Plant Extracellular Vesicles and Nanovesicles: Focus on Secondary Metabolites, Proteins and Lipids with Perspectives on Their Potential and Sources

While human extracellular vesicles (EVs) have attracted a big deal of interest and have been extensively characterized over the last years, plant-derived EVs and nanovesicles have earned less attention and have remained poorly investigated. Although a series of investigations already revealed promising beneficial health effects and drug delivery properties, adequate (pre)clinical studies are rare. This fact might be caused by a lack of sources with appropriate qualities. Our study introduces plant cell suspension culture as a new and well controllable source for plant EVs. Plant cells, cultured in vitro, release EVs into the growth medium which could be harvested for pharmaceutical applications. In this investigation we characterized EVs and nanovesicles from distinct sources. Our findings regarding secondary metabolites indicate that these might not be packaged into EVs in an active manner but enriched in the membrane when lipophilic enough, since apparently lipophilic compounds were associated with nanovesicles while more hydrophilic structures were not consistently found. In addition, protein identification revealed a possible explanation for the mechanism of EV cell wall passage in plants, since cell wall hydrolases like 1,3-β-glucosidases, pectinesterases, polygalacturonases, β-galactosidases and β-xylosidase/α-L-arabinofuranosidase 2-like are present in plant EVs and nanovesicles which might facilitate cell wall transition. Further on, the identified proteins indicate that plant cells secrete EVs using similar mechanisms as animal cells to release exosomes and microvesicles

The protective effect of tumor necrosis factor-alpha inhibitors in COVID-19 in patients with inflammatory rheumatic diseases compared to the general population: a comparison of two German registries

Objectives: To investigate, whether inflammatory rheumatic diseases (IRD) inpatients are at higher risk to develop a severe course of SARS-CoV-2 infections compared to the general population, data from the German COVID-19 registry for IRD patients and data from the Lean European Survey on SARS-CoV-2 (LEOSS) infected patients covering inpatients from the general population with SARS-CoV-2 infections were compared. Methods: 4310 (LEOSS registry) and 1139 cases (IRD registry) were collected in general. Data were matched for age and gender. From both registries, 732 matched inpatients (LEOSS registry: n = 366 and IRD registry: n = 366) were included for analyses in total. Results: Regarding the COVID-19 associated lethality, no significant difference between both registries was observed. Age > 65°years, chronic obstructive pulmonary disease, diabetes mellitus, rheumatoid arthritis, spondyloarthritis and the use of rituximab were associated with more severe courses of COVID-19. Female gender and the use of tumor necrosis factor-alpha inhibitors (TNF-I) were associated with a better outcome of COVID-19. Conclusion: Inflammatory rheumatic diseases (IRD) patients have the same risk factors for severe COVID-19 regarding comorbidities compared to the general population without any immune-mediated disease or immunomodulation. The use of rituximab was associated with an increased risk for severe COVID-19. On the other hand, the use of TNF-I was associated with less severe COVID-19 compared to the general population, which might indicate a protective effect of TNF-I against severe COVID-19 disease

Development and Validation of a Risk Score for Chronic Kidney Disease in HIV Infection Using Prospective Cohort Data from the D:A:D Study

Ristola M. on työryhmien DAD Study Grp ; Royal Free Hosp Clin Cohort ; INSIGHT Study Grp ; SMART Study Grp ; ESPRIT Study Grp jäsen.Background Chronic kidney disease (CKD) is a major health issue for HIV-positive individuals, associated with increased morbidity and mortality. Development and implementation of a risk score model for CKD would allow comparison of the risks and benefits of adding potentially nephrotoxic antiretrovirals to a treatment regimen and would identify those at greatest risk of CKD. The aims of this study were to develop a simple, externally validated, and widely applicable long-term risk score model for CKD in HIV-positive individuals that can guide decision making in clinical practice. Methods and Findings A total of 17,954 HIV-positive individuals from the Data Collection on Adverse Events of Anti-HIV Drugs (D:A:D) study with >= 3 estimated glomerular filtration rate (eGFR) values after 1 January 2004 were included. Baseline was defined as the first eGFR > 60 ml/min/1.73 m2 after 1 January 2004; individuals with exposure to tenofovir, atazanavir, atazanavir/ritonavir, lopinavir/ritonavir, other boosted protease inhibitors before baseline were excluded. CKD was defined as confirmed (>3 mo apart) eGFR In the D:A:D study, 641 individuals developed CKD during 103,185 person-years of follow-up (PYFU; incidence 6.2/1,000 PYFU, 95% CI 5.7-6.7; median follow-up 6.1 y, range 0.3-9.1 y). Older age, intravenous drug use, hepatitis C coinfection, lower baseline eGFR, female gender, lower CD4 count nadir, hypertension, diabetes, and cardiovascular disease (CVD) predicted CKD. The adjusted incidence rate ratios of these nine categorical variables were scaled and summed to create the risk score. The median risk score at baseline was -2 (interquartile range -4 to 2). There was a 1: 393 chance of developing CKD in the next 5 y in the low risk group (risk score = 5, 505 events), respectively. Number needed to harm (NNTH) at 5 y when starting unboosted atazanavir or lopinavir/ritonavir among those with a low risk score was 1,702 (95% CI 1,166-3,367); NNTH was 202 (95% CI 159-278) and 21 (95% CI 19-23), respectively, for those with a medium and high risk score. NNTH was 739 (95% CI 506-1462), 88 (95% CI 69-121), and 9 (95% CI 8-10) for those with a low, medium, and high risk score, respectively, starting tenofovir, atazanavir/ritonavir, or another boosted protease inhibitor. The Royal Free Hospital Clinic Cohort included 2,548 individuals, of whom 94 individuals developed CKD (3.7%) during 18,376 PYFU (median follow-up 7.4 y, range 0.3-12.7 y). Of 2,013 individuals included from the SMART/ESPRIT control arms, 32 individuals developed CKD (1.6%) during 8,452 PYFU (median follow-up 4.1 y, range 0.6-8.1 y). External validation showed that the risk score predicted well in these cohorts. Limitations of this study included limited data on race and no information on proteinuria. Conclusions Both traditional and HIV-related risk factors were predictive of CKD. These factors were used to develop a risk score for CKD in HIV infection, externally validated, that has direct clinical relevance for patients and clinicians to weigh the benefits of certain antiretrovirals against the risk of CKD and to identify those at greatest risk of CKD.Peer reviewe

SEC analysis of concentrations of species.

<p>Concentrations of the five species in <i>scheme 1</i> determined from SEC measurements as described in the text using different initial concentrations (1∶1:1 of Atox1:Cu:WD4) as indicated. Also, the % of the total copper found in hetero-complex is reported. The equilibrium concentrations established are used to derive K₁ and K₂ and from this the copper exchange factor K₁*K₂ is calculated. Two experiments with 150 µM starting concentrations are reported. For 75 µM, also the opposite reaction, mixing Cu-WD4 with apo-Atox1 was performed.</p

Thermodynamic parameters for steps 1 and 2, and overall reaction.

<p>The parameters are based on <i>scheme 1</i> and equilibrium constants from SEC (<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0036102#pone-0036102-t001" target="_blank">Table 1</a>) and ITC data, as described in the text.</p

Illustration of Scheme 1.

<p>Upon mixing Cu-loaded Atox1 (purple) and apo-WD (green), the proteins interact and form a hetero-complex, Atox1-Cu-WD4 (shown with Cu coordinating one Cys in Atox1 and both Cys in WD4; however, there are other possible Cu coordinations in the hetero-complex <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0036102#pone.0036102-RodriguezGranillo1" target="_blank">[17]</a>), and also products, apo-Atox1 and Cu-WD4, according to the equilibrium constants K₁ and K₂.</p

Computer simulations mimic ITC titration.

<p><b>A.</b> Concentrations of the five species as a function of progress of the titration experiment shown in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0036102#pone-0036102-g004" target="_blank">Figure 4A</a> using the K values that were derived from the SEC data. <b>B.</b> ITC experimental data (from <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0036102#pone-0036102-g004" target="_blank">Figure 4A</a>) together with the best fit to the data (see Supplement for details).</p

SEC probed at 2 wavelengths is used to separate equilibrium species in Scheme 1.

<p><b>A.</b> SEC analysis of a mixture of the two apo proteins (300 µM each) at 280 and 254 nm. <b>B.</b> SEC analysis of holo Atox1 (300 µM) at 280 and 254 nm. <b>C.</b> SEC analysis at 280 nm of a mixture of 300 µM Cu-Atox1 and 300 µM apo-WD4. For comparison, the 280 nm elution trace for the mixture of the two apo-proteins is shown. <b>D.</b> SEC analysis at 280 nm and 254 nm of a mixture of 300 µM Cu-Atox1 and 300 µM apo-WD4. <b>E.</b> De-convolution of the underlying peaks in the elution trace of the Cu-Atox1+apo-WD4 mixture.</p