Sirolimus-Coated Balloon Angioplasty of Infra-popliteal Lesions for the Treatment of Chronic Limb-Threatening Ischemia: Study Protocol for the Randomized Controlled LIMES Study.

PURPOSE: Evidence on efficacy and long-term safety of paclitaxel-coated devices is still conflicting. Therefore, this study aims to assess whether sirolimus-coated balloon angioplasty is safe and effective for the treatment of infra-popliteal occlusions in patients with chronic limb-threatening ischemia (CLTI). STUDY DESIGN: The randomized controlled, single-blinded, multicentre, investigator-initiated study aims to enrol 230 participants with CLTI and infra-popliteal occlusions at up to 25 centres. Patients will be randomized in a 1:1 ratio to either sirolimus-coated balloon angioplasty or to plain old balloon angioplasty (POBA). Bailout stenting in case of flow-limiting dissection or ≥ 50% residual diameter stenosis is permitted. OUTCOME MEASURES: Primary outcome is the Kaplan-Meier estimate of primary patency at 6 months, defined as the absence of target lesion occlusion with restoration of in-line flow to the ankle. Key secondary outcome is non-inferiority in the proportionate occurrence of major adverse limb events and perioperative all-cause death at 30 days. Overall, participants will be followed for 36 months to assess further secondary efficacy and safety outcomes. ASSUMED GAIN OF KNOWLEDGE: If sirolimus-coated balloon angioplasty turns out to be superior to uncoated-balloon angioplasty regarding patency of infra-popliteal lesions without safety signals, it could become a welcome treatment option for patients with CLTI. Trial Registration ClinicalTrial.gov Identifier: NCT04772300, German Clinical Trials Register: DRKS00024629. Level of Evidence Level 2a, randomized controlled trial.

Epigenetic responses to Phytophthora citrophthora gummosis in citrus.

Studies show that DNA methylation is associated with plant immunity but little is known as to how this epigenetic mechanism assists plants in adjusting their responses to biotic stress, especially when interacting with an hemibiotrophic pathogen such as citrus Phytophthora. The aim of the present study was to assess the effects of scion-rootstock interaction on plant resistance to P. citrophthora infection and DNA methylation patterns in 'Pera' sweet orange and 'Tahiti' acid lime grafted onto 'Rangpur' lime and 'Tropical' sunki rootstocks reinoculated with P. citrophthora. Results showed that reinoculated plants of the 'Pera' sweet orange/'Rangpur' lime and 'Tahiti' acid lime/'Tropical' sunki combinations with more and less sensitive varieties to Phytophthora, presented smaller stem lesions and increased frequency of full methylation and hemimethylation rates, compared to inoculated plants. In contrast, 'Tahiti' acid lime/'Rangpur' lime, two highly sensitive varieties, and 'Pera'/'Tropical' sunki, two much less sensitive varieties, showed high increases in the frequency of hemimethylation and non-methylation levels. Results suggest that in citrus, both the scion-rootstock interaction and DNA methylation affect the response to P. citrophthora infection. Reinoculated plants, depending on the combination, showed changes in intracellular hyphae growth through the formation of sets of fibers and crystal accumulation in the periderm, cortex, and phloem. In addition, starch grain concentration was higher in reinoculated plants in comparison to inoculated plants. These findings support the assumption that DNA methylation is a plant defense mechanism and therefore may be exploited to improve the response of plants to the gummosis of P. citrophthora in citrus.

Meta-analysis of the effects of liming on soil pH and cadmium accumulation in crops.

Increasing cadmium (Cd) contamination in agricultural fields has resulted in a higher risk of Cd accumulation in the food chain. Lime addition can mitigate soil acidification and reduce Cd accumulation in crops cultured in Cd-contaminated soil. To determine key factors controlling the outcomes of liming in reducing Cd accumulation and enhancing soil pH, we performed a meta-analysis using previously published data from field and pot experiments. The results indicated that the liming showed positive effect sizes on the soil pH but negative effect sizes on Cd accumulation in crops, indicating the addition of different lime materials could enhance soil pH and reduce Cd accumulation in crops. The effect sizes of liming on soil pH under pot experimental conditions were higher than that under field experimental conditions, however, the effect sizes of application types and amount of limes on soil pH did not significantly differ between their individual different levels. Under a low background value of soil pH, SOM, CEC and clay, the addition of limes showed a significantly higher effect size on soil pH when compared to their individual higher soil background value, suggesting that the lower background values of soil pH, SOM, CEC and clay might facilitate the outcomes of liming to enhance soil pH. The experiment patterns, crop types and lime application amounts showed a limit effect on the outcomes of liming to reduce the shoot and grain Cd concentrations in crops. The lime types only showed a significant effect size on the shoot Cd accumulation but not on the grain Cd accumulation, in which the CaCO3 had the highest effect size (absolute value, the same below) followed by Ca(OH)2 and CaO. The low soil background values of total Cd concentration and CEC content, but a high soil SOM background content might facilitate the outcomes of liming to reduce the shoot Cd concentration in crops. However, only the background value of soil clay content showed a significantly negative effect size on the grain Cd accumulation, where a high soil clay content had a higher effect size than a low soil clay content. These findings provide useful knowledge about the effects of experiment patterns, crop types, soil conditions, lime types and lime addition amounts on the efficiency of liming in enhancing soil pH and decrease crop Cd concentration.

[Evaluation on a Simultaneous Analytical Method of Pesticide Residues in Limes].

A LC-MS/MS simultaneous analytical method for screening 191 pesticide residues in limes had been developed and validated. Pesticides were extracted with acetonitrile from samples. Then mixed salts, which were anhydrous magnesium sulfate for dehydration, sodium carbonate for adjusting pH, and sodium chloride for salting out, were added to the sample. After centrifugation, supernatant was transferred to a tube. The sample solution was cleaned up using solid phase extraction (SPE) with C18/GC/PSA for the determination by LC-MS/MS. The developed method was improved the recovery rate of thiabendazole, which had a low recovery rate by the conventional method. Validation study, which was following the guidelines of the Ministry of Health, Labor and Welfare, were carried out at 0.01 and 0.1 µg g-1 to evaluate the method. The results of 175 pesticides out of 191 were with satisfactory. A total of 19 imported lime samples sold in Tokyo was analyzed to evaluate the method, then 18 samples contained pesticide residues below MRLs. The developed method is applicable for detection of pesticide residues in lime.

Control of septage sanitization by limes and lactic acid fermentation.

Considering low-cost and effective fecal sludge (FS) treatment alternatives is essential to avoid risk to human health and to ensure safe disposal in landfills and soils. This research assesses optimal pH adjustment of two techniques for sanitizing de-watered FS from a septic sewage-treatment plant. The preliminary analysis evaluated the efficiency of lactic acid fermentation (LAF) by two lactic acid strains: Lactobacillus casei and Lactobacillus acidophilus. The homofermentative strain was chosen to evaluate three supplementary sugars: smashed carrot, sugarcane-derived molasses, and anhydrous dextrose. Lime treatment was examined using two materials, CaO, 105% calcium carbonate equivalent (CCE), and Ca(OH)2, 75% CCE. Four samples were retrieved from a passive thermal drying bed, two characterized by its available nutrient content; all samples were analyzed for the pathogenic profile, and total coliforms (TC) were selected as indicators. For LAF, an inoculating rate of Lactobacillus casei 10 g/100 g sludge was found effective in decreasing the pH below 4.0 after 30 days of fermentation, using 22% w/w dextrose/septage and 20% w/w molasses/septage, where molasses contains 20.7% of soluble sugars. In the case of lime treatment, the pH was fitted by a power-law relationship to the rate of lime applied in a septage with an initial pH lower than 7.0. A Langmuir type equation fitted better the liming of two septages with initial pHs above 7.0. The rate of lime CaO 10% w/w was observed to increase the pH above critical value, 12, after 1 h and 24 h. Analysis confirmed the total elimination of TC in samples with pH < 4.0 and pH > 12, contrasting the respective controls. Rates of CaO considering the initial pH of the FS are recommended in order to reach pH 12. Septage sanitization can be completed using either CaO or lactic acid fermentation with molasses; selecting the ideal method will rely on cost-benefit analysis. Sanitization can be considered as well to improve safety soil nutrient recycling practices.

At-home methods for tenderizing meat using blade tenderization, lime juice and pineapple puree.

We evaluated at-home methods to tenderize flank steaks. Steaks sourced from a commercial beef processor were aged 28 d postmortem, prior to treatment application. Individual steaks were cut into four sampling portions and treated via blade tenderization (BT), and marination with lime juice (LJ) and/or pineapple puree (PP). Blade tenderization (2×) was conducted parallel and perpendicular to muscle fiber direction; LJ and PP were added at 25% of sample weight. Tenderness was assessed via Warner-Bratzler Shear Force (WBSF) and data was analyzed via mixed models. Neither the three-way interaction (P = 0.64), the BT x PP interaction (P = 0.19), nor LJ (P = 0.35) treatment altered WBSF. Blade tenderization x LJ (P = 0.09) and LJ x PP (P = 0.07) tended to alter WBSF. Blade tenderization (P = 0.03) and pineapple puree (P < 0.01) reduced WBSF by 2.40 and 4.50 N, respectively. WBSF was reduced by 7.3 to 24% via treatment combinations of LJ, BT, or PP.

Remediation by waste marble powder and lime of jarosite-rich sediments from Portman Bay (Spain).

We investigate the use of hydrated lime and calcite waste marble powder as remediation treatments of contaminated jarosite-rich sediments from Portman Bay (SE, Spain), one of the most contaminated points in the Mediterranean coast by mining-metallurgical activities. We tested two commercial hydrated limes with different Ca(OH)2 percentages (28 and 60% for Lime-1 and Lime-2 respectively) and two different waste marble powder, WMP, from the marble industry (60 and 96% of calcite for WMP-1 and WMP-2 respectively). Mixture and column experiments and modelling of geochemical reactions using PHREEQC were performed. Lime caused the precipitation of hematite, gypsum and calcite, whereas WMP treatments formed iron carbonates and hematite. The fraction of amorphous phases was mainly composed of iron oxides, hydroxides and oxyhydroxides that was notably higher in the lime treatment in comparison to the WMP treatment. The reactive surface area showed a positive trend with the amorphous phase concentration. Results highlighted the effectiveness of lime treatments, where Lime-2 showed a complete elimination of jarosite. Column experiments revealed a clear reduction of heavy metal concentration in the lixiviate for the treated sediments compared to the original sediments. Particularly, Lime-2 showed the highest reduction in the peak concentration of Fe, Mn, Zn and Cd. The studied treatments limited the stabilisation of Cr and Ni, whereas contrarily As increases in the treated sediment. PHREEQC calculations showed that the most concentrated heavy metals (Zn and Mn) are stabilized mainly by precipitation whereas Cu, Pb and Cd by a combination of precipitation and sorption processes. This chemical environment leads to the precipitation of stable iron phases, which sorb and co-precipitate considerable amounts of potentially toxic elements. Lime is significantly more effective than WMP, although it is recommended that the pH value of the mixture should remain below 9 due to the amphoteric behaviour of heavy metals.

LIMES: a tool for comparing species partition.

MOTIVATION: Species delimitation (SD) is on the verge of becoming a fully fledged research field in systematics, but the variety of available approaches tends to result in significant-sometimes striking-incongruences, when tested comparatively with a given taxonomic sampling. RESULTS: We present LIMES, an automatic calculation tool which qualitatively compares species partitions obtained by distinct SD approaches, regardless of their respective theoretical backgrounds, and even in absence of reference topology. The program implements four different previously published indexes, and allows their automated calculation. AVAILABILITY AND IMPLEMENTATION: LIMES is freely downloadable at www.limes.cnrs.fr. SUPPLEMENTARY INFORMATION: Supplementary data are available at Bioinformatics online.

Hormonal and metabolic responses of Mexican lime plants to CTV infection.

Plant viral infections alter gene expression and metabolism in infected host. To study the molecular responses of Mexican lime to CTV infection, an analysis of plant metabolome in response to infection with severe (T318) or mild (T385) isolates of CTV was performed. Healthy plants and those infected with any of the two virus strains showed different metabolite profiles, at different stages of new sprout development. Proline content increased in plants infected with CTV, proportionally to the virulence of the virus strain. Abscisic acid content decreased after virus infection whereas jasmonic and salicylic acid levels increased. CTV infection had an impact on plant secondary metabolism, by stimulating the synthesis of different metabolites such as l-methylhistidine, phenylpropanoid derivatives. These metabolites are common responses of different organisms, including higher mammals, to viral diseases, and its presence in this system points to the existence of universal responses to virus infection among different kingdoms.

Codon Usage Bias Analysis of Citrus tristeza Virus: Higher Codon Adaptation to Citrus reticulata Host.

Citrus tristeza virus (CTV), a member of the aphid-transmitted closterovirus group, is the causal agent of the notorious tristeza disease in several citrus species worldwide. The codon usage patterns of viruses reflect the evolutionary changes for optimization of their survival and adaptation in their fitness to the external environment and the hosts. The codon usage adaptation of CTV to specific citrus hosts remains to be studied; thus, its role in CTV evolution is not clearly comprehended. Therefore, to better explain the host⁻virus interaction and evolutionary history of CTV, the codon usage patterns of the coat protein (CP) genes of 122 CTV isolates originating from three economically important citrus hosts (55 isolate from Citrus sinensis, 38 from C. reticulata, and 29 from C. aurantifolia) were studied using several codon usage indices and multivariate statistical methods. The present study shows that CTV displays low codon usage bias (CUB) and higher genomic stability. Neutrality plot and relative synonymous codon usage analyses revealed that the overall influence of natural selection was more profound than that of mutation pressure in shaping the CUB of CTV. The contribution of high-frequency codon analysis and codon adaptation index value show that CTV has host-specific codon usage patterns, resulting in higheradaptability of CTV isolates originating from C. reticulata (Cr-CTV), and low adaptability in the isolates originating from C. aurantifolia (Ca-CTV) and C. sinensis (Cs-CTV). The combination of codon analysis of CTV with citrus genealogy suggests that CTV evolved in C. reticulata or other Citrus progenitors. The outcome of the study enhances the understanding of the factors involved in viral adaptation, evolution, and fitness toward their hosts. This information will definitely help devise better management strategies of CTV.

Lime powder regimen supplement alleviates urinary metabolic abnormalities in urolithiasis patients.

BACKGROUND: Urolithiasis has high recurrent rate after surgical removal within 3 years. Potassium citrate compound is used to prevent stone recurrence but it has intolerable gastrointestinal adverse effects. We conducted a phase 2 clinical study of lime power regimen (LPR), a limeade-based supplement containing potassium and citrate for 6 months period of treatment, to evaluate its effects on biochemical and clinical aspects of recurrent urolithiasis. METHODS: Seventy-four urolithiasis patients were randomly allocated to receive either LPR or placebo for 6 months in a double-blinded manner. Plasma and 24 h urine samples were collected to measure urinary pH, mineral excretion and urinary total antioxidant status , plasma for creatinine and plasma protein carbonyl, and stone for elemental analysis at the initiation and end-of-treatment (6 month). Adverse effects were recorded. RESULTS: Administration of LPR significantly increased urinary pH, citrate and potassium excretion. Urinary levels of calcium and oxalate, and plasma protein carbonyl content were reduced, while urinary total antioxidant status was elevated by LPR treatment. Urinary supersaturation was decreased and urinary protein excretion was ameliorated in LPR-treated patients. Gastrointestinal adverse effects were rarely observed. None of the participants developed stone recurrence for the duration of the trial. CONCLUSION: Lime power regimen is a potential drug to correct urinary metabolic disorders associated with urolithiasis in high risk stone recurrent patients. A phase 3 clinical trial is underway to validate anti-stone recurrence property of LPR in long-term treatment.

The Ancient Neapolitan Sweet Lime and the Calabrian Lemoncetta Locrese Belong to the Same Citrus Species.

"Neapolitan limmo" is an ancient and rare sweet Mediterranean lime, now almost extinct but used until a few decades ago for the production of a fragrant liqueur called the "four citrus fruits". The objective of this work was to compare, through the use of chemical (flavonoids, volatile organic compounds, and chiral compounds) and molecular (DNA fingerprint based on RAPD-PCR) markers, the residual population of Neapolitan limmo with other populations of sweet limes, identified in Calabria and known as "lemoncetta Locrese". We report for the first time specific botanical characteristics of the two fruits and unequivocally show that the ancient sweet Mediterranean limes Neapolitan limmo and lemoncetta Locrese are synonyms of the same Citrus species. Owing to the biodiversity conserved in their places of origin, it will now be possible to recover, enhance and implement the use of this ancient sweet lime for agro-industrial purposes.

Illumina-MiSeq analysis of fungi in acid lime roots reveals dominance of Fusarium and variation in fungal taxa.

A study was conducted to analyze fungal diversity in the roots of acid lime (Citrus aurantifolia) collected from Oman, a semi-arid country located in the South Eastern part of the Arabian Peninsula. MiSeq analysis showed the Ascomycota and Sordariomycetes were the most abundant phylum and class in acid lime roots, respectively. Glomeromycota, Basidiomycota and Microsporidia were the other fungal phyla, while Glomeromycetes and some other classes belonging to Ascomycota and Basidiomycota were detected at lower frequencies. The genus Fusarium was the most abundant in all samples, making up 46 to 95% of the total reads. Some fungal genera of Arbuscular mycorrhizae and nematophagous fungi were detected in some of the acid lime roots. Analysis of the level of fungal diversity showed that no significant differences exist among groups of root samples (from different locations) in their Chao richness and Shannon diversity levels (P < 0.05). Principle component analysis of fungal communities significantly separated samples according to their locations. This is the first study to evaluate fungal diversity in acid lime roots using high throughput sequencing analysis. The study reveals the presence of various fungal taxa in the roots, dominated by Fusarium species and including some mycorrhizae and nematophagous fungi.

Challenging cause of bullous eruption of the hands in the Arctic.

Phytophotodermatitis is caused by deposition of photosensitising compounds on the skin followed by ultraviolet exposure. We present an unusual case of a 29-year-old Australian male visiting Greenland who presented with severe itchy bullous eruption on his hands. The cause was a combination of exposure to lime fruit juice and prolonged sun exposure from the Arctic midnight sun.

Witch's Broom Disease of Lime (Candidatus Phytoplasma aurantifolia): Identifying High-Risk Areas by Climatic Mapping.

Biological invasions of vectorborne diseases can be devastating. Bioclimatic modeling provides an opportunity to assess and predict areas at risk from complex multitrophic interactions of pathogens, highlighting areas in need of increased monitoring effort. Here, we model the distribution of an economically critical vectorborne plant pathogen 'Candidatus Phytoplasma aurantifolia', the etiological agent of Witches' Broom Disease of Lime. This disease is a significant limiting factor on acid lime production (Citrus aurantifolia, Swingle) in the Middle East and threatens its production globally. We found that temperature, humidity, and the vector populations significantly determine disease distribution. Following this, we used bioclimatic modeling to predict potential novel sites of infections. The model outputs identified potential novel sites of infection in the citrus producing regions of Brazil and China. We also used our model to explore sites in Oman where the pathogen may not be infectious, and suggest nurseries be established there. Recent major turbulence in the citrus agricultural economy has highlighted the importance of this work and the need for appropriate and targeted monitoring programs to safeguard lime production.

Fitofotodermatosis por «mojitos», la otra enfermedad de «lime» / "Mojito'" phytophotodermatitis, the other "lime" disease

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Stability of lime essential oil microparticles produced with protein-carbohydrate blends.

The objective of this work was to analyze the influence of maltodextrin equivalent dextrose on the lime essential oil reconstitution, storage, release and protection properties. Four treatments were evaluated: whey protein concentrate (WPC), and blends of maltodextrin with dextrose equivalents of 5 (WM5), 10 (WM10) and 20 (WM20). The reconstitution and storage properties of the microparticles (solubility, wettability and density), water kinetics adsorption, sorption isotherms, thermogravimetric properties, controlled release and degradation kinetics of encapsulated lime essential oil were studied to measure the quality of the encapsulated materials. The results of the study indicated that the DE degree influences the characteristics of reconstitution, storage, controlled release and degradation characteristics of encapsulated bioactive compounds. The increase in dextrose equivalent improves microparticle solubility, wettability and density, mainly due to the size of the maltodextrin molecules. The adsorption kinetics and sorption isotherm curves confirmed the increase in the hygroscopicity of maltodextrins with higher degrees of polymerization. The size of the maltodextrin chains influenced the release and protection of the encapsulated lime essential oil. Finally, the maltodextrin polymerization degree can be considered a parameter that will influence the physicochemical properties of microencapsulated food.