Single Application Research Articles

Optimizing the methodology for the assessment of bulbar conjunctival lissamine green staining.

When using lissamine green for bulbar conjunctival evaluation, the most staining was observed with two applications of the same strip in the same eye, wetted with a drop of saline. The first application was 5 seconds after wetting the strip, and the second was 1 minute later, followed by immediate viewing. This suggests that bulbar staining should be viewed before the lid wiper region. This study aimed to optimize the assessment of lissamine green staining of the bulbar conjunctiva by investigating the application technique and subsequent observation period. Twenty-two participants with dry eye (as defined by the Tear Film and Ocular Surface Society Dry Eye Workshop II) were recruited to trial five application methods in randomized order: (1) application immediately after wetting a single strip, (2) application 5 seconds after wetting a single strip, (3) application 10 seconds after wetting a single strip, (4) consecutive single application of sodium fluorescein followed by lissamine green 5 seconds after wetting, and (5) two applications using the same lissamine green strip 5 seconds after wetting, 1 minute apart. Slit-lamp photography of the conjunctiva was performed immediately following application and at 30, 60, 90, and 300 seconds after application. Three experienced (masked) practitioners independently quantified the visible punctate spots and graded the staining intensity within the images in a random order. Values for punctate spot count (F = 6.29, p<0.0001) and lissamine green staining intensity (F = 6.29, p<0.0001) varied significantly between the different application methods. Using two applications of the same lissamine green strip, 1 minute apart, in the same eye resulted in the greatest values for both punctate spot count and lissamine green staining intensity. Lissamine green staining decreased with time for both spot count (F = 18.87, p<0.0001) and lissamine green staining intensity (F = 11.33, p<0.0001), with the most staining found immediately after application followed by a gradual decline. There was no evidence of any interaction effect between time and application method for either measure (p>0.05 for both). The optimal approach for assessing bulbar conjunctival lissamine green staining involves two applications of a whole infused drop resting on the same strip for 5 seconds, applied 1 minute apart. The ocular surface should be viewed immediately after application to maximize the conjunctival staining observed.

The emergence of new regional technological specialisations: exploring the role of organisations and their technological network structure

ABSTRACT Technological collaborations are crucial for shaping the emergence of new regional technological specialisations. This article explores the relationship between technological networks formed between organisations, within and across regions, and the ability of these regions to branch into new technological fields. To identify the organisations’ collaboration structure, we exploit patent co-applicant data and perform a network analysis of the organisations’ collaborations for 245 European NUTS-II regions from 1990 to 2016. Panel regression analyses reveal that (1) a more locally oriented and less dense region-internal network, (2) a more diverse set of region-external linkages, and (3) a higher share of single applicants in a region are positively associated with the emergence of new technological specialisations. These findings highlight the importance of maintaining a balance between the organisations’ inward and outward orientation to acquire new technological specialisations. Regions must leverage both internal technological capabilities and external networks to foster processes of technological renewal.

Optimal nitrogen management for high yield and N use efficiency of ratoon sorghum

Sorghum ratooning, a time and labor-saving cultivation practice, is increasingly being adopted by farmers in Southwest China as an alternative. Efficient N fertilizer management is critical for economical production of sorghum and the long-term protection of the environment. To investigate the impact of N management on grain yield and nitrogen use efficiencies (NUEs) of ratoon sorghum system, a three-year field experiment was conducted for Jinyunuo3 (a hybrid cultivar) and Guojiaohong1 (an inbred cultivar) using 12 combinations of N rates and splitting ratios. The results showed that increasing N rate and splitting application times led to improvements in various growth parameters such as dry matter weight, crop growth rate (CGR), leaf area index (LAI), and photosynthetic potential (PP). The main, ratoon, and annual yields increased with N rate increase, but there was no significant difference between 225 and 150 kg N ha−1 in the ratoon and annual yields. Splitting the application of N fertilizer enhanced grain yield compared to a single dose application method, especially three-split applications yielded higher than two-split applications. Compared with N rates of 225 and 150 kg ha−1, N rate of 75 kg ha−1 increased apparent recovery rate of applied nitrogen (REN), agronomic efficiency of applied nitrogen (AEN), and partial factor productivity from applied nitrogen (PFPN) in both main season and whole year. But through splitting application methods at high N rates could achieve similar or even higher levels of NUEs compared to all applied as basal fertilizer at low N rates. Therefore, it could be recommended that applying 150 kg N ha−1 with a basal-jointing-heading fertilizer ratio of 2:4:4 represented an efficient N management practice to synchronously obtain high grain yield and NUEs in ratoon sorghum system in Southwest China.

Application repetition and electrode-tissue contact result in deeper lesions using a pulsed-field ablation circular variable loop catheter.

Pulsed-field ablation (PFA) is a novel, myocardial-selective, non-thermal ablation modality used to target cardiac arrhythmias. Although prompt electrogram (EGM) signal disappearance is observed immediately after PFA application in the pulmonary veins, whether this finding results in adequate transmural lesions is unknown. The aim of this study is to check whether application repetition and catheter-tissue contact impact lesion formation during PFA. A circular loop PFA catheter was used to deliver repeated energy applications with various levels of contact force. A benchtop vegetal potato model and a beating heart ventricular myocardial model were utilized to evaluate the impact of application repetition, contact force, and catheter repositioning on contiguity and lesion depth. Lesion development occurred over 18 h in the vegetal model and over 6 h in the porcine model. Lesion formation was found to be dependent on application repetition and contact. In porcine ventricles, single and multiple stacked applications led to a lesion depth of 3.5 ± 0.7 and 4.4 ± 1.3 mm, respectively (P = 0.002). Furthermore, the greater the catheter-tissue contact, the more contiguous and deeper the lesions in the vegetal model (1.0 ± 0.9 mm with no contact vs. 5.4 ± 1.4 mm with 30 g of force; P = 0.0001). Pulsed-field ablation delivered via a circular catheter showed that both repetition and catheter contact led independently to deeper lesion formation. These findings indicate that endpoints for effective PFA are related more to PFA biophysics than to mere EGM attenuation.

Spray-induced gene silencing (SIGS) as a tool for the management of Pine Pitch Canker forest disease.

Global change is exacerbating the prevalence of plant diseases caused by pathogenic fungi in forests worldwide. The conventional use of chemical fungicides, which is commonplace in agricultural settings, is not sanctioned for application in forest ecosystems, so novel control strategies are imperative. SIGS (Spray-Induced Gene Silencing) is a promising approach that can modulate the expression of target genes in eukaryotes in response to double-stranded RNA (dsRNA) present in the environment that triggers the RNA interference (RNAi) mechanism. SIGS exhibited notable success in reducing virulence when deployed against some crop fungal pathogens, such as Fusarium graminearum, Botrytis cinerea and Sclerotinia sclerotiorum, among others. However, there is a conspicuous dearth of studies evaluating the applicability of SIGS for managing forest pathogens. This research aimed to determine whether SIGS could be used to control Fusarium circinatum, a widely impactful forest pathogen that causes Pine Pitch Canker disease. Through a bacterial synthesis, we produced dsRNA molecules to target fungal essential genes involved to vesicle trafficking (Vps51, DCTN1, and SAC1), signal transduction (Pp2a, Sit4, Ppg1, and Tap42), and cell wall biogenesis (Chs1, Chs2, Chs3b, Gls1) metabolic pathways. We confirmed that F. circinatum is able to uptake externally applied dsRNA, triggering an inhibition of the pathogen's virulence. Furthermore, this study pioneers the demonstration that recurrent applications of dsRNAs in SIGS are more effective in protecting plants than single applications. Therefore, SIGS emerges as an effective and sustainable approach for managing plant pathogens, showcasing its efficacy in controlling a globally significant forest pathogen subject to quarantine measures.

The Effect of Oil-based Polymer Coated CAN Fertilizer on the Yield and Quality of Triticum aestivum L. and Brassica napus L.

The fertilization with nitrogen plays a crucial role in today’s agriculture characteristic with a high demand for production. The utilization of coated fertilizers represents a possible option to lower the number of applications, lower the dose and limit the risk of environmental losses. The effect of conventional calcium ammonium nitrate (CAN) applied in a single application in blend (1:1 or 1:2) with oil-based polymer coated CAN in thicknesses of 4 (cCAN4) or 6% (cCAN6) by weight of the fertilizer on the yield and quality of winter wheat and oilseed rape was evaluate. The average yields of winter wheat were not significantly influenced by the examined fertilization in either the growing season (GS) or the experimental site. On the contrary, the qualitative parameters on average resulted in lower values in comparison with the control (split N fertilization), possibly due to the single application of nitrogen in early spring. The examined treatments significantly influenced the seed yield of oilseed rape in both GS and both experimental sites. On average, the highest yield of seed (2.8 t/ha) was observed on treatment cCAN4 in a 1:2 ratio. This represents a significant increase by 24.8% compared to the control (2.2 t/ha). Similarly, to winter wheat, the qualitative parameter of oilseed rape (oil content) was lower after the examined blends with coated CAN. The fertilization of oilseed rape with blends of uncoated and coated CAN applied in the single application is a validate alternative to commonly used split nitrogen doses applied repeatedly during vegetation.

A combined treatment for self-traumatic chronic skin lesions associated with post-surgical neuropathic pain in a domestic cat: a pharmacological and cold atmospheric plasma approach.

Cold atmospheric plasma (CAP) has been employed as a therapy against both acute and chronic skin lesions, contaminated or not, and has effects on angiogenesis and reepithelialization promoting healing. In this context, the present study aimed to evaluate the effects of a CAP jet associated with pharmacological treatment described by the 2015 AAHA/AAFP pain management guidelines and the 2022 WSAVA guidelines for the recognition, assessment, and treatment of pain, on the healing of chronic skin lesions caused by a pruritic reaction resulting from post-surgical neuropathic pain. To this end, a single CAP application was performed on a feline patient with a 6months old recurrent contaminated cervical skin lesions along with administration of ketamine (10µg/kg/min) following the prescription of prednisone (1mg/kg, SID, 6days), gabapentin (8mg/kg, BID, 60days) and amitriptyline (0.5mg /kg, SID, 60days). A single application of plasma associated with an NMDA antagonist, anti-inflammatory steroid, tricyclic antidepressant and gabapentinoid thus provided a significant improvement in the macroscopic appearance of the lesion within 10days, and the owner reported the cessation of intense itching within the first four hours after treatment and a consequent improvement in the animal's quality of life. The medical treatment was finished almost a year since the writing of this paper, without clinical or reported recurrent signs of the condition. Therefore, we observed that single dose CAP application associated with ketamine, gabapentin, amitriptyline and prednisone leads to significant healing of chronically infected skin lesions resulting from post-surgical neuropathic pain.

Restoration of the upper limbs of athletes in the Caucasian Mineral Waters resort using the combined application of electro- and peloidotherapy to the cervical– collar area

Objective. Of the work was to develop a method for restoring the upper limbs of athletes in the Caucasian Mineral Waters resort based on the combined use of electro- and peloid therapy on the cervical-collar area. Material and methods.Was carried out among highly qualified athletes during their stay at training camps in Kislovodsk. 30 athletes took part in the study (23 athletes — the main group and 7 athletes (CG). Before and after the application of the technique, the athletes were examined: the initial level and dynamics of the functional state of the upper extremities (according to electroneuromyography (ENMG) and rheography), as well as hemodynamics of cerebral vessels (according to rheoencephalography). The results of the study showed that a single and course application of electro- and peloid therapy procedures on the cervical-collar area contributes to a significant relaxing effect on the upper limbs of athletes. As a result, a decrease in neuromuscular activity and normalization of excessively increased blood flow, improvement of venous outflow. Conclusion. A method for restoring the upper extremities of athletes has been developed and scientifically substantiated, which is recommended for use especially for relaxation after a training day and during the transition (recovery) period of the training process in sanatorium-resort conditions.

Design and Development a Web Platform Portal for Mosque Financial Management

Transparency and accountability in the financial management of mosques is an area that requires significant attention because it builds the initial trust of the mosque in collecting funds through infaq and waqf. Currently, many mosques still use whiteboards as a tool to present their financial reports. This poses a problem for mosques when delivering financial reports to donors who live far away. Therefore, a tool that can serve as a medium for financial reporting is needed. Based on this need, the research aims to create a web-based mosque financial portal to record income (donations, alms), qurban records, and expenses. In other applications, each mosque uses an application designed for each mosque. This software has been developed to accommodate the needs of each mosque and is integrated into a single application portal. The development of this application uses the Waterfall method, which involves various stages such as user requirements, system design, implementation, and testing. This financial portal was successfully implemented in 11 mosques and prayer rooms across 5 provinces. This application features recording income (donations and alms), qurban records, and expenditure records. The test results show that many mosques can use this financial management portal to record income and expenses. Finally, we can conclude that this web-based mosque financial portal would be a viable solution to enhance the accountability services of mosques in managing their finances.

Modeling platform to assess the effectiveness of single and integrated Ixodes scapularis tick control methods

BackgroundLyme disease continues to expand in Canada and the USA and no single intervention is likely to curb the epidemic.MethodsWe propose a platform to quantitatively assess the effectiveness of a subset of Ixodes scapularis tick management approaches. The platform allows us to assess the impact of different control treatments, conducted either individually (single interventions) or in combination (combined efforts), with varying timings and durations. Interventions include three low environmental toxicity measures in differing combinations, namely reductions in white-tailed deer (Odocoileus virginianus) populations, broadcast area-application of the entomopathogenic fungus Metarhizium anisopliae, and fipronil-based rodent-targeted bait boxes. To assess the impact of these control efforts, we calibrated a process-based mathematical model to data collected from residential properties in the town of Redding, southwestern Connecticut, where an integrated tick management program to reduce I.xodes scapularis nymphs was conducted from 2013 through 2016. We estimated parameters mechanistically for each of the three treatments, simulated multiple combinations and timings of interventions, and computed the resulting percent reduction of the nymphal peak and of the area under the phenology curve.ResultsSimulation outputs suggest that the three-treatment combination and the bait boxes–deer reduction combination had the overall highest impacts on suppressing I. scapularis nymphs. All (single or combined) interventions were more efficacious when implemented for a higher number of years. When implemented for at least 4 years, most interventions (except the single application of the entomopathogenic fungus) were predicted to strongly reduce the nymphal peak compared with the no intervention scenario. Finally, we determined the optimal period to apply the entomopathogenic fungus in residential yards, depending on the number of applications.ConclusionsComputer simulation is a powerful tool to identify the optimal deployment of individual and combined tick management approaches, which can synergistically contribute to short-to-long-term, costeffective, and sustainable control of tick-borne diseases in integrated tick management (ITM) interventions.Graphical

The combination of a microbial and a non-microbial biostimulant increases yield in lettuce (Lactuca sativa) under salt stress conditions by up-regulating cytokinin biosynthesis.

Salinization poses a significant challenge in agriculture, exacerbated by anthropogenic global warming. Biostimulants, derived from living microorganisms or natural extracts, have emerged as valuable tools for conventional and organic agriculture. However, our understanding of the molecular mechanisms underlying the effects of biostimulants is very limited, especially in crops under real cultivation conditions. In this study, we adopted an integrative approach to investigate the effectiveness of the combined application of plant growth-promoting bacterium (Bacillus megaterium strain BM08) and a non-microbial biostimulant under control conditions (normal watering) and salt stress. After confirming the yield increase under both conditions, we investigated the molecular mechanisms underlying the observed effect by measuring a number of physiological parameters (i.e., lipid peroxidation, antioxidants, chlorophylls, total phenolics and phytohormone content), as well as RNA sequencing and primary metabolite analyses. Our findings reveal that the combined effect of the microbial and non-microbial biostimulants led to a decrease in the antioxidant response and an up-regulation of genes involved in cytokinin biosynthesis under salt stress conditions. This, in turn, resulted in a higher concentration of the bioactive cytokinin, isopentenyladenosine, in roots and leaves and an increase in γ-aminobutyric acid, a non-proteic amino acid related to abiotic stress responses. In addition, we observed a decrease in malic acid, along with an abscisic acid (ABA)-independent up-regulation of SR-kinases, a family of protein kinases associated with abiotic stress responses. Furthermore, we observed that the single application of the non-microbial biostimulant triggers an ABA-dependent response under salt stress; however, when combined with the microbial biostimulant, it potentiated the mechanisms triggered by the BM08 bacterial strain. This comprehensive investigation shows that the combination of two biostimulants is able to elicit a cytokinin-dependent response that may explain the observed yield increase under salt stress conditions.

The phytosiderophore analogue proline-2′-deoxymugineic acid is more efficient than conventional chelators for improving iron nutrition in maize

ABSTRACT Iron (Fe) is an essential nutrient but has poor bioavailability because of its low solubility under conditions of high pH, such as calcareous soils. Previously, we synthesized an analogue of the natural Fe(III) chelator 2′-deoxymugineic acid secreted by graminaceous plants for efficient Fe uptake, designated as proline-2′-deoxymugineic acid (PDMA). Soil application of Fe(III)-PDMA ameliorated symptoms of Fe deficiency in rice. In the present study, we explored the potential of PDMA as Fe nutrition supplement in maize, a major graminaceous crop, and rice for comparison. In calcareous soil pots, Fe deficiency chlorosis of maize was efficiently recovered by a single application of Fe(III)-PDMA. In contrast, maize plants treated with conventional Fe(III)-chelator complexes, such as Fe(III)-ethylenediaminetetraacetic acid (EDTA) or Fe(III)-N,N′-di(2-hydroxybenzyl)ethylenediamine-N,N′-diacetic acid monohydrochloride (HBED), showed little or no recovery. Similarly, rice Fe chlorosis in calcareous soil pots was efficiently recovered by a single application of Fe(III)-PDMA, but not by other conventional Fe(III)-chelator complexes. Application of Fe(III)-PDMA was also effective in ameliorating Fe deficiency chlorosis in hydroponically grown maize seedlings; other Fe(III)-chelator complexes showed minimal or no efficacy. Addition of low concentrations of metal-free PDMA recovered Fe chlorosis of maize or rice in the presence of Fe(III)-EDTA, suggesting possible ligand exchange from EDTA to PDMA for subsequent Fe(III)-PDMA uptake by ZmYS1 and OsYSL15 transporters. Similar effects were observed with Fe(III)-HBED, which has a higher stability constant, but to a lesser extent than Fe(III)-EDTA, suggesting that ligand exchange from HBED to PDMA might be less prone to occur than for chelators with moderate stability constants, such as EDTA. Ferric-chelate reductase assays of maize roots showed substantial reduction of Fe(III)-PDMA, but this reduction activity was not increased under Fe-deficient condition. These results suggested that PDMA is an efficient reagent for improvement of Fe nutrition in graminaceous crops, including maize, because of suitability of Fe(III)-PDMA as a substrate for chelation-based Fe uptake systems.

Impacts of Compost Amendment Type and Application Frequency on a Fire-Impacted Grassland Ecosystem

Composting organic matter can lower the global warming potential of food and agricultural waste and provide a nutrient-rich soil amendment. Compost applications generally increase net primary production (NPP) and soil water-holding capacity and may stimulate soil carbon (C) sequestration. Questions remain regarding the effects of compost nitrogen (N) concentrations and application rates on soil C and greenhouse gas dynamics. In this study, we explored the effects of compost with different initial N quality (food waste versus green waste compost) on soil greenhouse gas fluxes, aboveground biomass, and soil C and N pools in a fire-impacted annual grassland ecosystem. Composts were applied annually once, twice, or three times prior to the onset of the winter rainy season. A low-intensity fire event after the first growing season also allowed us to explore how compost-amended grasslands respond to burning events, which are expected to increase with climate change. After four growing seasons, all compost treatments significantly increased soil C pools from 9.5 ± 0.9 to 30.2 ± 0.7 Mg C ha−1 (0–40 cm) and 19.5 ± 0.9 to 40.1 ± 0.7 Mg C ha−1 (0–40 cm) relative to burned and unburned controls, respectively. Gains exceeded the compost-C applied, representing newly fixed C. The higher N food waste compost treatments yielded more cumulative soil C (5.2–10.9 Mg C ha−1) and aboveground biomass (0.19–0.66 Mg C ha−1) than the lower N green waste compost treatments, suggesting greater N inputs further increased soil stocks. The three-time green waste application increased soil C and N stocks relative to a single application of either compost. There was minimal impact on net ecosystem greenhouse gas emissions. Aboveground biomass accumulation was higher in all compost treatments relative to controls, likely due to increased water-holding capacity and N availability. Results show that higher N compost resulted in larger C gains with little offset from greenhouse gas emissions and that compost amendments may help mediate effects of low-intensity fire by increasing fertility and water-holding capacity.

Silver diamine fluoride in preventing enamel erosion: An in vitro study with salivary pellicle

ObjectivesTo compare the prevention of enamel erosion and discolouring effect with a single and two weekly topical applications of silver diamine fluoride (SDF) solution. MethodsHuman enamel blocks were divided into four groups. Group 1 (SDF2) received two weekly applications of SDF solution (Advantage Arrest: 260,000 ppm Ag, 44,300 ppm F, pH 9.1). Group 2 (SDF1) received a single application of SDF solution. Group 3 (SNF, Positive Control) received daily application of stannous-chloride/amine-fluoride/sodium-fluoride solution (Elmex® Enamel professional: 800 ppm Sn(II), 500 ppm F, pH 4.5). Group 4 (DW, Negative Control) received daily application of deionised water. The treated blocks were subjected to a 14-day erosive challenge. Crystal characteristics, elemental composition, surface morphology, percentage of surface microhardness loss (%SMHL), surface loss, and total colour change (ΔE) of the blocks were investigated using X-ray diffraction (XRD), energy-dispersive spectrometry (EDS) and scanning electron microscopy (SEM), Vickers’ hardness testing, non-contact profilometry, and digital spectrophotometry, respectively. ResultsXRD and EDS revealed precipitates of silver for SDF2 and SDF1 and tin for SNF. SEM showed prominent etched enamel pattern on DW than the other three groups. The%SMHL (%) of SDF2, SDF1, SNF, and DW were 26.6 ± 2.9, 33.6 ± 2.8, 38.9 ± 2.9, and 50.5 ± 2.8 (SDF2<SDF1<SNF<DW, p < 0.001). Surface loss (μm) of SDF2, SDF1, SNF, and DW were 3.1 ± 0.6, 3.9 ± 0.6, 3.9 ± 0.7, and 5.5 ± 0.8 (SDF2<SDF1=SNF<DW, p < 0.05). Colour change of SDF2, SDF1, SNF, and DW were 7.9 ± 3.9, 4.0 ± 3.6, 3.6 ± 3.5, and −4.4 ± 3.9 (SDF2>SDF1=SNF>DW, p < 0.05). ConclusionTwo weekly applications was more effective than a single application of SDF in preventing enamel erosion, though it caused more discolouration. Clinical significanceTopical application of 38 % SDF with two weekly applications protocol is effective in preventing enamel erosion.

The synergistic effect of biosynthesized CuONPs and phage (ϕPB2) as a novel approach for controlling Ralstonia solanacearum

BackgroundAs a vital soil-borne pathogenic bacterium, Ralstonia solanacearum can cause wilt disease in multiple Solanaceae plants. Several phages, such as ϕPB2, could infect R. solanacearum acting as a potential biological control agent in soil. In addition, some nanoparticles, especially copper preparation, also showed high toxicity on R. solanacearum with low toxicity on plant. However, whether they can be administered in combination and how effective they are in inhibiting the plant disease caused by R. solanacearum is known very little.ResultsIn this work, the characterization of CuONPs using scanning electron microscope, transmission electron microscope, X-ray photoelectron spectroscopy, Fourier transform infrared spectroscopy, and X-ray diffraction ascertained the presence of CuONPs which were nanometer particle of 83 nm. Then it was found that combined application of CuONPs with phage (ϕPB2) was superior to that of ϕPB2 or CuONPs alone in controlling tobacco bacterial wilt, with the CuONPs (250 mg/mL) and phage (106 PFU/mL) ratio being the best, at 79.1%. The combination of CuONPs and ϕPB2 also showed no obvious toxicity on tobacco growth than control like single application of CuONPs or ϕPB2. Furthermore, the transcriptome changes of R. solanacearum analysis indicated that the combination application and single allocation of CuONPs could inhibit “biofilm formation”, molecular function, biological processes, cellular components, metabolic process, and so on. In addition, the combination application showed higher inhibition of motility and biofilm, and better enhancement of cell membrane permeability, protein leakage, MDA concentration, and enzyme activity of their respiratory chain dehydrogenase than single application of CuONPs or phage (ϕPB2). Transcriptomes analysis also supported that the addition of ϕPB2 enhanced the toxicity of CuONPs by influencing the ABC transporters and quorum sensing, metabolic processes, and cellular biosynthetic processes of R. solanacearum.ConclusionIn total, our work not only proposed a novel way to increase the bactericidal effect of nanomaterials by adding phage, but also discovered the influence, synergistic effects, and mechanisms, which is useful to design novel way to combat phytopathogenic bacteria in the complicated environment.Graphical abstract

Remineralization of post-orthodontic white spot lesions with a fluoride varnish and a self-assembling P 11 - 4 peptides: a prospective in-vivo-study.

The objective was to evaluate the remineralization effects of fluoride varnish (Clinpro White varnish), self-assembling peptide (Curodont™ Repair) and their combined use on WSL after orthodontic treatment. Thirty-two subjects, aged of 10-18 (mean age 13.91 ± 2.92) with 107 post-orthodontic WSL were included in the study. Subjects were divided into four groups as control, tricalcium phosphate (TCP) containing fluoride varnish (Clinpro White varnish) group, self-assembling P11-4 peptides (Curodont™ Repair) group and combined application of the two products. At the beginning, each subjects' caries risk profile was assessed by evaluating diet cariogenicity, plaque index, gingival bleeding index and stimulated salivary flow rate. Before the application of the remineralization agents, WSL baseline demineralization values were determined with QLF Inspektor™ Pro, laser fluorescence using DIAGNOdent and color values were measured by Vita EasyShade. Remineralization data were obtained by measuring ΔF, ΔQ, and lesion area with QLF. The aesthetic improvement after the remineralization process was evaluated with a spectrophotometer at six weeks, three and six months. No statistically significant differences were found between the groups in terms of criteria determining patients' caries risk profiles, DIAGNOdent data, and plaque index scores (p > 0.05). Intra-group evaluation following remineralization revealed statistically significant increases in ΔF and ΔQ with a decrease in lesion area for the fluoride varnish group at six months, for the peptide group at three months, and for the combined application group at three and six months (p < 0.05). In inter-group comparisons, ΔF and ΔQ values were found to be statistically significant only in the fluoride group at six months compared to the other groups (p < 0.05). While the L* value decreased significantly in all groups at six months, a statistically significant difference in ΔE* values was observed only in the control group between three and six months. Fluoride varnish with TCP showed highest remineralization at 6 months, and the remineralization was positively affected in the short term (three months) after the use of self-assembling P11-4 peptides and their combined application. Remineralization obtained after single application of agents tested in six months in-vivo showed parallel results. In an attempt to trigger subsurface remineralization, the combined use of fluoride with self-assembling peptides as biomimetic remineralization agent needs further evaluation.

Abstract We105: cBIN1 Gene Therapy Improves Left Ventricular Filling Pressure in a Canine Model of Ischemic Dilated Cardiomyopathy

Introduction: In heart failure patients, increased left ventricular end diastolic pressure (LVEDP) is pathognomonic of myocardial dysfunction. Previously, we presented that gene therapy with exogeneous cardiac bridging integrator 1 (cBIN1), a membrane scaffolding protein that organizes T-tubule microdomains and is decreased in failing hearts, recovers systolic function in a canine model of chronic ischemic dilated cardiomyopathy (IDCM). The effect of catheter-based delivery of cBIN1 gene therapy on increased LVEDP is unclear. Hypothesis: We tested the hypothesis that cBIN1 gene therapy improves LVEDP in a canine model of IDCM. Methods: Twelve healthy dogs (30±2 kg) underwent left anterior descending artery ligation and developed IDCM with LVEF &lt;40% and NT-proBNP &gt;900 pmol/L over 3–4 months. Animals received a single application at 20 subendocardial LV sites of either AAV9-GFP (control, n=6) or AAV9-cBIN1 (n=6) using a MyoStar catheter-based injection. Pressure-volume loops were acquired in vivo, including measurement of LVEDP, end-systolic pressure-volume relation (ESPVR), isovolumic relaxation time (IVRT) and isovolumetric contraction time (IVCT). Measurements were performed on the day prior to administering the therapy ( pre-therapy ) and 5–8 weeks post-intramyocardial injection ( post-therapy ). For statistics, student t-test was used, and p&lt;0.05 refers as significant. Results are reported as mean±SD. Results: In AAV9-GFP control-treated dogs, compared with pre-therapy, post-therapy LVEDP elevated significantly (13±1 vs. 17±1 mmHg, p=0.002, Figure A ), whereas in the cBIN1-treated cohort, elevated LVEDP improved significantly (13±4 vs. 11±3 mmHg, p=0.042). With control therapy, ESPVR exhibited a significant decrease in contractility (1.40±0.44 vs. 1.81±0.49 mmHg/mL, p=0.001, Figure B ), whereas cBIN1 therapy caused a substantial improvement in ESPVR (1.54±0.71 vs. 1.34±0.43 mmHg/mL, p=0.022, Figure B ). At the post-therapy time-point, compared with control therapy, cBIN1 therapy improved IVRT (96±10 vs. 72±8 ms, p&lt;0.001, Figure C ) and IVCT (49±4 vs. 40±3 ms, p=0.009, Figure D ). Conclusion: cBIN1 gene therapy reduces elevated LV filling pressure and improves overall hemodynamics in a canine model of chronic IDCM.

Short occupational exposure to glyphosate and its biomonitoring via urinary levels of glyphosate and metabolite AMPA (Amino-MethylPhosphonic acid), in Italian vineyard workers

Glyphosate, an herbicide largely used in various contexts, can have adverse effects on human health. Although it is currently the most applied pesticide worldwide, few studies evaluated the extent of human exposure via biomonitoring. To expand such information, biological monitoring of exposure to glyphosate was conducted.The study has a before-and-after design to demonstrate the immediate impact of short-term interventions. Accordingly, the urine concentrations of glyphosate and its main biodegradation product (amino-methylphosphonic acid- AMPA) were measured before and the day after the single herbicide application in 17 male winegrowers. Urine samples were analyzed by high performance liquid chromatography coupled with a triple quadrupole mass spectrometer equipped with an electrospray ionization source.Glyphosate and AMPA were not detectable in pre-application urine samples (limit of quantification for glyphosate (LOQG) was 0.1 μg/L; limit of quantification for AMPA (LOQAMPA) was 0.5 μg/L). After application, glyphosate urinary levels were above LOQG in all workers. The median, min, and max values were 2.30, 0.51, and 47.2 μg/L, respectively. The same values were found for 50 %, 5 % and 95 % percentiles. After assigning numerical values, such as one half the LOQ, to each of the non-detects, the “z” of Wilcoxon matched-pairs signed-ranks test was −3.62 (p = 0.0003), suggesting the pre-application values being significantly lower than the post-application urinary glyphosate concentration. A similar analysis was not feasible with AMPA urinary levels, which were detectable only in 3 workers, after application. 12 (71 %) workers were significantly exposed to glyphosate, but adherence to the adoption of personal protective equipment was good: 14 (82 %) workers used gloves, 13 (76 %) used overalls and 13 (76 %) facial masks.Our data show that glyphosate can be absorbed by the workers after a single application and confirms the usefulness of biomonitoring in exposed workers. Further studies are needed in larger working populations and with multiple glyphosate applications, as well as to evaluate the correlations of glyphosate urine levels with exposure questionnaire data, in order to assess the actual relevance of risk and protection factors.

Evaluation of Pregabalin bioadhesive multilayered microemulsion IOP-lowering eye drops

In spite of available treatment options, glaucoma continues to be a leading cause of irreversible blindness in the world. Current glaucoma medications have multiple limitations including: lack of sustained action; requirement for multiple dosing per day, ocular irritation and limited options for drugs with different mechanisms of action. Previously, we demonstrated that pregabalin, a drug with high affinity and selectivity for CACNA2D1, lowered IOP in a dose-dependent manner. The current study was designed to evaluate pregabalin microemulsion eye drops and to estimate its efficacy in humans using in silico methods. Molecular docking studies of pregabalin against CACNA2D1 of mouse, rabbit, and human were performed. Pregabalin microemulsion eye drops were characterized using multiple in vivo studies and its stability was evaluated over one year at different storage conditions. Molecular docking analyses and QSPR of pregabalin confirmed its suitability as a new IOP-lowering medication that functions using a new mechanism of action by binding to CACNA2D1 in all species evaluated. Because of its prolonged corneal residence time and corneal penetration enhancement, a single topical application of pregabalin ME can provide an extended IOP reduction of more than day in different animal models. Repeated daily dosing for 2 months confirms the lack of any tachyphylactic effect, which is a common drawback among marketed IOP-lowering medications. In addition, pregabalin microemulsion demonstrated good physical stability for one year, and chemical stability for 3–6 months if stored below 25 °C. Collectively, these outcomes greatly support the use of pregabalin eye drops as once daily IOP-lowering therapy for glaucoma management.

A novel eco-friendly approach of combining vermicompost and effective microorganisms sustains wheat (Triticum aestivum L.) drought tolerance by modulating photosynthetic performance and nutrient acquisition

The most significant threat to global food security is water scarcity. Despite the fact that vermicompost (an effective organic fertilizer rich in humic substances, macro- and micro-nutrients, earthworm excretions, beneficial soil microbes, plant growth hormones, enzymes) and effective microorganisms (EM; photosynthetic bacteria, lactic acid bacteria, yeasts, actinomycetes, fermenting fungi) have been recognized as powerful strategies for alleviating environmental stresses, their combined effect has not been studied. Herein, as a first investigation, we aimed to enhance wheat’s drought tolerance using an eco-friendly approach that combined vermicompost and EM. The study employed twelve treatments in a completely randomized design. The treatments included control, as well as single and combined applications of vermicompost and EM at three different irrigation levels (100%, 70%, and 30% of field capacity). Vermicompost and EM, applied singly or in combination, ameliorated drought-induced reduction in wheat growth and productivity by elevating photosynthetic pigment content, photochemical processes, Calvin cycle enzyme activity, net photosynthetic rate, transpiration rate, stomatal conductance, maximum quantum efficiency of PSII photochemistry, actual photochemical efficiency of PSII, electron transport rate, photochemical quenching coefficient, and effective quantum yield of PSII photochemistry. Additionally, adding vermicompost and/or EM improved wheat drought tolerance by increasing nutrient (nitrogen, phosphorus, potassium, iron, zinc, copper) acquisition, roots’ ATP content, H+-pump activity, and membrane stability index while lowering hydrogen peroxide content, lipid peroxidation, and electrolyte leakage. The new evidence demonstrates that combining vermicompost with EM sustains wheat drought tolerance by regulating photosynthetic efficiency, nutrient acquisition, root H+-pump activity, and membrane stability. Overall, utilizing vermicompost/EM is a novel approach to improving plant physiological responses and overcoming drought-related challenges.