Notes

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In this study, we investigated the involvement of keratin 8 during human influenza A/NWS/33 virus (H1N1) infection in semi-permissive rhesus monkey-kidney (LLC-MK2) and permissive human type II alveolar epithelial (A549) cells. In A549 cells, keratin 8 showed major expression and phosphorylation levels. Influenza A/NWS/33 virus was able to subvert keratin 8 structural organization at late stages of infection in both cell models, promoting keratin 8 phosphorylation in A549 cells at early phases of infection. Accordingly, partial colocalizations of the viral nucleoprotein with keratin 8 and its phosphorylated form were assessed by confocal microscopy at early stages of infection in A549 cells. The employment of chemical activators of phosphorylation resulted in structural changes as well as increased phosphorylation of keratin 8 in both cell models, favoring the influenza A/NWS/33 virus's replicative efficiency in A549 but not in LLC-MK2 cells. In A549 and human larynx epidermoid carcinoma (HEp-2) cells inoculated with respiratory secretions from pediatric patients positive for, respectively, influenza A virus or respiratory syncytial virus, the keratin 8 phosphorylation level had increased only in the case of influenza A virus infection. The results obtained suggest that in A549 cells the influenza virus is able to induce keratin 8 phosphorylation thereby enhancing its replicative efficiency.Cherry rusty mottle-associated virus (CRMaV), which belongs the genus Robigovirus of the family Betaflexiviridae, is strongly associated with cherry rusty mottle disease of sweet cherry, Prunus avium. Here, we report on the successful development of an Agrobacterium-based inoculation system for a cloned CRMaV cDNA construct. Agro-inoculation of virus-free cherry rootstock 'Krymsk6' [P. cerasus x (P. cerasus x P. maackii)] resulted in the development of chlorotic yellow mottle symptoms on systemic leaves beginning at 50 days post inoculation. The presence of CRMaV in 'Krymsk6' agro-inoculated plants was confirmed by RT-PCR and ELISA. Subsequently, CRMaV from agro-inoculated 'Krymsk6' was graft-transmissible onto virus-free sweet cherry rootstock P. avium 'Mazzard' as evidenced by the production of typical cherry rusty mottle symptoms beginning at 35 days post grafting, and further confirmed by western blotting and RT-PCR. These results showed conclusively that CRMaV is the causal agent of cherry rusty mottle disease in sweet cherry. The reverse genetic system presented in this study can be used as a tool to investigate the molecular biology of CRMaV and also a template for infectious clone development for other viruses in the genus Robigovirus.
Sodium/glucose cotransporter 2 (SGLT2) inhibitors decrease plasma triglyceride levels and slightly increase low-density lipoprotein (LDL-c) and high-density lipoprotein cholesterol (HDL-c). However, the mechanisms underlying such changes in the blood lipid profile remain to be determined. We investigated how empagliflozin affects plasma markers of cholesterol absorption and synthesis, and evaluated the relationship between changes in these markers and blood lipids in patients with type 2 diabetes.

In a randomized, active-controlled, open-label trial, 51 patients were randomly allocated in 21 ratio to receive empagliflozin 10 mg/day (n = 32) or standard therapy (n = 19) for 12 weeks. We measured plasma levels of lathosterol as a marker of cholesterol synthesis, and campesterol and sitosterol as markers of cholesterol absorption, at baseline and 12 weeks after treatment. In the empagliflozin group, serum HDL-c, but not LDL-c, significantly increased between baseline and 12 weeks (54.4 ± 16.3 vs. 58.8 ± 19.6 This increase may be associated with SGLT2 inhibitor-induced increases in HDL cholesterol.
Small case series and case reports indicated atypical persistent pruritic eruptions (PPEs), another type of skin lesions in adult-onset Still's disease (AOSD), imply a worse prognosis than typical evanescent rashes.

To investigate clinical characteristics and
occurrence in AOSD with PPEs.

A retrospective cohort study analyzed 150 AOSD patients with rashes at the First Affiliated Hospital of Zhejiang University from January 2013 to December 2019.

AOSD with PPEs had higher lactate dehydrogenase (492.00 U/L vs 382.00 U/L, p＜0.0001) and ferritin (6944.10 ng/mL vs 4286.60 ng/mL, p=0.033), and lower fibrinogen (5.05 g/L vs 5.77 g/L, p=0.014) than those with evanescent rashes. AOSD with PPEs had a higher incidence (17.4% vs 3.1%, p=0.006) and cumulative event rate for MAS (p=0.008), who tended to receive high-dose glucocorticoid (36% vs 20.3%, p=0.036). Multivariate analysis indicated PPEs (HR=5.519, 95%CI=1.138-26.767, p=0.034), aspartate aminotransferase (AST) >120 U/L (HR=8.084, 95%CI=1.728-37.826, p=0.008) and splenomegaly (HR=21.152, 95%CI=2.263-197.711, p=0.007) were independent risk factors for MAS.

Single-center, retrospective nature, small sample size.

PPEs indicated increased severity and MAS occurrence than evanescent rashes. PPEs, AST >120 U/L and splenomegaly were risk factors for MAS in AOSD with skin involvement.
120 U/L and splenomegaly were risk factors for MAS in AOSD with skin involvement.Cyprinid herpesvirus 2 (CyHV-2) is the etiological agent of Goldfish herpesviral haematopoietic necrosis (GHVHN) in goldfish. In this study, three new cell lines including Fantail goldfish gill (FtGG), Fantail goldfish liver (FtGL) and Fantail goldfish brain (FtGB) had been established and characterized from the gill, liver and brain tissue of C. auratus respectively. Cell lines were optimally grown at 28 °C in Leibovitz-15 (L-15) medium supplemented with 10 % fetal bovine serum (FBS). The PDT during exponential growth of FtGG, FtGL and FtGB cells were determined to be 41.47 h, 63.43 h and 79.79 h respectively. Karyotyping analysis of cell lines remained diploid (2n = 100). The revival rate was 82 %, 72 % and 70 % in FtGG, FtGL and FtGB cells respectively after 6 months of cryopreservation. All the three cells showed similar cytopathic effect (CPE) between 3-5 days post-infection (dpi) with CyHV-2 and complete destruction of the monolayer was observed at 8-10 dpi. The viral titers of CyHV-2 in FtGG, FtGL and FtGB reached 107.375±0.35 TCID50 ml-1, 104·55±0.070 TCID50 ml-1 and 106.45±0.070 TCID50 ml-1 respectively. These newly established cell lines will be a useful diagnostic tool for viral diseases in this fish species and also for the isolation and study of goldfish viruses in future.
We aimed to assess the effects of six weeks of resistance training (RT) combined with saffron supplementation on markers implicated in depression as well as happiness levels in untrained young males.

Untrained young male participants were randomly assigned to one of two groups RT+saffron supplement (RS; n = 14) or RT+placebo (RP; n = 14). For 6 weeks, participants in the RS group took one 150mg pill of pure saffron immediately after each RT session and at the same time on non-training days. Those assigned to the RP group took a dextrose pill. Concentrations of Anandamide (AEA), 2-Arachidonoylglycerol (2-AG), serotonin, dopamine, β-endorphin (beta-endorphin), tryptophan, happiness levels (via questionnaire), and body composition were assessed before and after the 6 weeks of whole-body supervised RT (4x/week, 3 sets using 60-70% of 1-repetition maximum [1-RM]).

AEA (0.5ng/ml), 2-AG (0.04ng/ml), dopamine (0.7ng/ml), and β-endorphin (9.4pg/ml) concentrations significantly increased in the RS group (P<0.05) while no changes were detected in the RP group. Serotonin (RS=1.7ng/mL and RP=1ng/mL) concentrations and happiness levels significantly increased in both groups with greater changes in RS group while tryptophan concentrations remained unchanged (P> 0.05). In addition, both groups significantly increased muscular endurance with greater changes in RS group (P<0.05).

Six weeks of RT combined with saffron supplementation improved AEA, 2-AG, dopamine, β-endorphin, and serotonin concentrations. Moreover, the addition of saffron supplement to chronic RT results in greater improvements in happiness levels than RT alone.
Six weeks of RT combined with saffron supplementation improved AEA, 2-AG, dopamine, β-endorphin, and serotonin concentrations. Moreover, the addition of saffron supplement to chronic RT results in greater improvements in happiness levels than RT alone.This study aimed to examine the effect of transcranial direct current stimulation (tDCS) used as a recovery strategy, on heart rate (HR) measures and perceived well-being in 12 male professional soccer players. tDCS was applied in the days after official matches targeting the left dorsolateral prefrontal cortex (DLPFC) with 2 mA for 20 min (F3-F4 montage). Participants were randomly assigned to anodal tDCS (a-tDCS) or sham tDCS sessions. Players completed the Well-Being Questionnaire (WBQ) and performed the Submaximal Running Test (SRT) before and after tDCS. HR during exercise (HRex) was determined during the last 30 s of SRT. HR recovery (HRR) was recorded at 60 s after SRT. The HRR index was calculated from the absolute difference between HRex and HRR. A significant increase was observed for WBQ (effect of time; p0.05). selleck kinase inhibitor These results suggest that a-tDCS over the DLPFC may have a positive effect on enhancing well-being and parasympathetic autonomic markers, which opens up a possibility for testing tDCS as a promising recovery-enhancing strategy targeting the brain in soccer players. The findings suggest that brain areas related to emotional and autonomic control might be involved in these changes with a possible interaction effect of tDCS by placebo-related effects, but more research is needed to verify this effect.The coronavirus nucleocapsid protein (N) controls viral genome packaging and contains numerous phosphorylation sites located within unstructured regions. Binding of phosphorylated SARS-CoV N to the host 14-3-3 protein in the cytoplasm was reported to regulate nucleocytoplasmic N shuttling. All seven isoforms of the human 14-3-3 are abundantly present in tissues vulnerable to SARS-CoV-2, where N can constitute up to ~1% of expressed proteins during infection. Although the association between 14-3-3 and SARS-CoV-2 N proteins can represent one of the key host-pathogen interactions, its molecular mechanism and the specific critical phosphosites are unknown. Here, we show that phosphorylated SARS-CoV-2 N protein (pN) dimers, reconstituted via bacterial co-expression with protein kinase A, directly associate, in a phosphorylation-dependent manner, with the dimeric 14-3-3 protein, but not with its monomeric mutant. We demonstrate that pN is recognized by all seven human 14-3-3 isoforms with various efficiencies and deduce the apparent KD to selected isoforms, showing that these are in a low micromolar range. Serial truncations pinpointed a critical phosphorylation site to Ser197, which is conserved among related zoonotic coronaviruses and located within the functionally important, SR-rich region of N. The relatively tight 14-3-3/pN association could regulate nucleocytoplasmic shuttling and other functions of N via occlusion of the SR-rich region, and could also hijack cellular pathways by 14-3-3 sequestration. As such, the assembly may represent a valuable target for therapeutic intervention.
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