Mikania micrantha is a highly invasive vine, and its ability to sexually reproduce is a major obstacle to its eradication. The long-distance dissemination of M. micrantha depends on the distribution of seeds; therefore, inhibiting M. micrantha flowering and seed production is an effective control strategy. The number of blooms of M. micrantha differs at different altitudes . In this study, we used a combination of metabolomics and transcriptomics methods to study the patterns of metabolite accumulation in the flower buds of M. micrantha. Using LC-MS/MS, 658 metabolites were found in the flower buds of M. micrantha at three different altitudes . Flavonoids and phenolic acids were found to be the main differential metabolites, and their concentrations were lower at 900 m than at 200 m and 1300 m, with the concentrations of benzoic acid, ferulic acid, and caffeic acid being the lowest. The biosynthesis pathways for flavonoids and phenolic compounds were significantly enriched for differentially expressed genes (DEGs), according to the results of transcriptome analysis. The production of flavonoid and phenolic acids was strongly linked with the expressions of phenylalanine ammonia-lyase (PAL), caffeoyl-CoA O-methyltransferase (COMT), and 4-coumarate-CoA ligase (4CL), according to the results of the combined transcriptome and metabolome analysis. These genes’ roles in the regulation of distinct phenolic acids and flavonoids during M. micrantha bud differentiation are still unknown. This study adds to our understanding of how phenolic acids and flavonoids are regulated in M. micrantha flower buds at various altitudes and identifies regulatory networks that may be involved in this phenomenon, offering a new approach for the prevention and management of M. micrantha. Mikania micrantha Kunth (Asteraceae) is a perennial herbaceous or woody vine of the Mikania Willd in the Asteraceae family. It is one of the most destructive weeds in the world [M. micrantha is more adept in covering, diffusing, and rejecting than many other weeds. Through nutrient competition and covering, it can kill grass and shrub layers in the invaded region, as well as climb and cover trees and kill hosts [M. micrantha, as a noxious invasive plant, has seriously harmed crops by lowering their quality, which has resulted in decreased income for locals. This vine has impacted local economic development and the growth of agricultural and forestry crops [M. micrantha [M. micrantha, the flower biomass of which accounts for 38% to 43% of its total biomass [M. micrantha is capable of reproduction and can reproduce both asexually and sexually, with its seeds being the main means of long-distance dispersal [M. micrantha would be an effective method of controlling its rapid reproduction. Thus, controlling M. micrantha’s flowering and seed production is crucial for its management and defense.he world ,2. M. mill hosts . M. micrry crops ,5. For nicrantha ,7,8. The biomass . These f biomass ,11. M. mispersal . In thisChrysanthemum morifolium when waterlogging stress occurs in different stages of flower bud differentiation [During the transition from the nutritive to the reproductive growth stage of the plant, the apical meristematic tissue changes from the nutritive to the reproductive stem end, forming the flowering organ. The formation of plant flowering organs is a sign of plant flowering, and the time of plant flowering is crucial for plant growth, development, and reproduction ,14. Phenntiation . Accordintiation . Oota etntiation . The conntiation ,19.Using transcriptome sequencing (RNA-Seq) technology, genes associated with metabolite production can be identified . To furtM. micrantha at altitudes of 200, 900, and 1300 m. The ecological factors at these three altitudes substantially differed, but no notable differences were found among the three in terms of annual precipitation, relative humidity, or storm runoff. However, among the environmental factors related to altitude, temperature may considerably impact the ecological adaptability of M. micrantha. Light may also strongly affect the bud differentiation of M. micrantha [M. micrantha at different altitudes using transcriptomic and metabolomic analyses. We aimed to gain insights into the genetic associations among the small-molecule metabolites to understand the key metabolites affecting flower bud differentiation in M. micrantha. We examined the relationship between gene transcript levels and metabolite accumulation in an effort to identify metabolites that affect flower bud differentiation in order to discover new methods of stopping the spread of M. micrantha.In Yunnan Province, Liang et al. found differences in the distribution of icrantha . We founM. micrantha at different altitudes  was employed to assess the overall differences in the metabolite profiles among the groups and the variability within each group. The PCA results revealed that the first two principal components, PC1 and PC2, explained 44.2% and 32.9% of the total variance observed in the samples, respectively. Together, PC1 and PC2 accounted for a cumulative contribution rate of 77.1% A. Based ltitudes A. The anassified B. Among assified C,D. The M. micrantha buds at 900 m than at 200 and 1300 m, which suggests that the accumulation of most flavonoids and phenolic acids in M. micrantha buds is reduced at 900 m, possibly affecting the bud differentiation of M. micrantha. To gain deeper insight into the metabolic pathways involved, we mapped the enriched metabolites in the two comparison groups to the KEGG pathways. Specifically, the first 20 pathways were analyzed for each group  were identified in the comparison between E2 and E9, with 66 being upregulated and 41 being downregulated. In the comparison between E13 and E9, 53 DEMs were identified, with 22 upregulated and 31 downregulated. The identified DEMs were categorized into different groups, including phenolic acids B, flavonch group . The resM. micrantha flower buds sampled from the three altitudes obtained using RNA-Seq are shown in The transcriptome sequencing results of the M. micrantha is more active at 200 and 1300 m and suppressed at 900 m. At E2 and E13, the expression levels of the DEGs were higher than those at E9. Similar trends were observed in the expression levels of the DEGs involved in the phenylpropanoid biosynthesis pathway. Compared with 900 m, the plants at E2 and E13 exhibited higher levels of gene expression. The expression levels of the DEGs were higher at E2 and E13 than at E9. This indicates that phenylpropanoid biosynthesis is also more active at altitudes of 200 and 1300 m. The upregulation of DEGs in these pathways at 200 and 1300 m indicates the importance of their regulated metabolites in the bud differentiation process.To elucidate the function of the DEGs, two comparison groups of DEG enrichment in the 20 most significant KEGG metabolic pathways based on the FPKM values were analyzed. The DEGs were mostly concentrated in metabolic pathways, the biosynthesis of secondary metabolites, flavonoid biosynthesis, and phenylpropanoid biosynthesis pathways, according to the KEGG enrichment analysis of the total DEGs . Among tp-value associated with the differential gene, and green represents the enrichment p-value associated with the differential metabolite, which is represented by the −log. The higher the location on the vertical axis, the higher the degree of enrichment . Many metabolites were found to be positively or negatively regulated by multiple genes  [PAL, 4CL, F5H, and COMT. Multiple genes in the phenylpropanoid biosynthesis pathway play a role in driving metabolic flow towards phenolic acid synthesis. PAL, 4CL, and CMOT were significantly upregulated at E2 and E13 compared with those at E9, and the expression of PAL was significantly lower at 900 m than at the other two altitudes. As such, we speculated that PAL may positively regulate phenolic acid synthesis in the phenolic acid biosynthesis pathway  ,49. In o pathway A.Transcription factors are involved in the regulation of gene expression by binding to specific DNA sequences known as cis-acting elements, which are located in the promoter regions of genes, activating or repressing transcription through their interactions with each other and with other related proteins and playing a key role in the entire metabolic pathway in plants . MYB-likM. micrantha flower buds were collected on the same day from three 20 cm × 20 cm sample plots at three different altitudes: (1) E2: 200 m altitude; (2) E9: 900 m altitude; and (3) E13: 1300 m altitude. Three replicates were set up for each sample plot. During flower bud collection, a mixture of at least 10 flower buds of M. micrantha was used as one biological replicate representing three independent replicates at each altitude. The flower buds were immediately frozen in liquid nitrogen after collection and stored at −80 °C until the total RNA and metabolites were extracted. The M. micrantha plants collected at different altitudes were in the inflorescence primordia stage.The Tongbiguan Nature Reserve, Dehong Dai and Jingpo Autonomous Prefecture, Yunnan Province, China, has a subtropical monsoon climate with abundant light and heat, abundant rainfall, and a short frost period. https://www.metware.cn/Home11.html accessed on 6 March 2021) using a widely targeted metabolome method. The flower buds of M. micrantha from the three different altitude groups were dehydrated using a vacuum freeze-dryer and crushed into a powder to preserve the metabolites present in the samples. A portion of the lyophilized powder (100 mg) was weighed and extracted with 1.2 mL of 70% methanol. The extraction was performed overnight at 4 °C to ensure the efficient extraction of the metabolites from the sample. After extraction, the samples were centrifuged at 12,000 rpm for 10 min to separate the solid particles from the liquid supernatant, which contained the extracted metabolites. The supernatant was filtered . A scheduled multiple-reaction monitoring method was used to quantify the metabolites [https://www.kegg.jp/kegg/compound/ accessed on 6 May 2022). This analysis helped us to understand the biological pathways and processes associated with the differentially accumulated metabolites.The metabolome analysis was performed by Wuhan Maiwei Biotechnology Co., Ltd.  (abolites . The filabolites . Linear https://www.genome.jp/kegg accessed on 6 June 2022) database, a well-known and reliable database for interpreting the molecular-level details of genomes, enzymes, and chemicals in organisms [As with the transcriptome study, all samples used for the metabolome analysis were in the same stage and from the same location. A PureLink Plant RNA Kit  was used to extract the total RNA. An Agilent 2100 bioanalyzer was used to determine the quality of the RNA. mRNA was isolated from the total RNA through the use of magnetic beads with poly-T oligo attached. The cDNA library was constructed, and transcriptome sequencing was performed on an Illumina sequencing platform (Illumina HiSeqTM 2500)). The raw data obtained from sequencing were filtered to remove sequences containing only sequenced splice sequences. Sequences with an N content exceeding 10% of the number of bases in the read and sequences with low-quality (Q ≤ 20) bases exceeding 50% of the number of bases in the read were excluded. The raw sequences were quality-controlled to obtain clean sequences by removing splice sequences, low-quality sequences, multiple N sequences, and sequences that were too short. Then, the sequencing base error rate and GC base content were calculated. The expression levels of the genes were calculated using fragments per kilobase per million mapped fragments (FPKM) , and DESrganisms .From the quantitative analysis, specific genes and metabolites were identified as differentially expressed, meaning their expression levels significantly differed between the compared conditions or groups. These DEGs and DEMs were the focus of further analysis, as they may be important for understanding the underlying biological processes. The metabolites and their associated transcripts were then mapped to the relevant metabolic pathways in the KEGG database. The cor function in the R statistical programming language was used to calculate the Pearson correlation coefficients between the differentially expressed genes (DEGs) and the differentially expressed metabolites (DEMs). Finally, the network diagrams representing the correlations between the DEGs and DEMs were plotted using Cytoscape 3.8.2 software. The network diagrams helped us to understand the interconnectedness and potential regulatory relationships between the DEGs and DEMs.M. micrantha during flower bud differentiation at different altitudes, and we identified a total of 658 differential metabolites, including 161 flavonoids and 78 phenolic acids. Candidate genes affecting the synthesis of flavonoids and phenolic acids in M. micrantha buds, including 23 structural genes in the phenylpropane synthesis pathway and a transcription factor gene-E3N88_30635 (MYB 111), were also explored, providing a new avenue for the study of the molecular mechanism of the accumulation of phenolic acids and flavonoids in M. micrantha buds. This study also has reference value for the subsequent use of bioengineering methods to regulate the flowering of M. micrantha.In this study, based on a combined transcriptomic and metabolomic approach, we investigated the differences in the metabolites and genes of


MET, ALK, EGFR, ERBB2, and RET , predicted response rates to target therapy, resistance mechanisms to first line treatments, in addition to how next generation sequencing (NGS) might predict tumor response. In fact, we’ve witnessed a revolution in the way we understand, diagnose, and treat non-small cell lung cancer (NSCLC) – particularly adenocarcinoma.In this Research Topic of Frontiers in Oncology, six interesting papers covering molecular targets, including Yang et al.), allowing fast and accurate molecular profiling. Thirdly those advances coupled to the chemistry advances led to the development of safe and effective tyrosine kinase inhibitors which were validated in well-designed pivotal registration trials. As a result, we have observed clear improvements in tumor response and survival upon the precision medicine framework in lung cancer.The development of molecular diagnosis and the resulting precision medicine have been based on three pillars. First, decades of advances in basic and translational research led to tumor biology clear up, with new signaling pathways and molecular targets being discovered, resulting in a wave of novel treatments for cancer patients based on crucial genetic alterations, the so-called actionable mutations. Secondly clinical practice underwent significant modifications with the adoption of NGS multigene panels in NSCLC, including both DNA and RNA-sequencing .in situ hybridization (FISH), and polymerase chain reaction (PCR) turned NGS as a resourceful tool for physicians . Previously, DNA-sampling itself was considered enough to identify potential targets, however, it is becoming highly recommended to include RNA-sequencing on those tests . Not to mention that ideally molecular testing must be performed in both diagnostic sample and in a disease progression sample , fluorescence n sample , aiming n sample , 3.Concerning liquid biopsy, its use may enhance molecular profiling and assist in a more feasible treatment, as well as analysis of circulating tumor DNA, which, for instance, may guide duration of treatment , as wellAffordability is indeed an issue in LMIC as incorporation of novel test and drugs exponentially increase costs. The advent of precision medicine highlighted how inequity lung cancer care currently is, with disparities along several subsets of patients, especially among those who live in rural areas, Black patients, and patients derived from low and middle-income countries, in which molecular testing is almost nonexistent in poorer countries , 7. IneqIn continuum, with the advancements in precision medicine, the use of targeted therapy in early-stage NSCLC has been increasingly explored. The most recent example is the overall survival results from the ADAURA trial showing benefits with adjuvant osimertinib for EGFR-mutated NSCLC . Many prIn sum, the era of precision medicine in lung cancer has changed the landscape of the lung cancer treatment. There is no doubt that combined efforts to amplify access to these tests must include police-makers, collaborations between both diagnostic and pharmaceutical companies, and health care providers. Extending molecular testing to broader communities also depends on cost-effectiveness evaluations, high sensitivity rates, and real-world evidence on how affordable they can be. As technology advances, we must focus on overcoming the current existing barriers and increase access to molecular diagnosis, so that patients can yield better outcomes and quality of life .CF: Writing – original draft, Writing – review & editing. MR: Writing – original draft, Writing – review & editing. GV: Writing – original draft, Writing – review & editing.


The aim of this study was to evaluate the nanoleakage and microtensile bond strength (μTBS) of an ethanol based-adhesive containing Titanium dioxide (TiO2) nanofibers to dentin. TiO2 nanofiber was produced by electrospinning and it was inserted in an ethanol-based adhesive in 0.5, 1.5 and 2.5% by weight. The original adhesive did not receive nanofiber. The middle dentin was exposed by diamond saw under water-cooling and dentin was polished with wet 600-grit SiC abrasive paper. Resin composite build-ups were applied incrementally to the dentin after adhesive application. After storage in distilled water (24 hours/37°C) the teeth were sectioned perpendicularly to the bonded interface and sticks were obtained. Twenty-five sticks per group were tested by μTBS with a crosshead speed of 0.5mm/minute. The average values (MPa) obtained in each substrate were subjected to one-way ANOVA  with the tooth being considered the experimental unit. The nanoleakage pattern was observed in ten sticks per group and analyzed by Chi-square test . p=0,028) in relation to nanoleakage. There was no difference in μTBS among the experimental groups. However, there was a statistically significant difference among 2.5 % nanofiber adhesive, 0.5 % nanofibers and control groups, (TiO2 nanofibers in 2.5% of weight inserted in dental adhesive reduced the nanoleakage, but did not improve the μTBS to dentin. Key words:Dentin-bonding agents, nanoleakage, tensile bond strength. The advances in dental materials and adhesive technology have enabled the dentists to make esthetic anterior restorations in a simple and economical way . NowadaySimplified etch-and-rinse adhesives have reduced clinical steps, but they have showed permeability of water from the oral environment and from the underlying bonded dentin -6, leadiDifferent laboratorial approaches have been proposed to improve monomer infiltration, reduce the rate of water sorption, reduce collagen degradation and qualify the adhesion. Effects of primer/adhesive placement agitation and drying time for five seconds on dentin have showed improvement of the shear bond strength to dentin . AnotherStaphylococcus aureus  and nanoleakage. The hypothesis is that the performance of a total etch adhesive system with and without titanium dioxide nanofibers will be similar.-Experimental designin vitro study involving the incorporation of titanium dioxide nanofibers in an ethanol-based total etch adhesive systems (in four levels). Bond strength and nanoleakage were the main response variables for comparison purposes.It was an -Specimen preparationTwenty extracted, caries-free human third molars were used in this study. The teeth were disinfected in 0.5% chloramine, stored in distilled water and used within 6 months after extraction. A ﬂat dentin surface was exposed after wet grinding the occlusal enamel on a # 180 grit SiC paper. The exposed dentin surfaces were further polished on wet #600-grit silicon-carbide paper for 60 seconds to standardize the smear layer.One solvent-based, etch-and-rinse adhesive systems without inorganic particles were tested: Ambar ® . All the inorganic nanofibers were prepared by sol-gel processing and electrospinning technique using a viscous solution , and it has an antimicrobial function as well. Each nanofiber has about 200 to 300 nanometers of diameter. The adhesive received a nanofiber insert by weight, at 0.5, 1.5 and 2.5%, it is forming three groups respectively . The control group was the original bonding agent – G4.After acid etching, the surfaces were rinsed with distilled water for 15 s and air-dried for 15 s. The surfaces were then rewetted with water . Two coa-Bond strength testThe restored teeth were longitudinally sectioned perpendicularly to the bonded interfaces with a diamond saw  to obtain the sticks. Specimens areas were measured with a digital caliper and recorded . Each stick had a cross-sectional area of approximately 0.96 mm2.Half of the specimens, from each tooth, were tested in microtensile testing, and they were randomly assigned. The other half was analyzed by SEM, in relation to nanoleakage. Each bonded stick was attached to a device for microtensile bond strength test (μTBS), with cyanoacrylate resin  and subjected to a tensile force in a Microtensile tester  at a crosshead speed of 0.5 mm/min (n=25).The mean BS for every testing group was expressed as the average of the ﬁve teeth used per group. The micro tensile bond strength data was subjected to one-way analysis of variance, with a significant level set at 0.05.-Nanoleakage evaluationThis study utilized ten sticks from each group for nanoleakage evaluation (n=40). None of the sticks evaluated by nanoleakage were tested by μTBS. The sticks were coated with two layers of fast-setting nail varnish applied up to within 1 mm of the bonded interfaces. The specimens were re-hydrated in distilled water for 10 min prior to immersion in the tracer solution. Conventional silver nitrate  was prepared in a 50 wt% silver nitrate solution (pH=4.2) The sticks were placed in the conventional silver nitrate in darkness for 24 h, rinsed thoroughly in distilled water, and immersed in photo developing solution for 8 h under a ﬂuorescent light to reduce silver ions into metallic silver grains within voids along the bonded interface.All these sticks were wet-polished with 600-grit SiC paper to remove the nail varnish. Then, the specimens were placed inside an acrylic ring, which was attached to a double-sided adhesive tape, and embedded in epoxy resin. After the epoxy resin set, the thickness of the embedded specimens was reduced to approximately half by grinding with silicon carbide papers under running water. Specimens were polished with a 600, 1000, 2000 and 2400 grit SiC paper and 6, 3, 1 and 0.25 mm diamond paste  using a polish cloth. They were ultrasonically cleaned, silica dried, mounted on stubs, and coated with carbon-gold . Resin–dentin interfaces were analyzed in a ﬁeld-emission scanning electron microscope operated in the backscattered electron mode . Presence or absence of nanoleakage was observed, and this data were analyzed by Chi-square test, with a significant level set at 0.05.-Bond Strength Test (µTBS)p=0,607).The range of cross-sectional area was 0.96mm2 for all sticks. The different values of original adhesive (G4) and modified adhesives (Titanium dioxide nanofiber insert by weight) G1, G2, G3 respectively are summarized in -Nanoleakage evaluationp=0,028).The presence or absence of nanoleakage was noted and it is shown in Representative SEM backscattered images of the resin tags Fig.  and resiRegarding the resin-tags, Figure in vitro . We believe that the microtensile testing and nanoleakage analysis are convenient methods for screening and testing the strength and quality of adhesive interfaces in vitro ,9.The higher performance of this kind of actual adhesives has been previously reported and they are used to make esthetic anterior restorations in all patients . The aimThe adhesive used in the current study was intentionally chosen because it is not filled with inorganical particles. Although the incorporation of titanium dioxide nanofibers did not improve the bond strength of Ambar, it did not decrease its performance. The hypothesis must be accepted in terms of bond strength measurement.However, the application of the nanofibers in adhesive system significantly affected the quality of hybrid layer in relation to nanoleakage. The incorporation of titanium dioxide nanofibers at 1.5 and 2.5% levels resulted in the lowest levels of nanoleakage of the adhesive system when compared to the original bonding agent without nanofibers. For this reason, the hypothesis must be accepted, since differences were observed among the groups. It was also noted the presence of nanofibers with crosslink with tags at the same groups, and the collagen probably. If this really occurs, it is important to analyses the durability of bonding in these adhesive compositions, because the crosslink is a form to stabilize the composite adhesive and the TiO2 nanofibers are insoluble in water, and this fact can also contribute to reduce the nanoleakage ,10. The This current study verified that the resin-dentin bond strength values were similar for those different amounts of nanofibers. This finding could be attributed to the fact that the same material was applied to build the nanofiber. The high performance of the hybrid layer formed without nanoleakage in almost all specimens was probably because the nanofiber net reduced the polymerization stress, polymerization shrinkage and elastic modulus . AccordiThe tags penetration was not influenced by nanofibers in every test groups, view by scanning electronic microscopy. Unfortunately, this issue has not yet been evaluated in other adhesives ethanol and alcohol based and should be further investigated. We believed that this paper is the first step to open the doors for titanium dioxide nanofibers in adhesive systems; however, clinical trials must be conducted to verify these results in a clinical situation.Within the limitation of this study, it was possible to conclude that titanium dioxide nanofibers in 2,5% of weight inserted in a dental bonding agent reduced the nanoleakage within the hybrid layer, and did not improve the μTBS of composite restorations.


Mycobacterium bovis. The Test and Vaccinate or Remove (TVR) project was a 5-year intervention (2014–2018) applied to Eurasian badgers (Meles meles) in a 100 km2 area of County Down, Northern Ireland. This observational study used routine bTB surveillance data of cattle to determine if the TVR intervention had any effect in reducing the infection at a herd level. The study design included the TVR treatment area (Banbridge) compared to the three adjacent 100 km2 areas  which did not receive any badger intervention. Results showed that there were statistically lower bTB herd incidence rate ratios in the Banbridge TVR area compared to two of the other three comparison areas, but with bTB herd history and number of bTB infected cattle being the main explanatory variables along with Year. This finding is consistent with other study results conducted as part of the TVR project that suggested that the main transmission route for bTB in the area was cattle-to-cattle spread. This potentially makes any wildlife intervention in the TVR area of less relevance to bTB levels in cattle. It must also be noted that the scientific power of the TVR study (76%) was below the recommended 80%, meaning that results must be interpreted with caution. Even though statistical significance was achieved in two cattle-related risk factors, other potential risk factors may have also demonstrated significance in a larger study.Bovine tuberculosis (bTB) is a chronic, zoonotic infection of domestic and wild animals caused mainly by  M. bovis infection in cattle (bovine tuberculosis (bTB)) and Eurasian badgers (M. meles) is a persistent and costly problem for the farming industries and governments of the United Kingdom (UK) and Republic of Ireland (ROI) [M. bovis infection in cattle and badgers exists from a variety of sources. In ROI, the role of the badger in maintaining persistent infection in cattle herds was examined in the east Offaly (1989–1995) and in four area (1997–2002) field trials, where proactively culled areas were compared to matched reference areas in which reactive culling was carried out [M. bovis infection of cattle and badgers in a locality is molecular genotyping, where GB and Irish studies show that cattle and badgers tend to share the same genotypes in the same areas [nd (ROI) . Evidencrea 1997–002 field 1989–199 and in fried out . The overied out . More reme areas . In relame areas –16. Currme areas –20. Rathme areas . It mustme areas –15.2 area of County Down, Northern Ireland [M. bovis prevalence in badgers from 14% to 1.9% [The Test and Vaccinate or Remove (TVR) project was a 5-year badger intervention (2014–2018) in a 100 km Ireland . During  Ireland . A Bayes to 1.9% .2 core area had an impact on the level of bTB in the local cattle herds compared with similar-sized non-treatment areas in the same locality over the same time period. It was also an objective of this study to determine if the application of TVR intervention led to any increase in bTB among herds adjacent to the treatment area (perturbation effect). This part of the objective was met by comparing the bTB herd incidence in a 2 km buffer zone around the TVR intervention with similar buffer zones around the comparison areas.The primary objective of this study was to assess if the application of a TVR intervention in the 100 kmThis observational study investigated incidence rates for cattle herd bTB breakdowns disclosed in the TVR treatment area (Banbridge) compared to three areas  that did not receive any badger intervention . Also inThe three comparison areas were chosen using the same methodology  as outlined for the TVR treatment area . These aEach cattle herd in Northern Ireland has its location coordinates stored in a national database representative of its Centre of Activity (COA) along with details of its bTB testing history and cattle movements, including purchases . The COAIn terms of descriptive statistics, farm demographics within eight separate areas, the Banbridge TVR study area, the three comparison areas, and the four 2 km buffer zones during the years 2011 to 2019 have been presented in The response variable in this study was the IRR, determined by modelling the count of newly confirmed bTB herd breakdowns and using log time at risk as an offset explanatory variable. As the IRR values quoted in the results and discussion are in all cases adjusted for at least one or more of the study variables, they are adjusted IRRs. At a herd level, confirmation of bTB infection was deemed to have occurred when a Comparative Intradermal Tuberculin Test (CITT) positive animal was disclosed with visible bTB-like lesions at slaughter or there was disclosure of greater than one CITT-positive animal independent of lesion status. Confirmation was also deemed to have occurred when lymph nodes from an animal were confirmed as histological and/or bacteriological positive for bTB on laboratory examination. Time at risk was the total time in years when all cattle herds within a particular area were not restricted due to bTB. This was calculated by subtracting the aggregation of time periods during which confirmed bTB herd breakdowns had their Officially Tuberculosis Free (OTF) status removed from the total time in years for all herds within a particular area. Removal of OTF status meant a herd had its official capacity to sell animals to other herds withdrawn, its re-establishment only occurring when there was statutory compliance with bTB scheme rules.2 (Supplementary Table 1).There were 11 explanatory factors included; area (as described above); year (2011 to 2019 inclusive); number of farm businesses ; total CITT herd tests ; total number of CITT reactors and Lesions at Routine Slaughter (LRS) where bTB was confirmed, measured in a unit of per 50 animals; median herd size determined from annual bTB herd test data; percentage of herds which were defined as dairy enterprises (presence of a milk licence); percentage of farms which disclosed a confirmed bTB herd breakdown in the previous 2 years; percentage of farms which purchased any cattle in the previous year; and percentage of farms with a COA in a particular area which have all their associated ground claimed on BPS in that area. Also included as an explanatory variable for each area was the average number of active main badger setts per km2 , which wP < 0.05, or in situations where more than one of the models had a P < 0.05, the model with the lowest P-value. In any case, where none of the three model structures were significant, the most parsimonious model (area only) was chosen.Analysis of the data in this study was carried out in two parts. The first part used Poisson regression  comparing the TVR area (as baseline) individually to each of the three comparison areas along with an amalgamation of the three comparison areas , and eacSupplementary Tables 2 and 3) and then forward stepwise regression , with those assumed to equate to badger social group densities [ensities .P < 0.01), 1.39 (P < 0.01), and 1.20 (P = 0.011), respectively. However, the Castlewellan area had an adjusted IRR which was not statistically significantly different from the Banbridge TVR area: 0.99 (P = 0.865).With the first part of the analysis, each of the three non-intervention areas  were compared in terms of adjusted IRR using Banbridge as the reference level . The modP = 0.1), Dromore adjusted IRR = 1.03 (P = 0.768), Ballynahinch adjusted IRR = 1.04 (P = 0.627), and Castlewellan adjusted IRR = 1.13 (P = 0.2).In each of the four study areas , the adjusted IRR was also calculated relative to their individual buffer zone (used as the reference level) in order to determine if rate differences between them were statistically significant at the 5% level . The besSupplementary Tables 2 and 3) showed that the two variables with the strongest statistically significant association were the percentage of farms with a bTB herd breakdown in the previous 2 years and the total number of CITT reactors and LRSs . Multivariable results from the Poisson forward stepwise regression resulted in a model containing three of the original variables: percentage of farms with a bTB herd breakdown in the previous 2 years, total number of CITT reactors, and LRSs  and year . Dispersion for this model was calculated at a ratio value of 1.366 (P = 0.081), indicating that the level of dispersion was not statistically significant at the 5% level [In the second part of the analysis, the univariate results  at the start to 1.9% (95% CrI: 0.8–3.8) after 5 years of TVR application . PreviouThe TVR project took place in the Banbridge area  during tP < 0.05) in Dromore, Ballynahinch, and the combined areas, compared to Banbridge, but it did not differ from the adjusted IRR in Castlewellan. In terms of herd demographics, the area most analogous to Banbridge in terms of the number of herds, herd size, percentage dairy, purchasing practices, and percentage of land contained in the area was Dromore, with an adjusted IRR relative to Banbridge of 1.30 . Castlewellan herd demographics were least similar to Banbridge for the three comparison areas used in this study as it contained a greater number of smaller herds and fewer dairy enterprises  with only sporadic inter-species transitions (2/37 direct transmission events) and no badger-to-badger direct transmission events being observed [In this study, potential explanatory variables were also applied in both univariate and multivariable models in order to determine which had the strongest association with the IRR . Resultobserved , 13. Whiobserved , 30, andobserved , 31, 32.2 and three local comparison areas  of similar size where no treatment was applied. The median number of herds in the Banbridge treatment area during the study period was 214 (IQR: 210–220). Given these demographics, a simulation study was carried out to explore the power of routine bTB surveillance to assess the impact of the TVR intervention on bTB incidence in cattle herds [This study had one treatment area (Banbridge) of approximately 100 kmle herds . This wole herds , the lacM. bovis transmission [Each of the areas (study and comparison) had a 2 km buffer zone allocated around its edge, and a relative bTB herd adjusted IRR was calculated for each buffer zone. One of the objectives of this study was to determine if TVR led to any perturbation effect . In Englsmission . A studysmission . Furthersmission . ResultsWhile these findings This study used routine bTB cattle herd surveillance data to determine if the TVR intervention had any effect in reducing the infection at a herd level. Even though results showed that there was a statistically significant lower adjusted IRR in the Banbridge TVR area compared to two of the other three comparison areas, results from a multivariable model in this study and those obtained from other studies , 13, 35


The diagnostic performance of the SYNTAX score 2020 (SS-2020) when calculated using CCTA remains unknown. This study aimed to compare treatment recommendations based on the SS-2020 derived from coronary computed tomography angiography (CCTA) versus invasive coronary angiography (ICA). This interim analysis included 57 of the planned 114 patients with de-novo three-vessel disease, with or without left main coronary artery disease, enrolled in the ongoing FASTTRACK CABG trial. The anatomical SYNTAX scores derived from ICA or CCTA were evaluated by two separate teams of blinded core-lab analysts. Treatment recommendations were based on a maximal individual absolute risk difference in all-cause mortality between percutaneous coronary intervention (PCI) and coronary artery bypass graft (CABG) of 4.5% . The level of agreement was evaluated with Bland–Altman plots and Cohen’s Kappa. The mean age was 66.2 ± 9.2 years and 89.5% of patients were male. Mean anatomical SYNTAX scores derived from ICA and CCTA were 35.1 ± 11.5 and 35.6 ± 11.4 (p = 0.751), respectively. The Bland–Altman analysis showed mean differences of − 0.26 and − 0.93, with standard deviation of 3.69 and 5.23, for 5- and 10-year all-cause mortality, respectively. The concordance in recommended treatment for 5- and 10-year mortalities were 84.2% (48/57 patients) and 80.7% (46/57 patients), with Cohen’s κ coefficients of 0.672 and 0.551. There was moderate to substantial agreement between treatment recommendations based on the SS-2020 derived using CCTA and ICA, suggesting that CCTA could be used as an alternative to ICA when making decisions regarding the modality of revascularization.The online version contains supplementary material available at 10.1007/s10554-023-02884-0. Coronary computed tomography angiography (CCTA) is a non-invasive diagnostic tool that is increasingly being used as an alternative to invasive coronary angiography (ICA) in patients with suspected coronary artery disease (CAD), which can provide important information on the extent and severity of CAD, as well as on the composition of atherosclerotic plaque and can identify plaque with high-risk features for adverse outcomes , 2. The This study was the interim analysis which included all 57 enrolled patients out of the planned 114 patients in the FASTTRACK CABG trial (NCT04142021) . BrieflyCCTA was performed exclusively at all four academic sites using the Revolution CT scanner  which has a nominal spatial resolution of 230 microns along the X–Y planes, a rotational speed of 0.28 s, and a Z-plane coverage of 16 cm enabling imaging of the whole heart in one heartbeat , 10. In Anatomical SYNTAX score calculations derived from ICA and CCTA were performed blindly by two independent groups of experienced analysts of the academic core laboratory team. Coronary segments with a visual diameter stenosis > 50% in vessels ≥ 1.5 mm using CCTA or ICA were assessed and weighted according to their location in the coronary tree . The SS-Treatment recommendation was made based on the individual absolute risk difference (ARD) between PCI and CABG for all-cause mortality at 5 and 10 years calculated by subtracting the predicted CABG mortality from the predicted PCI mortality. According to an external validation in a large contemporary registry, an individual predicted ARD in all-cause death at 5 years of < 4.5% and ≥ 4.5% offers a sensible cut-off for “equipoise of PCI and CABG” or “CABG better,” respectively . A repreContinuous variables were expressed as mean ± standard deviation and were compared with the use of Paired t-test. Categorical variables were reported as numbers and percentages and were compared using the Chi-square test. Individual absolute risk differences (ARDs) between predicted PCI and CABG for mortality at 5- and 10-year were shown by scatterplot in descending order of magnitude according to the predicted ARD in mortality  for each patient. The dots in the scatter plots were connected with the use of locally estimated smoothing (LOESS) spline curves . The Kap2, and 26.3% of patients had diabetes mellitus, but only one patient was insulin dependent. The mean LVEF was 55.9 ± 7.8%.Baseline characteristics are displayed in Table . The mean anatomical SYNTAX scores derived from ICA and CCTA were comparable , despite the total number of lesions per patient being significantly higher with CCTA . The presence of a trifurcation lesion was significantly higher in ICA than CCTA . The number of heavily calcified lesions was comparable among the two modalities , however the presence of severe tortuosity  and the frequency of lesions > 20 mm long  were both significantly higher with ICA than CCTA. Despite the above-mentioned individual differences, the global average anatomical SYNTAX scores were similar. Furthermore, the incorporation of clinical characteristics to construct the SS-2020 confirmed the comparability of the SS-2020 and the predicted risk of MACE/all-cause mortality following surgical or percutaneous treatment.Lesion characteristics and the results of SS-2020 calculations between ICA and CCTA are shown in Table 2The predicted mortality with PCI or CABG according to the SS-2020, as well as the predicted individual ARD in all-cause mortality at 5- and 10-year, ranked in order of magnitude based on CCTA and ICA, is shown in a scatter plot indicating 5-year  and conventional ICA.Although, a significant difference was identified in predicted CABG mortality, the Bland–Altman analysis demonstrated acceptable agreement between the two modalities with no significant differences in individual ARDs as predicted by the SS-2020.The main findings from the present study, which to the best of our knowledge is the first report assessing the diagnostic performance of SS-2020 based on CCTA in patients with 3VD with or without LMCAD are:Several studies have compared the anatomical SYNTAX score derived from CCTA and ICA and reaffirmed that its calculation using CCTA is reasonably accurate and reproducible, especially in non-complex patients , 18–20, In the future CCTA may play a pivotal role as a “one-stop-shop” in screening, diagnosis, decision making, and treatment planning . RecentlIn the present study, we used the SS-2020 derived from CCTA to predict the best treatment considering personalized risk and treatment benefit and compared that selection with the one derived from ICA. Since the anatomical SYNTAX score (effect modifier) and the presence of LMCAD/3VD type of disease (effect modifier) were comparable between the two imaging modalities, the ARDs according to the SYNTAX 2020 were not significantly different, resulting in agreements of treatment recommendations for 5- and 10-year mortality of 84.2% and 80.7%, respectively. These results indicate that CCTA could be used as an alternative to ICA to make decisions regarding the modality of revascularization and to predict MACE and all-cause mortality. We acknowledge that 15.8% and 19.3% of the proportions had discordance in treatment recommendation between the two modalities of imaging. In the probabilistic formula predicting mortality based on the SS-2020 (Appendix), the difference in predicted mortality was partially determined by the two so-called “effect modifiers”—the presence of LMCAD/3VD and anatomical SYNTAX score when comparing the two imaging modalities in the same patient. In the present study, anatomical SYNTAX score and the occurrence of LMCAD/ 3VD were not statistically significant between the two modalities; however, there was a tendency for LMCAD to be more frequently diagnosed in ICA than CCTA, . This difference in imaging modalities might be implicated in the discordance in predicted mortality and thereby in treatment decision-making. In the original SYNTAX III REVOLUTION trial, the agreement in treatment recommendation based on the SYNTAX score II between the two modalities reached 93%, which was higher than our current results  and can There were several limitations to this study. First, clinical outcomes were not available. Only probabilistic prediction was calculated; therefore, we could not assess the calibration and discrimination of the two modalities versus MACE or mortality. Secondly, several subgroups were under-represented, such as female or patients with reduced LVEF. Thirdly, although external validation of the SS-2020 in the most contemporary cohort of the CREDO-Kyoto registry has shown the persisting actuality and accuracy of the probabilistic model , cliniciIn patients with 3VD, with or without LMCAD, there was moderate to substantial agreement between treatment recommendations based on the SS-2020 derived using CCTA and ICA, suggesting that CCTA could be used as an alternative to ICA when making decisions regarding the modality of revascularization.Supplementary file1 (DOCX 17409 kb)Below is the link to the electronic supplementary material.


Conclusions: We should pay attention to interpreting ReDS values when assessing the degree of pulmonary congestion in patients with PE, because ReDS values may be inappropriately elevated in this cohort.Background: The remote dielectric sensing (ReDS) system is a recently introduced non-invasive technology used to easily estimate the degree of lung fluid volume without any expert techniques. In the previous literature, ReDS values had a moderate correlation with invasively measured pulmonary artery wedge pressure (PAWP), the gold standard for representing left heart preload. Considering the mechanism of ReDS technology, ReDS values may be inappropriately elevated in the presence of pleural effusion (PE), and the ability of the ReDS system to estimate PAWP may decrease in such a situation. Methods: In-hospital patients with cardiovascular diseases underwent computed tomography, and the presence of pleural effusion (PE) was evaluated. The measurement of ReDS values using the ReDS system and the measurement of PAWP using invasive right heart catheterization were also performed simultaneously. The impact of the presence of PE on the relationship between the ReDS values and PAWP was evaluated. Results: A total of 59 patients  were included. The median ReDS value was 25%  and the median PAWP level was 13  mmHg. Of these patients, 11 had PE. PAWP was not significantly different between the two groups, whereas the ReDS values in the PE group were significantly higher than in the no-PE group. The impact of the presence of PE on the ReDS values was significant, with a beta value of 6.61 (95% confidence interval: 4.80–8.42,  Accurate assessment of the degree of pulmonary congestion is key to the successful management of congestive heart failure and to improving clinical outcomes in patients with congestive heart failure by adjusting diuretic therapy . The detSeveral clinical modalities are used to assess pulmonary congestion, including chest X-ray, plasma B-type natriuretic peptide levels, lung ultrasound, computed tomography, and invasive right heart catheterization . Of thesThe remote dielectric sensing (ReDS) system is a new technology that non-invasively estimates lung fluid volume based on electromagnetic energy and displays a percentage within one minute on a monitor . Its abiSeveral factors appear to influence ReDS values. ReDS values are overestimated in patients with chronic pulmonary obstructive disease . ObesityIn this cross-sectional observational study, patients who had undergone the following procedures were included: chest computed tomography, ReDS measurements, and right heart catheterization. The impact of the presence of PE on the difference between ReDS values and PAWP was evaluated.Hospitalized patients with cardiovascular diseases who had undergone the following three tests simultaneously in a clinically stable situation were retrospectively included in this study: (1) ReDS measurements, (2) right heart catheterization, and (3) chest computed tomography . PatientThe ReDS system is a recently developed non-invasive electromagnetic-energy-based technology used to quantify lung fluid levels within a minute of application . ReDS emReDS values were measured in 45–60 s with the patients in a seated resting position, breathing normally, using two sonar sensors worn on the right shoulder . The disInvasive right heart catheterization was performed via the right jugular vein under local anesthesia to directly measure intra-cardiac pressure and estimate cardiac function by board-certified cardiologists in a specialized catheter laboratory. Hemodynamic parameters were measured in a standard manner during breath-holding in an expiratory state, including right atrial pressure, pulmonary artery pressure, PAWP, and cardiac output. Raw data were double-checked and a consensus was established by multiple cardiologists who were blinded to the measured ReDS values in each patient.Chest computed tomography images without enhancement were obtained upon admission to semi-quantify the degree of right thoracic PE. An anteroposterior quartile was used to define the degree of PE by a researcher (TI) who was blinded to the results of the ReDS assessment and right heart catheterization at the time of measurement . The antp-values < 0.05 were considered statistically significant.Continuous variables were expressed as median and interquartile range (IQR) and compared between the two groups using a Mann–Whitney U test. The normality of their distribution was confirmed using the Shapiro–Wilk test. Categorical variables were expressed as numbers and percentages and compared between the two groups using Fischer’s exact test. The correlation between ReDS values and PAWP was evaluated using Pearson’s correlation coefficient. Linear regression analysis was performed to assess the relationship between ReDS values and PAWP following confirmation of the normality of residuals. The impact of the presence of PE on ReDS values was evaluated via analysis of covariance, in which PAWP was used as a covariant. Statistical analyses were performed using SPSS Statistics 26 . Two-sided A total of 59 patients were included . The medp < 0.05 for both; p > 0.05).A total of 11 patients had PE between the first and second interquartile lines, indicating the presence of significant PE (PE group). No patients had PE greater than the second interquartile line . The othp < 0.001, N = 59; The ReDS values and PAWP had a moderate correlation in all cohorts ) = 0.47 × (PAWP (mmHg)) + 26.0. The linear regression equation in the no-PE group is as follows: ) = 0.73 × (PAWP (mmHg)) + 15.1. According to the linear regression equation of the no-PE group, the ReDS values under normal conditions without PE could be estimated. All actually measured ReDS values in the PE group were higher than the estimated ReDS values, except for one patient  versus 30% ) = (PAWP (mmHg) + 12.3)/1.03. The dotted line represents the following: ) = ).p = 0.005 according to Wilcoxon’s signed-rank test). In other words, the actually measured ReDS values were overestimated in the presence of PE. The impact of the presence of PE on the ReDS values was significant, with a beta value of 6.61  upon adjusting for the levels of PAWP. All actually measured ReDS values were higher than the estimated ReDS values, except for one patient , the ReDS value will be underestimated ,17. We dThe presence of excessive extra-thoracic subcutaneous fat would inappropriately increase ReDS values . The manTheoretically, another disease that may inappropriately increase ReDS values is PE. PAWP was not significantly different between the patients with and without PE, whereas ReDS values were inappropriately elevated in patients with PE compared with those without PE. The actually measured ReDS values in the PE group were higher than expected .It is no surprise that the ReDS system cannot distinguish extra-lung fluid  from intra-lung fluid . When patients have PE, we should pay special attention to estimating the degree of pulmonary congestion by using the ReDS system alone; ReDS values may be inappropriately elevated and the severity of pulmonary congestion may be overestimated. Other multiple modalities may be recommended to accurately interpret the severity of pulmonary congestion in patients with PE, instead of the ReDS system alone.This is a proof-of-concept study with a small sample size . We included patients who had received right heart catheterization. This is generally performed under relatively stable clinical conditions to avoid hemodynamic deterioration during/after the procedure . Thus, wThe ReDS values appeared to be overestimated approximately 5% more than expected in patients with PE. The clinical implication of such a degree of overestimation of ReDS values may vary depending on each clinical scenario and requires further validation studies. For example, such an overestimation of ReDS values could have great implications in patients with relatively higher ReDS values.All participants in the PE group had mild PE. The amount was between the first interquartile and the second interquartile lines. We could not evaluate the impact of severe PE, which may have a greater impact on ReDS values compared with mild PE. Right heart catheterization in such patients is challenging and has the risk of hemodynamic deterioration. Also, none of the patients with PE had obvious causes for their PE, such as malignancy or pneumonia. In other words, all PE cases in this study were idiopathic. The impact of PE on ReDS values may vary in each etiology.In patients with PE, ReDS values were inappropriately elevated. The severity of pulmonary congestion may be overestimated when we use the ReDS system in patients with PE. Other modalities, such as right heart catheterization, are recommended for assessing the severity of pulmonary congestion in patients with PE.


Neither specific uptake of dopamine and binding of [3H]dihydrotetrabenazine, nor mean values of the calculated ratios of dopamine uptake and VMAT2 binding, a measure of uptake rate per transport site, were significantly different between ILBD and controls. ATP-dependence of [3H]dopamine uptake revealed significantly higher rates in putamen than in caudate at saturating concentrations of ATP in controls, a subregional difference lost in ILBD. Our findings support a loss of the normally higher VMAT2 activity in putamen as a contributing factor to the higher susceptibility of the putamen to dopamine depletion in idiopathic PD. Moreover, we suggest ILBD postmortem tissue as a valuable source for testing hypotheses on processes in idiopathic PD.Incidental Lewy body disease (ILBD) is a neuropathological diagnosis of brains with Lewy bodies without clinical neuropsychiatric symptoms. Dopaminergic deficits suggest a relationship to preclinical Parkinson’s disease (PD). We now report a subregional pattern of striatal dopamine loss in ILBD cases, with dopamine found significantly decreased in the putamen (−52%) and only to a lower extent in the caudate ; this is similar to the pattern in idiopathic PD in various neurochemical and in vivo imaging studies. We aimed to find out if our recently reported impaired storage of dopamine in striatal synaptic vesicles prepared from striatal tissue of cases with idiopathic PD might be an early or even causative event. We undertook parallel measurements of [ However, there is a range of neurological syndromes with striatal loss of dopamine that are distinct from idiopathic PD, such as multiple system atrophy and progressive supranuclear palsy6, summarized as atypical PD. In neurochemical studies on postmortem brains of patients with idiopathic PD, a characteristic pattern of dopamine loss has been found in the striatum with a nearly complete depletion of dopamine in all subdivisions of the putamen and substantial levels of dopamine remaining in the caudate7. With the progress of in vivo imaging methods, this pattern has also been detected in live patients with various stages of idiopathic PD12 and allowed differentiation between idiopathic PD and atypical PD17. Although the incidence of idiopathic PD increases with age, and there is an age-dependent reduction in striatal dopamine innervation18, the regional pattern of striatal dopamine loss in normal aging differs substantially from the pattern typically observed in idiopathic PD19. By contrast, 18F-dopa uptake for asymptomatic co-twins of patients with PD already revealed the typical pattern with a small but significant reduction of the dopamine marker in putamen but not in caudate20.Individuals with Lewy bodies or Lewy neurites at autopsy but without clinical findings of parkinsonism or dementia during their lifetime can be subsumed under cases with incidental Lewy body disease (ILBD). They are considered pre-symptomatic Parkinson’s disease (PD) by their nigrostriatal pathological features that are intermediate between those in morphologically normal control persons and those with overt PD21 and the vesicular monoamine transporter 21. However, all striatal findings are based on analysis of the putamen, and there are neither data on the caudate in ILBD in the literature nor on striatal dopamine tissue levels. We therefore set out for analysis of monoamine neurotransmitter tissue levels in both caudate and putamen of cases with ILBD. In a recent study on postmortem striatal tissue of cases with idiopathic PD, we observed a loss of 56% and 90% dopamine transport efficiency in synaptic vesicles of dopamine nerve terminals in the caudate and in the putamen, respectively22. Therefore, we also performed vesicular uptake on the caudate and putamen of our cases in this presumptive preclinical phase of PD.In ILBD, two neurochemical markers of striatal dopamine nerve terminals have been reported to be reduced, the immunoreactivity of tyrosine hydroxylase25. Whereas noradrenaline and serotonin were either unchanged or insignificantly increased in ILBD . The calculated ratio of dopamine uptake to [3H]DTBZ binding reflects the number of transported dopamine molecules per unit time per VMAT2 transport site, a measure of turnover or transport efficiency. There was no significant difference between the putaminal or caudate tissue of controls and ILBD cases neither in dopamine uptake and [3H]DTBZ binding nor in turnover  since dopamine tissue levels essentially stem from dopamine within synaptic vesicles where dopamine depends on continuous accumulation by the vesicular monoamine transporter. The loss of the higher activity of the vesicular monoamine transporter in putamen than in caudate in ILBD might be an early event in the development of neurodegeneration in idiopathic PD. Since transport was determined by an accumulation assay, differences in energy-dependent storage might, in principle, contribute to the measured pmol/mgprot/min, although the short incubation time of 5 min, aiming at initial rates, makes this contribution less likely. Storage may, however, be a factor in dopamine tissue levels. Findings on reduced NADH-ubiquinone reductase and NADH cytochrome c reductase activities in the substantia nigra45 and down-regulated genes of subunits of the respiratory chain complex I in nigral dopamine neurons47 suggest disrupted energy metabolism in dopamine neurons in PD which might have a higher impact on dopamine in putamen with its higher levels and therefore higher energy-dependence. However, another very recent laser capture microdissection study on neuromelanin-positive cells revealed an upregulation of oxidative phosphorylation pathways in ILBD48, suggesting that reduced vesicular uptake and storage in putamen might be more related to changes in vesicular monoamine transporter 2 or vesicular H+-ATPase.Since active transport into storage vesicles is driven by a transmembrane pH and electrochemical gradient generated by the vesicular H49, activation of NADPH oxidase 250, or LRRK2 kinase51 in tyrosine hydroxylase positive neurons. ILBD postmortem tissue could be a valuable source for testing these hypotheses in idiopathic PD at an early stage with more validity for a causal role in the progression of the disease.In conclusion, although our findings cannot rule out that ILBD is only an associated factor of PD, they give new support to the assumption that ILBD might be a preclinical stage of idiopathic PD. Obviously, this stage is only available for examinations postmortem since it is based on a neuropathological analysis. However, quite a few novel hypotheses about the neurodegenerative process in PD have been tested in postmortem tissue from PD patients, such as the increased expression of the vesicular glutamate transporter52  pathology and Consortium to Establish a Registry for Alzheimer’s Disease (CERAD) scoring of the density of neuritic plaques) and McKeith Lewy body stage53, LBD-related neuropathological findings of the subjects, diagnosed according to the Lewy Pathology Consensus Criteria54 are shown in Table Tissue for this study was provided by the Newcastle Brain Tissue Resource, with full informed consent of donors. Brain tissue was obtained at autopsy and stored within the Newcastle Brain Tissue Resource (NBTR), with the full written informed consent of the donors, in accordance with the Newcastle University Ethics Board . Samples from the midcaudate (head) and midputamen  in 1 M Tris-HCl, pH 8.6, and after washing with H2O and desorption with 0.2 ml 0.1 M perchloric acid containing 0.4 mM sodium bisulfite; 100 µl was injected into a high-performance liquid chromatography system with electrochemical detection (HPLC/ED) with a sodium phosphate mobile phase with l-octane sulfonic acid as described previously55. For determination of serotonin, 3,4-dihydroxyphenylacetic acid, and homovanillic acid, the supernatant was diluted 1:4 with 0.1 M perchloric acid containing 0.4 mM sodium bisulfite, and 100 µl was injected directly into an HPLC/ED system with a sodium phosphate mobile phase with l-heptane sulfonic acid as described previously55.Distinct samples of 20–50 mg from caudate and putamen frozen and kept at −80 °C were ultrasonicated from all subjects with an ultrasonic probe sonicator in 25 volumes of ice-cold perchloric acid, sodium bisulfite, and 3,4-dihydroxybenzylamine as internal standard . For determination of noradrenaline and dopamine, homogenates were centrifuged at 16,100 × 3HDTBZ]-binding from the same preparation, about 140–180 mg of striatal tissue were homogenized in 8 volumes of ice-cold 0.3 M sucrose containing 25 mM Tris (pH 7.4) and 10 µM pargyline, using seven up-down strokes in a glass Teflon Potter-type homogenizer and the whole further procedure was performed at 4 °C. The sucrose-homogenate was centrifuged at 1000 × g for 15 min, and the supernatant was recentrifuged at 20,000 × g for 30 min. The resulting pellet (“P2”) was subjected to osmotic shock by resuspension in 1 ml H2O, frozen for about 15 min, and, after thawing, homogenized in a total of 2 ml H2O. The aqueous samples were centrifuged at 22,000 × g for 15 min, and the osmolarity of the supernatant was readjusted by the addition of 1.3 M potassium phosphate buffer (pH 7.4) in 1/10 the volume. The supernatant of the 20,000 × g centrifugation (see above) was centrifuged at 100,000 × g for 30 min, and the resulting pellet was resuspended in the about 2.17 ml of the 22,000 × g supernatant readjusted to 0.13 M potassium phosphate, thus combining vesicles in the supernatant of the P2 pellet and in the hypoosmotically shocked P2 pellet. After incubation in a total volume of 3 ml assay buffer KP  in the presence of 2 mM MgATP in a 30 °C water bath for 4 min and centrifugation at 4 °C at 100,000 × g for 45 min the resulting pellet was resuspended in 4.8 ml KP. On each preparation, 1.5 ml were centrifuged at 4 °C at 153,000 × g for 45 min, and the pellet was resuspended in 100 µl of binding buffer SP  and frozen at −80 °C until the [3H]DTBZ binding experiment and the rest of the preparation was used for vesicular uptake. Protein content was determined according to the Bradford method by a Bio-Rad assay kit.For performing vesicular uptake and dopamine , various concentrations of MgATP  and 1 µM reserpine for determination of unspecific uptake as described previously22. Transport was initiated by placing the tubes in a 30 °C water bath and adding 0.5 ml vesicle suspension  for 5 min. Uptake was terminated by the addition of 2.5 ml ice-cold KP and immediate filtration under vacuum onto Whatman GF/B filter paper pre-soaked in 1% polyethylenimine using a Brandel harvester. The filters were washed twice with an additional 3 ml of cold KP and, after shaking in a liquid scintillation cocktail at 55 °C for 1 h, analyzed for tritium radioactivity in a liquid scintillation counter.Uptake was performed in a total volume of 1.5 ml KP containing 0.1 µM [22.Binding of (±)-a-dihydrotetrabenazine [2-3H]  was performed on 25 µl thawed preparation in SP at a total volume of 25 µl for 90 min at 30 °C as described previouslyData are reported as mean ± SE. Statistical analysis was done for neurotransmitter tissue levels using two-way analysis of variance for the factors disease (ILBD or control) and region (caudate or putamen) and for comparison of uptake in the putamen and caudate in the various control and ILBD cases using two-way repeated measures ANOVA (two factor repetition) for the factors region (caudate or putamen) and ATP , both tests followed by pairwise multiple comparison procedures (Holm–Sidak method) after data passed the normality test (Shapiro–Wilk) and the equal variance test (Brown–Forsythe).Further information on research design is available in the Supplementary Figure 1Reporting Summary


Molecularly imprinted polymers (MIPs) have gained significant attention as artificial receptors due to their low cost, mild operating conditions, and excellent selectivity. To optimize the synthesis process and enhance the recognition performance, various support materials for molecular imprinting have been explored as a crucial research direction. Yeast, a biological material, offers advantages such as being green and environmentally friendly, low cost, and easy availability, making it a promising supporting substrate in the molecular imprinting process. We focus on the preparation of different types of MIPs involving yeast and elaborate on the specific roles it plays in each case. Additionally, we discuss the advantages and limitations of yeast in the preparation of MIPs and conclude with the challenges and future development trends of yeast in molecular imprinting research. Molecular imprinting ,2,3 is aOver the past few years, researchers have focused on addressing these challenges through the exploration of different supporting substrates in molecular imprinting, aiming to optimize the synthesis process of MIPs and enhance their recognition performance. Conventional carriers used in molecular imprinting encompass membranes, silica particles, graphene, as well as other organic and inorganic materials ,14,15,16Numerous comprehensive reviews published in recent years have thoroughly discussed the application capabilities of different MIPs in the molecular imprinting field ,22,23,24This study is an analytical bibliographic review related to the preparation of different types of MIPs involving yeast and elaborates on the specific roles it plays in each case. Data collection was carried out from May 2023 to July 2023, using the following databases: PubMed, Science Direct from Elsevier, Wiley Online Library and Springer-Nature, ACS—American Chemical Society, and Google Scholar, as well as databases of scientific articles and patents “The LENS” and “ORBIT Intelligence”.The inclusion criteria for this work included original articles exclusive to the yeast and MIPs studied, with the full text available in Portuguese, English, and other languages. Exclusion criteria included abstracts, online sites without scientific sources, incomplete texts, and unrelated and repeated articles.Molecular imprinting; Imprinting Technique, Molecular; Molecular Imprinting Techniques; Techniques, Molecular Imprinting; Yeast; Fungus. The articles were selected by reading the titles and abstracts of the publications, associated using the Boolean descriptor “AND”, to refine the samples.As for the search strategy, the descriptive words used in this work were as follows: The review is based primarily on articles published after 2010. However, some older articles were also mentioned to provide relevant background or well-documented information. As seen from Yeast, a fungus, is a relatively large eukaryotic and single-celled organism . Yeast is widely distributed in nature and possesses advantages such as a short growth cycle, fast reproduction rate, simple structure, easy cultivation, relatively simple genome, and convenience of operation. These advantages make it a widely used model organism for research in the fields of biology and genetics ,26. ThesFirst and foremost, as the first extensively studied and stable model organism, yeast can be used as a target for studying the mechanisms of molecular imprinting. For example, Derick et al.  used yea2), hydroxyl (–OH), and phosphate (–P=O) groups. These characteristics make yeast suitable for molecular imprinting without further modification. Additionally, yeast has the advantages of low cost and ease of cultivation compared to other imprinting carriers such as silica gel, chitosan, nanotube membranes, and graphene. As seen from Over the years, a large number of molecularly imprinted materials (MIMs), including materials at the nanoscale, have been carefully designed and applied in molecular imprinting . Among tThe utilization of different materials provides the basis for the separation or detection of numerous compounds, including large molecules and cellular structures, thereby promoting rapid development in the field of molecular imprinting. In 2012, Li et al.  describeIn order to achieve the selective recognition and capture of target molecules, yeast is commonly employed as a supporting substrate in molecular imprinting techniques. The general principle involves a simple pretreatment of the yeast and creating an imprinted polymer layer on its surface. However, the precise role of yeast varies across different methods utilized for synthesizing MIPs.According to the interactions between the template molecules and functional monomers, MIPs can be classified into three types: covalent MIPs, non-covalent MIPs, and semi-covalent MIPs . In the Precipitation polymerization is a widely used method for the preparation of spherical particles. The basic principle involves adding reactants including template molecules, functional monomers, and cross-linking agents to a large amount of solvent. Initially, pre-assembly occurs, followed by the addition of an initiator to initiate the polymerization reaction at a specific temperature. After template removal using an elution agent, MIPs with specific imprinting cavities are obtained. This method typically yields uniformly sized spherical particles with a large surface area and significant selective recognition capabilities . PrecipiIn recent years, reversible addition–fragmentation chain transfer (RAFT) ,49,50 an3O4 nanoparticles and used an in situ precipitation polymerization method to prepare thermo-responsive magnetic molecularly imprinted polymers (TMMIPs) on the yeast surface, which was proved by FT-IR spectra: the peaks at 609 cm−1 were assigned to Fe–O bond vibrations of Fe3O4, which demonstrates that Fe3O4 was successfully introduced to mag-yeast and TMMIPs. The TMMIPs exhibited excellent stability and regenerability. They were successfully applied to the selective adsorption and release of tetracycline from aqueous solutions. The use of yeast as a support substrate offers advantages such as low cost, easy availability, and the presence of abundant active biomolecules on the cell wall, eliminating the need for further modification. More importantly, yeast provides a platform on which Fe3O4 nanoparticles can be evenly distributed, or else the Fe3O4 nanoparticles would be messy and could easily agglomerate. This study highlighted the promising role of yeast as a supporting substrate in the molecular imprinting process.To address these issues, Ma et al.  developeIn molecular imprinting, emulsion polymerization is another commonly used technique for preparing MIPs. In this method, the reactants including template molecules, functional monomers, and cross-linking agents are dissolved in a certain amount of organic solvent. The mixed solution is then transferred to an aqueous phase and emulsified by stirring, followed by the addition of an initiator to initiate the polymerization reaction. The MIPs obtained through this method have a controllable and uniform particle size, large surface area, and the ability to imprint water-soluble target analytes ,54,55. HCommon solid stabilizers in Pickering emulsions include inorganic materials such as silica, iron oxide nanoparticles, and calcium carbonate, as well as clays such as laponite and montmorillonite ,58,59,60Inspired by the aforementioned studies, the involvement of yeast as external and internal support substrates in emulsion polymerization has been well-reported in multiple studies. Taking advantage of yeast’s non-toxicity and excellent biocompatibility, Zhu et al.  utilizedIn the emulsion polymerization of MIPs involving yeast, yeast not only functions as a stabilizer but also participates as an internal support substrate. Wang et al.  utilizedWith the in-depth research on molecular imprinting, it has been observed that traditional embedding methods suffer from issues such as deep embedding of imprint recognition sites, difficulties in template molecule removal leading to limited effective recognition sites, and slow responses to template molecules. These problems are still challenging to address, and merely changing the polymerization method does not result in a qualitative transformation. Consequently, surface molecular imprinting has emerged and quickly become a new research hotspot .Surface molecular imprinting refers to the utilization of specific carriers  that undergo polymerization reactions on their surfaces, making it easily accessible to control the distribution of imprint sites on the surfaces. This facilitates the removal of template molecules and subsequent response and detection processes . The MIP−1 at 298 K. Similarly, Pan et al. [−1, and the maximum release proportion for the MIPs at 293 K in water for 24 h was 71.08%. The selective recognition experiments demonstrated a high affinity and selectivity of the MIPs towards CFX over competitive compounds. Yeast-based MIPs have demonstrated potential applications in the fields of selective separation and enrichment.The selection of support substrates with excellent morphology and mechanical properties is crucial for the preparation of surface MIPs. The use of yeast in molecular imprinting has been proven advantageous through multiple research studies. Li et al.  reportedn et al.  also uti3O4). Guan et al. [3·6H2O and FeCl2·4H2O. This method achieved selective adsorption of trace amounts of β-cypermethrin in contaminated wastewater. The MIPs exhibited an adsorption capacity of 39.64 mg g−1 at 298 K. The MIP data fit well in the Langmuir isotherm model compared to the Freundlich model, whereas the kinetic properties of the MMIPs were well described by the pseudo-second-order equation. Qiu et al. [2, magnetic yeast particles as carriers did not require intermediate chemical modification steps and significantly improved the grafting efficiency. These findings suggest that surface imprinting polymers based on composite materials combining yeast and magnetite hold great promise as one of the most viable candidates for the separation of environmental pollutants.In addition to using yeast as a standalone solid support for surface molecular imprinting, the inclusion of magnetic materials enables the separation of MIPs after use. Magnetic solid support substrates have the advantage of easy removal from the medium through simple magnetic separation and can be reused. Therefore, researchers have achieved the production of magnetic adsorbents by combining yeast with magnetite , allows for the removal of environmental pollutants from the medium through simple magnetic separation enabling their recyclability. Various methods have demonstrated the advantages of yeast involvement in MIP synthesis, as seen from Easy availability and low cost: Yeast is easily obtainable and can be cultivated at a low cost in a short time.Rich surface chemical functional groups: The yeast surface contains abundant chemical functional groups, which greatly simplify modification steps and reduce secondary pollution. The presence of hydroxyl, amino, carboxyl, and other functional groups allows for interactions with template molecules, enhancing the recognition and selectivity of MIPs.Controllable morphology and structure: By adjusting cultivation conditions and synthesis parameters, MIPs with controllable morphology and structure can be synthesized on the yeast surface. This tunability enables the design and fabrication of MIPs with specific shapes, pore sizes, and surface properties to meet diverse application requirements.Excellent biocompatibility: Yeast is a natural biological material with good biocompatibility and biodegradability. Using yeast as a carrier for MIPs reduces adverse environmental impacts.Regulation of inorganic material growth: Yeast can regulate the growth of inorganic materials, providing a rich template for the synthesis of nanomaterials through template-assisted synthesis.Diverse microbial cell structures: Microbial cells, including yeast, possess various structures that can serve as templates for the synthesis of nanomaterials, offering a wide range of templates.Addition of magnetic materials: The inclusion of magnetic materials allows for easy separation of the material after use. Magnetic solid supports have the advantage of easy removal from the medium through simple magnetic separation and can be reused.The application of yeast provides new avenues and possibilities for the development and application of molecular imprinting. However, as a material in molecular imprinting, yeast still has some potential drawbacks such as (1) limitations in reusability. Yeast may suffer damage or deactivation during the molecular imprinting process, limiting its reusability. This could increase the preparation cost and resource consumption of MIMs. (2) A limited applicability range as the chemical and physical properties of yeast may restrict its recognition ability for specific molecules. This limitation narrows the applicability range of yeast as a molecular imprinting material, rendering it ineffective in recognizing certain target molecules.Despite these potential drawbacks, appropriate optimization and improvement strategies can address these issues to some extent. Moreover, yeast-containing imprint materials show tremendous potential in areas such as adsorption and separation of antibiotics, pesticides, and antibacterial agents ,41. ThisThis review explored the utilization of yeast in the field of MIPs, with specific attention to the fabrication methods and applications of MIPs utilizing yeast as the supporting substrate. It provided valuable insights into the role and prospects of yeast in the development of molecular imprinting. However, due to the absence of research on the comparative adsorption performance of yeast and traditional substrates using equivalent ratios of template, monomer, and cross-linker compositions under equi-volume and solvent conditions, definitive conclusions regarding the differential adsorption performance between yeast and conventional substrates cannot be established. We firmly believe that future investigations aiming to enhance yeast properties and expand its applications will continue to expand. Prospective research could focus on various domains, especially green strategies, taking into account the numerous advantages offered by yeast as a biomaterial and the increasing emphasis on green chemistry. Exploring more environmentally sustainable research methodologies will be an intriguing challenge.