Mammary cistern size during the dry period in healthy dairy cows: A preliminary study for an ultrasonographic evaluation

We evaluated the udder cistern (UC) size during the dry period using ultrasound. Forty healthy quarters were evaluated in both the longitudinal and cross-section of the UC. Quarters were evaluated at the drying-off (T0) and 24 h later (T1), then regularly until the end of the dry period (T7–T58), during the colostrum production phase (TCPP) and at 7 days in milking (T7PP). The Spearman test was applied to find the correlation between the ultrasonographic UC size (UUCS) assessment and time. The Friedman test and Dunn’s test for multiple comparisons as a post-hoc test were performed to compare the forequarter and hindquarter cross-sections (FQCSs and HQCSs, respectively) and the forequarter and hindquarter longitudinal sections (FQLSs and HQLSs, respectively) at T0 vs. T58 vs. TCPP vs. T7PP. A total of 440 images were evaluated. A negative linear correlation between time and FQCS and FQLS (r = −0.95; p < 0.0004) and between time and HQCS and HQLS (r = −0.90; p < 0.002) was found. The UUCS decreased throughout the dry period, starting to increase at the beginning of the next lactation. Measuring the UUCS provides useful information for monitoring the dry period

Essential oils as post-harvest crop protectants against the fruit fly drosophila suzukii: Bioactivity and organoleptic profile

The essential oils extracted from mandarin (Citrus reticulata Blanco) fruits, and from tea tree (Maleleuca alternifolia (Maiden and Betche) Cheel) leaves have been chemically analyzed and tested for their bioactivity against D. suzukii. Besides, to estimate consumers’ acceptability of the essential oil (EO) treatments, we evaluated their impact on the organoleptic characteristics of the EO-treated fruits. The main chemical constituents of the two EOs were 1,8-cineole and 4-terpineol for M. alternifolia (22.4% and 17.6% of the total components, respectively), and limonene (83.6% of the total components) for C. reticulata. The behavioral tests indicate that the two EOs are able to deter D. suzukii oviposition and that D. suzukii shows positive chemotaxis to low concentrations of the EOs and negative chemotaxis when the EO concentration increases. While no negative effects on the organoleptic profiles were detected for fruits treated with C. reticulata EO, the olfactory profile of fruits treated with M. alternifolia EO was so negative that they were defined as “not suitable for consumption” by panellists. Overall, our findings indicate that the use of EOs for the post-harvest protection of small fruits is feasible, provided that the essential oils are selected not only for their bioactivity against the insect pest but also for their affinity with the consumers’ sensorial system

SINEUPs: A novel toolbox for RNA therapeutics

RNA molecules have emerged as a new class of promising therapeutics to expand the range of druggable targets in the genome. In addition to 'canonical' protein-coding mRNAs, the emerging richness of sense and antisense long non-coding RNAs (lncRNAs) provides a new reservoir of molecular tools for RNA-based drugs. LncRNAs are composed of modular structural domains with specific activities involving the recruitment of protein cofactors or directly interacting with nucleic acids. A single therapeutic RNA transcript can then be assembled combining domains with defined secondary structures and functions, and antisense sequences specific for the RNA/DNA target of interest. As the first representative molecules of this new pharmacology, we have identified SINEUPs, a new functional class of natural antisense lncRNAs that increase the translation of partially overlapping mRNAs. Their activity is based on the combination of two domains: An embedded mouse inverted SINEB2 element that enhances mRNA translation (effector domain) and an overlapping antisense region that provides specificity for the target sense transcript (binding domain). By genetic engineering, synthetic SINEUPs can potentially target any mRNA of interest increasing translation and therefore the endogenous level of the encoded protein. In this review, we describe the state-of-the-art knowledge of SINEUPs and discuss recent publications showing their potential application in diseases where a physiological increase of endogenous protein expression can be therapeutic

SINEUP non-coding RNA activity depends on specific N6-methyladenosine nucleotides

SINEUPs are natural and synthetic antisense long non-coding RNAs (lncRNAs) selectively enhancing target mRNAs translation by increasing their association with polysomes. This activity requires two RNA domains: an embedded inverted SINEB2 element acting as effector domain, and an antisense region, the binding domain, conferring target selectivity. SINEUP technology presents several advantages to treat genetic (haploinsufficiencies) and complex diseases restoring the physiological activity of diseased genes and of compensatory pathways. To streamline these applications to the clinic, a better understanding of the mechanism of action is needed. Here we show that natural mouse SINEUP AS Uchl1 and synthetic human miniSINEUP-DJ-1 are N6-methyladenosine (m6A) modified by METTL3 enzyme. Then, we map m6A-modified sites along SINEUP sequence with Nanopore direct RNA sequencing and a reverse transcription assay. We report that m6A removal from SINEUP RNA causes the depletion of endogenous target mRNA from actively translating polysomes, without altering SINEUP enrichment in ribosomal subunit-associated fractions. These results prove that SINEUP activity requires an m6A-dependent step to enhance translation of target mRNAs, providing a new mechanism for m6A translation regulation and strengthening our knowledge of SINEUP-specific mode of action. Altogether these new findings pave the way to a more effective therapeutic application of this well-defined class of lncRNAs

Degradation of exogenous caffeine by Populus alba and its effects on endogenous caffeine metabolism

This is the first study reporting the presence of endogenous caffeine, theobromine, and theophylline in all organs of poplar plants. Liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS) was used in order to evaluate the uptake, translocation, and metabolism of caffeine-(trimethyl-(13)C) in Populus alba L. Villafranca clone grown in hydroponic conditions. We investigated the remediation of caffeine since it is one of the most widely consumed drugs and it is frequently detected in wastewater treatment plant effluents, surface water, and groundwater worldwide. Our results demonstrated that poplar can absorb and degrade exogenous caffeine without negative effects on plant health. Data showed that concentrations of all endogenous compounds varied depending on caffeine-(trimethyl-(13)C) treatments. In particular, in control conditions, endogenous caffeine, theobromine, and theophylline were mainly distributed in roots. On the other hand, once caffeine-(trimethyl-(13)C) was provided, this compound and its dimethy-(13)C metabolites are mainly localized at leaf level. In conclusion, our results support the possible use of Villafranca clone in association with other water treatment systems in order to complete the process of caffeine remediation

ESI and APCI LC-MS/MS in Model Investigations on the Absorption and Transformation of Organic Xenobiotics by Poplar Plants (Populus alba L.)

Populus alba Villafranca clone was chosen for a proof of concept study to determine the potential uptake and accumulation of xenobiotics by trees. Caffeine, erythromycin, and sodium dodecyl sulphate (SDS) were studied as representative elements of, respectively, widely present organic pollutants, antibiotics, and surfactants. Plants were grown hydroponically and irrigated with a recirculating Hoagland's nutrient solution (control) and Hoagland's nutrient solution fortified with the desired pollutant. Different plantsâ\u80\u99 tolerance was observed in relation to xenobiotics applied. Under caffeine and erythromycin treatments, plants showed good health all over the period of experiments, and no differences in poplar growth were observed. On the contrary diffused necrosis at leavesâ\u80\u99 level was observed when SDS was applied to nutrient solution. Xenobiotics uptake was investigated in roots, stem, and leaves, and quantification was performed using liquid chromatography, coupled with tandem mass spectrometry using selected reaction ion monitoring on a triple quadrupole mass spectrometer, with electrospray or atmospheric pressure chemical ionization, depending on the analyte. All the pollutants were detected in all organs analyzed. Poplar plants also showed an active role in the elimination of the pollutants as some of them were metabolized by the tree

Surfactant and heavy metal interaction in poplar: A focus on SDS and Zn uptake

Surfactants are widely used detergent ingredients and, thanks to their chemical properties, they are applied for remediation of sites polluted by heavy metals and organic contaminants, both in soil flushing and in phytoremediation. However, their direct effects on tree physiology especially in consociation with heavy metal pollution, as well as their possible absorption by plants, have not been appropriately investigated. In order to evaluate plant uptake/translocation of the surfactant sodium dodecyl sulfate (SDS) and the heavy metal zinc (Zn) in Populus alba L. Villafranca clone, SDS was applied alone (0.5 mM) or in combination with Zn (1 mM). Physiological effects on plant growth and photosynthetic performance were investigated. An increasing trend of Zn translocation towards basal leaves as a consequence of SDS co-treatment (1 mM Zn + 0.5 mM SDS; P = 0.03) was observed, proving the ability of SDS to improve heavy metals translocation. However, SDS exposure (both in 0.5 mM SDS and 1 mM Zn + 0.5 mM SDS treated plants) resulted in the appearance of foliar necrosis that expanded with an acropetal trend and finally led to leaf abscission. This phenotype may be caused by the emergence of an additional stress during the experimental trial, which could be related to the dissociation of sodium (Na) ions from the dodecyl sulfate molecules in the hydroponic system. In fact, while liquid chromatography-tandem mass spectrometry measurements revealed that dodecyl sulfate is mainly retained at root levels, Na is translocated to the aerial parts of the plant

Poplar and diclofenac pollution: A focus on physiology, oxidative stress and uptake in plant organs.

Poplar plants may have an important role in the removal of pharmaceuticals from contaminated waters. In this context, plant uptake of the non-steroidal anti-inflammatory drug diclofenac, as well as physiological response in terms of growth traits and stress enzymes activity was assessed in Populus alba Villafranca clone, in order to establish the effectiveness of this species against pharmaceutical active compounds pollution.This evaluation was conducted in mesocosms with 1 mg L-1 of this pharmaceutical for a maximum period of 28 days. Root appears to be the organ with clear uptake of diclofenac (14.76 +/- 2.42 ng g(-1) fresh weight after 1 day of treatment), and presence of products derived from its metabolism. Indeed, 4-OH-diclofenac metabolite was detected in root tissues, indicating diclofenac uptake and metabolism inside the plants, already after 1 day of treatment.Regarding enzyme activities, glutathione-S-transferases increased in roots after long-term exposure to diclofenac, while an increase in activity of ascorbate peroxidase and glutathione reductase was detected in short and medium-term exposure, as a result of abiotic stress caused by diclofenac. Results suggest the ability of poplar to actively participate in the removal of diclofenac from water when used for phytoremediation purpose. (C) 2018 Elsevier B.V. All rights reserved