Effects of blood sample storage time, temperature, anti-coagulants and blood stabiliser on lymphocyte phenotyping.

Medical diagnostic laboratories have come under further scrutiny to ensure quality standards of their service and external quality assurance (EQA) programs involving multiple laboratories have been used to gauge this quality based on a consensus. However, because of the geographical distances within a country or internationally, cell surface marker expressions may change due to time delays and transport temperatures. Attention was given to this issue some decades ago and hence requires a re-evaluation in consideration of updated methods, reagents and instruments for flow cytometry and phenotyping. We have undertaken an extensive study to examine the effects of various conditions on blood storage akin to that experienced by patient samples as well as EQA programs, examining expression of lymphocyte surface markers, CD3, CD4, CD8, CD2, CD19, CD20, CD16/56 and HLA-DR. Assessment of lithium-heparin anticoagulated whole blood showed an increase in percentage of CD3+ and CD8+ T cells and a decrease in CD16/56+ NK cells after storage at room temperature (RT) for 24 and/or 48 h. In comparison, storage at 4°C led to a decrease in percentage of CD4+ and increase in percentage of CD8+ cells. The low temperature also caused an increase in percentage of B cells (CD19+, CD20+). While storage at RT did not alter levels of HLA-DR+ CD3+ T cells, there was a significant increase in percentage of these cells after 48 h. Changes were also seen at both temperatures when EDTA was used as an anti-coagulant. Assessment of blood treated with a stabiliser, normally used in the EQA samples (Streck Cell Preservative), reduced the range of lymphocyte subsets affected, with only CD2+ and CD20+ cells being significantly different at both temperatures, We conclude that 24-48 h storage/transport can affect the percentage of CD3+, CD4+ T cells, CD8+ T cells, B cells, NK cells and HLADR+ T cells which can be minimised by using the blood stabiliser as per EQA programs and we emphasise the need to adopt this in the processing of patients' blood samples.

Molecular and physiological responses to salt stress in salinity-sensitive and tolerant Hibiscus rosa-sinensis cultivars.

Ornamental plants are used to decorate urban and peri-urban areas, and during their cultivation or utilisation, they can be exposed to abiotic stress. Salinity is an abiotic stress factor that limits plant growth and reduces the ornamental value of sensitive species. In this study, transcriptomic analysis was conducted to identify genes associated with tolerance or sensitivity to salinity in two hibiscus (Hibiscus rosa-sinensis L.) cultivars, 'Porto' and 'Sunny wind'. The physiological and biochemical parameters of plants exposed to 50, 100, or 200 mM NaCl and water (control) were monitored. Salinity treatments were applied for six weeks. After four weeks, differences between cultivars were clearly evident and 'Porto' was more tolerant than 'Sunny wind'. The tolerant cultivar showed lower electrolyte leakage and ABA concentrations, and higher proline content in the leaves. Accumulation of Na in different organs was lower in the flower organs of 'Porto'. At the molecular level, several differential expressed genes were observed between the cultivars and flower organs. Among the highly expressed DEGs, coat protein, alcohol dehydrogenase, and AP2/EREBP transcription factor ERF-1. Among the downregulated genes, GH3 and NCED were the most interesting. The differential expression of these genes may explain the salt stress tolerance of 'Porto'.

Identification and expression profile of the SMAX/SMXL family genes in chickpea and lentil provide important players of biotechnological interest involved in plant branching.

MAIN CONCLUSION: SMAX/SMXL family genes were successfully identified and characterized in the chickpea and lentil and gene expression data revealed several genes associated with the modulation of plant branching and powerful targets for use in transgenesis and genome editing. Strigolactones (SL) play essential roles in plant growth, rooting, development, and branching, and are associated with plant resilience to abiotic and biotic stress conditions. Likewise, karrikins (KAR) are "plant smoke-derived molecules" that act in a hormonal signaling pathway similar to SL playing an important role in seed germination and hairy root elongation. The SMAX/SMXL family genes are part of these two signaling pathways, in addition to some of these members acting in a still little known SL- and KAR-independent signaling pathway. To date, the identification and functional characterization of the SMAX/SMXL family genes has not been performed in the chickpea and lentil. In this study, nine SMAX/SMXL genes were systematically identified and characterized in the chickpea and lentil, and their expression profiles were explored under different unstressless or different stress conditions. After a comprehensive in silico characterization of the genes, promoters, proteins, and protein-protein interaction network, the expression profile for each gene was determined using a meta-analysis from the RNAseq datasets and complemented with real-time PCR analysis. The expression profiles of the SMAX/SMXL family genes were very dynamic in different chickpea and lentil organs, with some genes assuming a tissue-specific expression pattern. In addition, these genes were significantly modulated by different stress conditions, indicating that SMAX/SMXL genes, although working in three distinct signaling pathways, can act to modulate plant resilience. Most CaSMAX/SMXL and partner genes such as CaTiE1 and CaLAP1, have a positive correlation with the plant branching level, while most LcSMAX/SMXL genes were less correlated with the plant branching level. The SMXL6, SMXL7, SMXL8, TiE1, LAP1, BES1, and BRC1 genes were highlighted as powerful targets for use in transgenesis and genome editing aiming to develop chickpea and lentil cultivars with improved architecture. Therefore, this study presented a detailed characterization of the SMAX/SMXL genes in the chickpea and lentil, and provided new insights for further studies focused on each SMAX/SMXL gene.

Biochemical and molecular changes in peach fruit exposed to cold stress conditions.

Storage or transportation temperature is very important for preserving the quality of fruit. However, low temperature in sensitive fruit such as peach can induce loss of quality. Fruit exposed to a specific range of temperatures and for a longer period can show chilling injury (CI) symptoms. The susceptibility to CI at low temperature varies among cultivars and genetic backgrounds. Along with agronomic management, appropriate postharvest management can limit quality losses. The importance of correct temperature management during postharvest handling has been widely demonstrated. Nowadays, due to long-distance markets and complex logistics that require multiple actors, the management of storage/transportation conditions is crucial for the quality of products reaching the consumer.Peach fruit exposed to low temperatures activate a suite of physiological, metabolomic, and molecular changes that attempt to counteract the negative effects of chilling stress. In this review an overview of the factors involved, and plant responses is presented and critically discussed. Physiological disorders associated with CI generally only appear after the storage/transportation, hence early detection methods are needed to monitor quality and detect internal changes which will lead to CI development. CI detection tools are assessed: they need to be easy to use, and preferably non-destructive to avoid loss of products.

Method for selecting ornamental species for different shading intensity in urban green spaces.

In urban areas, ornamental plants face different constraints, such as the shading of buildings and trees. Therefore, the selection of suitable species and their integration or combination with pre-existing plants is very important. Trees, shrubs, and herbaceous plant species must be distributed according to plant light requirements and shading intensity. Ornamental plants are classified into two groups based on their light intensity or shade tolerance: sun and shade species. To properly position the plants, especially in the immediate vicinity of buildings, it is necessary to study the projection of shadows during the year and the most critical periods, such as July and August. The position of ornamental species with different shading tolerances can be obtained by characterizing the leaf gas exchange for each species. Among the physiological parameters, the most important is the light compensation point, which is the light intensity corresponding to a net photosynthesis equal to zero. This means that the assimilation of carbon dioxide through photosynthesis is equal to the carbon dioxide emitted by respiration. This steady state represents the most critical condition for plants to endure the summer. The distribution of species inside a green area should be determined by considering the minimum light intensity that allows sufficient photosynthesis to compensate for the respiration rate. In this context, non-destructive leaf gas exchange, chlorophyll a fluorescence, and chlorophyll content can be useful tools for selecting suitable ornamental plants under diverse shading conditions.

New Strategies to Increase the Abiotic Stress Tolerance in Woody Ornamental Plants in Mediterranean Climate.

The native flora of different Mediterranean countries, often woody species, was widely recognized for its ornamental potential. The shrubs, in particular, are a typology of plants very widespread in the Mediterranean environment and constituent the 'Macchia', the typical vegetation of this ecosystem. The use of native shrubs for the realization of ornamental green areas has been recently examined for their adaptability to abiotic stress. Abiotic stresses, in fact, are the major limiting growth factor in urban and peri-urban areas. The identification and use of tolerant ornamental species allow the reduction of management costs and preserve the aesthetical value of green areas. Tolerance to drought stress, for instance, in the Mediterranean climate can improve the ecosystem services of these plants in the urban environment. In this review, the possibility to early individuate different plant species' mechanisms to tolerate or avoid the stresses is analysed, as well as the possibility to increase abiotic stress tolerance through genetic and agronomic strategies. The exploration of wild or spontaneous species can be a good source for selecting tolerant plants to be used as ornamental plants in urban areas. Among agronomic strategies, biostimulants, mulching, and plant combination can provide a useful solution to counteract abiotic stress in the urban environment.

The Role of Blue and Red Light in the Orchestration of Secondary Metabolites, Nutrient Transport and Plant Quality.

Light is a fundamental environmental parameter for plant growth and development because it provides an energy source for carbon fixation during photosynthesis and regulates many other physiological processes through its signaling. In indoor horticultural cultivation systems, sole-source light-emitting diodes (LEDs) have shown great potential for optimizing growth and producing high-quality products. Light is also a regulator of flowering, acting on phytochromes and inducing or inhibiting photoperiodic plants. Plants respond to light quality through several light receptors that can absorb light at different wavelengths. This review summarizes recent progress in our understanding of the role of blue and red light in the modulation of important plant quality traits, nutrient absorption and assimilation, as well as secondary metabolites, and includes the dynamic signaling networks that are orchestrated by blue and red wavelengths with a focus on transcriptional and metabolic reprogramming, plant productivity, and the nutritional quality of products. Moreover, it highlights future lines of research that should increase our knowledge to develop tailored light recipes to shape the plant characteristics and the nutritional and nutraceutical value of horticultural products.

Expression profile of the NCED/CCD genes in chickpea and lentil during abiotic stress reveals a positive correlation with increased plant tolerance.

Carotenoid cleavage dioxygenase (CCD) gene family is organized in two subfamilies: (i) 9-cis epoxycarotenoid dioxygenase (NCED) genes and (ii) CCD genes. NCED genes are essential for catalyzing the first step of the abscisic-acid (ABA) biosynthesis, while CCD genes produce precursors of the strigolactones hormone. The functional characterization of these gene subfamilies has not been yet performed in chickpea and lentil. Herein, were identified and systematically characterized two NCED and five CCD genes in the chickpea and two NCED and six CCD genes in lentil. After in silico sequence analysis and phylogeny, the expression profile of the NCED/CCD genes was determined by meta-analysis and real-time PCR in plants under different stress conditions. Sequence data revealed that NCED/CCD genes are highly conserved between chickpea and lentil. This conservation was observed both at gene and protein sequence levels and phylogenetic relationships. Analysis of the promoter sequences revealed that all NCED/CCD genes have a considerable number of cis-regulatory elements responsive to biotic and abiotic stress. Protein sequence analysis evidenced that NCED/CCD genes share several conserved motifs and that they have a highly interconnected interaction network. Furthermore, the three-dimensional structure of these proteins was determined and indicated that some proteins have structures with considerable similarity. The meta-analysis revealed that NCED/CCD genes are dynamically modulated in different organs and under different stress conditions, but they have a positive correlation with plant tolerance. In accordance, real-time PCR data showed that both NCED and CCD genes are differentially modulated in plants under drought stress. In particular, CaNCED2, CaCCD5, LcNCED2, LcCCD1, and LcCCD2 genes have a positive correlation with improved plant tolerance to drought stress. Therefore, this study presented a detailed characterization of the chickpea and lentil NCED/CCD genes and provided new insights to improve abiotic stress tolerance in these two important crops.

PKCζ activation promotes maturation of cord blood T cells towards a Th1 IFN-γ propensity.

A significant number of babies present transiently with low protein kinase C zeta (PKCζ) levels in cord blood T cells (CBTC), associated with reduced ability to transition from a neonatal Th2 to a mature Th1 cytokine bias, leading to a higher risk of developing allergic sensitisation, compared to neonates whose T cells have 'normal' PKCζ levels. However, the importance of PKCζ signalling in regulating their differentiation from a Th2 to a Th1 cytokine phenotype propensity remains undefined. To define the role of PKCζ signalling in the regulation of CBTC differentiation from a Th2 to a Th1cytokine phenotype we have developed a neonatal T cell maturation model which enables the cells to develop to CD45RA- /CD45RO+ T cells while maintaining the Th2 immature cytokine bias, despite having normal levels of PKCζ. The immature cells were treated with phytohaemagglutinin, but in addition with phorbol 12-myristate 13-acetate (PMA), an agonist which does not activate PKCζ. This was compared to development in CBTC in which the cells were transfected to express constitutively active PKCζ. The lack of PKCζ activation by PMA was monitored by western blot for phospho-PKCζ and translocation from cell cytosol to the membrane by confocal microscopy. The findings demonstrate that PMA fails to activate PKCζ in CBTC. The data show that CBTC matured under the influence of the PKC stimulator, PMA, maintain a Th2 cytokine bias, characterised by robust IL-4 and minimal interferon gamma production (IFN-γ), and lack of expression of transcriptional factor, T-bet. This was also reflected in the production of a range of other Th2/Th1 cytokines. Interestingly, introduction of a constitutively active PKCζ mutant into CBTC promoted development towards a Th1 profile with high IFN-γ production. The findings demonstrate that PKCζ signalling is essential for the immature neonatal T cells to transition from a Th2 to a Th1 cytokine production bias.

Preharvest melatonin foliar treatments enhance postharvest longevity of cut tuberose via altering physio-biochemical traits.

Introduction: Melatonin (MLT) is a bioactive molecule involved in the physiological functioning of plants. Reports related to preharvest applications of melatonin on the postharvest performance of cut flowers are not available in the literature. Materials & methods: This study evaluated the effects of different concentrations of exogenous MLT [0 mM (MT0), 0.5 mM (MT1), 0.7 mM (MT2), 1 mM (MT3)] applied preharvest on the physiological characteristics and postharvest performance of cut tuberose, a globally demanded cut flower. Results & discussion: The results revealed that all treatments increased postharvest vase life by up to 4 d. The MT1, MT2, and MT3 treatments increased total soluble proteins (TSP) by 25%, 41%, and 17%, soluble sugars (SS) by 21%, 36%, and 33%, an+d postharvest catalase (CAT) activity by 52%, 66%, and 70%, respectively. Malondialdehyde (MDA) and hydrogen peroxide (H2O2) decreased in all preharvest treatments by up to 23% and 56%, respectively. Proline concentration decreased in all treatments, particularly MT3 (38%). These findings suggest that preharvest MLT treatment is a promising strategy for improving the postharvest quality of cut tuberose.

LED Lighting to Produce High-Quality Ornamental Plants.

The flexibility of LED technology, in terms of energy efficiency, robustness, compactness, long lifetime, and low heat emission, as well as its applications as a sole source or supplemental lighting system, offers interesting potential, giving the ornamental industry an edge over traditional production practices. Light is a fundamental environmental factor that provides energy for plants through photosynthesis, but it also acts as a signal and coordinates multifaceted plant-growth and development processes. With manipulations of light quality affecting specific plant traits such as flowering, plant architecture, and pigmentation, the focus has been placed on the ability to precisely manage the light growing environment, proving to be an effective tool to produce tailored plants according to market request. Applying lighting technology grants growers several productive advantages, such as planned production (early flowering, continuous production, and predictable yield), improved plant habitus (rooting and height), regulated leaf and flower color, and overall improved quality attributes of commodities. Potential LED benefits to the floriculture industry are not limited to the aesthetic and economic value of the product obtained; LED technology also represents a solid, sustainable option for reducing agrochemical (plant-growth regulators and pesticides) and energy inputs (power energy).

Biofortification of baby leafy vegetables using nutrient solution containing selenium.

BACKGROUND: Biofortification of vegetables is an important innovation technique in the horticultural sector. Vegetables can be a vector of different minor elements that have beneficial effects on human health. Selenium (Se) is an important element for human nutrition and plays a significant role in defence mechanisms. The aim of this work was to investigate the effect of Se in the nutrient solutions on the crop biofortification ability, yield, and quality parameters of four baby leafy vegetables destined to the minimally processed industry. Experiments were performed on lamb's lettuce, lettuce, wild rocket, and spinach. These crops were cultivated in the floating systems with nutrient solution enriched with 0, 2.6, 3.9, and 5.2 µmol L-1 Se provided as sodium selenate. RESULTS: At harvest, Se concentrations, yield, nitrate concentration, sugars, and some mineral elements were measured. Data collected and analyses showed that yield, nitrate, sucrose, and reducing sugars were not affected by Se treatments, even if varied among species. Se concentrations linearly increased in leaves of different species by increasing the Se concentration in the nutrient solution. Rocket was the species with the highest accumulation ability and reached a concentration of 11 µg g-1 fresh weight Se in plants grown with 5.2 µmol L-1 Se. CONCLUSION: A floating system with Se-enriched nutrient solution is an optimal controlled growing biofortification system for leafy vegetables. The accumulation ability decreased in different species in the order wild rocket, spinach, lettuce, and lamb's lettuce, highlighting a crop-dependent behaviour and their attitude to biofortification. © 2023 The Authors. Journal of The Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Molecular Responses of Vegetable, Ornamental Crops, and Model Plants to Salinity Stress.

Vegetable and ornamental plants represent a very wide group of heterogeneous plants, both herbaceous and woody, generally without relevant salinity-tolerant mechanisms. The cultivation conditions-almost all are irrigated crops-and characteristics of the products, which must not present visual damage linked to salt stress, determine the necessity for a deep investigation of the response of these crops to salinity stress. Tolerance mechanisms are linked to the capacity of a plant to compartmentalize ions, produce compatible solutes, synthesize specific proteins and metabolites, and induce transcriptional factors. The present review critically evaluates advantages and disadvantages to study the molecular control of salt tolerance mechanisms in vegetable and ornamental plants, with the aim of distinguishing tools for the rapid and effective screening of salt tolerance levels in different plants. This information can not only help in suitable germplasm selection, which is very useful in consideration of the high biodiversity expressed by vegetable and ornamental plants, but also drive the further breeding activities.

Can C-reactive protein be used as a surrogate marker of IL-6 in a broad array of clinical entities?

Background: C-reactive protein (CRP) is commonly performed, whereas cytokine testing is limited to research. Aims: To determine CRP correlation to cytokines IL-6, IL-1ß and TNF-α. Results: Consecutive samples (n = 307) were collected over 24 h. Ninety-six patients (31%) had acute infections, and 23 patients (7.5%) had autoimmune or inflammatory disease presentations. A strong correlation between CRP and two IL-6 assays (r = 0.74 and r = 0.71; p < 0.001) was present. CRP did not correlate with IL-1ß and TNF-α across the data set. Bacterial infection had a significantly higher CRP and IL-6 (p < 0.001), while only CRP was elevated in inflammatory and autoimmune diseases (p < 0.001). Discussion: CRP may be used as a surrogate marker of IL-6 levels in the routine diagnostic laboratories.

Biochar: An emerging recipe for designing sustainable horticulture under climate change scenarios.

The interest in sustainable horticulture has recently increased, given anthropogenic climate change. The increasing global population will exacerbate the climate change situation induced by human activities. This will elevate global food demands and the vulnerability of horticultural systems, with severe concerns related to natural resource availability and usage. Sustainable horticulture involves adopting eco-friendly strategies to boost yields while maintaining environmental conservation. Biochar (BC), a carbon-rich material, is widely used in farming to improve soil physical and chemical properties and as an organic substitute for peat in growing media. BC amendments to soil or growing media improve seedling growth, increase photosynthetic pigments, and enhances photosynthesis, thus improving crop productivity. Soil BC incorporation improves abiotic and biotic stress tolerance, which are significant constraints in horticulture. BC application also improves disease control to an acceptable level or enhance plant resistance to pathogens. Moreover, BC amendments in contaminated soil decrease the uptake of potentially hazardous metals, thus minimizing their harmful effects on humans. This review summarizes the most recent knowledge related to BC use in sustainable horticulture. This includes the effect of BC on enhancing horticultural crop production and inducing resistance to major abiotic and biotic stresses. It also discuss major gaps and future directions for exploiting BC technology.

Comparative transcriptomic profiling of peach and nectarine cultivars reveals cultivar-specific responses to chilled postharvest storage.

Introduction: Peach (Prunus persica (L.) Batsch,) and nectarine fruits (Prunus persica (L.) Batsch, var nectarine), are characterized by a rapid deterioration at room temperature. Therefore, cold storage is widely used to delay fruit post-harvest ripening and extend fruit commercial life. Physiological disorders, collectively known as chilling injury, can develop typically after 3 weeks of low-temperature storage and affect fruit quality. Methods: A comparative transcriptomic analysis was performed to identify regulatory pathways that develop before chilling injury symptoms are detectable using next generation sequencing on the fruits of two contrasting cultivars, one peach (Sagittaria) and one nectarine, (Big Top), over 14 days of postharvest cold storage. Results: There was a progressive increase in the number of differentially expressed genes between time points (DEGs) in both cultivars. More (1264) time point DEGs were identified in 'Big Top' compared to 'Sagittaria' (746 DEGs). Both cultivars showed a downregulation of pathways related to photosynthesis, and an upregulation of pathways related to amino sugars, nucleotide sugar metabolism and plant hormone signal transduction with ethylene pathways being most affected. Expression patterns of ethylene related genes (including biosynthesis, signaling and ERF transcription factors) correlated with genes involved in cell wall modification, membrane composition, pathogen and stress response, which are all involved later during storage in development of chilling injury. Discussion: Overall, the results show that common pathways are activated in the fruit of 'Big Top' nectarine and 'Sagittaria' peach in response to cold storage but include also differences that are cultivar-specific responses.

Fruitomics: The Importance of Combining Sensory and Chemical Analyses in Assessing Cold Storage Responses of Six Peach (Prunus persica L. Batsch) Cultivars.

Cold storage is used to extend peach commercial life, but can affect quality. Quality changes are assessed through the content of nutritionally relevant compounds, aroma, physical characters and/or sensorially. Here, six peach and nectarine cultivars were sampled at commercial harvest and after 7 days of 1 °C storage. A trained panel was used to evaluate sensorial characters, while carotenoids, phenolics, vitamin C, total sugars, and qualitative traits including firmness, titrable acidity and soluble solid content were integrated with volatile organic compound (VOC) analysis previously reported. The different analyses reveal interesting patterns of correlation, and the six cultivars responded differently to cold storage. Sensory parameters were correlated with 64 VOCs and seven intrinsic characters. Acidity, firmness, and 10 VOCs were strongly negatively correlated with harmony and sweetness, but positively correlated with bitterness, astringency, and crunchiness. In contrast, Brix, b-carotene, and six VOCs were positively correlated with harmony and sweetness.

Chemical priming enhances plant tolerance to salt stress.

Salt stress severely limits the productivity of crop plants worldwide and its detrimental effects are aggravated by climate change. Due to a significant world population growth, agriculture has expanded to marginal and salinized regions, which usually render low crop yield. In this context, finding methods and strategies to improve plant tolerance against salt stress is of utmost importance to fulfill food security challenges under the scenario of the ever-increasing human population. Plant priming, at different stages of plant development, such as seed or seedling, has gained significant attention for its marked implication in crop salt-stress management. It is a promising field relying on the applications of specific chemical agents which could effectively improve plant salt-stress tolerance. Currently, a variety of chemicals, both inorganic and organic, which can efficiently promote plant growth and crop yield are available in the market. This review summarizes our current knowledge of the promising roles of diverse molecules/compounds, such as hydrogen sulfide (H2S), molecular hydrogen, nitric oxide (NO), hydrogen peroxide (H2O2), melatonin, chitosan, silicon, ascorbic acid (AsA), tocopherols, and trehalose (Tre) as potential primers that enhance the salinity tolerance of crop plants.