for achieving the nonreciprocal transport.

Application of protection motivation theory in epidemic prevention in patients with respiratory diseases under the COVID-19 pandemic: A cross-sectional study.

OBJECTIVE: This study aimed to investigate the effects of nursing intervention based on protection motivation theory (PMT) on patients with respiratory diseases in the context of the Coronavirus Disease 2019 (COVID-19) pandemic. METHODS: A total of 74 patients with respiratory diseases who were hospitalized from June 2020 to February 2021 were enrolled and stratified into a control group (n = 37) and an experimental group (n = 37) according to a stratified random sampling method. The control group adopted a routine nursing intervention program of the respiratory department, whereas the experimental group received a PMT-based nursing intervention program on the basis of the control group. Chronic Disease Self-Management Study Measures (CDSMS) and Self-Efficacy for Managing Chronic Diseases 6-item Scale (SECD6) were used to evaluate the effect of PMT intervention before intervention, after 1 week, and after 4 weeks of intervention. The levels of forced vital capacity (FVC), forced expiratory volume in 1 second (FEV1), FEV1/FVC and peak expiratory flow (PEF) were measured to evaluate pulmonary function. RESULTS: Before the intervention, there were no significant differences in the scores of CDSMS and SECD6 scales and liver function indexes between the two groups (p > 0.05). After 1 and 4 weeks of intervention, the scores of CDSMS and SECD6 scales of the experimental group were significantly higher than those of the control group (p < 0.0001). The indexes of pulmonary function of the experimental group were improved, but there was no significant difference compared with the control group (p > 0.05). CONCLUSION: Nursing intervention based on PMT contributes to the improvement of self-management behaviors and self-efficacy, which is conducive to the prognoses of patients.

Composition, Properties, and Utilization of Fumaric Acid Sludge By-Produced from Industrial Phthalic Anhydride Wastewater Treatment.

To understand fumaric acid sludge (FAS) systematically and comprehensively and find out how to utilize it, we conducted a series of characterization analyses on FAS. Fourier transform infrared (FT-IR) Spectra shows that the main component of FAS is fumaric acids and also contains a small amount of silicate. The nuclear magnetic resonance hydrogen (1H-NMR) spectrum also shows that fumaric acid accounted for a large proportion of FAS. The X-ray diffraction (XRD) shows that the main phase in FAS is fumaric acid, and there is also a small amount of Kaliophilite. After gas chromatography and mass spectrometry (GC-MS) and pyrolysis gas chromatography and mass spectrometry (Py-GC-MS) analysis, it indicates that the possible volatiles and pyrolysis products in FAS are fumaric acid, maleic acid, maleic anhydride, phthalic acid, etc. In the test of Liquid chromatography and mass spectrometry (LC-MS), we determined the contents of phthalic acid, fumaric acid, and maleic acid in FAS. The detailed mass content of each component in FAS is as follows: phthalic acid is about 0.10-0.15%; maleic anhydride is about 0.40-0.80%; maleic acid is about 18.40-19.0%; fumaric acid is about 55.00-56.90%; succinic anhydride is about 0.06-0.08%; acrylic acid is about 0.06-0.08%; malic acid is about 0.90-1.00%; acetic acid is about 0.10-0.20%; silicate is about 0.25-0.30%; phthalic anhydride is about 0.20-0.30%; water is about 24.30-24.80%. The filtrate loss reducer (PAAF) used in oilwell drilling fluids synthesized by FAS not only has excellent temperature and complex saline resistance, the API filtration loss (FL) was only 13.2 mL/30 min in the complex saline based mud, but is also cost-effective.

Detection of SARS-CoV-2 antibodies in serum and dried blood spot samples of vaccinated individuals using a sensitive homogeneous proximity extension assay.

A homogeneous PCR-based assay for sensitive and specific detection of antibodies in serum or dried blood spots (DBS) is presented and the method is used to monitor individuals infected with or vaccinated against SARS-CoV-2. Detection probes were prepared by conjugating the recombinant spike protein subunit 1 (S1), containing the receptor binding domain (RBD) of SARS-CoV-2, to each of a pair of specific oligonucleotides. The same was done for the nucleocapsid protein (NP). Upon incubation with serum or DBS samples, the bi- or multivalency of the antibodies (IgG, IgA or IgM) brings pairs of viral proteins with their conjugated oligonucleotides in proximity, allowing the antibodies to be detected by a modified proximity extension assay (PEA). Anti-S1 and anti-NP antibodies could be detected simultaneously from one incubation reaction. This Antibody PEA (AbPEA) test uses only 1 µl of neat or up to 100,000-fold diluted serum or one ø1.2 mm disc cut from a DBS. All 100 investigated sera and 21 DBS collected prior to the COVID-19 outbreak were negative, demonstrating a 100% specificity. The area under the curve, as evaluated by Receiver Operating Characteristic (ROC) analysis reached 0.998 (95%CI: 0.993-1) for samples taken from 11 days after symptoms onset. The kinetics of antibody responses were monitored after a first and second vaccination using serially collected DBS from 14 individuals. AbPEA offers highly specific and sensitive solution-phase antibody detection without requirement for secondary antibodies, no elution step when using DBS sample in a simple procedure that lends itself to multiplex survey of antibody responses.

A Review on the Effect of the Mechanism of Organic Polymers on Pellet Properties for Iron Ore Beneficiation.

Iron ore pellets not only have excellent metallurgical and mechanical properties but are also essential raw materials for improving iron and steel smelting in the context of the increasing global depletion of high-grade iron ore resources. Organic polymers, as important additive components for the production of high-quality pellets, have a significant impact on the formation as well as the properties of pellets. In this review, the mechanisms of organic polymers on the pelletizing properties, bursting temperature, and pellet strength at low and high temperatures, as well as the existing measures and mechanisms to improve the high-temperature strength of the organic binder pellets are systematically summarized. Compared with traditional bentonite additives, the organic polymers greatly improve the pelletizing rate and pellet strength at low temperatures, and significantly reduces metallurgical pollution. However, organic binders often lead to a decrease in pellet bursting temperature and pellet strength at high temperatures, which can be significantly improved by compounding with a small amount of low-cost inorganic minerals, such as bentonite, boron-containing compounds, sodium salts, and copper slag. At the same time, some industrial solid wastes can be rationally used to reduce the cost of pellet binders.

A review on the industrial solid waste application in pelletizing additives: Composition, mechanism and process characteristics.

Reducing the cost of pellet additives as a substitute for reducing bentonite binder is an important research direction of new pellet additives. There are some industrial solid wastes that have the similar physical and chemical properties to bentonite, and SiO2 content of them may be much lower than bentonite, but also contains a lot of Fe2O3, Al2O3, MgO, B2O3 and other components beneficial to the quality of pellets, which have been paid more attention by many pellet workers. In this review, the effect mechanism of Fe2O3, Na2O/K2O, Al2O3, SiO2, CaO, MgO and B2O3 in the industrial solid wastes on the fired strength and reduction expansion of pellets were systematically summarized. At the same time, the influences of five representative large scale modified industrial solid waste additives including iron tailings, bauxite tailings, fly ash, red mud and boron sludge on the properties of green pellets and finished pellets were described in detail. It can be seen that the applications of industrial solid waste in pellet additives can partially or completely replace bentonite binder, especially fly ash, red mud and boron sludge, which can not only improve the quality of pellets, but also decrease the cost, save energy and reduce pollution, with significant economic benefits.

[Retracted] In vitro antitumor activity of the ethyl acetate extract Potentilla chinensis in osteosarcoma cancer cells.

Following the publication of the above paper, a concerned reader drew to the Editor's attention that several figures bore striking similarities to other papers that were published at around the same time written by different authors based in different research institutions. Fig. 3 (in colour) was essentially the same as a greyscale figure (Fig. 4) in a paper published in Oncology Reports, which has now been retracted [Wan G, Tao J­G, Wang G­D, Liu S­P, Zhao H­X and Liang Q­D: 3­ß­Εrythrodiol isolated from Conyza canadensis inhibits MKN­45 human gastric cancer cell proliferation by inducing apoptosis, cell cycle arrest, DNA fragmentation, ROS generation and reduces tumor weight and volume in mouse xenograft mode. Oncol Rep 35: 2328­2338, 2016]. Furthermore, Figs. 5 and 6 in the above paper appeared to share data with Figs. 7 and 11, respectively, in a paper published in Phytomedicine [Sui C­G, Meng F­D and Jiang Y­h: Antiproliferative activity of rosamultic acid is associated with induction of apoptosis, cell cycle arrest, inhibition of cell migration and caspase activation in human gastric cancer (SGC­7901) cells. Phyomedicine 22: 796­806, 2015]. After having conducted an independent investigation in the Editorial Office, the Editor of Molecular Medicine Reports has determined that the above paper should be retracted from the Journal on account of a lack of confidence concerning the originality and the authenticity of the data. The authors were asked for an explanation to account for these concerns, but the Editorial Office never received any reply. The Editor regrets any inconvenience that has been caused to the readership of the Journal. [the original article was published in Molecular Medicine Reports 14: 3634­3640, 2016; DOI: 10.3892/mmr.2016.5679].

The Human Adenovirus Type 2 Transcriptome: An Amazing Complexity of Alternatively Spliced mRNAs.

We have used the Nanopore long-read sequencing platform to demonstrate how amazingly complex the human adenovirus type 2 (Ad2) transcriptome is with a flexible splicing machinery producing a range of novel mRNAs both from the early and late transcription units. In total we report more than 900 alternatively spliced mRNAs produced from the Ad2 transcriptome whereof more than 850 are novel mRNAs. A surprising finding was that more than 50% of all E1A transcripts extended upstream of the previously defined transcriptional start site. The novel start sites mapped close to the inverted terminal repeat (ITR) and within the E1A enhancer region. We speculate that novel promoters or enhancer driven transcription, so-called eRNA transcription, is responsible for producing these novel mRNAs. Their existence was verified by a peptide in the Ad2 proteome that was unique for the E1A ITR mRNA. Although we show a high complexity of alternative splicing from most early and late regions, the E3 region was by far the most complex when expressed at late times of infection. More than 400 alternatively spliced mRNAs were observed in this region alone. These mRNAs included extended L4 mRNAs containing E3 and L5 sequences and readthrough mRNAs combining E3 and L5 sequences. Our findings demonstrate that the virus has a remarkable capacity to produce novel exon combinations, which will offer the virus an evolutionary advantage to change the gene expression repertoire and protein production in an evolving environment.IMPORTANCE Work in the adenovirus system led to the groundbreaking discovery of RNA splicing and alternative RNA splicing in 1977. These mechanisms are essential in mammalian evolution by increasing the coding capacity of a genome. Here, we have used a long-read sequencing technology to characterize the complexity of human adenovirus pre-mRNA splicing in detail. It is mindboggling that the viral genome, which only houses around 36,000 bp, not being much larger than a single cellular gene, generates more than 900 alternatively spliced mRNAs. Recently, adenoviruses have been used as the backbone in several promising SARS-CoV-2 vaccines. Further improvement of adenovirus-based vaccines demands that the virus can be tamed into an innocent carrier of foreign genes. This requires a full understanding of the components that govern adenovirus replication and gene expression.

Phosphorylation Time-Course Study of the Response during Adenovirus Type 2 Infection.

PTMs such as phosphorylations are usually involved in signal transduction pathways. To investigate the temporal dynamics of phosphoproteome changes upon viral infection, a model system of IMR-90 cells infected with human adenovirus type 2 (Ad2) is used in a time-course quantitative analysis combining titanium dioxide (TiO2 ) particle enrichment and SILAC-MS. Quantitative data from 1552 phosphorylated sites clustered the highly altered phosphorylated sites to the signaling by rho family GTPases, the actin cytoskeleton signaling, and the cAMP-dependent protein kinase A signaling pathways. Their activation is especially pronounced at early time post-infection. Changes of several phosphorylated sites involved in the glycolysis pathway, related to the activation of the Warburg effect, point at virus-induced energy production. For Ad2 proteins, 32 novel phosphorylation sites are identified and as many as 52 phosphorylated sites on 17 different Ad2 proteins are quantified, most of them at late time post-infection. Kinase predictions highlighted activation of PKA, CDK1/2, MAPK, and CKII. Overlaps of kinase motif sequences for viral and human proteins are observed, stressing the importance of phosphorylation during Ad2 infection.

Study on Extraction Separation of Thioarsenite Acid in Alkaline Solution by CO 3 2 - -Type Tri-n-Octylmethyl-Ammonium Chloride.

To overcome the problem of arsenic separation and enrichment from an alkaline leaching solution in arsenic-containing dust, a CO 3 2 - -type tri-n-octylmethyl-ammonium chloride (TOMAC) method for extracting thioarsenite is proposed in this paper. Considering an alkaline leaching solution as the research object, after vulcanization pretreatment, TOMAC transformation and organic phase saturated extraction capacity were measured, and the extraction mechanism was preliminarily studied. First, Cl--type quaternary ammonium salt was effectively transformed to HCO 3 - -type by treating organic phase with saturated NaHCO3five times. TOMAC was effectively transformed from HCO 3 - to CO 3 2 - type by alkaline washing with 1.0 mol/l NaOH solution; this washing was repeated thrice. Thereafter, the effects of organic phase composition, phase ratio, extraction time, and temperature on the extraction and separation of arsenic were investigated. The results show that under the conditions of 30% CO 3 2 - -type TOMAC + 15% sec-octanol + 55% sulfonated kerosene, VO/VA = 1/1, and 5 min extraction at room temperature, the single-stage extraction rate of AsIII is 85.2%. The AsIII concentration in raffinate can be reduced to less than 1.33 × 10-3 mol/l by four-stage countercurrent extraction, and the extraction rate of AsIII can exceed 98.4%.

Depression and anxiety among HIV-positive men who have sex with men and men who have sex with women in China.

China is experiencing an emerging HIV epidemic among men who have sex with men (MSM). Minority stress theory posits that marginalized populations experience additional stress, which influences experiences of psychological distress and health outcomes. This study aimed to understand psychological distress of MSM relative to men who have sex with women (MSW) in an urban Chinese setting. Cross-sectional survey data were collected from 162 HIV-positive Chinese men receiving HIV treatment at Beijing's Ditan Hospital. Multiple linear regression with imputation was used to identify correlates of psychological distress. Relative to MSW, MSM were younger, more educated, and less likely to be in a relationship or have children. While both groups reported clinically elevated levels of depression and anxiety, sexual behavior was not associated with either outcome. Higher endorsement of depression symptomology was associated with worse reported physical health (ß = -1.37, p < .05) and greater endorsement of maladaptive coping (ß = 2.39, p < .05), whereas higher endorsement of anxiety symptomology was associated with greater endorsement of adaptive coping (ß = 0.78, p < .05), diminished physical health (ß = -0.86, p < .05), and a high school or greater level of education (ß = 4.13, p < .05). These findings suggest that interventions targeting coping strategies may address psychological distress among HIV-positive Chinese men.

Adenovirus in the omics era - a multipronged strategy.

Human adenoviruses (HAdVs) are common pathogens associated with a wide variety of respiratory, ocular, and gastrointestinal diseases. To achieve its effective lytic mode of replication, HAdVs have to reprogram host-cell gene expression and fine-tune viral gene expression in a temporal manner. In two decades, omics revolution has advanced our knowledge about the HAdV and host-cell interplay at the RNA and protein levels. This review summarizes the current knowledge from large-scale datasets on how HAdV infections adjust coding and noncoding RNA expression, as well as how they reprogram host-cell proteome during the lytic course of infection.

Preparation of bioactive peptides with antidiabetic, antihypertensive, and antioxidant activities and identification of α-glucosidase inhibitory peptides from soy protein.

In this study, the peptides of soy protein obtained by enzymatic digestion with proteases were analyzed for their antidiabetic, antihypertensive, and antioxidant activities. Peptides prepared with alkaline proteinase (AP) exhibited the highest α-glucosidase inhibitory activity compared with those from papain and trypsin digestion. AP hydrolysates also exhibited dipeptidyl peptidase IV (DPP-IV) inhibitory, angiotensin-converting enzyme (ACE) inhibitory, and antioxidant activities. Gastrointestinal digestion of peptides enhanced α-glucosidase, DPP-IV, and ACE inhibitory activities compared with AP hydrolysates. AP peptides showing highest α-glucosidase inhibitory activity were purified by anion-exchange and size-exclusion chromatography， and identified using tandem MS. We found three novel α-glucosidase inhibitory peptides with sequences LLPLPVLK, SWLRL, and WLRL with IC50 of 237.43 ± 0.52, 182.05 ± 0.74, and 162.29 ± 0.74 µmol/L, respectively. Therefore, peptides hydrolyzed from soy protein are promising natural ingredients for nutraceutical applications assisting in the management of diabetes.

Long Non-Coding RNA AWPPH Promotes Postoperative Distant Recurrence in Resected Non-Small Cell Lung Cancer by Upregulating Transforming Growth Factor beta 1 (TGF-ß1).

BACKGROUND Postoperative recurrence of cancers is responsible for a large portion of deaths in cancer patients. Our study investigated the involvement of lncRNA AWPPH in recurrence of resected non-small cell lung cancer (NSCLC). MATERIAL AND METHODS A total of 128 patients were followed up for 3 years. Blood was extracted from each patient on the day of discharge, the day of the diagnosis of recurrence, or at the end of follow-up. Blood from 30 healthy controls was used as a control group. Patient were divided into 3 groups - a non-recurrence group (NR, n=54), a local recurrence group (LR, n=42), and a distant recurrence (DR, n=32) group - according to the follow-up results. Blood AWPPP was detected by qRT-PCR. AWPPH expression vectors were transfected into cells of human NSCLC cell lines. Cell migration and invasion were detected by Transwell migration and invasion assay, respectively. TGF-ß1 expression was detected by Western blot analysis. RESULTS Blood AWPPH levels were the highest in the DR group, followed by the LR and NR groups. The lowest blood AWPPH levels were observed in the control group. Blood AWPPH levels increased significantly in the DR group but not in the NR and LR groups during follow-up. Blood AWPPH levels were positively correlated with TGF-ß1 mRNA levels in the DR group but not in the NR and LR groups during follow-up. AWPPH overexpression promoted cell migration and invasion and upregulated TGF-ß1 expression. CONCLUSIONS lncRNA AWPPH can promote postoperative distant recurrence in resected NSCLC by upregulating TGF-ß1.

Transcriptomic and proteomic analyses reveal new insights into the regulation of immune pathways during adenovirus type 2 infection.

BACKGROUND: Human adenovirus (Ad) infection leads to the changes of host cell gene expression and biosynthetic processes. Transcriptomics in adenovirus type 2 (Ad2)-infected lung fibroblasts (IMR-90) cells has previously been studied using RNA sequencing. However, this study included only two time points (12 and 24 hpi) using constrained 76 bp long sequencing reads. Therefore, a more detailed study of transcription at different phases of infection using an up-graded sequencing technique is recalled. Furthermore, the correlation between transcription and protein expression needs to be addressed. RESULTS: In total, 3556 unique cellular genes were identified as differentially expressed at the transcriptional level with more than 2-fold changes in Ad2-infected cells as compared to non-infected cells by using paired-end sequencing. Based on the kinetics of the gene expression changes at different times after infection, these RNAs fell into 20 clusters. Among them, cellular genes involved in immune response were highly up-regulated in the early phase before becoming down-regulated in the late phase. Comparison of differentially expressed genes at transcriptional and posttranscriptional levels revealed low correlation. Particularly genes involved in cellular immune pathways showed a negative correlation. Here, we highlight the genes which expose inconsistent expression profiles with an emphasis on key factors in cellular immune pathways including NFκB, JAK/STAT, caspases and MAVS. Different from their transcriptional profiles with up- and down-regulation in the early and late phase, respectively, these proteins were up-regulated in the early phase and were sustained in the late phase. A surprising finding was that the target genes of the sustained activators failed to show response. CONCLUSION: There were features common to genes which play important roles in cellular immune pathways. Their expression was stimulated at both RNA and protein levels during the early phase. In the late phase however, their transcription was suppressed while protein levels remained stable. These results indicate that Ad2 and the host cell use different strategies to regulate cellular immune pathways. A control mechanism at the post-translational level must thus exist which is under the control of Ad2.

Upregulated BMP-Smad signaling activity in the glucuronyl C5-epimerase knock out MEF cells.

Glucuronyl C5-epimerase (Hsepi) catalyzes the conversion of glucuronic acid to iduronic acid in the process of heparan sulfate biosynthesis. Targeted interruption of the gene, Glce, in mice resulted in neonatal lethality with varied defects in organ development. To understand the underlying molecular mechanisms of the phenotypes, we used mouse embryonic fibroblasts (MEF) as a model to examine selected signaling pathways. Our earlier studies found reduced activities of FGF-2, GDNF, but increased activity of sonic hedgehog in the mutant cells. In this study, we focused on the bone morphogenetic protein (BMP) signaling pathway. Western blotting detected substantially elevated endogenous Smad1/5/8 phosphorylation in the Hsepi mutant (KO) MEF cells, which is reverted by re-expression of the enzyme in the KO cells. The mutant cells displayed an enhanced proliferation and elevated alkaline phosphatase activitywhen cultured in osteogenic medium. Analysis of the genes involved in the BMP signaling pathway revealed upregulation of a number of BMP ligands, but reduced expression of several Smads and BMP antagonist (Grem1) in the KO MEF cells. The high level of Smad1/5/8 phosphorylation was also found in primary calvarial cells isolated from the KO mice. The results suggest that Hsepi expression modulates BMP signaling activity, which, at least partially, is associated with defected molecular structure of heparan sulfate expressed in the cells.

Time-resolved proteomics of adenovirus infected cells.

Viral infections cause large problems in the world and deeper understanding of the disease mechanisms is needed. Here we present an analytical strategy to investigate the host cell protein changes during human adenovirus type 2 (HAdV-C2 or Ad2) infection of lung fibroblasts by stable isotope labelling of amino acids in cell culture (SILAC) and nanoLC-MS/MS. This work focuses on early phase of infection (6 and 12 h post-infection (hpi)) but the data is combined with previously published late phase (24 and 36 hpi) proteomics data to produce a time series covering the complete infection. As many as 2169 proteins were quantitatively monitored from 6 to 36 hpi, while some proteins were time-specific. After applying different filter criteria, 2027 and 2150 proteins were quantified at 6 and 12 hpi and among them, 431 and 544 were significantly altered at the two time points. Pathway analysis showed that the De novo purine and pyrimidine biosynthesis, Glycolysis and Cytoskeletal regulation by Rho GTPase pathways were activated early during infection while inactivation of the Integrin signalling pathway started between 6 and 12 hpi. Moreover, upstream regulator analysis predicted MYC to be activated with time of infection and protein and RNA data for genes controlled by this transcription factor showed good correlation, which validated the use of protein data for this prediction. Among the identified phosphorylation sites, a group related to glycolysis and cytoskeletal reorganization were up-regulated during infection. The results show specific aspects on how the host cell proteins, the final products in the genetic information flow, are influenced by Ad2 infection, which would be overlooked if only knowledge derived from mRNA data is considered.

Tespa1 Deficiency Dampens Thymus-Dependent B-Cell Activation and Attenuates Collagen-Induced Arthritis in Mice.

Thymocyte-expressed, positive selection-associated 1 (Tespa1) plays an important role in both T cell receptor (TCR)-driven thymocyte development and in the FcεRI-mediated activation of mast cells. Herein, we show that lack of Tespa1 does not impair B cell development but dampens the in vitro activation and proliferation of B cells induced by T cell-dependent (TD) antigens, significantly reduces serum antibody concentrations in vivo, and impairs germinal center formation in both aged and TD antigen-immunized mice. We also provide evidence that dysregulated signaling in Tespa1-deficient B cells may be linked to CD40-induced TRAF6 degradation, and subsequent effects on 1-phosphatidylinositol-4,5-bisphosphate phosphodiesterase gamma-2 (PLCγ2) phosphorylation, MAPK activation, and calcium influx. Furthermore, we demonstrate that Tespa1 plays a critical role in pathogenic B cells, since Tespa1-deficient chimeric mice showed a lower incidence and clinical disease severity of collagen-induced arthritis. Overall, our study demonstrates that Tespa1 is essential for TD B cell responses, and suggests an important role for Tespa1 during the development of autoimmune arthritis.

Temporal characterization of the non-structural Adenovirus type 2 proteome and phosphoproteome using high-resolving mass spectrometry.

The proteome and phosphoproteome of non-structural proteins of Adenovirus type 2 (Ad2) were time resolved using a developed mass spectrometry approach. These proteins are expressed by the viral genome and important for the infection process, but not part of the virus particle. We unambiguously confirm the existence of 95% of the viral proteins predicted to be encoded by the viral genome. Most non-structural proteins peaked in expression at late time post infection. We identified 27 non-redundant sites of phosphorylation on seven different non-structural proteins. The most heavily phosphorylated protein was the DNA binding protein (DBP) with 15 different sites. The phosphorylation occupancy rate could be calculated and monitored with time post infection for 15 phosphorylated sites on various proteins. In the DBP, phosphorylations with time-dependent relation were observed. The findings show the complexity of the Ad2 non-structural proteins and opens up a discussion for potential new drug targets.