Plant Tissues Research Articles

Morphological and molecular preservation through universal preparation of fresh-frozen tissue samples for multimodal imaging workflows.

The landscape of tissue-based imaging modalities is constantly and rapidly evolving. While formalin-fixed, paraffin-embedded material is still useful for histological imaging, the fixation process irreversibly changes the molecular composition of the sample. Therefore, many imaging approaches require fresh-frozen material to get meaningful results. This is particularly true for molecular imaging techniques such as mass spectrometry imaging, which are widely used to probe the spatial arrangement of the tissue metabolome. As high-quality fresh-frozen tissues are limited in their availability, any sample preparation workflow they are subjected to needs to ensure morphological and molecular preservation of the tissues and be compatible with as many of the established and emerging imaging techniques as possible to obtain the maximum possible insights from the tissues. Here we describe a universal sample preparation workflow, from the initial step of freezing the tissues to the cold embedding in a new hydroxypropyl methylcellulose/polyvinylpyrrolidone-enriched hydrogel and the generation of thin tissue sections for analysis. Moreover, we highlight the optimized storage conditions that limit molecular and morphological degradation of the sections. The protocol is compatible with human and plant tissues and can be easily adapted for the preparation of alternative sample formats (e.g., three-dimensional cell cultures). The integrated workflow is universally compatible with histological tissue analysis, mass spectrometry imaging and imaging mass cytometry, as well as spatial proteomic, genomic and transcriptomic tissue analysis. The protocol can be completed within 4 h and requires minimal prior experience in the preparation of tissue samples for multimodal imaging experiments.

Comparative transcriptomic analysis revealed important processes underlying the static magnetic field effects on Arabidopsis.

Static magnetic field (SMF) plays important roles in various biological processes of many organisms including plants, though the molecular mechanism remains largely unclear. Here in this study, we evaluated different magnetic setups to test their effects on growth and development on Arabidopsis (Arabidopsis thaliana), and discovered that plant growth was significantly enhanced by inhomogeneous SMF generated by a regular triangular prism magnet perpendicular to the direction of gravity. Comparative transcriptomic analysis revealed that auxin synthesis and signal transduction genes were upregulated by SMF exposure. SMF also facilitated plants to maintain the iron homeostasis. The expression of iron metabolism-related genes was downregulated by SMF, however, the iron content in plant tissues remains relatively unchanged. Furthermore, SMF exposure also helped the plants to reduce ROS level and synergistically maintain the oxidant balance by enhanced activity of antioxidant enzymes and accumulation of nicotinamide. Taken together, our data suggested that SMF is involved in regulating the growth and development of Arabidopsis thaliana through maintaining iron homeostasis and balancing oxidative stress, which could be beneficial for plant survival and growth. The work presented here would extend our understanding of the mechanism and the regulatory network of how magnetic field affects the plant growth, which would provide insights into the development of novel plant synthetic biology technologies to engineer stress-resistant and high-yielding crops.

Dual Infrared 2‐Photon Microscopy Achieves Minimal Background Deep Tissue Imaging in Brain and Plant Tissues

AbstractTraditional deep fluorescence imaging has primarily focused on red‐shifting imaging wavelengths into the near‐infrared (NIR) windows or implementation of multi‐photon excitation approaches. Here, the advantages of NIR and multiphoton imaging are combined by developing a dual‐infrared two‐photon microscope that enables high‐resolution deep imaging in biological tissues. This study first computationally identifies that photon absorption, as opposed to scattering, is the primary contributor to signal attenuation. A NIR two‐photon microscope is constructed next with a 1640 nm femtosecond pulsed laser and a NIR PMT detector to image biological tissues labeled with fluorescent single‐walled carbon nanotubes (SWNTs). Spatial imaging resolutions are achieved close to the Abbe resolution limit and eliminate blur and background autofluorescence of biomolecules, 300 µm deep into brain slices and through the full 120 µm thickness of a Nicotiana benthamiana leaf. NIR‐II two‐photon microscopy can also measure tissue heterogeneity by quantifying how much the fluorescence power law function varies across tissues, a feature this study exploits to distinguish Huntington's Disease afflicted mouse brain tissues from wildtype. These results suggest dual‐infrared two‐photon microscopy can accomplish in‐tissue structural imaging and biochemical sensing with a minimal background, and with high spatial resolution, in optically opaque or highly autofluorescent biological tissues.

In-silico analysis of ribosome inactivating protein (RIP) of the Cucurbitaceae family.

Ribosome-inactivating proteins (RIPs) are highly active N-glycosidases that depurinate both bacterial and eukaryotic rRNAs, halting protein synthesis during translation. Found in a diverse spectrum of plant species and tissues, RIPs possess antifungal, antibacterial, antiviral, and insecticidal properties linked to plant defense. In this study, we investigated the physiochemical properties of RIP peptides from the Cucurbitaceae family through bioinformatics approaches. Molecular weight, isoelectric point, aliphatic index, extinction coefficient, and secondary structures were analyzed, revealing their hydrophobic nature. The novelty of this work lies in the comprehensive examination of RIPs from the Cucurbitaceae family and their potential therapeutic applications. The study also elucidated the binding interactions of Cucurbitaceae RIPs with key biological targets, including Interleukin-6 (IL-6). Strong hydrogen bond interactions between RIPs and these targets suggest potential for innovative insilico drug design and therapeutic applications, particularly in cancer treatment. Comprehensive analysis of bond lengths using Ligpolt + software provides insights for optimizing molecular interactions, offering a valuable tool for drug design and structural biology studies.

Charting the Cannabis plant chemical space with computational metabolomics

IntroductionThe chemical classification of Cannabis is typically confined to the cannabinoid content, whilst Cannabis encompasses diverse chemical classes that vary in abundance among all its varieties. Hence, neglecting other chemical classes within Cannabis strains results in a restricted and biased comprehension of elements that may contribute to chemical intricacy and the resultant medicinal qualities of the plant.ObjectivesThus, herein, we report a computational metabolomics study to elucidate the Cannabis metabolic map beyond the cannabinoids.MethodsMass spectrometry-based computational tools were used to mine and evaluate the methanolic leaf and flower extracts of two Cannabis cultivars: Amnesia haze (AMNH) and Royal dutch cheese (RDC).ResultsThe results revealed the presence of different chemical compound classes including cannabinoids, but extending it to flavonoids and phospholipids at varying distributions across the cultivar plant tissues, where the phenylpropnoid superclass was more abundant in the leaves than in the flowers. Therefore, the two cultivars were differentiated based on the overall chemical content of their plant tissues where AMNH was observed to be more dominant in the flavonoid content while RDC was more dominant in the lipid-like molecules. Additionally, in silico molecular docking studies in combination with biological assay studies indicated the potentially differing anti-cancer properties of the two cultivars resulting from the elucidated chemical profiles.ConclusionThese findings highlight distinctive chemical profiles beyond cannabinoids in Cannabis strains. This novel mapping of the metabolomic landscape of Cannabis provides actionable insights into plant biochemistry and justifies selecting certain varieties for medicinal use.

The Transformation of Per- and Polyfluoroalkyl Substances in the Aquatic Environment of a Fluorochemical Industrial Park

Jiangsu High-Tech Fluorochemical Industrial Park in Changshu City, Jiangsu Province, is the largest fluorochemical industrial park in Asia. The occurrence of per- and polyfluoroalkyl substances (PFASs) in surface water and widespread local plants was investigated in Jiangsu High-Tech Fluorochemical Industrial Park. Thirty-two target PFASs were detected in dissolved-phase, particle-phase and plant samples. The concentrations of total PFASs ranged from 1650 to 8250 ng/L in the dissolved-phase samples, 132 to 6810 ng/g dw in the particle-phase samples and 25.8 to 9460 ng/g dw in different plant tissues. Perfluorohexanoic acid (PFHxA), Perfluorooctanoate acid (PFOA) and 6:2 fluorotelomer carboxylic acid (6:2 FTCA) were predominant PFASs and contributed 80−91% to ΣPFAS in water samples. A total of 67 emerging PFAS were identified in all samples using nontargeted analysis. Typha orientalis showed better accumulation ability, with an average ΣPFAS concentration of 3450 ng/g dw and the highest root concentration factor (RCF) of 171. Typha orientalis, Eichhornia crassipes and Alternanthera sessilis have potential for use in PFAS phytostabilization.

Effect of BvAgNP on growth, development, and glyoxalase gene expression analysis in Mammillaria bombycina and Selenicereus undatus.

Silver nanoparticles (AgNPs) have been used in plant tissue culture as growth stimulants, promoting bud initiation, germination, and rooting. In prior studies, AgNPs were synthesized and characterized by green synthesis using extracts from Beta vulgarisvar. cicla (BvAgNP), and their functionality as seed disinfectant and antimicrobial was verified. In this study, we evaluated the effect of BvAgNP on the growth and development of Mammillaria bombycina and Selenicereus undatus in vitro, as well as the expression of glyoxalase genes. Explants from M. bombycina and S. undatus in vitro were treated with 25, 50, and 100mg/L of BvAgNP. After 90 days, morphological characteristics were evaluated, and the expression of glyoxalase genes was analyzed by qPCR. All treatments inhibited rooting for M. bombycina and no bud initiation was observed. S. undatus, showed a maximum response in rooting and bud generation at 25mg/L of BvAgNP. Scanning electron microscopy (SEM) results exhibited a higher number of vacuoles in stem cells treated with BvAgNP compared to the control for both species. Expression of glyoxalase genes in M. bombycina increased in all treatments, whereas it decreased for S. undatus, however, increasing in roots. This study presents the effects of BvAgNP on the growth and development of M. bombycina and S. undatus, with the aim of proposing treatments that promote in vitro rooting and bud initiation.

Caveolae Disassembly using Methyl-β-cyclodextrin Causes the Abolition of Coupling of the Caveolae and the Sarcoplasmic Reticulum in the Rat Femoral Artery

Background: Caveolae are essential in regulating signal transduction mechanisms of ion channels in vascular tissue, including BKCa channels (maxi-K). The current study investigated the localization of maxi-K channels within caveolae. Methods: Sixteen rats were divided into two groups: A control group and a treated group, where arteries in the treated group were incubated with methyl-β-cyclodextrin (MβCD) to disassemble caveolae from artery tissue. Immunohistochemistry (IHC), immunocytochemistry (ICC), transmission electron microscopy (TEM) and Western blot techniques were used in this study. Result: IHC of intact arteries showed colocalization of maxi-K channels with caveolin-1 in smooth muscle and endothelial cells and colocalized of maxi-K channels with caveolin-3 in smooth muscle cells only. These findings were also corroborated with ICC in a single smooth muscle cell. TEM revealed caveolae covering most plasma membranes of smooth muscle and endothelial cells and showed that caveolae sit close to the sarcoplasmic reticulum only in smooth muscle cells. TEM showed incubating arteries with MβCD led to the disassembly of caveolae from artery tissue. This study concluded that maxi-K channels localize to caveolae and that caveolae abolishment by MβCD led to the abolition of the coupling of caveolae and the sarcoplasmic reticulum.

Effects of Magnetized, Chelated Iron Foliage Treatments, and Metal Halide Lamps on Plant Water Structure, Water Vapor Dynamics, and Resilience for Legumes under Water Stress

A greenhouse study was conducted to determine the effects of foliar applications of magnetized, chelated liquid iron fertilizer for increasing the drought tolerance of two legumes. The study objectives were to determine the drought tolerance effects of four treatments on foliar gas exchange, soil moisture, and plant growth for soybean (Glycine max) and velvet bean (Mucuna pruriens) plants. The plant treatments included applications with chelated liquid iron fertilizer (2.5 and 5%) with a conventional boom sprayer, with and without magnets in the spray lines, and metal halide lamps. Three gas exchange measurements were collected before applying the foliage treatments and after two water stress treatments. A foliage and metal halide lamp treatment deactivated or unlinked nine interconnected gas exchange parameters that are correlated with plant defense activities during water stress conditions. The deactivation of interconnected regulatory gas exchange functions improved metabolic efficiency, reduced stress levels, and boosted plant resilience to abiotic stressors. Also, the study findings suggest that the study treatments maintained or increased the level of biologically structured water in plant tissues and vascular systems.

Sustainable strategy for rehabilitating phosphate mining sites and valorisation of phosphate industry by-products and sludge using pistachio tree (Pistacia atlantica), false pepper (Schinus molle), and eucalyptus (Eucalyptus globulus) trees

The development of anthroposols has been proposed as a new environmentally friendly approach to ensuring the successful revegetation of phosphate mining sites. The phosphate industry's by-products, including phosphogypsum (PG), phosphate sludge (PS), and sewage sludge (SS), can be valuable resources in restoring the ecological balance of mined soil areas. The aim of this study was to safely and sustainably restore the ecological integrity of the phosphate mining site through the evaluation of nutrients and heavy metals dynamics in soil and plant tissues of three tree species and treated by-products containing 65 % PG, 30 % PS, and 5 % SS. The tree species used were Pistacia atlantica, Schinus molle, and Eucalyptus globulus. The experimental layout was a randomised complete block design with six replicates and three treatments. Growth diameter, height, nutrient uptakes and heavy metal dynamic were evaluated from the rhizosphere soils and plant tissues over two years. Hierarchical head maps of correlations between the measured growth parameters, soil and nutrient uptakes of the tree species were analysed using a phylogenetic generalised linear mixed model. S. molle and E. globulus had higher average diameter and height than P. atlantica plants. P. atlantica and S. molle showed greater nitrogen, phosphorus, potassium, calcium, and magnesium concentrations than E. globulus trees. Tree growth parameters were closely linked to soil nutrient bioavailability. The heavy metal accumulation ratio was higher in the E. globulus and S. molle leaves than in stems. Using by-products could be valorised for rehabilitating mine sites together with E. globulus and S. molle species.

Sodium nitroprusside, a donor of nitric oxide, enhances arbuscular mycorrhizal fungi symbiosis with corn plant and mitigates Cd bioavailability in the rhizosphere

Arbuscular mycorrhizal fungi (AMF) have been demonstrated to influence the bioavailability of heavy metals (HM) in the rhizosphere and their absorption by plants. Nitric oxide (NO) has been shown to increase plant tolerance to environmental stresses and promote the plant-fungal symbiotic relationship. However, to date, no research has been conducted to investigate the impact of utilizing living organisms with specific molecules that are effective in promoting plant growth in conjunction with the chemical alterations of HM in the rhizosphere. Therefore, this study was conducted as a completely randomized factorial design using the rhizobox technique to investigate whether the use of Funneliformis mosseae fungi and sodium nitroprusside (SNP, 100 mM) as a donor of NO, alone or in combination, can improve corn plant growth and affect the Cd fractions in the rhizosphere of Cd-contaminated soil. The results show that the inoculation of AMF and the application of SNP, either alone or in combination, significantly increased plant growth. This was achieved by reducing the bioavailable form of Cd in the rhizosphere, which in turn led to a decrease in the concentration of Cd in corn plant tissues. The inoculation of AMF, either alone or in combination with SNP, resulted in a decrease in the concentration of Cd in its exchangeable (EXCH), carbonate-bound (CAR), and Fe–Mn oxides-bound (MnOX and FeOX) forms, and an increase in its organic matter-bound (ORG) and residual (RES) forms in the rhizosphere. The study found that the rhizosphere had lower concentration of bioavailable form of Cd (EXCH-Cd (43%), CAR-Cd (9%), MnOX-CD (18%), FeOX-Cd (33%) and ORG-Cd (30%)) and higher concentration of low-toxic form of Cd (RES-Cd (56%)). This indicates the role of root exudates in the redistribution of Cd fractions in soil. The study also revealed that AMF colonization, in combination with SNP, affected the biogeochemical fractions of Cd and reduced Cd mobility in the rhizosphere. This improvement in Cd mobility led to reduced Cd accumulation in the plant tissues, resulting in improved corn plant growth.

Organic petrographic and mineralogical composition of the No. 6 coal seam of the Soutpansberg Coalfield, South Africa: Insights into paleovegetation and depositional environment

This study investigates the paleodepositional conditions of the No. 6 Seam of the Madzaringwe Formation in Makhado and Voorburg south area of the Soutpansberg Coalfield (Limpopo Province, South Africa) utilizing organic petrography and inorganic geochemical proxies. The coals are predominantly high-volatile bituminous B-A rank with high ash yields (avg. 36.1 wt%), characterized by high-vitrinite (~ 41.5 vol%), moderate-to-high inertinite (9.8 vol%–33.7 vol%) and low liptinite (~ 2.3 vol%). The distribution of inertinite varies among different coal horizons (from bottom-lower to middle-upper), suggesting differential oxidation conditions and/or paleofire occurrence. Vitrinite-to-inertinite (V/I) ratio, tissue preservation–gelification index (TPI–GI), and groundwater–vegetation index (GWI–VI) plots, indicate that the peat-forming forest-swamp vegetation accumulated under mesotrophic-to-rheotrophic hydrological conditions. The presence of structured macerals (i.e., telinite, collotelinite, fusinite, and semifusinite) suggests well-preserved plant tissues, whereas framboidal pyrite and sulphur content (0.24 wt%–2.16 wt%) point to brackish-water influence at the peat stage. The coals contain quartz, kaolinite, siderite, muscovite, dolomite, calcite, and pyrite minerals, most of which were likely sourced from felsic igneous rocks. The Al/(Al+Fe+Mn) and (Fe+Mn)/Ti ratios for the studied samples range between 0.24–0.97 and 0.57–70.10, respectively. The ratios, Al–Fe–Mn plot, and presence of massive botryoidal-type pyrite imply some influence of meteoric waters or fluids from hydrothermal activity post-deposition. Moreover, the chemical index of alteration (CIA: 98.25–99.67), chemical index of weathering (CIW: 92.04–97.66), and A–CN–K ternary diagram suggest inorganic matter suffered strong chemical weathering, indicating warm paleoclimatic conditions during the coal formation.

Seasonal phenology and starch allocation patterns in populations of Oxycaryum cubense forma cubense and paraguayense in Mississippi, Louisiana, and Florida

Abstract Phenological studies for Cuban bulrush [Oxycaryum cubense (Poepp. & Kunth) Lye] have been limited to the monocephalous form in Lake Columbus (Mississippi). Accordingly, there is little available information on potential phenological differences among Cuban bulrush forms (monocephalous vs. polycephalous) and populations in other geographic locations in the United States. Therefore, seasonal patterns of biomass and starch allocation in Cuban bulrush were quantified from two populations in Lake Columbus on the Tennessee-Tombigbee Waterway in Mississippi (monocephalous), two populations from Lake Martin in Louisiana (polycephalous), and two populations from Orange Lake in Florida (polycephalous). Monthly samples of Cuban bulrush inflorescence, emergent, and submersed tissue were harvested from two plots per state from October 2021 to September 2022. During monthly data collection, air temperature and photoperiod were recorded. Starch allocation patterns were similar among all sites, with starch storage being less than 1.5% dry weight for all plant tissues. Biomass was greatest in Lake Columbus (monocephalous; 600.7 g DW m-2) followed by Lake Martin (polycephalous; 392.3 g DW m-2) and Orange Lake (polycephalous; 233.85 g DW m-2). Peak inflorescence biomass occurred in the winter for the Lake Martin and Orange Lake populations, and in the summer for the Lake Columbus population. Inflorescence biomass in Lake Columbus had a positive relationship (r2=0.53) with warmer air temperatures. Emergent and submersed biomass generally had negative relationships with both photoperiod and temperature (r2=0.022-0.7789) in all sites. Peak biomass was also negatively related to temperature and photoperiod. Results from this study indicate that there are differences in biomass allocation between the two growth forms of Cuban bulrush and that growth can occur at temperatures below freezing. Low temperature tolerance may allow this species to expand its range further north than previously suspected.

GpEF1A: a novel lysine methyltransferase gene from Gypsophila perfoliata L. involved in boron homeostasis.

Rapid accumulation of boron (B) leads to toxicity in plant tissues, and the narrow gap between deficiency and toxicity makes it difficult to adjust essential B levels in soil for plant productivity. Therefore, understanding different aspects of B tolerance is necessary to provide new and valid solutions to B toxicity. Gypsophila perfoliata stands out as a remarkable example of a B-tolerant plant, with a natural propensity to thrive in environments such as B mines and soils enriched with high levels of B. In this study, a yeast functional screening experiment was conducted using cDNA libraries from G. perfoliata leaf and root cells for B tolerance. Ten colonies from the leaf library grew in 80 mm boric acid, while none emerged from the root library. Analysis of isolated cDNAs showed identical sequences and a unique motif related to B tolerance. The gene GpEF1A was identified in the tolerant yeast colonies, with predicted structural features suggesting its role, and RT-qPCR indicating increased expression under B stress. A regulatory role for EF1A lysine methylation was proposed in mammalian cells and fungi because of its dynamic and inducible nature under environmental constraints. This could also be relevant for plant cells, as the high similarity of the GpEF1A gene in some salt-tolerant plants might indicate the upregulation of EF1A as a conserved way to cope with abiotic stress conditions. This report represents the first instance of involvement of GpEF1A in B tolerance, and further detailed studies are necessary to understand other components of this tolerance mechanism.

Plants as effective bioindicators for heavy metal pollution monitoring

This study investigated the bioindicator potential of Amaranthus retroflexus L., Plantago lanceolata L., Rumex acetosa L., and Trifolium pratense L. including the use of Lolium multiflorum L. as a reference species, for heavy metal pollution monitoring, in particular Zinc (Zn), Cadmium (Cd), Nickel (Ni), and Lead (Pb). Controlled heavy metal contamination was applied through irrigation with metal nitrate solutions two levels of contamination (low and high). The study also focused on analyzing heavy metals concentration in plant tissues and related physiological responses. Distinct physiological responses to heavy metal stress were observed among the investigated species, highlighting unique variations in their reactions. Hydrogen peroxide, malondialdehyde content, and enzymatic activities emerged as reliable indicators of plant stress induced by heavy metal solutions. P. lanceolata displayed elevated Zn concentrations in both roots and leaves (3271 ± 337 and 4956 ± 82 mg kg−1). For Pb, L. multiflorum and P. lanceolata showed highest root concentrations (2964 ± 937 and 1605 ± 289 mg kg−1), while R. acetosa had higher leaf concentration (1957 ± 147 mg kg−1). For Ni, L. multiflorum had the highest root concentration (1148 ± 93 mg kg−1), and P. lanceolata exhibited the highest leaf concentration (2492 ± 28 mg kg−1). P. lanceolata consistently demonstrated the highest Cd concentrations in both roots (126 ± 21 mg kg−1) and leaves (163 ± 12 mg kg−1). These results provide valuable insights for selecting effective bioindicator species to establish control strategies for heavy metal pollution.

Pseudoplectania mystica (Ascomycota, Pezizales), a new cup fungus with an endophytic habit of a broad range of host plants

A cup fungus producing ascomata on senescing to dead rhizomes of bamboo in East China is described as Pseudoplectania mystica sp. nov. (Ascomycota, Pezizales) based on morphological, ecological and phylogenetic evidence. Some sequences of unidentified endophytic fungi uploaded to the GenBank database by other researchers were also identified as P. mystica in our phylogeny. By checking the original literature of these sequences, P. mystica was further found to act as an endophyte in a broad range of host plants including Aegiceras, Cinnamomum, Dendrobium and Lindera, probably lurking in the tissues of the host plants as invisible mycelium in most cases and only producing visible ascomata following senescence of the host tissues. Such ecology and a combination of all its morphological characteristics make P. mystica recognizable from other known species of Pseudoplectania. This study contributes to the understanding of the species diversity and lifecycle of Pseudoplectania.

#2034 CKD-MBD management: results from a national survey on bone turnover biomarkers practice and therapeutic strategies among Italian nephrologists

Abstract Background and Aims Advanced stages of chronic kidney disease (CKD) are constantly characterized by a mineral and bone disorders (MBD) syndrome concerning a complex systemic condition that includes laboratory abnormalities of bone and mineral metabolism involving calcium, phosphorus, parathyroid hormone (PTH), or vitamin D; abnormalities in bone turnover, mineralization, volume, or strength; extra-skeletal calcifications, such as vascular or other soft tissue. CKD-MBD syndrome determines relevant consequences in terms of fragility fractures, cardiovascular events and higher mortality. The nephrologist's awareness of CKD-MBD diagnostic and management tools and therapeutic strategies is key to improving CKD patients’ prognosis and outcomes. Method A new national survey (composed of 17 online questions) was conducted among Italian nephrologists to investigate the reference laboratory availability of bone turnover markers (BTMs), the clinical attitude on secondary hyperparathyroidism (sHPT) management, and the CKD-MBD therapeutic approach in different stages of CKD and dialysis patients. Results 89 Italian nephrologists participated in the survey. The reference laboratories largely fulfill the biomarkers request of ionized calcium (97.7%), phosphorus (100%), PTH (100%), alkaline phosphatase (ALP) (100%), magnesium (100%), 25-OH and 1,25-OH vitamin D levels (92.1%; 53.9%); while most of the hospital laboratories do not regularly support the availability of specific BTMs, both for diagnosis and monitoring the bone resorption and formation (Fig. 1)—such as bone ALP (75.3%), calcifediol (37.1%), calcitriol (40.5%), fibroblast growth factor-23 (FGF-23) (intact 9% and c-terminal 4.5%), vitamin K (33.7%), Klotho (6.75%, soluble 4.5%), osteocalcin (38.2%), matrix gla protein (10.1%), tartrate-resistant acid phosphatase 5b (TRAP-5b) (6.75%), cross-linked collagen type I peptide (CTX) (32.6%) and procollagen type 1 n-terminal propeptide (P1NP) (10.1%). The nephrologist's attitude to start management and treatment of sHPT is revealed to be mainly led by KDOQI (n = 42, 47%) and KDIGO (n = 39, 44%) guidelines considering their PTH cut-off values; in 53.9% of clinicians the measure of PTH levels is then performed every 3 months according to KDOQI guidelines. In relation to the definition of high turnover metabolic bone disease by BTMs, this metabolic pattern is identified in > 50% and 30-40% of patients with CKD 4-5D by 31.5% and 21.4% of nephrologists, respectively. 65% of clinicians currently considered ALP of equal importance as alterations of PTH as a predictor of fracture events. Other biomarkers, such as FGF-23, P1NP or TRAP-5b, emerge to not be still used in regular clinical practice. In patients diagnosed with CKD-MBD and skeletal fragility, 76.4% of nephrologists consider the AIFA (Italian Medicines Agency) note 79 to support osteoporosis treatment. The therapeutic strategies of CKD-MBD include the administration of vitamin D (e.g. cholecalciferol 27-37.1%, calcifediol 9-12.4%, calcitriol 42.7-55.1%) and its analogues (e.g. paricalcitol 23.6-52.8%) and the use of antiresorptive agents (e.g. alendronate 3.4%-23.6%, denosumab 22.5%-30.3%) with variable frequencies among different advanced CKD stages and for those patients on peritoneal or hemodialysis (Fig. 2). Conclusion Results present a suboptimal use of BTMs and a current heterogeneous therapeutic management of CKD-MBD syndrome and renal osteodystrophy in Italian clinical practice. The survey represents a starting point of work to further implement awareness of diagnostic and therapeutic issues of CKD-MBD.

Fabrication of a Triple-Layer Bionic Vascular Scaffold via Hybrid Electrospinning.

Tissue engineering aims to develop bionic scaffolds as alternatives to autologous vascular grafts due to their limited availability. This study introduces a novel wet-electrospinning fabrication technique to create small-diameter, uniformly aligned tubular scaffolds. By combining this innovative method with conventional electrospinning, a bionic tri-layer scaffold that mimics the zonal structure of vascular tissues is produced. The inner and outer layers consist of PCL/Gelatin and PCL/PLGA fibers, respectively, while the middle layer is crafted using PCL through Wet Vertical Magnetic Rod Electrospinning (WVMRE). The scaffold's morphology is analyzed using Scanning Electron Microscopy (SEM) to confirm its bionic structure. The mechanical properties, degradation profile, wettability, and biocompatibility of the scaffold are also characterized. To enhance hemocompatibility, the scaffold is crosslinked with heparin. The results demonstrate sufficient mechanical properties, good wettability of the inner layer, proper degradability of the inner and middle layers, and overall good biocompatibility. In conclusion, this study successfully develops a small-diameter tri-layer tubular scaffold that meets the required specifications.

First Report of Stem Canker Disease Caused by Neoscytalidium dimidiatum on Euphrates Poplar in Xinjiang, China.

Euphrates poplar (Populus euphratica Oliv.) constitutes about 61% of the global poplar population, thriving in arid regions of western China (Wu et al. 2023). It plays a crucial role in maintaining ecological balance, securing oasis agriculture, and driving socio-economic progress in the region. During a June 2023 investigation in the P. euphratica forest within the Hotan area of Xinjiang (37°20'21″N, 79°21'15″E), over 12% of the P. euphratica trees displayed branch withering symptoms. The affected trees exhibited cracked bark, trunk decay, darkened coloration, and an eventual black coal-smoke-like appearance. Fungal spores were notably present beneath peeling bark on trunks and main branches. The deep ulcers extended longitudinally into the cambium, leading to tree mortality. In some cases, lateral spread into the sapwood caused dark discoloration of vascular tissue. Twenty diseased branches from various locations were collected and 5-10 mm2 lesions were excised from the edges. These were then surface-disinfected with 75% ethanol for 30 s and 1% sodium hypochlorite for 2 min. After three rinses with sterile distilled water, excess moisture was removed using sterile filter paper, followed by incubating the samples on Potato Dextrose Agar (PDA) medium. Cultures were subsequently grown at 25 ± 1 ℃ under a 12-h photoperiod for three days, thus resulting in the isolation of 25 fungal strains with similar morphological characteristics. All strains displayed rapid colony growth (40 mm/d). On PDA medium, the mycelium initially presented as a white colony, transitioning to an olive-green to greyish color, finally turning dark-grey to black. Colonies generated mycelia that disintegrated into 0- to 1-septate, cylindrical to round, hyaline to brown arthroconidia, occurring singly or in arthric chains, averaging 8.9 ± 2.1 μm × 4.9 ± 1.3 μm, with a length/width ratio of 1.79. Based on morphological characteristics, the isolates were identified as Neoscytalidium dimidiatum (Penz.) Crous & Slippers (Crous et al. 2006). Molecular characterization involved amplifying the partial internal transcribed spacer (ITS) region and translation elongation factor 1-α (TEF1-α) and β-tubulin (TUB2) genes using ITS1/ITS4 (White et al. 1990), EF1-728F/EF1-986R (Carbone and Kohn 1999), and BT2a/BT2b primers (Glass and Donaldson 1995). Sequences, available in GenBank (ITS: PP033096, PP033097, PP033098; TUB2: PP032812, PP032813, PP032814; TEF1-α: PP032815, PP032816, PP032817), exhibited 99-100% identity with the epitype N. dimidiatum Arp2-D (ITS, MK813852; TUB2, MK816354; TEF1-α, MK816355). Phylogenetic analysis, employing maximum likelihood and Bayesian inference on concatenated ITS-TEF1-TUB, was constructed using IQ-Tree and MrBayes3.2.7, revealing isolates clustering within the N. dimidiatum clade. Three isolates (HY01, HY02, and HY05) from different collection points were chosen for pathogenic investigation. Pathogenicity testing on one-year-old healthy P. euphratica seedlings involved removing a 4-mm-diameter bark plug using a cork borer. A 3-day-cultured N. dimidiatum plug of the same size was inoculated, with a blank PDA as control. The wound was covered with moistened sterile absorbent cotton and finally sealed with parafilm for three days. Experiments were repeated thrice. Symptoms manifested by day 2 post-inoculation, resembling the original symptoms by day 7. In the control group, plants remained healthy. N. dimidiatum was exclusively re-isolated from lesions on inoculated stems, confirmed as N. dimidiatum through morphological characteristics and sequence analysis, aligning with Koch's hypothesis. To our knowledge, this is the first report of N. dimidiatum inducing stem canker on P. euphratica in China. This pathogen has been reported on many tree hosts including citrus (Alananbeh et al., 2020), common fig (Güney et al., 2022), dragon fruit (Salunkhe et al., 2023), and Almond (Nouri et al., 2018). Therefore our findings will serve as a warning for authorities to a potential threat in China's P. euphratica and other trees cultivation. Thus, further epidemiological studies are essential for devising effective management strategies.

Design and fabrication of dual-layer PCL nanofibrous scaffolds with inductive influence on vascular cell responses

Confronted with the profound threat of cardiovascular diseases to health, vascular tissue engineering presents potential beyond the limitations of autologous and allogeneic grafts, offering a promising solution. This study undertakes an initial exploration into the impact of a natural active protein, elastin, on vascular cell behavior, by incorporating with polycaprolactone to prepare fibrous tissue engineering scaffold. The results reveal that elastin serves to foster endothelial cell adhesion and proliferation, suppress smooth muscle cell proliferation, and induce macrophage polarization. Furthermore, the incorporation of elastin contributes to heightened scaffold strength, compliance, and elongation, concomitantly lowering the elastic modulus. Subsequently, a bilayer oriented polycaprolactone (PCL) scaffold infused with elastin is proposed. This design draws inspiration from the cellular arrangement of native blood vessels, leveraging oriented fibers to guide cell orientation. The resulting fiber scaffold exhibits commendable mechanical properties and cell infiltration capacity, imparting valuable insights for the rapid endothelialization of vascular scaffolds.