Knowledge gaps on grape sour rot inferred from a systematic literature review.

Sour rot (SR) is one of the major diseases affecting grapevine berries, causing severe yield losses and deterioration of wine quality. SR is caused by an etiologic complex of microorganisms, including yeasts, bacteria, and filamentous fungi. This systematic review focuses on the etiology, epidemiology, and control of SR. A total of 74 papers published between 1986 and 2023 were assessed in this review. Description of disease symptoms was quite consistent across the papers, including oxidation of the grape skin, disaggregation of the internal tissues, and detachment of the rotten berries from the pedicel. The affected bunches are characterized by the smell of acetic acid and ethyl acetate that attracts fruit flies (Drosophila spp.). However, several knowledge gaps and/or inconsistencies were identified with respect to SR etiology, epidemiology, and control. Overall, 146 microorganisms were isolated from the affected berries (44.5% yeasts, 34.3% bacteria, and 21.2% filamentous fungi); however, the selected papers could not definitively clarify which species are primarily involved in the etiology of the disease. A general inconsistency was also observed in the methods used to assess the incidence and severity of SR in vineyards, making inter-study comparisons extremely challenging. Inconsistencies were also found in the methods used for pathogenicity assessment in artificial inoculation studies. Furthermore, gaps were detected in terms of SR epidemiology, with a focus on environmental conditions affecting the disease development. The SR management options are limited, and efficacy trials often result in poor, variable, and inconsistent levels of control, which might be attributed to the lack of knowledge on disease epidemiology. These knowledge gaps and inconsistencies were analyzed in this review to inform future research activities.

Impact of Inducible Nitric Oxide Synthase Activation on Endothelial Behavior under Magnesium Deficiency.

Endothelial dysfunction is a crucial event in the early pathogenesis of cardiovascular diseases and is linked to magnesium (Mg) deficiency. Indeed, in endothelial cells, low Mg levels promote the acquisition of a pro-inflammatory and pro-atherogenic phenotype. This paper investigates the mechanisms by which Mg deficiency promotes oxidative stress and affects endothelial behavior in human umbilical vascular endothelial cells (HUVECs). Our data show that low Mg levels trigger oxidative stress initially by increasing NAPDH oxidase activity and then by upregulating the pro-oxidant thioredoxin-interacting protein TXNIP. The overproduction of reactive oxygen species (ROS) activates NF-κB, leading to its increased binding to the inducible nitric oxide synthase (iNOS) promoter, with the consequent increase in iNOS expression. The increased levels of nitric oxide (NO) generated by upregulated iNOS contribute to disrupting endothelial cell function by inhibiting growth and increasing permeability. In conclusion, we provide evidence that multiple mechanisms contribute to generate a pro-oxidant state under low-Mg conditions, ultimately affecting endothelial physiology. These data add support to the notion that adequate Mg levels play a significant role in preserving cardiovascular health and may suggest new approaches to prevent or manage cardiovascular diseases.

Integration of mathematical modeling and target-based application of biocontrol agents for the control of Botrytis cinerea in vineyards.

BACKGROUND: Biocontrol agents (BCAs) are alternatives to synthetic fungicides with low risk to the environment and human health. Although several studies on the biocontrol of gray mold in vineyards have been performed, it is necessary to improve the usage of BCAs in fields conditions. Therefore, in the present study, BCAs were used both in calendar-based [based on four growth stages (GSs), i.e., flowering, pre-bunch closure, veraison, and before harvest] and predictive model-based strategies (only when Botrytis cinerea infection risk was predicted by the model). The BCAs applied during the seasons were selected considering the grapevine GSs. Treatments performed with BCAs were compared with synthetic fungicide treatments and an untreated control. The trials were conducted in three experimental vineyards with four epidemics. To evaluate the level of gray mold control of each treatment, disease severity was assessed at harvest and the presence of latent infection was evaluated. RESULTS: The integrative use of the predictive model and BCAs provided satisfactory levels of gray mold control, with gray mold severity levels significantly lower (P < 0.001) than those of the untreated control, which had severity values (< 7%) similar to those observed with synthetic fungicides following both calendar and model-based strategies. CONCLUSIONS: The integrative use of the predictive model and BCAs represents a valid alternative to conventional methods of gray mold control in vineyards, with more than 75% reduction in fungicide usage. © 2024 Society of Chemical Industry.

Biocontrol of Botrytis cinerea as Influenced by Grapevine Growth Stages and Environmental Conditions.

The growth of four commercial biocontrol agents (BCAs: Bacillus amyloliquefaciens (BAD), Aureobasidium pullulans (APD), Metschnikowia fructicola (MFN), and Trichoderma atroviride (TAS)) was evaluated using turbidimetric assays on artificial substrates mimicking the chemical berry composition at four stages: pea-sized berries, veraison, softening, and ripe berries. The response of BCA growth differed among BCAs. Subsequently, the BCAs' population size was assessed after 1 to 13 days of incubation on the substrate mimicking ripe berries at 15 to 35 °C. The population size of BAD increased with temperatures, while that of MFN decreased; the population sizes of APD and TAS showed bell-shaped patterns with lower growth at 15 or 35 °C. Finally, the BCAs were applied to ripe berries and then incubated at 15 to 30 °C. After 1 to 13 days, the berries were inoculated with B. cinerea and incubated for 7 days, after which the BCA control efficacy was assessed. The highest control was observed at 25 °C for BAD and APD, at 15 to 20 °C for MFN, and at 25 to 30 °C for TAS. The results confirm that the plant substrates and temperature affect the population size of the BCA following application; temperature also affects the preventative efficacy of BCA against B. cinerea.

The Role of Txnip in Mediating Low-Magnesium-Driven Endothelial Dysfunction.

Magnesium deficiency is associated with a greater risk of developing cardiovascular diseases since this cation is fundamental in regulating vascular function. This clinical evidence is sustained by in vitro studies showing that culturing endothelial cells in low concentrations of magnesium promotes the acquisition of a pro-oxidant and pro-inflammatory phenotype. Here, we show that the increase in reactive oxygen species in endothelial cells in low-magnesium-containing medium is due to the upregulation of the pro-oxidant protein thioredoxin interacting protein (TXNIP), with a consequent accumulation of lipid droplets and increase in endothelial permeability through the downregulation and relocalization of junctional proteins. Silencing TXNIP restores the endothelial barrier and lipid content. Because (i) mitochondria serve multiple roles in shaping cell function, health and survival and (ii) mitochondria are the main intracellular stores of magnesium, it is of note that no significant alterations were detected in their morphology and dynamics in our experimental model. We conclude that TXNIP upregulation contributes to low-magnesium-induced endothelial dysfunction in vitro.

The Interplay between TRPM7 and MagT1 in Maintaining Endothelial Magnesium Homeostasis.

The transient receptor potential cation channel subfamily M member 7 (TRPM7) is an ubiquitous channel fused to an α-kinase domain involved in magnesium (Mg) transport, and its level of expression has been proposed as a marker of endothelial function. To broaden our present knowledge about the role of TRPM7 in endothelial cells, we generated stable transfected Human Endothelial Cells derived from the Umbilical Vein (HUVEC). TRPM7-silencing HUVEC maintain the actin fibers' organization and mitochondrial network. They produce reduced amounts of reactive oxygen species and grow faster than controls. Intracellular Mg concentration does not change in TRPM7-silencing or -expressing HUVEC, while some differences emerged when we analyzed intracellular Mg distribution. While the levels of the plasma membrane Mg transporter Solute Carrier family 41 member 1 (SLC41A1) and the mitochondrial channel Mrs2 remain unchanged, the highly selective Magnesium Transporter 1 (MagT1) is upregulated in TRPM7-silencing HUVEC through transcriptional regulation. We propose that the increased amounts of MagT1 grant the maintenance of intracellular Mg concentrations when TRPM7 is not expressed in endothelial cells.

Efficacy of preharvest application of biocontrol agents against gray mold in grapevine.

The use of biocontrol agents (BCAs) represents a promising alternative to conventional methods for the management of gray mold in vineyards during the berry ripening stage. The main advantages of BCAs are the short preharvest interval and lack of chemical fungicide residues in wine. In this study, eight commercial BCAs (based on different Bacillus or Trichoderma species and strains, Aureobasidium pullulans, Metschnikowia fructicola, and Pythium oligandrum) and a reference fungicide (boscalid) were applied to a vineyard during berry ripening over three seasons to evaluate the dynamics over time in terms of their relative efficacies in gray mold control. At 1-13 days after application of BCAs to the berry surfaces in field conditions, the berries were collected and artificially inoculated with conidia of Botrytis cinerea under controlled laboratory conditions, and gray mold severity was observed after 7 days of incubation. Significant differences were observed in gray mold severity among years, according to the number of days the BCAs grew on the berry surface before B. cinerea inoculation, and the season by day interaction (altogether accounting for >80% of the experimental variance). The variability in BCA efficacy was closely related to the environmental conditions at the time of application and in the following days. Overall, the BCA efficacy increased with the degree days accumulated between BCA application in the vineyard and B. cinerea inoculation in the dry (no rain) periods (r = 0.914, P = 0.001). Rainfall and the associated drop in temperature caused a relevant reduction of BCA efficacy. These results demonstrate that BCAs are an effective alternative to conventional chemicals for the preharvest control of gray mold in vineyards. However, environmental conditions can considerably affect the BCA efficacy.

The Effects of Sirolimus and Magnesium on Primary Human Coronary Endothelial Cells: An In Vitro Study.

Drug eluting magnesium (Mg) bioresorbable scaffolds represent a novel paradigm in percutaneous coronary intervention because Mg-based alloys are biocompatible, have adequate mechanical properties and can be resorbed without adverse events. Importantly, Mg is fundamental in many biological processes, mitigates the inflammatory response and is beneficial for the endothelium. Sirolimus is widely used as an antiproliferative agent in drug eluting stents to inhibit the proliferation of smooth muscle cells, thus reducing the occurrence of stent restenosis. Little is known about the potential interplay between sirolimus and Mg in cultured human coronary artery endothelial cells (hCAEC). Therefore, the cells were treated with sirolimus in the presence of different concentrations of extracellular Mg. Cell viability, migration, barrier function, adhesivity and nitric oxide synthesis were assessed. Sirolimus impairs the viability of subconfluent, but not of confluent cells independently from the concentration of Mg in the culture medium. In confluent cells, sirolimus inhibits migration, while it cooperates with Mg in exerting an anti-inflammatory action that might have a role in preventing restenosis and thrombosis.

Plant Disease Models and Forecasting: Changes in Principles and Applications over the Last 50 Years.

This review gives a perspective of selected advances made since the middle of the 20th century in plant disease modeling, and the associated increase in the number of models published during that time frame. This progress can be mainly attributed to advances in (i) sensors and automatic environmental data collection technology, (ii) instrumentation and methods for studying botanical epidemiology, and (iii) data analytics and computer science. We review the evolution of techniques for developing data-based (empirical) models and process-based (mechanistic) models using the wheat rusts as a case study. We also describe the increased importance of knowledge about biological processes for plant disease modeling by using apple scab as a second case study. For both wheat rusts and apple scab, we describe how the models have evolved over the last 50 years by considering certain milestones that have been achieved in disease modeling. Finally, we describe how plant disease models are used as part of a multi-modeling approach to develop decision-making tools in the application of integrated pest management.

The presence of BBB hastens neuronal differentiation of cerebral organoids - The potential role of endothelial derived BDNF.

Despite remaining the best in vitro model to resemble the human brain, a weakness of human cerebral organoids is the lack of the endothelial component that in vivo organizes in the blood brain barrier (BBB). Since the BBB is crucial to control the microenvironment of the nervous system, this study proposes a co-culture of BBB and cerebral organoids. We utilized a BBB model consisting of primary human brain microvascular endothelial cells and astrocytes in a transwell system. Starting from induced Pluripotent Stem Cells (iPSCs) we generated human cerebral organoids which were then cultured in the absence or presence of an in vitro model of BBB to evaluate potential effects on the maturation of cerebral organoids. By morphological analysis, it emerges that in the presence of the BBB the cerebral organoids are better organized than controls in the absence of the BBB. This effect might be due to Brain Derived Neurotrophic Factor (BDNF), a neurotrophic factor released by the endothelial component of the BBB, which is involved in neurodevelopment, neuroplasticity and neurosurvival.

The Presence of Blood-Brain Barrier Modulates the Response to Magnesium Salts in Human Brain Organoids.

Magnesium (Mg) is fundamental in the brain, where it regulates metabolism and neurotransmission and protects against neuroinflammation. To obtain insights into the molecular basis of Mg action in the brain, we investigated the effects of Mg in human brain organoids, a revolutionary 3D model to study neurobiology and neuropathology. In particular, brain organoids derived from human induced pluripotent stem cells were cultured in the presence or in the absence of an in vitro-generated blood-brain barrier (BBB), and then exposed to 1 or 5 mM concentrations of inorganic and organic Mg salts (Mg sulphate (MgSO4); Mg pidolate (MgPid)). We evaluated the modulation of NMDA and GABAergic receptors, and BDNF. Our data suggest that the presence of the BBB is essential for Mg to exert its effects on brain organoids, and that 5 mM of MgPid is more effective than MgSO4 in increasing the levels of GABA receptors and BDNF, and decreasing those of NMDA receptor. These results might illuminate novel pathways explaining the neuroprotective role of Mg.

A Systematic Map of the Research on Disease Modelling for Agricultural Crops Worldwide.

In this work, we developed a systematic map to identify and catalogue the literature pertaining to disease modelling for agricultural crops worldwide. Searches were performed in 2021 in the Web of Science and Scopus for papers reporting any type of disease model for 103 crops. In total, 768 papers were retrieved, and their descriptive metadata were extracted. The number of papers found increased from the mid-1900s to 2020, and most of the studies were from North America and Europe. More disease models were retrieved for wheat, potatoes, grapes, and apples than for other crops; the number of papers was more affected by the crop's economic value than by its cultivated area. The systematic map revealed an underrepresentation of disease models for maize and rice, which is not justified by either the crop economic value or by disease impact. Most of the models were developed to understand the pathosystem, and fewer were developed for tactical disease management, strategic planning, or scenario analysis. The systematic map highlights a variety of knowledge gaps and suggests questions that warrant further research.

Magnesium and the Brain: A Focus on Neuroinflammation and Neurodegeneration.

Magnesium (Mg) is involved in the regulation of metabolism and in the maintenance of the homeostasis of all the tissues, including the brain, where it harmonizes nerve signal transmission and preserves the integrity of the blood-brain barrier. Mg deficiency contributes to systemic low-grade inflammation, the common denominator of most diseases. In particular, neuroinflammation is the hallmark of neurodegenerative disorders. Starting from a rapid overview on the role of magnesium in the brain, this narrative review provides evidences linking the derangement of magnesium balance with multiple sclerosis, Alzheimer's, and Parkinson's diseases.

A Comparison of Doxorubicin-Resistant Colon Cancer LoVo and Leukemia HL60 Cells: Common Features, Different Underlying Mechanisms.

Chemoresistance causes cancer relapse and metastasis, thus remaining the major obstacle to cancer therapy. While some light has been shed on the underlying mechanisms, it is clear that chemoresistance is a multifaceted problem strictly interconnected with the high heterogeneity of neoplastic cells. We utilized two different human cell lines, i.e., LoVo colon cancer and promyelocytic leukemia HL60 cells sensitive and resistant to doxorubicin (DXR), largely used as a chemotherapeutic and frequently leading to chemoresistance. LoVo and HL60 resistant cells accumulate less reactive oxygen species by differently modulating the levels of some pro- and antioxidant proteins. Moreover, the content of intracellular magnesium, known to contribute to protect cells from oxidative stress, is increased in DXR-resistant LoVo through the upregulation of MagT1 and in DXR-resistant HL60 because of the overexpression of TRPM7. In addition, while no major differences in mitochondrial mass are observed in resistant HL60 and LoVo cells, fragmented mitochondria due to increased fission and decreased fusion are detected only in resistant LoVo cells. We conclude that DXR-resistant cells evolve adaptive mechanisms to survive DXR cytotoxicity by activating different molecular pathways.

High Magnesium and Sirolimus on Rabbit Vascular Cells-An In Vitro Proof of Concept.

Drug-eluting bioresorbable scaffolds represent the last frontier in the field of angioplasty and stenting to treat coronary artery disease, one of the leading causes of morbidity and mortality worldwide. In particular, sirolimus-eluting magnesium-based scaffolds were recently introduced in clinical practice. Magnesium alloys are biocompatible and dissolve in body fluids, thus determining high concentrations of magnesium in the local microenvironment. Since magnesium regulates cell growth, we asked whether high levels of magnesium might interfere with the antiproliferative action of sirolimus. We performed in vitro experiments on rabbit coronary artery endothelial and smooth muscle cells (rCAEC and rSMC, respectively). The cells were treated with sirolimus in the presence of different concentrations of extracellular magnesium. Sirolimus inhibits rCAEC proliferation only in physiological concentrations of magnesium, while high concentrations prevent this effect. On the contrary, high extracellular magnesium does not rescue rSMC growth arrest by sirolimus and accentuates the inhibitory effect of the drug on cell migration. Importantly, sirolimus and magnesium do not impair rSMC response to nitric oxide. If translated into a clinical setting, these results suggest that, in the presence of sirolimus, local increases of magnesium concentration maintain normal endothelial proliferative capacity and function without affecting rSMC growth inhibition and response to vasodilators.

A Weather-Driven Model for Predicting Infections of Grapevines by Sporangia of Plasmopara viticola.

A mechanistic model was developed to predict secondary infections of Plasmopara viticola and their severity as influenced by environmental conditions; the model incorporates the processes of sporangia production and survival on downy mildew (DM) lesions, dispersal and deposition, and infection. The model was evaluated against observed data (collected in a 3-year vineyard) for its accuracy to predict periods with no sporangia (i.e., for negative prognosis) or with peaks of sporangia, so that growers can identify periods with no/low risk or high risk. The model increased the probability to correctly predict no sporangia [P(P-O-) = 0.67] by two times compared to the prior probability, with fewer than 3% of the total sporangia found in the vineyard being sampled when not predicted by the model. The model also correctly predicted peaks of sporangia, with only 1 of 40 peaks unpredicted. When evaluated for the negative prognosis of infection periods, the model showed a posterior probability for infection not to occur when not predicted P(P-O-) = 0.87 with only 9 of 108 real infections not predicted; these unpredicted infections were mild, accounting for only 4.4% of the total DM lesions observed in the vineyard. In conclusion, the model was able to identify periods in which the DM risk was nil or very low. It may, therefore, help growers avoid fungicide sprays when not needed and lengthen the interval between two sprays, i.e., it will help growers move from calendar-based to risk-based fungicide schedules for the control of P. viticola in vineyards.

Magnesium Deficiency Induces Lipid Accumulation in Vascular Endothelial Cells via Oxidative Stress-The Potential Contribution of EDF-1 and PPARγ.

BACKGROUND: Magnesium deficiency contributes to atherogenesis partly by promoting the dysfunction of endothelial cells, which are critical in vascular homeostasis and diseases. Since EDF-1 and PPARγ regulate crucial endothelial activities, we investigated the modulation of these proteins involved in lipogenesis as well the deposition of lipids in human endothelial cells cultured in different concentrations of magnesium. METHODS: Human endothelial cells from the umbilical vein were cultured in medium containing from 0.1 to 5 mM magnesium for 24 h. The levels of EDF-1 and PPARγ were visualized by Western blot. Reactive oxygen species (ROS) were measured by DCFDA. Lipids were detected after O Red Oil staining. RESULTS: Magnesium deficiency leads to the accumulation of ROS which upregulate EDF-1. Further, PPARγ is increased after culture in low magnesium, but independently from ROS. Moreover, lipids accumulate in magnesium-deficient cells. CONCLUSIONS: Our results suggest that magnesium deficiency leads to the deposition of lipids by inducing EDF-1 and PPARγ. The increase in intracellular lipids might be interpreted as an adaptive response of endothelial cells to magnesium deficiency.

Use of LAMP for Assessing Botrytis cinerea Colonization of Bunch Trash and Latent Infection of Berries in Grapevines.

A real-time loop-mediated isothermal amplification (LAMP) assay was evaluated for the detection of Botrytis cinerea in grapevine bunch trash, immature berries, and ripening berries. A simple method for the preparation of crude extracts of grape tissue was also developed for on-site LAMP analysis. When tested with 14 other fungal species frequently found in grapevines, the LAMP assay was specific and sensitive to a B. cinerea DNA quantity of 0.1 ng/µL. The sensitivity was further tested using bunch trash samples with B. cinerea colonization levels between 6 and 100% and with bulk-berry samples composed of 4 pathogen-free berries or 4 berries among which 25 to 100% had been inoculated with B. cinerea. The LAMP assay detected the lowest B. cinerea colonization level tested in bunch trash and in immature and mature berries in less than 20 min. In single-berry experiments, LAMP amplified B. cinerea DNA from all artificially inoculated individual immature and mature berries. No amplification occurred in B. cinerea-free material. The real-time LAMP assay has the potential to be used as a rapid on-site diagnostic tool for assessing B. cinerea colonization in bunch trash and B. cinerea latent infections in berries, which represent critical stages for decision-making about disease management.

Biocontrol of Botrytis cinerea on Grape Berries as Influenced by Temperature and Humidity.

Six commercial biocontrol agents (BCAs, containing Aureobasidium pullulans, Bacillus amyloliquefaciens, Bacillus amyloliquefaciens plantarum, Bacillus subtilis, Pythium oligandrum, or Trichoderma atroviride) were applied to ripening berries that were then incubated at one of four temperatures (T, 15, 20, 25, and 30°C) and one of four relative humidity levels (RH, 60, 80, 90, and 100%). After 1 to 13 days of incubation (BCA colonization period), the berries were inoculated with conidia of Botrytis cinerea and kept at 25°C and 100% RH for 7 days, at which time Botrytis bunch rot (BBR) was assessed. The response of BBR control to T/RH conditions and BCA colonization period differed among BCAs; the coefficients of variation among the BCAs ranged from 44.7 to 72.4%. An equation was developed that accounted for the combined effects of T, RH, and BCA colonization period on BBR control. The equation, which had an R2>0.94, could help farmers select the BCA to be used for a specific application based on weather conditions at the time of treatment and in the following days.

Consideration of Latent Infections Improves the Prediction of Botrytis Bunch Rot Severity in Vineyards.

The current study validated a mechanistic model for Botrytis cinerea on grapevine with data from 23 independent Botrytis bunch rot (BBR) epidemics (combinations of vineyards × year) that occurred between 1997 and 2018 in Italy, France, and Spain. The model was operated for each vineyard by using weather data and vine growth stages to anticipate, at any day of the vine-growing season, the disease severity (DS) at harvest (severe, DS ≥ 15%; intermediate, 5 < DS < 15%; and mild, DS ≤ 5%). To determine the ability of the model to account for latent infections, postharvest incubation assays were also conducted using mature berries without symptoms or signs of BBR. The model correctly classified the severity of 15 of 23 epidemics (65% of epidemics) when the classification was based on field assessments of BBR severity; when the model was operated to include BBR severity after incubation assays, its ability to correctly predict BBR severity increased from 65% to >87%. This result showed that the model correctly accounts for latent infections, which is important because latent infections can substantially increase DS. The model was sensitive and specific, with the false-positive and false-negative proportion of model predictions equal to 0.24 and 0, respectively. Therefore, the model may be considered a reliable tool for decision-making for BBR control in vineyards.