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Abstract Humic susbtances (HS) are increasingly applied as biostimulants in agriculture, though their mechanism of action is not yet completely understood due to their complex and heterogeneous composition. Here, we isolated thirteen different humic and fulvic acids from different sources, such as soils, composts, peat, leonardite and lignite, and characterized the distribution of their carbon components by 13C-CPMAS–NMR spectroscopy. Callus of Pinus Laricio, as a model plant species, was treated with different humic extracts and its growth and content of carbohydrates, phenols, and enzyme related to the nitrogen metabolism (invertase, glutamine synthetase, glutamate synthase, phosphoenoly pyruvate carboxylase, malate dehydrogenase) and stress resistance (catalase) were monitored. While a multivariate statistical analysis of NMR results well-separated the HS characteristics based on their origin, humic materials generally increased callus growth, as expected, with largest effects being exerted by the mostly polar humic acids from composts. However, the rest of measured parameters were not linearly related to hydrophobicity and aromaticity of humic isolates as well as their origin, but their conformational dynamics had to be advocated to explain their effects on callus cellular components. This work confirms that HS of various origin can act as sustainable biostimulants of plant growth, though the comprehension of their effects on plants biochemical activities requires further research and additional understanding of their molecular composition and conformational behaviour. Graphical Abstract

Abstract Background We applied Fourier-Transformed Infrared Spectroscopy (FTIR) techniques in combination with selective humus extractions to investigate in situ the chemical features of organic matter in three soil profiles recently developed from Dutch sand dunes, under well-documented vegetation sequence and containing a relatively simple mineral-organic matter system. Water-soluble and alkaline-soluble extracts were also studied to match the changes of humus in the soil residues. Results Diffuse Reflectance Infrared Fourier Transform (DRIFT) and transmission spectra differed in resolution and sensitivity. DRIFT was superior in revealing structural information on the organic matter present in the soil samples whereas transmission spectra showed higher resolution for the bands of inorganic material at lower spectral frequencies. Differences between H and B horizons were due to the amount of hydrophilic organic acids, partly unsaturated, that were extracted by alkali. Extractable carboxylic acids and other hydrophilic compounds such as peptides and carbohydrates were larger in the less developed soils under pine and crowberry than in the more advanced profiles under beech. Humin residues from both H and B horizons retained unextractable aliphatic components and carboxylic groups involved in strong complexes with minerals, thereby confirming other findings that showed the largely aliphatic character of the unextractable humic fraction. Accumulation of poorly-soluble organic materials in these soils occur by protection from biodegradation due both to complexation with soil minerals and to a process by which apolar humic constituents form a separate hydrophobic phase where no biological activity can take place. Transport of poorly soluble compounds from H to B horizons, noticed especially in the more developed profiles, may have occurred through humic aggregates containing hydrophobic phases. Conclusions This work shows that the DRIFT technique in combination with soil chemical treatments can be suitably employed to investigate the changes of organic matter in whole soil samples and represents a valid tool to investigate the role of organic matter in soil profile development.

A vineyard of the cv. Trebbiano d’Abruzzo located in Abruzzo, Italy, was monitored for more than ten years to distinguish healthy vines not only from vines with the visual leaf symptoms of esca, but also from those vines that were esca-infected but did not produce any visual symptoms for one or more growing seasons. In the period 2004–2006, leaves and berries were collected at four phenological growth stages from three groups of vines: healthy vines, infected vines showing esca symptoms, and infected vines that did not show symptoms. The macro and micro-elements of the leaves and berries, and the quality parameters of the must were determined. Esca did not seem to affect nutrient uptake in the vines. Nevertheless there were some differences in the nutrient levels of the leaves between healthy and diseased vines consistent with the degradation of the leaf blade caused by esca. Berries from symptomatic vines were less ripe at the time of harvesting and therefore had higher levels of mineral elements. These berries also had higher levels of nitrogen, which are thought to be associated with the defence response of diseased vines to esca, as are higher levels of iron in the leaves of diseased vines. The study confirmed earlier findings that fruit composition did not differ greatly between healthy and diseased-but-asymptomatic vines. In the three-year study period there were differences in the incidence of leaf symptoms and differences in nutrient levels attributable to fertiliser applications and rainfall. These differences suggested that the amount of mineral nutrients affected the onset of esca symptoms: a higher availability of nutrients in a growing season increased the proportion of diseased vines with symptoms and lowered the proportion of diseased vines without symptoms, whereas in a growing season with the lower levels of water and potassium, the yield was reduced, but this was accompanied by an increase in the proportion of diseased vines without symptoms. It is suggested that a higher availability of nutrients for diseased vines lowers the resistance of these vines and, by improving the nutrition not only of the vines themselves but also of the esca fungi, increase fungal virulence, as a result of which there is a greater incidence of diseased vines showing leaf symptoms.

Oxidative stress plays a pivotal role in driving immunosenescence by disrupting cellular homeostasis and impairing immune function. Humic substances exhibit scavenging activity against reactive oxygen species (ROS), inhibit ROS generation via metal chelation, and modulate endogenous antioxidant enzyme activity. Additionally, humic substances display anti-inflammatory effects, further supporting cellular redox balance. Given their antioxidant activity, humic substances hold promise as natural compounds for mitigating oxidative stress-associated immunosenescence. Here we describe the evaluation of antioxidant capacities of humic products by ABTS spectrophotometric assay., (© 2025. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.)

Coffee husks have been valorised by isolating humic materials before (HLS-Raw) or after (HS-Comp) composting them. Such substrates were reacted with chitosan at different ratios to synthesize novel nanoparticles (NP) with radical scavenging properties. Size and antioxidant activity of nanomaterials increased at higher HS/chitosan ratio, while zeta potential decreased. Our results furthermore highlighted that he molecular features of raw humic materials, as studied by solid-state NMR spectroscopy, affected the studied NP properties. Indeed, at equal reactant ratio, NP from HS-Comp had lower zeta potential due to their higher amount of carboxyl moieties contained, which counterbalanced a greater number of chitosan positive charges. Moreover, these NP showed lower antioxidant activity despite the greater phenolic content of the original HS. We hypothesized that the higher hydrophobicity of HS-Comp led to the partial partitioning of their phenolic components into the inner domains of the derived NP, lowering their antioxidant capacity. Additionally, in a short-term stability assay, NP coalescence was observed, especially at higher humic/chitosan ratio, with nanomaterial precipitation and consequent decrease in antioxidant capacity. Overall, we demonstrated that humic materials from raw or composted coffee husks could successfully crosslink chitosan chains to synthesize sustainable NP with tuneable physical-chemical and antioxidant properties., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

The development of new natural antibiotics is considered as the heart of several investigations in the nutraceutical field. In this work, leaves of Quercus ilex L. treated by tropospheric ozone (O 3 ) and nitrogen (N) deposition, exhibited a clear antimicrobial efficacy against five multi-drug resistant (MDR) bacterial strains (two gram-positive and three gram-negative). Under controlled conditions, it was studied how simulated N deposition influences the response to O 3 and the antibacterial and antioxidant activity, and antioxidant performance. The extraction was performed by ultra-pure acetone using two different steps. A higher antioxidant activity was measured in the presence of interaction between O 3 and N treatments on Quercus leaves. At the same time, all organic extracts tested have shown bacteriostatic activity against all the tested strains with a MIC comprised between 9 and 4 micrograms/mL, and a higher antioxidant efficacy shown by spectrophotometric assay. Stronger antimicrobial activity was found in the samples treated with O 3 , whereas N-treated plants exhibited an intermediate antibacterial performance. This performance is related to the stimulation of the non-enzymatic antioxidant system induced by the oxidative stress, which results in an increase in the production of antimicrobial bioactive compounds.

Biostimulants (Bio-effectors, BEs) comprise plant growth-promoting microorganisms and active natural substances that promote plant nutrient-acquisition, stress resilience, growth, crop quality and yield. Unfortunately, the effectiveness of BEs, particularly under field conditions, appears highly variable and poorly quantified. Using random model meta-analyses tools, we summarize the effects of 107 BE treatments on the performance of major crops, mainly conducted within the EU-funded project BIOFECTOR with a focus on phosphorus (P) nutrition, over five years. Our analyses comprised 94 controlled pot and 47 field experiments under different geoclimatic conditions, with variable stress levels across European countries and Israel. The results show an average growth/yield increase by 9.3% (n=945), with substantial differences between crops (tomato > maize > wheat) and growth conditions (controlled nursery + field (Seed germination and nursery under controlled conditions and young plants transplanted to the field) > controlled > field). Average crop growth responses were independent of BE type, P fertilizer type, soil pH and plant-available soil P (water-P, Olsen-P or Calcium acetate lactate-P). BE effectiveness profited from manure and other organic fertilizers, increasing soil pH and presence of abiotic stresses (cold, drought/heat or salinity). Systematic meta-studies based on published literature commonly face the inherent problem of publication bias where the most suspected form is the selective publication of statistically significant results. In this meta-analysis, however, the results obtained from all experiments within the project are included. Therefore, it is free of publication bias. In contrast to reviews of published literature, our unique study design is based on a common standardized protocol which applies to all experiments conducted within the project to reduce sources of variability. Based on data of crop growth, yield and P acquisition, we conclude that application of BEs can save fertilizer resources in the future, but the efficiency of BE application depends on cropping systems and environments., Competing Interests: PN is employed since 01.10.2022 at Yara GmbH Co & KG., Germany. NM is employed since 01.06.2021 at Yara GmbH Co & KG., Germany. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest. The author(s) declared that they were an editorial board member of Frontiers, at the time of submission. This had no impact on the peer review process and the final decision., (Copyright © 2024 Nkebiwe, Stevens Lekfeldt, Symanczik, Thonar, Mäder, Bar-Tal, Halpern, Biró, Bradáčová, Caniullan, Choudhary, Cozzolino, Di Stasio, Dobczinski, Geistlinger, Lüthi, Gómez-Muñoz, Kandeler, Kolberg, Kotroczó, Kulhanek, Mercl, Tamir, Moradtalab, Piccolo, Maggio, Nassal, Szalai, Juhos, Fora, Florea, Poşta, Lauer, Toth, Tlustoš, Mpanga, Weber, Weinmann, Yermiyahu, Magid, Müller, Neumann, Ludewig and de Neergaard.)

Aqueous solutions of humic acid (HA) derivatized by a catalyzed O-alkylation reaction with methyl, pentyl, and benzyl groups at 40, 60, and 80% of total HA acidity were used to wash off polycyclic aromatic hydrocarbons (PAHs) from two contaminated soils. The enhanced surfactant properties enabled the alkylated HA to remove phenanthrene, anthracene, fluoranthene, and pyrene from both soils more extensively than the original unmodified HA, the 60% benzylation generally showing the greatest soil washing efficiency. For both soils, all alkylated HA revealed greater PAH removals than Triton X-100 nonionic surfactant, while the benzylated and methylated HA nearly and fully matched pollutants release by the anionic SDS in the coarse- and fine-textured soils, respectively. A consecutive second washing with 60% benzylated HA removed additional PAHs, in respect to the first washing, from the coarser-textured soil, except for fluoranthene, while removal from the finer-textured soil incremented even more for all PAHs. These findings indicate that the enhanced hydrophobicity obtained by a simple and unexpensive chemical derivatization of a natural humic surfactant can be usefully exploited in the washing of polluted soils, without being toxic to the soil biota and by potentially promoting the subsequent bio-attenuation of organic pollutants., (© 2024. The Author(s).)

A hydrotropic solution of maleic acid (MA) was exploited to fractionate giant reed (AD) and eucalypt (EUC). The pre-treatment was successful for AD, while it was unsatisfactory for EUC, likely due to unoptimized reaction conditions. Interestingly, lignin nanoparticles (LNP) were produced via spontaneous aggregation following spent liquor dilution. LNP were studied by a plethora of analytical techniques, such as thermogravimetry, electron microscopy, and Nuclear Magnetic Resonance spectroscopy (NMR). Notwithstanding LNP from both AD and EUC showed similar thermal behaviour and morphology, a greater content of aliphatic hydroxyl, carboxyl, guaiacyl and p-hydroxyphenyl moieties was reported for AD-LNP, whereas EUC-LNP had a larger amount of syringyl groups and a higher S/G ratio. Also, the 1 H-DOSY NMR indicated the lower size of AD-LNP. Moreover, the LNP were found to negatively impact on the development of several human or plant pathogens, and their bioactivity was related to the occurrence of guaiacyl and p-hydroxyphenyl moieties and a lower the LNP size. We therefore found that MA delignification allows both to achieve high delignification efficiency and to obtain LNP with promising antibacterial effect. Such LNP may help counteracting the antibiotics resistance and sustain the quest for finding sustainable agrochemicals., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier B.V.)

Background: In the last few years, considerable attention has been focused on the plastic-degrading capability of insects and their gut microbiota in order to develop novel, effective, and green strategies for plastic waste management. Although many analyses based on 16S rRNA gene sequencing are available, an in-depth analysis of the insect gut microbiome to identify genes with plastic-degrading potential is still lacking., Results: In the present work, we aim to fill this gap using Black Soldier Fly (BSF) as insect model. BSF larvae have proven capability to efficiently bioconvert a wide variety of organic wastes but, surprisingly, have never been considered for plastic degradation. BSF larvae were reared on two widely used plastic polymers and shotgun metagenomics was exploited to evaluate if and how plastic-containing diets affect composition and functions of the gut microbial community. The high-definition picture of the BSF gut microbiome gave access for the first time to the genomes of culturable and unculturable microorganisms in the gut of insects reared on plastics and revealed that (i) plastics significantly shaped bacterial composition at species and strain level, and (ii) functions that trigger the degradation of the polymer chains, i.e., DyP-type peroxidases, multicopper oxidases, and alkane monooxygenases, were highly enriched in the metagenomes upon exposure to plastics, consistently with the evidences obtained by scanning electron microscopy and 1 H nuclear magnetic resonance analyses on plastics., Conclusions: In addition to highlighting that the astonishing plasticity of the microbiota composition of BSF larvae is associated with functional shifts in the insect microbiome, the present work sets the stage for exploiting BSF larvae as "bioincubators" to isolate microbial strains and enzymes for the development of innovative plastic biodegradation strategies. However, most importantly, the larvae constitute a source of enzymes to be evolved and valorized by pioneering synthetic biology approaches. Video Abstract., (© 2023. BioMed Central Ltd., part of Springer Nature.)

An O-alkylation reaction catalyzed by tetrabutylammonium hydroxide (TBAH) as a phase-transfer agent was applied to a humic acid (HA) to modify its hydrophobic properties. The carboxyl and hydroxyl functional groups of HA acted as nucleophiles in substitution reactions (Sn 2 ) with methyl iodide, pentyl bromide and benzyl bromide added in amounts equimolar to 20, 60 and 80% of HA total nucleophilic sites. The occurrence of O-alkylation was shown by DRIFT spectrometry, NMR spectroscopy, High Performance Size Exclusion Chromatography (HPSEC) and elemental analysis of reaction products. DRIFT spectra showed changes in C-H stretching and bending regions following the insertion of methyl and pentyl groups, while the incorporation of benzyl groups revealed the characteristics aromatic C-H stretching bands. Both liquid- and solid-state NMR spectra revealed characteristic signals for alkyl/aryl esters and ethers. HPSEC chromatograms of alkylated materials invariably displayed an increase in hydrodynamic volume in respect to the original HA, thereby suggesting that the enhanced hydrophobicity conveyed further associations among humic molecules. Analytical, HPSEC and spectroscopic results suggest that benzylation was the most effective reaction at all percentages of HA total nucleophilicity, followed, in the order, by pentylation and methylation, The benzylation reaction was used to improve reaction and work-up conditions and show that HA could be efficiently alkylated also with substantial reduction of TBAH amount, with no THF addition, increase of reaction time and of washing cycles to remove catalyst impurities. These findings indicate that the hydrophobicity of humic substances can be modulated through a mild O-alkylation reaction under a phase-transfer catalysis according to the extent of exposed HA nucleophilic sites. Such a structural modification of humic matter may have multiple chemical, environmental and biological applications., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

Background: An important goal of modern medicine is the development of products deriving from natural sources to improve environmental sustainability. In this study, humic substances (HS) and compost teas (CTs) extracted from artichoke (ART) and coffee grounds (COF) as recycled biomasses were employed on Ocimum basilicum plants to optimize the yield of specific metabolites with nutraceutical and antibacterial features by applying sustainable strategies., Methods: The molecular characteristics of compost derivates were elucidated by Nuclear Magnetic Resonance spectroscopy to investigate the structure-activity relationship between organic extracts and their bioactive potential. Additionally, combined untargeted and targeted metabolomics workflows were applied to plants treated with different concentrations of compost extracts., Results: The substances HS-ART and CT-COF improved both antioxidant activity (TEAC values between 39 and 55 μmol g -1 ) and the antimicrobial efficacy (MIC value between 3.7 and 1.3 μg mL -1 ) of basil metabolites. The metabolomic approach identified about 149 metabolites related to the applied treatments. Targeted metabolite quantification further highlighted the eliciting effect of HS-ART and CT-COF on the synthesis of aromatic amino acids and phenolic compounds for nutraceutical application., Conclusions: The combination of molecular characterization, biological assays, and an advanced metabolomic approach, provided innovative insight into the valorization of recycled biomass to increase the availability of natural compounds employed in the medical field.

Objective: The aetiologic diagnosis of non-traumatic acute myelopathies (AMs), and their differentiation from other mimicking conditions (i.e. 'mimics'), are clinically challenging, especially in the emergency setting. Here, we sought to identify: (i) red flags suggesting diagnoses alternative to AMs and (ii) clinical signs and magnetic resonance imaging (MRI) features differentiating non-compressive from compressive AMs., Materials and Methods: We retrospectively retrieved MRI scans of spinal cord dictated at emergency room from January 2016 to December 2020 in the suspicion of AMs. Patients with traumatic myelopathies and those with subacute/chronic myelopathies (i.e. MRI scans acquired >48 h from symptom onset) were excluded from analysis., Results: Our search retrieved 105 patients; after excluding 16 cases of traumatic myelopathies and 14 cases of subacute/chronic myelopathies, we identified 30 cases with non-compressive AMs, 30 cases with compressive AMs and 15 mimics. The presence of pyramidal signs ( p = 0.012) and/or pain ( p = 0.048) correctly identified 88% of cases with AMs. We failed to identify clinical indicators for distinguishing non-compressive and compressive AMs, although cases with inflammatory AMs were younger than cases with all the remaining conditions ( p < 0.05). Different MRI patterns could be described according to the final diagnosis: among non-compressive AMs, inflammatory lesions were more often posterior or central; vascular malformation had a fairly widespread distribution; spine ischaemia was more often central. Anterior or lateral compression were more often associated with neoplasms and disc herniation , whereas hemorrhages and infections produced spine compression on all sides., Conclusion: We propose a simple clinical indicator (i.e. pyramidal signs and/or pain) to distinguish AMs from their mimics in an emergency setting. Urgent spinal cord MRI remains essential to discriminate compressive and non-compressive aetiologies.

Over the years, several authors have discussed the possibility of considering somatic dysfunction (SD) as a "nosological element" detectable on palpation. There are many aspects to consider regarding the etiology and diagnosis of SD, and the literature on osteopathic issues provides details on physiological signs that characterize it, including tissue texture changes. Recent knowledge suggests that how tissue and, in particular, connective tissue, responds to osteopathic treatment may depend on the modulation of the inflammation degree. Low-grade inflammation (LGI) may act on the extracellular matrix (ECM) and on cellular elements; and these mechanisms may be mediated by biological water. With its molecules organized in structures called exclusion zones (EZ), water could explain the functioning of both healthy and injured tissues, and how they can respond to osteopathic treatment with possible EZ normalization as a result. The relationship between inflammation and DS and the mechanisms involved are described by several authors; however, this review suggests a new model relating to the characteristics of DS and to its clinical implications by linking to LGI. Tissue alterations detectable by osteopathic palpation would be mediated by body fluids and in particular by biological water which has well-defined biophysical characteristics. Research in this area is certainly still to be explored, but our suggestion seems plausible to explain many dynamics related to osteopathic treatment. We believe that this could open up a fascinating scenario of therapeutic possibilities and knowledge in the future.

Background: Developing eco-friendly antioxidant and antimicrobial substances originating from biomass residues has recently attracted considerable interest. In this study, two lignosulfonates and various oxidized water-soluble lignins were investigated for their antioxidant properties, as assessed by ABTS, DPPH and Folin-Ciocalteu methods, and their antimicrobial activity against some bacterial strains responsible for human pathologies., Results: The lignosulfonates showed the largest antiradical/antimicrobial capacity, whereas the other substrates were less effective. The observed antioxidant/antibacterial properties were positively correlated with lignin aromatic/phenolic content. The positive correlation between antiradical and antimicrobial activities suggests that lignin scavenging capacity was also involved in its antibacterial activity. A greater antimicrobial performance was generally observed against Gram-positive bacterial strains, and it was attributed to the intrinsic larger susceptibility of Gram-positive bacteria to lignin phenols. A significant though lesser inhibitory activity was also found against Escherichia coli., Conclusion: Our results confirmed the dependence of lignin antioxidant/antibacterial power on its extraction method and chemical structure, as well as on the type of bacterial strains. Identifying the relationship between lignin molecular composition and its antioxidant/antibacterial features represents an advance on the potential future use of renewable and eco-compatible lignin materials in nutraceutical, pharmaceutical and cosmetic sectors. © 2021 Society of Chemical Industry., (© 2021 Society of Chemical Industry.)

A wide range of acute neurological disorders may present with symptoms similar to a stroke, so-called 'stroke mimics'. Migraine aura and seizures account for the most extensive stroke mimics population. A large number of patients with a definite stroke mimics diagnosis (most commonly those with psychiatric disorders or seizures) had been treated with IV alteplase without adverse related events. We report a case of a man aged 70 years admitted to the emergency room because of acute onset of delirium and a loss of strength in the left arm (National Institutes of Health Stroke Scale {NIHSS}: 10), severe hyponatremia (127 mEq/L), and no evidence of intracranial arterial occlusion at CT scan. He was eligible for intravenous thrombolysis and, after treatment, neurological symptoms improved (NIHSS: 2). The subsequent appearance of "clonus" in the left lower limb, the persistence of hyponatremia, and the presence of electroencephalogram (EEG) abnormalities led to the clinical suspicion of focal motor-onset seizure with impaired awareness. The patient was treated successfully with anti-seizure medications (ASMs): lacosamide 200 mg IV during the acute setting care, followed by oral lacosamide 200 mg bis in die (BID). Since two other focal seizures occurred, brivaracetam 25 mg BID has been added in therapy with subsequent clinical discontinuance and EEG normalization. Two consecutive magnetic resonance imaging (MRI) examinations showed several cortical lesions restricted in high signal in diffusion-weighted imaging (DWI) which corresponding to T2-weighted and fluid-attenuated inversion recovery (FLAIR) hyperintensities, but without lesions evidence in apparent diffusion coefficient (ADC) map. These radiological changes disappeared at a follow-up MRI performed 20 days after the symptoms' onset. The patient fully recovered was discharged home without developing pharmacological adverse events. In this case, MRI provided an opportunity for early identification of seizure-related alterations. Hence, we discuss how prospective MRI studies during seizures and interictal period would contribute to defining the relationship between the electroclinical characteristics and MRI alteration patterns, and therefore, the potential role of MRI in the differential diagnosis between seizures and stroke mimic., Competing Interests: The authors have declared that no competing interests exist., (Copyright © 2021, Mangiardi et al.)

Background: The major endovascular mechanic thrombectomy (MT) techniques are: Stent-Retriever (SR), aspiration first pass technique (ADAPT) and Solumbra (Aspiration + SR), which are interchangeable (defined as switching strategy (SS)). The purpose of this study is to report the added value of switching from ADAPT to Solumbra in unsuccessful revascularization stroke patients., Methods: This is a retrospective, single center, pragmatic, cohort study. From December 2017 to November 2019, 935 consecutive patients were admitted to the Stroke Unit and 176/935 (18.8%) were eligible for MT. In 135/176 (76.7%) patients, ADAPT was used as the first-line strategy. SS was defined as the difference between first technique adopted and the final technique. Revascularization was evaluated with modified Thrombolysis In Cerebral Infarction (TICI) with success defined as mTICI ≥ 2b. Procedural time (PT) and time to reperfusion (TTR) were recorded., Results: Stroke involved: Anterior circulation in 121/135 (89.6%) patients and posterior circulation in 14/135 (10.4%) patients. ADAPT was the most common first-line technique vs. both SR and Solumbra (135/176 (76.7%) vs. 10/176 (5.7%) vs. 31/176 (17.6%), respectively). In 28/135 (20.7%) patients, the mTICI was ≤ 2a requiring switch to Solumbra. The vessel's diameter positively predicted SS result (odd ratio (OR) 1.12, confidence of interval (CI) 95% 1.03-1.22; p = 0.006). The mean number of passes before SS was 2.0 ± 1.2. ADAPT to Solumbra improved successful revascularization by 13.3% (107/135 (79.3%) vs. 125/135 (92.6%)). PT was superior for SS comparing with ADAPT (71.1 min (CI 95% 53.2-109.0) vs. 40.0 min (CI 95% 35.0-45.2); p = 0.0004), although, TTR was similar (324.1 min (CI 95% 311.4-387.0) vs. 311.4 min (CI 95% 285.5-338.7); p = 0.23)., Conclusion: Successful revascularization was improved by 13.3% after switching form ADAPT to Solumbra (final mTICI ≥ 2b was 92.6%). Vessel's diameter positively predicted recourse to SS.

The acknowledgement of bioactive functions of compost teas promotes the research on characteristics and potential application of these heterogeneous water-soluble extracts from recycled biomasses. In this work, compost teas were isolated from on-farm composts made with agro-industrial residues of artichoke, pepper and coffee husks with the aim to evaluate the structural-activity relationship of dissolved bioactive molecules. The molecular features of compost teas were determined by 13 C-CPMAS NMR spectroscopy, Infrared spectroscopy, and off-line pyrolysis-Gas Chromatography/Mass Spectrometry. Bioactivity of different compost teas was tested on Basil seeds germination, while the antioxidant capacity was measured by ABTS and DDPH spectrophotometric assays. The antimicrobial activity was measured against some pathogenic human bacterial strains. The seed germination experiment showed no phytotoxic effects and a significant increase of both root and epicotyls upon application of coffee husks and pepper CT samples. The same compost teas revealed the largest antioxidant activity and a clear antimicrobial effect determined by MIC (Minimal Inhibitory Concentration) against some gram-negative bacterial strains such as Pseudomonas aeruginosa, Escherichia coli, and Klebsiella pneumoniae. The bioactivity of CT-samples was related to their general hydrophobic features and to specific molecular composition. In particular, 13 C-CPMAS NMR spectra and off-line thermochemolysis GC-MS highlighted a close correlation between radical scavenger activity and antibacterial bioactive functions with bio-available soluble aromatic compounds, such as lignin and phenols derivatives. The antioxidant and, antibacterial properties of compost teas from green composts encourages an innovative potential application of these eco-friendly products not only in agricultural applications but also in nutraceutical and pharmaceutical fields., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

The aim of this study was to assess the impact of osteopathic manipulative treatment (OMT) on newborn babies admitted at a neonatal intensive care unit (NICU). This was an observational, longitudinal, retrospective study. All consecutive admitted babies were analyzed by treatment (OMT vs. usual care). Treatment group was randomly assigned. Between-group differences in weekly weight change and length of stay (LOS) were evaluated in the overall and preterm populations. Among 1249 babies (48.9% preterm) recorded, 652 received usual care and 597 received OMT. Weight increase was more marked in the OMT group than in the control group (weekly change: +83 g vs. +35 g; p < 0.001). Similar trends were found in the subgroup of preterm babies. A shorter LOS was found in the OMT group vs. the usual care group both in overall population (average mean difference: -7.9 days, p = 0.15) and in preterm babies (-12.3 days; p = 0.04). In severe preterm babies, mean LOS was more than halved as compared to the control group. OMT was associated with a more marked weekly weight increase and, especially in preterm babies, to a relevant LOS reduction: OMT may represent an efficient support to usual care in newborn babies admitted at a NICU.

Humic-like substances (HLSs) isolated by alkaline oxidative hydrolysis from lignin-rich agro-industrial residues have been shown to exert biostimulant activity toward maize ( Zea mays L.) germination and early growth. The definition of a quantitative structure-activity relationship (QSAR) between HLS and their bioactivity could be useful to predict their biological properties and tailor plant biostimulants for specific agronomic and industrial uses. Here, we created several projection on latent structure (PLS) regression by using published analytical data on the molecular composition of lignin-derived HLS obtained by both 13 C-CPMAS-NMR spectra directly on samples and 31 P-NMR spectra after derivatization of hydroxyl functions with a P-containing reagent (2-chloro-4,4,5,5-tetramethyl-1,3,2-dioxaphospholane). These spectral data were used to model the effect of HLS on the elongation of primary root, lateral seminal roots, total root apparatus, and coleoptile of maize. The 13 C-CPMAS-NMR data suggested that methoxyl and aromatic moieties positively affected plant growth, while the carboxyl/esterified functions showed a negative impact on the overall seedling development. Alkyl C seems to promote Col elongation while concomitantly reducing that of the root system. Additionally, 31 P-NMR-derived spectra revealed that the elongation of roots and Col were enhanced by the occurrence of aliphatic hydroxyl groups, and guaiacyl and p -Hydroxyphenyl lignin monomers. The PLS models based on raw dataset from 13 C-CPMAS-NMR spectra explained more than 74% of the variance for the length of lateral seminal roots, total root system and coleoptile, while other parameters derived from 13 C-CPMAS-NMR spectra, namely the Hydrophobicity and Hydrophilicity of materials were necessary to explain 83% of the variance of the primary root length. The results from 31 P-NMR spectra explained the observed biological variance by 90, 96, 96, and 93% for the length of primary root, lateral seminal roots, total root system and coleoptile, respectively. This work shows that different NMR spectroscopy techniques can be used to build up PLS models which can predict the bioactivity of lignin-derived HLS toward early growth of maize plants. The established QSAR may also be exploited to enhance by chemical techniques the bioactive properties of HLS and enhance their plant stimulation capacity., (Copyright © 2020 Savy, Brostaux, Cozzolino, Delaplace, du Jardin and Piccolo.)

Constipation is frequent amongst infants who have undergone surgery for anorectal malformation (ARM). Faecal retention, due to a dysfunctional enteric reflex of defaecation, can cause abdominal cramps, pseudoincontinence and, in the worst cases, megacolon. Prokinetic protocols are used to stimulate at least 1 bowel movement per day, including laxatives, enema, stools softeners and dietary schedules. While osteopathic manipulative treatment is adopted in adults for functional constipation, it has not been described for infants. Herein, we report the case of an infant undergoing anorectoplasty for a low ARM who was referred to the osteopath 2 years after the onset of severe constipation associated with pseudoincontinence and abdominal cramps and was refractory to the prokinetic protocol. In a child with a good ARM prognosis, autonomous daily bowel movements should be achieved. In this child, the imbalanced tension of the pelvic floor and immaturity of the parasympathetic plexus led to a functional alteration of the defaecation reflex. After adjunction of osteopathic manipulative treatment (OMT) to the therapeutic panel, the constipation showed gradual remission, with acquisition of autonomous defaecation 4 months after the therapy began. This suggests the importance of investigating the efficacy of OMT inclusion in the postsurgical prokinetic protocols for ARM patients with a good prognosis., Competing Interests: The authors declare that they have no competing interests., (Copyright © 2020 by S. Karger AG, Basel.)

Background : To evaluate the effectiveness/side-effects of osteopathic manipulation treatment (OMT) performed on the 7th post-natal day, on cerebro-splanchnic oximetry, tissue activation and hemodynamic redistribution in late preterm (LP) infants by using near infrared spectroscopy (NIRS). Methods : Observational pretest-test study consisting in a cohort of 18 LPs who received OMT on the 7th post-natal day. NIRS monitoring was performed at three different time-points: 30 min before (T0), (30 min during (T1) and 30 min after OMT (T2). We evaluated the effects of OMT on the following NIRS parameters: cerebral (c), splanchnic (s) regional oximetry (rSO2), cerebro-splanchnic fractional tissue oxygen extraction (FTOE) and hemodynamic redistribution (CSOR). Results : crSO2 and cFTOE significantly ( P < 0.001) improved at T0-T2; srSO2 significantly ( P < 0.001) decreased and sFTOE increased at T0-T1. Furthermore, srSO2 and sFTOE significantly improved at T1-T2. Finally, CSOR significantly ( P < 0.05) increased at T0-T2. Conclusions : The present data show that OMT enhances cerebro-splanchnic oximetry, tissue activation and hemodynamic redistribution in the absence of any adverse clinical or laboratory pattern. The results indicate the usefulness of further randomized studies in wider populations comparing the effectiveness of OMT with standard rehabilitation programs.

Phosphate-(P)-solubilizing microorganisms (PSM) are important drivers of P cycling in natural and agro-ecosystems. Their use as plant inoculants to improve P acquisition of crops has been investigated for decades. However, limited reproducibility of the expected effects, particularly under field conditions, remains a major challenge. This study demonstrates that the form of nitrogen fertilization has a significant impact on the performance of various fungal and bacterial PSM inoculants in maize grown on neutral to alkaline soils with limited P availability. Under these conditions, a high soil pH-buffering capacity frequently limits the efficiency of nutrient mobilization, mediated by plant roots and microorganisms via rhizosphere acidification. In a soil pH range between 7.0 and 8.0, nitrate fertilization promoting rhizosphere alkalinisation further aggravates this problem. Accordingly, in greenhouse experiments, six strains of Pseudomonas , Bacillus , Paenibacillus , Streptomyces , and Penicillium with proven P-solubilizing potential, completely failed to promote P acquisition in maize grown on a calcareous Loess sub-soil pH 7.6 with nitrate fertilization and rock phosphate (Rock-P) as a sparingly soluble P source. However, after replacement of nitrate fertilization by ammonium, stabilized with the nitrification inhibitor 3,4-dimethylpyrazole-phosphate (DMPP), five out of seven investigated PSM inoculants (comprising 12 fungal and bacterial PSM strains) exerted beneficial effects on plant growth and reached up to 88% of the shoot biomass production of a control supplied with soluble triple-superphosphate (TSP). Stabilized ammonium combined with PSM-inoculants improved P acquisition ( Trichoderma harzianum T22, Pseudomonas sp. DMSZ 13134), while other strains particularly stimulated root growth ( T. harzianum OMG16, Bacillus amyloliquefaciens FZB42), which promoted the acquisition also of other mineral nutrients, such as N, K, and Mn. A similar effect was recorded under field conditions on an alkaline clay-loam soil pH 8.6. The combination of stabilized ammonium with a range of consortium products based on T. harzianum OMG16, B. amyloliquefaciens , micronutrients, and humic acids completely compensated the effect of a TSP fertilization on field establishment, nutrient acquisition, and yield formation in maize, while non-stabilized urea-di-ammonium phosphate fertilization was largely ineffective. These findings suggest that the efficiency of PSM-plant interactions can be influenced by the form of N fertilization, offering promising perspectives for synergistic effects with stabilized ammonium fertilizers.

The ligno-cellulose residues from biorefinery production of bio-ethanol were used as woody structuring material within an on-farm composting system, with the aim to obtain bioactive water soluble and humic fractions from composted materials. The molecular characterization of initial biomasses and final products revealed a transformation towards more stable compounds during composting and showed the selective incorporation of specific phenolic derivatives of ligno-cellulose in both bulk samples and corresponding extracts. While the use of the stable bulk composts as organic fertilizer resulted in a decrease of nitrogen and phosphorous assimilation in maize tissues, a bio-stimulation was shown by water soluble organic compounds and humic substances in germination tests and pot experiments, respectively. The differential responses obtained in maize seedlings and plants were related to the molecular composition and concentration of the applied water extracts and humic substances, thus suggesting a role of phenols and lignin derivatives in the stimulation of maize roots and shoots and the enhancement of P uptake. The results confirm that ligno-cellulose residues may be effectively recycled as composting additives in order to enrich mature compost in aromatic and lignin compounds. A preliminary knowledge on the molecular characteristics and biological properties of composted materials is an essential requirement to select the most suitable derivatives from composted ligno-cellulose wastes in sustainable agricultural practices., (Copyright © 2018. Published by Elsevier B.V.)

The ability of Trichoderma harzianum (strain OMG-08) as plant growth promoting fungus (PGPF), was tested on Zea mays plants grown in soil pots added with different inorganic (triple superphosphate and rock phosphate) and organic (cow and horse manure composts) P fertilizers. The effect of treatments was evaluated by following the variations of plants dry biomass and nutrient content, as well as the metabolic changes in plant leaves by both GC-MS and NMR spectroscopy. A synergic effect was observed in treatments with both composts and fungus inoculation, in which not only plant growth and P uptake were enhanced, but also the expression of different metabolites related to an improved photosynthetic activity. Conversely, the combination of Trichoderma with inorganic fertilizers was less effective and even showed a reduction of plants shoot biomass and N content. The corresponding plant metabolome revealed metabolic compounds typical of biotic or abiotic stresses, which may be attributed to a reduced capacity of inorganic fertilizers to provide a sufficient P availability during plant growth. Our findings also indicate that the molecular composition of compost differentiated the Trichoderma activity in sustaining plant growth. The positive effects of the combined Trichoderma and compost treatment suggest that it may become an alternative to the phosphorus mineral fertilization., Competing Interests: The authors have declared that no competing interests exist.

Lignosulfonates are important by-products of the paper industry and may be transformed into different commodities. We studied the molecular properties of ammonium (LS-AM) and calcium Lignosulfonates (LS-C) and evaluated their bioactivity towards the early development of maize plantlets. The FT-IR, 13 C NMR and 1 H- 13 C-HSQC-NMR spectra showed that the two lignosulfonates varied in hydroxyl, sulfonate and phenolic content, while DOSY-NMR spectroscopy suggested a similar diffusivity. High Performance Size Exclusion Chromatography (HPSEC) was used to simulate the effects of root-exuded acids and describe the conformational dynamics of both LS substrates in acidic aqueous solutions. This technique showed that LS-C was stabilized by the divalent Ca 2+ counterion, thus showing a greater conformational stability than LS-AM, whose components could not be as efficiently aggregated by the monovalent NH4 + counter-ion. The plant bioassays revealed that LS-AM enhanced the elongation of the root system, whereas LS-C significantly increased both total and shoot plant weights. We concluded that the lignosulfonate bioactivity on plant growth depended on the applied concentrations, their molecular properties and conformational stability., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Scanning electron microscopy evaluation of root surfaces after ultrasonic instrumentation was performed with 2 different metallic tips on piezoelectric devices. Fresh extracted teeth were collected for experimental observation and randomly divided into 2 groups: Test Group, where the root surfaces were treated using an iron, rough, double nano-structural coated (T-Black), corindone-treated tip, and Control Group where the root surfaces were treated with a conventional iron smooth tip. A scanning electronic microscope analysis was performed and the surface roughness and the amount of residual debris were evaluated. Descriptive and inferential statistics were performed. Twenty specimens were analyzed, 10 per group and a total of 21.4 × 10 μm has been observed. On treated area percentage of debris after ultrasonic scaling in Test Group was 1.9 ± 1.8%, while in Control Group it was 5.7 ± 4.3%. Within the limits of the study, it seems that the efficacy of the novel iron, rough, double nano-structural coated (T-Black), corindone-treated structure tip showed greater performance in terms of root surface debridement than the conventional iron smooth tip. The possibility to use a single tool (ultrasonic device with a specific tip) for the root planing procedure within the nonsurgical mechanical therapy may represent a significant advantage for the clinicians. The tested novel tip seems to be able to show the requested ideal characteristics. However, further clinical studies are needed to demonstrate the in vitro results.

Both high-resolution magic-angle-spinning (HRMAS) and magnetic resonance imaging (MRI) NMR spectroscopies were applied here to identify the changes of metabolome, morphology, and structural properties induced in seeds (caryopses) of maize plants grown at field level under either mineral or compost fertilization in combination with the inoculation by arbuscular mycorrhizal fungi (AMF). The metabolome of intact caryopses was examined by HRMAS-NMR, while the morphological aspects, endosperm properties and seed water distribution were investigated by MRI. Principal component analysis (PCA) was applied to evaluate 1 H CPMG (Carr-Purcel-Meiboom-Gill) HRMAS spectra as well as several MRI-derived parameters ( T 1 , T 2 , and self-diffusion coefficients) of intact maize caryopses. PCA score-plots from spectral results indicated that both seeds metabolome and structural properties depended on the specific field treatment undergone by maize plants. Our findings show that a combination of multivariate statistical analyses with advanced and nondestructive NMR techniques, such as HRMAS and MRI, enables the evaluation of the effects induced on maize caryopses by different fertilization and management practices at field level. The spectroscopic approach adopted here may become useful for the objective appraisal of the quality of seeds produced under a sustainable agriculture.

Four water extractable organic matter (WEOM) were obtained from composts made out of residues of: 1. artichoke (C-CYN), 2. artichoke/fennel (C-CYNF), 3. tomato/woodchips (C-TOM), 4. Municipal solid waste (C-MSW), and their bioactivity was tested for maize seed germination and maize seedling growth. The molecular properties of both original composts and their WEOM were characterized by spectroscopic ( 13 C-CPMAS- and 1 H NMR, FTIR-ATR), thermochemolysis-GC/MS, and thermal methods (TGA, DSC). While all WEOM had significant effects on plant growth, CYN-WEOM was the only material that concomitantly increased germination rate and primary and lateral root length of maize seedlings. The lignin-rich WEOM from green composts were generally more effective than those obtained from equally hydrophobic, but mainly alkyl-rich municipal organic wastes. A flexible conformational structure, due to the balanced content of aromatic compounds and carbohydrates, appeared to facilitate the release of bioactive molecules from WEOM suprastructures and stimulate plant growth., (Copyright © 2017 Elsevier B.V. All rights reserved.)

Introduction . Adding stem cells to biodegradable scaffolds to enhance bone regeneration is a valuable option. Different kinds of stem cells with osteoblastic activity were tested, such as bone marrow stromal stem cells (BMSSCs). Aim . To assess a correct protocol for osteogenic stem cell differentiation, so BMSSCs were seeded on a bone porcine block (BPB). Materials and Methods . Bone marrow from six minipigs was extracted from tibiae and humeri and treated to isolate BMSSCs. After seeding on BPB, critical-size defects were created on each mandible of the minipigs and implanted with BPB and BPB/BMSSCs. After three months, histomorphometric analysis was performed. Results . Histomorphometric analysis provided percentages of the three groups. Tissues present in control defects were 23 ± 2% lamellar bone, 28 ± 1% woven bone, and 56 ± 4% marrow spaces; in BPB defects were 20 ± 5% BPB, 32 ± 2% lamellar bone, 24 ± 1% woven bone, and 28 ± 2% marrow spaces; in BPB/BMSSCs defects were 17 ± 4% BPB/BMSSCs, 42 ± 2% lamellar bone, 12 ± 1% woven bone, and 22 ± 3% marrow spaces. Conclusion . BPB used as a scaffold to induce bone regeneration may benefit from the addition of BDPSCs in the tissue-engineered constructs.