Differences in mineral and osmotic balances enhance zinc translocation in an aquaporin overexpressing poplar

: Zinc (Zn) is an essential micronutrient for plants, but it is toxic beyond a certain threshold. Populus alba (L.) 'Villafranca' clone is known for its good tolerance to high Zn concentration compared to other poplar species. A line of this species overexpressing the tonoplast intrinsic aquaporin AQUA1 gene has showed an improved tolerance to Zn excess in comparison to the wild-type (wt) line. The aims of this work were to: 1) verify if AQUA1 plants can uptake Zn more efficiently after a longer period of exposure; 2) evaluate if a higher Zn uptake in transgenic lines can have negative effects; 3) assess Zn competing elements (iron and manganese), soluble sugars, osmolytes, and potassium to investigate differences in water and osmotic homeostasis between lines. Under Zn excess, AQUA1 plants showed a twofold Zn translocation factor and a higher xylem sap Zn concentration than the wt plants. Transgenic plants preferentially allocated Zn in aerial biomass and this different behaviour matched with modified manganese and iron balances suggesting that the increased Zn uptake might be related to a decrease in iron transport in the transgenic line. Moreover, a higher instantaneous water use efficiency in control conditions and an increase in bark soluble sugars under Zn excess could allow a higher resistance of AQUA1 plants to the water and osmotic perturbations caused by Zn. Indeed, the Zn excess increased the xylem osmolyte content only in wt plants. Further investigations are required to understand the role of AQUA1 in osmotic regulation

Removal of micro-pollutants from urban wastewater by constructed wetlands with Phragmites australis and Salix matsudana

This study assessed the ability to remove micro-pollutants from wastewater using herbaceous species (Phragmites australis L.) and trees (Salix matsudana Koidz.) in constructed wetland (CW) systems. The targets of the study were as follows: (i) pharmaceuticals like diclofenac, ketoprofen, and atenolol; (ii) 4-n-NP (4-n-nonylphenol) and the ethoxylated derivatives monoethoxylated nonylphenol (NP1EO) and diethoxylated nonylphenol (NP2EO); (iii) triclosan, a bactericide used in personal care products. The 12 CW systems, filled with clay and gravel, were irrigated with wastewater from municipal area of Pagnana (Tuscany, Italy) and influent and effluent water samples analyzed periodically by gas chromatography-mass spectrometry (GC-MS/MS). The removal efficiency of CWs planted with willow and common red ranged from 8.4 up to 100%, with the higher removal efficiency for triclosan. On the contrary, the removal efficiency of NPs and NPEOs appears lower than pharmaceuticals. Data demonstrated that P. australis efficiently removed NP, diclofenac, and atenolol, while S. matsudana preferentially removed NP1EO, NP2EO, ketoprofene, and triclosan. A specific selection of plants used in CWs could be exploited for the removal of specific xenobiotics from wastewater

Cocoa Bar Antioxidant Profile Enrichment with Underutilized Apples Varieties

The impact of dried apples (Malus x domestica Borkh.) addition on improving the antioxidant characteristics of dark chocolate was evaluated. The antioxidant activity was measured through DPPH scavenging activity and showed an increase in the cocoa bar with 'Nesta' dry apple (17.3% vs. 46.8%) in comparison to cocoa mass. The 15 polyphenols analyzed by UHPLC-ESI-MS/MS indicated great variability among the apple varieties. Quercetin was detected in the highest concentrations (ranged from 753.3 to 1915.5 mu g g(-1)), while the lowest were for kaempferol 7-O-glucoside, measured only in 'Mora' and 'Nesta' cocoa bars (from 0.034 to 0.069 mu g g(-1), respectively). P-coumaric acid, trans-ferulic acid, and chlorogenic acid contribute largely to the antioxidant activity in cocoa bars. Principal component analysis shows that a cocoa bar with the addition of 'Nesta' dry apple differ from others due to its higher content of polyphenols (1614 +/- 61.8 mg gallic acid equivalents per 100 g). In conclusion, data confirm that cocoa bars with dry apples might be considered as a polyphenol-enriched food

Olive Fruit Ripening Degree and Water Content Relationships with Phenolic Acids and Alcohols, Secoiridoids, Flavonoids and Pigments in Fruit and Oil

: Olive drupe traits (i.e., ripening index and pericarp water content) and minor components (i.e., phenols and pigments in both fruit and oil) are important for human health and are affected by agronomic background. The aim of this study was to investigate the relationship between fruit traits, phenols, and pigments in samples derived from different soil and water management practices. Chromatographic (UHPLC-MS/MS) and spectroscopic (1HNMR and near UV-Vis spectroscopy) techniques were employed for the characterization of olive fruits and oils. The use of various techniques allowed the identification of interesting trace compounds. We observed that most of the fruit phenols (a total of 29 compounds) were correlated with the degree of ripening: most of the phenolic acids (and their derivatives), phenolic alcohols, and secoiridoids were negatively correlated, whereas the majority of the studied flavonoids were positively correlated. The relationship between the ripening index and fruit phenolic compounds appears to be dependent on the metabolic pathway that controls the synthesis of each individual compound. Conversely, the secoiridoids and pigments in olive oil showed a negative correlation with pulp moisture, probably because of the influence of the water content on the extractability and transfer in the oil phase of these minor components

Porcine circovirus 3 is highly prevalent in serum and tissues and may persistently infect wild boar (Sus scrofa scrofa)

Porcine circovirus 3 (PCV‐3) prevalence has been minimally investigated in wild boar; dynamics of infection and viral tissue distribution are currently unknown. In this study, serum samples from 518 wild boar (from years 2004 to 2018) were used to study frequency of infection. Also, serum samples from 19 boar captured and recaptured at least two times for a period of time from 1 month to 1 year were collected to determine PCV‐3 infection dynamics. Finally, to elucidate PCV‐3 DNA organic distribution, sera, different tissues and faeces were obtained from 35 additional wild boar. PCV‐3 DNA was extracted and amplified with a conventional PCR. For the PCV‐3 PCR‐positive sera from the longitudinally sampled and different tissue types, a quantitative PCR was performed. Genome sequence was obtained from a number of PCV‐3 PCR‐positive samples from different years, different time‐points of infection and tissues. Obtained results confirmed the susceptibility of wild boar to the virus, showing high frequency of PCV‐3 detection (221 out of 518, 42.66%) and demonstrating circulation at least since 2004. Compiled data indicate the possibility of long‐term infections, since 5 out of 10 PCV‐3 PCR‐positive boars longitudinally sampled showed positivity in samplings separated for more than 5 months. All tested tissue types' harboured PCV‐3 genome, with the highest percentage of PCR positivity in submandibular lymph node, tonsil, lung, liver, spleen and kidney. The amount of DNA in all tested PCV‐3 PCR‐positive samples was moderate to low. All partial and complete PCV‐3 sequences obtained from wild boar displayed high nucleotide identity, higher than 98%. In conclusion, this study further confirms that wild boar is susceptible to PCV‐3 infection, showing high frequency of detection in this animal species. Furthermore, PCV‐3 can be found in different tissues of wild boar and is apparently able to cause persistent infection.Instituto Nacional de Investigación y Tecnologia Agraria y Alimentaria. Grant Number: E‐RTA2017‐00007‐00‐0

Prospective individual patient data meta-analysis of two randomized trials on convalescent plasma for COVID-19 outpatients

Data on convalescent plasma (CP) treatment in COVID-19 outpatients are scarce. We aimed to assess whether CP administered during the first week of symptoms reduced the disease progression or risk of hospitalization of outpatients. Two multicenter, double-blind randomized trials (NCT04621123, NCT04589949) were merged with data pooling starting when = 50 years and symptomatic for <= 7days were included. The intervention consisted of 200-300mL of CP with a predefined minimum level of antibodies. Primary endpoints were a 5-point disease severity scale and a composite of hospitalization or death by 28 days. Amongst the 797 patients included, 390 received CP and 392 placebo; they had a median age of 58 years, 1 comorbidity, 5 days symptoms and 93% had negative IgG antibody-test. Seventy-four patients were hospitalized, 6 required mechanical ventilation and 3 died. The odds ratio (OR) of CP for improved disease severity scale was 0.936 (credible interval (CI) 0.667-1.311); OR for hospitalization or death was 0.919 (CI 0.592-1.416). CP effect on hospital admission or death was largest in patients with <= 5 days of symptoms (OR 0.658, 95%CI 0.394-1.085). CP did not decrease the time to full symptom resolution

High-titre methylene blue-treated convalescent plasma as an early treatment for outpatients with COVID-19: a randomised, placebo-controlled trial.

BACKGROUND: Convalescent plasma has been proposed as an early treatment to interrupt the progression of early COVID-19 to severe disease, but there is little definitive evidence. We aimed to assess whether early treatment with convalescent plasma reduces the risk of hospitalisation and reduces SARS-CoV-2 viral load among outpatients with COVID-19. METHODS: We did a multicentre, double-blind, randomised, placebo-controlled trial in four health-care centres in Catalonia, Spain. Adult outpatients aged 50 years or older with the onset of mild COVID-19 symptoms 7 days or less before randomisation were eligible for enrolment. Participants were randomly assigned (1:1) to receive one intravenous infusion of either 250-300 mL of ABO-compatible high anti-SARS-CoV-2 IgG titres (EUROIMMUN ratio ≥6) methylene blue-treated convalescent plasma (experimental group) or 250 mL of sterile 0·9% saline solution (control). Randomisation was done with the use of a central web-based system with concealment of the trial group assignment and no stratification. To preserve masking, we used opaque tubular bags that covered the investigational product and the infusion catheter. The coprimary endpoints were the incidence of hospitalisation within 28 days from baseline and the mean change in viral load (in log10 copies per mL) in nasopharyngeal swabs from baseline to day 7. The trial was stopped early following a data safety monitoring board recommendation because more than 85% of the target population had received a COVID-19 vaccine. Primary efficacy analyses were done in the intention-to-treat population, safety was assessed in all patients who received the investigational product. This study is registered with ClinicalTrials.gov, NCT04621123. FINDINGS: Between Nov 10, 2020, and July 28, 2021, we assessed 909 patients with confirmed COVID-19 for inclusion in the trial, 376 of whom were eligible and were randomly assigned to treatment (convalescent plasma n=188 [serum antibody-negative n=160]; placebo n=188 [serum antibody-negative n=166]). Median age was 56 years (IQR 52-62) and the mean symptom duration was 4·4 days (SD 1·4) before random assignment. In the intention-to-treat population, hospitalisation within 28 days from baseline occurred in 22 (12%) participants who received convalescent plasma versus 21 (11%) who received placebo (relative risk 1·05 [95% CI 0·78 to 1·41]). The mean change in viral load from baseline to day 7 was -2·41 log10 copies per mL (SD 1·32) with convalescent plasma and -2·32 log10 copies per mL (1·43) with placebo (crude difference -0·10 log10 copies per mL [95% CI -0·35 to 0·15]). One participant with mild COVID-19 developed a thromboembolic event 7 days after convalescent plasma infusion, which was reported as a serious adverse event possibly related to COVID-19 or to the experimental intervention. INTERPRETATION: Methylene blue-treated convalescent plasma did not prevent progression from mild to severe illness and did not reduce viral load in outpatients with COVID-19. Therefore, formal recommendations to support the use of convalescent plasma in outpatients with COVID-19 cannot be concluded. FUNDING: Grifols, Crowdfunding campaign YoMeCorono