Input: Claim: 3D printing is used to create custom prosthetics and implants tailored to individual patients. [SEP] Text: a Carbon fiber socket of partial hand amputee was CT scanned at 1 mm slice increments and then reconstructed. STL file was exported. b The CT reconstruction was used to create socket overlay. Lifting hook was designed in CAD. c Overlay and hook were merged into one piece. d Design was 3D printed in titanium alloy. 3D Print was fit tested on carbon fiber socket. e Caron fiber was weaved through titanium overlay and laminated into the socket. Prosthetist added a ratchet system with strap extending around the elbow for pulling exercises. f & g The attachment was successful in allowing the service member the ability perform both pushing and pulling weight lifting exercisesFull size image Example 2: Hockey skate adaptersA bilateral below the knee amputee who was formerly an avid skater wanted to resume ice skating. The patient felt that they would not be able to tighten the skates enough around the prosthetic foam foot, which would reduce control and energy transfer to the skate and limited the desired sensory feedback to their residual limbs while skating. The blade mount was scanned with CT and reconstructed at 0. 625 mm slice thickness (Fig. 3a). The digital reconstruction was then exported as an STL file. Note that due to the complexity and glossy finish of the surface, it would have been difficult to obtain a 3D surface scan of the blade mount. The articulating surfaces were then offset from the blade holder, and a small lip was maintained for better seating of the device. An assembly was then designed in Solidworks to hold the prosthetic ankle joint and allow for adjustability of the device (Fig. 3b). The CAD assembly was also exported as an STL file. The assembly was aligned with the articulating surface of the device and connected using numerous posts (Fig. 3c & d). The parts were merged together to create the final product. The final design was then mirrored to produce the componentry for the contralateral side. Special attention was given to the adjustability of the ankle joint and the center of gravity as the patient did not have access to the biomechanical benefit of an active ankle joint. The device was first 3D Printed on a (SLA 7000), from AccuraSL 7570 (3D systems) for test fitting. Once the fit was confirmed, the device was manufactured on an Arcam(ArcamAB) from titanium alloy (Fig. 3e). The device was assembled by the prosthetist.

Output: corroborate


Input: Claim: Everyone's microbiome composition is identical. [SEP] Text: Here, we observed larger volumes in the yy control genotype relative to the WW control genotype, and a general increase in both genotypes after frass transplantation. Different diets may partly explain the discrepancies between the studies. For instance, laboratory-reared moths were fed dandelion (Taraxacum spp. ) in the previous study, whereas an artificial diet was used here. Likewise, the larval diet of wild-caught adults is unknown. Hence, it is difficult to draw comparisons between absolute volumes of defensive secretions. However, it can be inferred that diet is not a major contributor to the defensive secretions, because the frass transplantation increased the volume of defensive secretions in both genotypes, which fed on the same artificial diet mixed with frass of the other genotype. Differences in antipredator efficacy have also been reported, where the abdominal secretions of yellow moths are more deterrent against ants than secretions from white individuals [87]. The exact chemical composition of the abdominal fluids has not been characterized. However, the wood tiger moth can sequester hepatoxic organic compounds, such as pyrrolizidine alkaloids (PAs), from its diet [123], and these compounds have been detected in the abdominal secretions of wild-caught adults (Winters et al. , unpublished). PAs are well-known for providing protection to plants against herbivores, which in turn can host bacterial associates with detoxifying abilities [46, 107]. Some bacteria have even been suggested to synthesize PAs, as inferred by the presence of alkaloid biosynthesis gene clusters [91]. Manipulative experiments are needed to study PA sequestration-detoxification processes in the wood tiger moth. The great diversity of bacteria found in the abdominal secretions holds great potential to help elucidate the mechanism underlying these processes and to better understand differences in protection between the colour morphs. The great and largely unexplained variation of bacteria among individuals in the defensive secretions represents an example of how the drivers of bacterial community composition can be decoupled in larvae vs. adults [34]. We have previously shown that the wood tiger moth's pre- and post-metamorphosis life stages are only partly decoupled [26]. Our across-life stage analysis provides additional evidence for this partial de-coupling.

Output: contrast


Input: Claim: Some studies suggest that olfactory receptors may function through quantum effects. [SEP] Text: But referring to my comment above, couldn't it be that IGF worked only through an RTK already before it gave rise to GSS, then GSS too would act on the RTK. Minor comments In a couple of places the author writes "genes coding peptides" or similary. It should "encoding peptides" or "coding for peptides", as indeed written correctly on line 263. Please scan the manuscript for this. 61 It is advisable to delete "closely" as some ligands bind to receptor subtypes that arose by duplication some 500 million years ago or more and have diverged considerably. I suggest that it is emphasized that this discussion concerns peptide ligands by inserting the word "peptide" before ligand(s) in this paragraph. 66 Correct the spelling of "coupled" 72 The sentence says "One of these" which seems to refer to dilp7 in the preceding sentence, but it should be "One of the irps". 86-87 The part after the comma is confusing. It does not necessarily have anything to do with the fact that the IGF gene is part of a local triplet. 112 Should be: number of Amulacraria species. 128 For those unfamiliar with these species, please write: "transcriptome data from the three crinoid species Antedon…. " 154 should be "orthologous" 178 shold be "cleave any secreted protein with appropriate cleavage site". 206-211 This peptide organization would be good to illustrate with a figure. 216 "other echinoderms" should be "other ambulacrarians" 223 should be "cleavage sites" 258 it looks grammatically odd to see "are single copy", I suggest "is a single copy gene", this cannot be misunderstood. 260 Why write genus name for one and species name for the other? 308 should be "shows" 310 should be "where" 316-317: This sounds like a circular argument. Of course the tree based on sequence alignment and similarity will reflect the way they "look". Please rephrase. 324: should be NCBI 335: should be "multiple" 338-9: should be "from time to time" 339: "assembled constructs" is probably not the correct term here, I believe "assembled sequeces" would be more suitable. 340: "both" makes this sentence a bit difficult, please rephrase.

Output: pass


Input: Claim: 3D printing is used to create custom prosthetics and implants tailored to individual patients. [SEP] Text: The prosthetic attachment gave a bilateral amputee the ability to skate again (Fig. 3f & g). A special tool was also manufactured to allow the prosthetist or amputee to tighten bolts with minimal access. Fig. 3Adapter. a Blade mount was CT scanned at 0. 625 mm slice increments and then reconstructed. STL file was exported. b Design and freeform articulating surfaces created. c Files were aligned with the reconstruction of the skate blade mount. d Connecting geometry (posts) added between entities. e Three part assembly manufactured from titanium alloy using an electron beam melting machine. f & g Prosthetic attachments allow patients to enjoy skating or play stand up hockeyFull size image Example 3: Wine glass holderA 34-year old man status post upper extremity amputation had difficulty holding a standard wine glass due to the limitation of their terminal device. The prosthetic hook either applied too much pressure to the stem of the glass or the glass would wobble in the empty space between the two prongs. In order to resolve this issue, a mold of the empty space was created by placing lab putty in the open hook. The hook was then closed, allowing the putty to fill the void between the two prongs, creating an impression of both prongs (Fig. 4a). The set mold was then removed and scanned using CT at 5 mm slice increments. While 5 mm slice thickness is larger than the typical requirements, this level of detail was sufficient to serve as a basis for design. The scan was reconstructed in v17 and exported as an STL file (Fig. 4b). Note that due to the undercuts of the object, an accurate 3D surface scan would be difficult to obtain. The STL file was verified and imported into FreeFormPlus 2013 (3D Systems). A cup was added to support the bowl of the wine glass, and a 9 mm hole and slit were added for wine glass insertion (Fig. 4c). The final device was manufactured on an Objet500 Connex2 3D printer (Stratasys). The device was made from a combination of Plus and Plus to obtain the appropriate durometer for the device (Fig. 4d). This prosthetic device enabled the amputee to enjoy wine again (Fig. 4e). Fig. 4Holder. a Lab putty was used to make impression of the space between the prosthetic hook.

Output: corroborate


Input: Claim: Everyone's microbiome composition is identical. [SEP] Text: For instance, the genotype-specificity of Erysipelotrichaceae remained in the abdominal fluids, which showed that their bacterial community was not completely uncoupled from the previous life stages. A curious observation was a higher diversity of Enterococcus ASVs in the abdominal fluids compared to larvae, frass or gut. Together with the signs of phylogenetic clustering in the fluids, this finding suggests that distinct and partly unidentified drivers govern the bacterial community dynamics in the adult’s defensive secretions vs. in the larvae. ConclusionsWe show for the first time that genotype-specific life-history traits in a lepidopteran host can be reversed with a reciprocal frass transplantation. Our results help clarify bacterial community assembly in Lepidoptera at different life-stages and relate the bacterial community composition to the genotype and growth of the wood tiger moth. With nearly full-length 16S rRNA gene amplicons, we were able to discern the dynamics of closely related enterococci and recovered a novel cluster of insect-associated, genotype-specific Erysipelotrichaceae. Our findings indicate that an insect host can fine-tune its bacterial community in a genotype-specific manner even against a highly variable bacterial community background with stochastic community assembly. The strong selection of bacteria in larval frass appeared to be relaxed in adults, which shows that previous findings of consistent bacterial community in adult Lepidoptera [33] may not apply to species that do not feed as adults. Thus, not only life stage but also species-specific adult feeding habits and host genotype can influence bacterial community assembly contributing to the high variation and inconsistencies observed in Lepidoptera microbiomes. Overall, our results suggest that the digestive processes of slow- and fast-growing host genotypes differ in filtering or retaining specific bacterial groups. Looking deeper into the functional, chemical and genomic background of these differences could reveal the molecular and ecological mechanism of the filtering. Availability of data and materials The sequence data are available in the for Archive under accession code PRJNA804133. R scripts and data files for microbiome analysis are available at R scripts and data files for life history analysis will be available at 31. 46. 9. 2. Nat Biotechnol. 2019;37:852–7. 0081. 34. 300. 3. 103. 63. 9. 1335. 9505. 118. 15.

Output: contrast


Input: Claim: Some studies suggest that olfactory receptors may function through quantum effects. [SEP] Text: 343: should be and adverb: "particularly" 352: "subbranches" should be followed by a comma. 357: "it is interesting to see" comments on something that cannot be seen in this clade in Fig. 9. Please describe differently or refer to a supplementary tree that contains this information. 363: What is "The second branch"? Please describe in an unambiguous way. 371: "are" should be "is" 372: "it are the glycoprotein hormone and relaxin receptors that are identified" can be corrected to "the glycoprotein hormone and relaxin receptors are identified" 373: add comma after GRL101. 386: it is more common to say "echinoderms" 391: "This" has unclear refrence and the sentence is dangerously close to circular reasoning: "pressure is weak…because it is poorly maintained" - please rephrase. 394: should be "suggest" 427: "with" should be "by" 430: As the sentence says "a second insulin RTK gene", it will have to be: "none was found". 431: "neither" does not fit here, better to write "were unsuccessful" 435-436: why can't binding to a second receptor have arisen later in evolution? See my comments at the beginning of my referee report. 463: "it were the…that were identified" can be simplified to: "the…receptors were identified…" 492: "evolved" can be replaced by "diverged" Fig. 6. It would be very useful to have lines showing the organization of the prepro-peptides in relation to these schematic mRNA/cDNA outlines, and not only the Cys positions. Probably is would suffice with the prepropeptide outlines for the top and bottom sequences. Fig. S11: Is TM1 correctly marked? I am not totally familiar with LRR TM regions but I think it might be shifted right-wards. All six sequences have an R in the TM1 region and four of them have a K a few positions further upstream. This would be a bit unusual to have within a TM region, but the LRR:s are a bit unusual so it may be correct. Please double check the boundaries.

Output: pass


Input: Claim: 3D printing is used to create custom prosthetics and implants tailored to individual patients. [SEP] Text: b Putty impression was scanned using computed tomography (CT) at 5 mm slice increment and then reconstructed. STL was exported. c The cup, hole, and slit added to the design. d Device was 3D printed using multi-material jetting machine utilizing flexible material. e Prosthetic attachment enables an amputee to enjoy wine againFull size image Results and discussionIn this manuscript, we researched and presented several methodologies to construct personalized examples highlighting the benefits CT reconstructions and the 3D printing process provide in successfully manufacturing prosthetic devices in limited quantities. These are but a few examples of the applications of this technique. Three-dimensional reconstructions from computed tomography create an accurate starting geometry for designing custom prosthetic attachment and devices when barriers to traditional design processes and production methods exist. Once 3D reconstructions were obtained anywhere from thirty minutes (Holder) to three hours (Adapters) were spent on the digital design process. 3D Printing has created new opportunities for the production of prosthetic devices and allows for the creation of unique, customized devices. Some limiting factors of these developed methodologies include the availability of equipment and materials, software, scanners, CT scan expenses, 3D printers, and experienced staff. Materials for these prosthetic components were chosen based on the technologies and materials available at the institution. Preliminary results show that the titanium alloy used has allowed for large safety factors, greater than 2. 5, on prosthetic attachments analyzed. Future endeavors include 3D printing a mold for to manufacture the Holder from silicone rubber. For less complex designs conventional machining could also be utilized if the facility had in-house capabilities or outsourcing could be more economical. For these particular project and other similar projects, the cost of outsourcing these components would have been higher than the cost of in-house production. CT scanning cost will differ by the institution; however, these objects do not require the radiologist reading. ConclusionUsing CT in conjunction with digital design and 3D printing can be utilized to create custom rehabilitation devices. Facility resources and knowledge can be limiting factors. 3D printing has created new opportunities previously unavailable to prosthetics, occupational therapy, and assistive technology departments. This methodology continues to be utilized when conventional techniques are limiting or suboptimal. Interprofessional collaboration, imaging, and digital manufacturing expertise are vital to the successful form, fit, and function of these devices. 2012. 15. F2F. MA. FAAST86.

Output: corroborate


Input: Claim: Everyone's microbiome composition is identical. [SEP] Text: Deciphering composition and function of the root microbiome of a legume plant access Published: 17 January 2017 Deciphering composition and function of the root microbiome of a legume plant Kyle Hartman1,2, Marcel GA van der Heijden1,2,3, Valexia Roussely-Provent4, Jean-Claude Walser5 & …Klaus Schlaeppi1 Show authors Microbiome 0. 1% relative abundance) operational taxonomic units (OTUs) in Arabidopsis thaliana roots. However, it required considerable effort including large-scale isolation using serial dilutions (seven different bacteria isolation media were used! ) and subsequent high-throughput taxonomy identification. Experimental manipulation of the microbiome and assays with plants require contained systems in which host-microbiota interaction experiments can be conducted without outside microbial contamination. Recently, microcosm systems have been used in combination with bacteria reference stocks to examine the dynamic process of root microbiome assembly from a defined input community under microcosm conditions [21, 22]. In these experiments, stable and reproducible community assembly was observed. However, these experiments were not designed to clarify how root communities compare to plants grown in artificial substrate in microcosms or in natural soil conditions. Here, we addressed some of the aforementioned research gaps and report a detailed characterization of the Trifolium root bacteria microbiome. We sampled the whole-root system including nodules, removed the rhizosphere and investigated the entire root bacterial communities consisting of rhizoplane and endosphere habitats. We utilized a multi-step approach to investigate the composition and culturable fraction of its root microbiome (Fig. 1). We also move towards a functional understanding of specific members of the Trifolium root microbiome and developed a microcosm system (Additional file 1: Figure S1a–d) in which we conducted multi-strain inoculation experiments with Trifolium germinated from surface-sterilized seeds and investigated the inoculation-induced effects on plant growth. Fig. 1Characterization of the root microbiome. We collected a natural field soil and used it in a series of Trifolium growth experiments. (I) We investigated the composition of the root bacteria microbiome using 16S rRNA sequencing of root samples.

Output: contrast


Input: Claim: Some studies suggest that olfactory receptors may function through quantum effects. [SEP] Text: Cite this review as (2021) #2 of "Ambulacrarian insulin-related peptides and their putative receptors suggest how insulin and similar peptides may have evolved from insulin-like growth factor (v0. 1)". PeerJ Submission (PDF) - submitted Apr 16, 2021 All text and materials provided via this peer-review history page are made available under a Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

Output: pass


Input: Claim: 3D printing is used to create custom prosthetics and implants tailored to individual patients. [SEP] Text: Custom-made 3D printed subperiosteal titanium implants for the prosthetic restoration of the atrophic posterior mandible of elderly patients: a case series Download PDF Download eeOpen access Published: 08 January 2020 Custom-made 3D printed subperiosteal titanium implants for the prosthetic restoration of the atrophic posterior mandible of elderly patients: a case series Carlo Mangano1, Andrea Bianchi2, Mangano3, Jessica Dana4, Marco Colombo5, Ivan Solop3 & …Oleg Admakin3 Show authors 3D Printing in Medicine 69. 6, SD ± 2. 8, median 69, 95% CI 67. 9–71. 6) were included in the study. The fit of the implants was satisfactory, with a mean rating of 7 out of 10 (SD ± 1. 6, median 7, 95% CI 6–8). Only two implants had insufficient fit, because of the presence of scattering in the CBCT; however, they were adapted to the sites during the interventions. The mean duration of the intervention was 44. 3 min (SD ± 19. 4, median 37, 95% CI 32. 3–56. 3). At the one-year follow-up, no implants were lost (survival rate 100%). One implant presented immediate postoperative complications with pain, discomfort and swelling, and two patients experienced late complications, having their provisional restorations fractured during the temporisation phase. All these complications were minor in nature, but the final complication rate amounted to 30% (three of ten patients). ConclusionsAlthough this study has limits (small patient sample and short follow-up), DMLS has proven to be an effective method for fabricating accurate subperiosteal implants, with high survival rates. This may represent an alternative treatment procedure in elderly patients with a severely atrophic posterior mandible, since it allows avoidance of regenerative bone therapies. Further studies are needed to confirm these outcomes. BackgroundSubperiosteal dental implants appeared in Sweden and the in the middle of the last century [1, 2]. Subperiosteal implants were custom-made fixtures, inserted below the periosteum, and stabilised by contact with the underlying bone, by means of fixation screws and the fibro-mucous tissue that covered them [2,3,4].

Output: corroborate


Input: Claim: Everyone's microbiome composition is identical. [SEP] Text: (II) We utilized the same root material for an isolation effort to explore the culturable fraction of root bacteria microbiome and assembled a reference stock of bacteria isolates. (III) We subsequently developed a microcosm system to explore plant-microbiota interactions and (IV) investigated the composition of the Trifolium root microbiome in the system by inoculating microbiota extracted from the field soil. (V) We conducted microbiota manipulation experiments in which we inoculated culturable, abundant members of the root microbiome and scored their effects on plant growthFull size image ResultsComposition of the Trifolium root microbiomeThe 16S amplicon sequencing of 24 Trifolium root samples and 15 soil samples from climate chamber and natural site growth experiments (Fig. 1, Table 1, Additional file 1: Figure S2,) yielded 9,923,925 high-quality, non-chimeric sequences across all samples, with a median of 153,072 (range 21,731–981,922) sequences per sample (Additional file 2). We rarefied the dataset to an even sequencing depth of 20,000 sequences and identified 3495 bacteria OTUs and one archaea OTU. Table 1 Overview of the number of replicate samples by sample type, experiment, and experimental replicate, or plotFull size table We confirmed in the Trifolium root microbiome the typical patterns that are often observed in microbial ecology. The soil microbiome is richer and phylogenetically more diverse than the root microbiome (Additional file 1: Figure S3; Table S1). We quantified the major components driving differences between samples (ß-diversity) using unconstrained principal coordinates analysis (PCoA) on weighted UniFrac distances and found a clear separation along axis 1 (explaining 69. 7% of the overall variation) and confirmed the general pattern that soil and roots harbor distinct microbiomes (Fig. 2). Axis 2 explained 15. 5% of the variation overall and separated mainly the root but not the soil samples, and we did not notice an obvious clustering whether the plants were grown in the same soil in a climate chamber or in the field, suggesting negligible effects of the growth condition on β-diversity. We detected a significant effect of growth condition on OTU richness only (Additional file 1: Figure S3; Table S1).

Output: contrast


Input: Claim: Some studies suggest that olfactory receptors may function through quantum effects. [SEP] Text: Peer review processEditorsReviewer #1Reviewer #2Reviewer #3Author ResponsePeer review processNot revised: Preprint includes the authors’ original preprint (without revision), an eLife assessment, public reviews, and a provisional response from the authors. Read more about eLife’s peer review process. of of Cambridge, Cambridge, United KingdomReviewer #1 (Public Review): Summary: Both flies and mammals have D1-like and D2-like dopamine receptors, yet the role of D2-like receptors in Drosophila learning and memory remains underexplored. The paper by Qi et al. investigates the role of the D2-like dopamine receptor D2R in single pairs of dopaminergic neurons (DANs) during single-odor aversive learning in the Drosophila larva. First, they use confocal imaging to screen driver strains with expression in only single pairs of dopaminergic neurons. Next, they use thermogenetic manipulations of one pair of DANs (DAN-c1) to implicate DAN-c1 activity during larval aversive learning. They then use confocal imaging to demonstrate expression of D2R in the DANs and mushroom body of the larval brain. Finally, they show that optogenetic activation during training phenocopies D2R knockdown in these neurons: aversive learning is impaired when DAN-c1 is targeted, while appetitive and aversive learning are impaired when the mushroom body is manipulated. Qi et al. thus propose a model in which D2R limits excessive dopamine release to facilitate successful olfactory learning. Strengths: The paper reproduces prior findings by Qi and Lee (2014), which demonstrated that D2R knockdown in DL1 DANs or the mushroom body impairs aversive olfactory learning in Drosophila larvae. The authors extended this previous work by screening 57 GAL4 drivers to identify tools that drive expression in individual DANs and used one of the tools, the R76F02-AD; R55C10-DBD driver, to manipulate DAN-c1 neurons with greater specificity. They also show that GFP-tagged D2R is expressed in most DANs and the mushroom body.

Output: pass


Input: Claim: 3D printing is used to create custom prosthetics and implants tailored to individual patients. [SEP] Text: With the advent of digital technology, a new era in dentistry has begun [25]. From acquisition methods such as cone beam computed tomography (CBCT), which have considerably reduced the number of x-rays given to patients [26], to intraoral scanners [27], digital software, 3D printers and many other methods and materials [28], these technologies have simplified, improved and substantially sped up several procedures. Such technological advancement allows clinicians to see the world of dentistry in a completely different way, which is developing exponentially [25]. This digital revolution opens up new horizons, such as 3D printing and in particular direct metal laser sintering (DMLS) [29], which allows fabrication of custom-made meshes [30, 31] and even implants [32, 33] perfectly adapted to the patient’s specific anatomy. This allows the opportunity to revisit some old concepts, such as the placement of subperiosteal implants, and reinterpret them in a new technological context based on consolidated anatomical and physiological principles [34,35,36]. The reduction of treatment to a single surgical session, lower costs for the patient and, above all, the precision that makes the method more predictable and safer in the short term have brought the attention of clinicians back to the use of subperiosteal implants, particularly for the management of complex atrophies such as in the posterior mandible of elderly patients [36, 37]. In severe posterior mandible bone resorption, when the patient does not want to undergo bone regeneration, modern digital technologies may represent a viable solution, with the possibility to fabricate custom-made subperiosteal implants perfectly adapted to their local morphology and anatomy [36, 37]. This is of particularly interest for elderly patients with special needs, who do not want or cannot undergo complex regenerative surgeries, but need a fixed prosthetic restoration [37]. The purpose of this case series was to show the clinical application of custom-made 3D-printed subperiosteal titanium implants for fixed prosthetic restoration of the atrophic posterior mandible of elderly patients. MethodsData acquisition2017 and June 2018, all partially edentulous patients with missing teeth in the posterior mandible, who were considered for possible inclusion in this study, received a radiographic evaluation.

Output: corroborate


Input: Claim: Everyone's microbiome composition is identical. [SEP] Text: However, experiment-to-experiment variation (especially climate chamber experiment 2) largely explained the variability between root samples (Additional file 1: Figure S4). Possible effects due to differences in climatic conditions were generally not detected and would have an effect size smaller than replicate experimental variation. Fig. 2Sample type, growth conditions, and experiment explain much of the variation in soil and root bacteria communities. Unconstrained principal coordinates analysis (PCoA) of weighted UniFrac distances of root and soil samples from climate chamber () and natural site growth experiments (), as well as the unplanted experimental field soil (Exp. Soil). file 1: Figure S4 for points colored by the replicate experimentFull size image In the following, we break down the dissimilarities between soil and root samples to compositional patterns evident in the taxonomic profiles of the samples. Soil samples contained abundant Proteobacteria, Actinobacteria, and Acidobacteria accounting for a mean of 54. 7, 24. 7, and 6. 9%, respectively (Additional file 1: Figure S5). root microbiome was dominated by Proteobacteria that accounted for a mean abundance of 90. 7% across both experimental conditions (Additional file 1: Figure S5). For the detailed characterization of the Trifolium root microbiome (Fig. 1, step I), we first identified the OTUs that were significantly higher in relative abundance in root compared to soil samples and discovered a total of 61 OTUs significantly enriched in root samples (Fig. 3), 15 of which were abundant with a mean relative abundance of at least 0. 1% across all root samples. These 15 OTUs accounted for 74. 5% of rarefied sequences, and we termed them “RootOTUs”—referring to the abundant and root-specific members of the Trifolium root microbiome. The RootOTUs consisted mostly of Proteobacteria (14 OTUs, Additional file 1: Table S2) and represented six different orders: Rhizobiales (6), Sphingomonadales (3), Enterobacteriales (2), Burkholderiales (1), Caulobacterales (1), and Rhodospirillales (1). The remaining non-Proteobacteria RootOTU belonged to the Firmicutes and was classified in the genus Syntrophomonas.

Output: contrast


Input: Claim: Some studies suggest that olfactory receptors may function through quantum effects. [SEP] Text: Although the authors only train larvae with a single odor, they demonstrate that driving D2R knockdown in DAN-c1 neurons impairs aversive learning, as do other loss-of-function manipulations of DAN-c1 neurons. Weaknesses: The authors claim to have identified drivers that label single DANs in Figure 1, but their confocal images in Figure S1 suggest that many of those drivers label additional neurons in the larval brain. It is also not clear why only some of the 57 drivers are displayed in Figure S1. Critically, R76F02-AD; R55C10-DBD labels more than one neuron per hemisphere in Figure S1c, and the authors cite Xie et al. (2018) to note that this driver labels two DANs in adult brains. Therefore, the authors cannot argue that the experiments throughout their paper using this driver exclusively target DAN-c1. Missing from the screen of 57 drivers is the driver MB320C, which typically labels only PPL1-γ1pedc in the adult and should label DAN-c1 in the larva. If MB320C labels DAN-c1 exclusively in the larva, then the authors should repeat their key experiments with MB320C to provide more evidence for DAN-c1 involvement specifically. The authors claim that the SS02160 driver used by Eschbach et al. (2020) labels other neurons in addition to DAN-c1. Could the authors use confocal imaging to show how many other neurons SS02160 labels? Given that both Eschbach et al. and Weber et al. (2023) found no evidence that DAN-c1 plays a role in larval aversive learning, it would be informative to see how SS02160 expression compares with the driver the authors use to label DAN-c1. The claim that DAN-c1 is both necessary and sufficient in larval aversive learning should be reworded. Such a claim would logically exclude any other neuron or even the training stimuli from being involved in aversive learning (see Yoshihara and Yoshihara (2018) for a detailed discussion of the logic), which is presumably not what the authors intended because they describe the possible roles of other DANs during aversive learning in the discussion.

Output: pass


Input: Claim: 3D printing is used to create custom prosthetics and implants tailored to individual patients. [SEP] Text: a Mandible, occlusal view; b maxilla, occlusal view; c occlusion, lateral viewFull size imageInclusion and exclusion criteriaThe inclusion criteria for enrolment in the study were: age over 65 yearsgood systemic and oral healthacceptable oral hygienepartially edentulous mandible, with two or more teeth missing in the posterior sectors and marked atrophy that disallowed insertion of standard size implants (length ≥ 10 mm)willingness not to undergo regenerative bone surgerywillingness to attend the follow-up control visits. The exclusion criteria for this study were: age under 65 yearssystemic pathologies or pharmacological therapies that could contraindicate the intervention (such as immunocompromised states, non-compensated diabetes, tumours of the head and neck, or treatment with oral or parenteral bisphosphonates)inadequate oral hygienesmoking habitbruxismcompletely edentulous mandible, or partially edentulous mandible in the posterior sectors with bony bases allowing insertion of implants of standard dimensions (length ≥ 10 mm) without the risk of damaging nervous structureslack of willingness to undergo the necessary control visits. The study was approved by the local at (Moscow) with number #8819 and carried out in full compliance with the 1975 Declaration of Helsinki on patient rights (2008 revision). Implant designdata obtained from the CBCT were extracted and imported into software where the residual anatomy of the patient’s bone was reconstructed in 3D (Mimics®, Materialise, Leuven, Belgium), and the file saved as. STL. In this phase, care was taken to investigate the position of noble structures (such as the inferior alveolar nerve), and to select the proper threshold values, to best define the cortical walls of the residual bone. The best position for the fixation screw was also evaluated. The. STL file of the 3D bone reconstruction was then imported to reverse engineering software (Studio 2012®, ) where a cleaning operation was carried out, with elimination of scattering where present and sharp edges or mesh errors; furthermore, the. STL density was reduced. This 3D reconstruction was then aligned with the.

Output: corroborate


Input: Claim: Everyone's microbiome composition is identical. [SEP] Text: We noted that one RootOTU (OTU1, matching Rhizobium leguminosarum) dominated the Trifolium root microbiome and explained the high prevalence of Proteobacteria (Additional file 1: Figure S5). OTU1 ranged from 35. 4 to 89. 7% in samples from both growth conditions and accounted for a median of 73. 5% of the root community (Fig. 3b). We confirmed that the high abundance of OTU1 in the overall root community was due to the rhizobia bacteria present in root nodules (Additional file 1: Supplementary methods), and we noted a few non-OTU1 sequences inside the nodules, suggesting additional within-nodule bacteria diversity (Additional file 1: Supplementary results, Figure S6). Fig. 3Abundant and root-specific OTUs of the Trifolium root microbiome. a The plot reports the mean relative abundance and the log2 fold change between root and soil samples of all OTUs present in the rarefied community (open black circles). Filled red circles indicate the 61 OTUs significantly enriched (P < 0. 05, FDR corrected) in root samples. Dark red circles indicate the 15 OTUs present in the RootOTUs (see text). b Box plot (overplotted with individual data points) showing the median relative abundance of OTU1 (Rhizobium leguminosarum) in sequenced climate chamber (blue triangles) and natural site (green circles) root samplesFull size image In summary, root bacterial communities did not differ substantially whether the plants were grown under controlled or field conditions, thereby validating our approach using climate chamber experiments. The abundant and root-specific members of the Trifolium root microbiome consisted mainly of Proteobacteria and nodule-inhabiting rhizobia bacteria accounted for ~70% of the root microbiome. Isolated members of the Trifolium root microbiomeWe isolated bacteria from Trifolium roots of two climate chamber experiments and from plants grown at the natural site (Table 1) and characterized a total of 200 cultured bacteria (Fig. 1, step II). Proteobacteria dominated the culture collection, being represented by 78. 5% isolates while Actinobacteria, Firmicutes, and Bacteroidetes accounted for 8, 8, and 5. 5% of isolates, respectively (Fig. 4a).

Output: contrast


Input: Claim: Some studies suggest that olfactory receptors may function through quantum effects. [SEP] Text: Moreover, if DAN-c1 artificial activation conveyed an aversive teaching signal irrespective of the gustatory stimulus, then it should not impair aversive learning after quinine training (Figure 2k). While the authors interpret Figure 2k (and Figure 5) to indicate that artificial activation causes excessive DAN-c1 dopamine release, an alternative explanation is that artificial activation compromises aversive learning by overriding DAN-c1 activity that could be evoked by quinine. The authors should not necessarily expect that D2R enhancer driver strains would reflect D2R endogenous expression, since it is known that TH-GAL4 does not label p(PAM) dopaminergic neurons. Their observations of GFP-tagged D2R expression could be strengthened with an anti-D2R antibody such as that used by Lam et al. , (1999) or Love et al. , (2023). Finally, the authors could consider the possibility other DANs may also mediate aversive learning via D2R. Knockdown of D2R in DAN-g1 appears to cause a defect in aversive quinine learning compared with its genetic control (Figure S4e). It is unclear why the same genetic control has unexpectedly poor aversive quinine learning after training with propionic acid (Figure S5a). The authors could comment on why RNAi knockdown of D2R in DAN-g1 does not similarly impair aversive quinine learning (Figure S5b). #2 (Public Review): Summary: The study wanted to functionally identify individual DANs that mediate larval olfactory learning. Then search for DAN-specific driver strains that mark single dopaminergic neurons, which subsequently can be used to target genetic manipulations of those neurons. 56 GAL4 drivers identifying dopaminergic neurons were found (Table 1) and three of them drive the expression of GFP to a single dopaminergic neuron in the third-instar larval brain hemisphere. The DAN driver R76F02-AD;R55C10-DBD appears to drive the expression to a dopaminergic neuron innervating the lower peduncle (LP), which would be DAN-c1. Split-GFP reconstitution across synaptic partners (GRASP) technique was used to investigate the "direct" synaptic connections from DANs to the mushroom body.

Output: pass


Input: Claim: 3D printing is used to create custom prosthetics and implants tailored to individual patients. [SEP] Text: STL files obtained from the intraoral scan of the patient’s arch, and with the diagnostic wax-up, with the ultimate aim of having a file with the whole information of the patient. This allowed better understanding of the ideal prosthetic emergence profile, and therefore how to properly design the implant. All these files were imported to another CAD software (Meshmixer®, ), where the prosthetic abutments and osteosynthesis screws for implant stabilisation were designed. The subperiosteal implant framework was drawn, connecting all the structures through a series of Boolean operations (Fig. 3). The. STL of the final modelling was then exported again to Studio 2012® () for a final edge correction, final quality control and file regularisation (Fig. 4). The implant file was then ready for fabrication. Fig. 3The custom-made subperiosteal implant is designed in a CAD software (Meshmixer®, ). a The implant with the integral abutments and the fixation screws, buccal view; b occlusal view; c lingual view; d detail of the axes for the fixation screwsFull size image Fig. 4Detail of the mesh of the implant, ready for fabrication via DMLSFull size imageImplant fabricationA DMLS machine was used (ProX-DMP100®, 3) to print the subperiosteal implants. This machine was able to build the custom-made subperiosteal implant exactly as designed, starting from titanium grade 5 micro-powders, layer by layer, using a powerful laser beam (50 W fibre laser with a wavelength of 1070 nm), with layer size of 20 μm. The build envelope capacity of the machine was 100 × 100 × 80 mm. The porous and chemically pure implant was washed with organic acids, decontaminated and sterilized. At the same time, the 3D bone reconstruction and a replica of the subperiosteal implant were printed in resin using a 3D printer (ProJet 3510 MP®, 3D system, ). The 3D model of the bone, printed with a stereolithographic (SLA) printer (3500PD®, DWS systems, Thiene, Vicenza, Italy), was used to verify the anatomy, and to control the fit of a replica of the DMLS subperiosteal implant (Fig.

Output: corroborate


Input: Claim: Everyone's microbiome composition is identical. [SEP] Text: The isolates were assigned to 34 different genera (Fig. 4b). The 19 genera of the Proteobacteria (157 isolates) included abundant Pseudomonas (83 isolates), Janthinobacterium (19), and Stenotrophomonas (9). We found seven genera in the phylum Actinobacteria (16 isolates) with Microbacterium (7), Micrococcus (3), and Micromonospora (2) having more than one representative isolate. In the Firmicutes (16 isolates), we noted five different genera, with Bacillus (9), Staphylococcus (3), and Paenibacillus (2) being the most abundant. Finally, we found three genera in the Bacteroidetes (11 isolates): Flavobacterium (8), Mucilaginibacter (2), and Pedobacter (1). Fig. 4Taxonomic diversity of the Trifolium bacteria reference stock. a Taxonomic composition of the isolate collection at the Phylum level. b The phylogenetic diversity of the isolates at the genus level and the number of isolates assigned to each genus is indicated in parentheses. Isolates are labeled at the genus level and color-coded by phylum in a Full size image We clustered the bacteria isolate sequences to the representative sequences of the OTUs of the Trifolium root community profiles at ≥97% sequence similarity (see Additional file 1: Supplementary methods) and determined whether a bacteria isolate constituted an abundant and root-enriched member of the Trifolium microbiome. Overall, out of the 200 bacteria isolates, 181 (90. 5%) isolates clustered to 34 OTUs of the root community profile while for 19 (8. 5%) isolates, we did not find a matching community member. All of the 34 isolated OTUs were present in the rarefied root community (2426 OTUs), corresponding to an isolation rate of 1. 4% (Fig. 5). The isolation rate increased to 23. 6% when comparing to the abundant community members: 55 abundant OTUs had a mean relative abundance of ≥0. 1% across all root samples, and for 13 of these, we were able to culture bacteria strains. We identified 11 bacteria isolates for 2 of the 15 RootOTUs (Fig. 3; Additional file 1: Table S2).

Output: contrast


Input: Claim: Some studies suggest that olfactory receptors may function through quantum effects. [SEP] Text: Activation of MBNs during training impairs both larval aversive and appetitive learning. Finally, based on the data the authors propose a model where the effective learning requires a balanced level of activity between D1R and D2R (Figure 7). Strengths: The work is well written, clear, and concise. They use well documented strategies to examine GAL4 drivers with expression in a single DAN, behavioral performance in larvae with distinct genetic tools including those to do thermo and optogenetics in behaving flies. Altogether, the study was able to expand our understanding of the role of D2R in DAN-c1 and MB neurons in the larva brain. Weaknesses: Is not completely clear how the system DAN-c1, MB neurons and Behavioral performance work. We can be quite sure that DAN-c1;Shits1 were reducing dopamine release and impairing aversive memory (Figure 2h). Similarly, DAN-c1;ChR2 were increasing dopamine release and also impaired aversive memory (Figure 5b). However, is not clear what is happening with DAN-c1;TrpA1 (Figure 2K). In this case the thermos-induction appears to impair the behavioral performance of all three conditions (QUI, DW and SUC) and the behavior is quite distinct from the increase and decrease of dopamine tone (Figure 2h and 5b). The study successfully examined the role of D2R in DAN-c1 and MB neurons in olfactory conditioning. The conclusions are well supported by the data, with the exception of the claim that dopamine release from DAN-c1 is sufficient for aversive learning in the absence of unconditional stimulus (Figure 2K). Alternatively, the authors need to provide a better explanation of this point. The study provides insight into the role of D2R in associative learning expanding our understanding and might be a reference similar to previous key findings (Qi and Lee, 2014, #3 (Public Review): It is a strength of the paper that it analyses the function of dopamine neurons (DANs) at the level of single, identified neurons, and uses tools to address specific dopamine receptors (DopRs), exploiting the unique experimental possibilities available in larval Drosophila as a model system.

Output: pass


Input: Claim: 3D printing is used to create custom prosthetics and implants tailored to individual patients. [SEP] Text: 5); the implant replica was printed in biocompatible resin, to help the surgeon in preparation of the flap and access for fixation of the implants intraoperatively. Fig. 5A replica of the implant (Iuxta3D®, ), fabricated via DMLS, is tested in a 3D printed copy of the patient’s mandible, printed with a stereolithographic (SLA) 3D printer (3500PD®, ). The adaptation looks perfect. a Buccal view; b occlusal viewFull size imageSurgeryOnce all materials were ready, the surgical phase could start, using local anaesthesia (4% articaine, 1:100,000 adrenaline). A crestal incision was performed, delimited by mesial and distal release incisions, and a full-thickness flap was raised, for a complete view of the implant site. The implant replica was used to prepare the flap, to verify the perfect adaptation of the implant intraoperatively, and for preparation (drilling) of access for the miniscrews to be used for fixation of the grid (Fig. 6). Once the site was prepared, the DMLS implant (Iuxta3D®, ) was removed from its sterile pack, placed on the site to verify its adaptation on the residual bone, and then fixed with the aid of osteosynthetic miniscrews. The surgical site was sutured, and using periosteum release incisions, flap passivation was performed to cover the whole section and obtain first-intention healing. In suturing, particular attention was paid to avoid excessive tension, maintaining a correct amount of keratinised gingiva around the emerging abutments, which is necessary for clinical success, both surgically and prosthetically (Fig. 7). At the end of surgery, antibiotic therapy was prescribed (amoxicillin plus clavulanic acid 1 g every 12 h for 6 days), along with painkillers (ibuprofen 600 mg to be taken for the following two to 3 days) and antibacterial therapy (mouthrinses of 0. 12% chlorhexidine, two to three times per day, for five to 6 days). Fig. 6Surgery on patient.

Output: corroborate


Input: Claim: Everyone's microbiome composition is identical. [SEP] Text: The cultured RootOTUs included the dominant OTU1 (R. leguminosarum; 5 isolates), as well as OTU48 (Pantoea agglomerans; 6 isolates). Fig. 5Mapping of reference stock bacteria to root microbiome OTUs. The upper bar graph represents the relative abundance of the 2426 OTUs in the root-associated bacteria community of Trifolium, with the 500 most abundant OTUs shown in gray bars. The dark gray bars indicate the 55 most abundant root OTUs (mean RA >0. 1%). The blue bars indicate OTUs for which at least one isolate is present in the reference stock. The lower, inverted bar graph indicates the number of isolates in the reference stock mapping to an OTU in the community profile. Bars are shaded the same as in the upper graph to indicate the relative abundance of each OTU. Bars are labeled with the representative OTU name and its total number of sequences in the community profile in parenthesesFull size image We concluded that almost a quarter of the abundant root community members can be obtained in culture, and we achieved this with a manageable effort (200 strains) and straightforward microbiological techniques. By linking to the information of the root community profiles, we have characterized the bacteria strains of the reference stock with rank and relative abundance in the Trifolium root microbiome, and thereby the reference stock represents a toolbox for future microbiota manipulation experiments. Towards functional investigations of the Trifolium root microbiotaFinally, we developed microcosms (Fig. 1, step III) and evaluated their potential to conduct plant-microbiota interaction experiments. Recent microbiota inoculation experiments [21, 22] revealed that approximately half of the inoculated bacteria strains previously isolated from roots of soil-grown Arabidopsis either completely failed or failed to robustly colonize the roots of their host plant under microcosm conditions. We speculate that this could partly be due to the different physical and chemical conditions in the microcosms compared to soil and that these conditions are unfavorable for certain isolates. Therefore, we performed a soil extract experiment to pre-screen for possible microcosm-adapted bacteria strains. For this, we characterized the root microbiome of Trifolium that assembled after inoculation of a diverse soil microbiota extracted from the experimental field soil (Additional file 1: Figure S7a, b, Figure S8; Supplementary methods and results).

Output: contrast


Input: Claim: Some studies suggest that olfactory receptors may function through quantum effects. [SEP] Text: Indeed, the result of their screening for transgenic drivers covering single or small groups of DANs and their histological characterization provides the community with a very valuable resource. In particular the transgenic driver to cover the DANc1 neuron might turn out useful. However, I wonder in which fraction of the preparations an expression pattern as in Figure 1f/ S1c is observed, and how many preparations the authors have analyzed. Also, given the function of DANs throughout the body, in addition to the expression pattern in the mushroom body region (Figure 1f) and in the central nervous system (Figure S1c) maybe attempts can be made to assess expression from this driver throughout the larval body (same for Dop2R distribution). A first major weakness is that the main conclusion of the paper, which pertains to associative memory (last sentence of the abstract, and throughout the manuscript), is not justified by their evidence. Why so? Consider the paradigm in Figure 2g, and the data in Figure 2h (22 degrees, the control condition), where the assay and the experimental rationale used throughout the manuscript are introduced. Different groups of larvae are exposed, for 30min, to an odour paired with either i) quinine solution (red bar), ii) distilled water (yellow bar), or iii) sucrose solution (blue bar); in all cases this is followed by a choice test for the odour on one side and a distilled-water blank on the other side of a testing Petri dish. The authors observe that odour preference is low after odour-quinine pairing, intermediate after odour-water pairing and high after odour-sucrose pairing. The differences in odour preference relative to the odour-water case are interpreted as reflecting odour-quinine aversive associations and odour-sucrose appetitive associations, respectively. However, these differences could just as well reflect non-associative effects of the 30-min quinine or sucrose exposure per se (for a classical discussion of such types of issues see Rescorla 1988, Neurosci, or regarding 1988, Behav Genetics, or with some reference to the original paper by Honjo & Furukubo-Tokunaga 2005, J Neurosci that the authors reference, also Gerber & Stocker 2007, Chem Sens).

Output: pass


Input: Claim: 3D printing is used to create custom prosthetics and implants tailored to individual patients. [SEP] Text: The definitive zirconia framework was produced by milling with a powerful five-axis milling machine (Roland DWX-50®, ), subsequently sintered in an oven (Tabeo®, Mihm-Vogt, Stutensee, Germany), characterized and ready for ceramic stratification. The technician generated 3D models of the mandible and maxilla and printed them using a powerful 3D printer (3500PD®, ), and was able to add ceramic on the zirconia framework. The final zirconia-ceramic bridge was then delivered to the patient (Fig. 14), carefully controlled in occlusion and for the quality of interproximal contact points and colour. Cementation was carried out with a temporary cement (Tempbond®, ). The patient was enrolled in an annual recall program, based on two- to three-yearly appointment for professional oral hygiene. Fig. 8CAD design (DentalCad®, Darmstad, Exocad, Germany) of the pre-milled restoration. a Buccal view; b occlusal view; c perspective view; d the pre-milled restorationFull size image Fig. 9The pre-milled provisional restoration. a Buccal view; b the pre-milled restoration relined and adapted in occlusionFull size image Fig. 10Control after 2 months, at the end of the provisionalizationFull size image Fig. 11Second intraoral scan, taken at the end of the provisionalization. a Scan with the provisionals in situ, useful as a reference for occlusion; b direct intraoral scan of the emerging abutmentsFull size image Fig. 12The final scan was sent to the dental technician, who once again replaced the meshes of the abutments with the original CAD files of the implant in the same aforementioned CAD software (DentalCad®, Darmstad, Exocad, Germany). a The meshes of the emerging abutments are replaced by the original CAD design; b the original CAD files of the abutments in position, after alignmentFull size image Fig. 13Final CAD project of the zirconia framework. a Buccal view of the framework; b the zirconia structure, buccal view; c the zirconia structure, lingual viewFull size image Fig.

Output: corroborate


Input: Claim: Everyone's microbiome composition is identical. [SEP] Text: We defined the root bacteria community (Fig. 1, step IV) and determined which bacteria isolates (from the reference stock, Fig. 5) corresponded to abundant OTUs on the roots under microcosm conditions (Additional file 1: Figure S8; Supplementary methods). See the Additional file 1: Supplementary results for a comparison between microcosm and soil-grown root communities (Additional file 1: Figure S9a, b; Supplementary methods and results). We then conducted microcosm experiments in which we inoculated Trifolium in the microcosms with bacteria strains isolated from its root microbiome. The goal was not to screen strains or to test specific functions but instead to combine all our tools (reference stock, microcosms, community sequencing, and soil extract information) and validate the overall experimental approach for future microbiota inoculation experiments. We assembled a simplified community, choosing strains from the reference stock that corresponded to abundant OTUs on the roots under microcosm conditions and belonged to well-represented bacterial genera in the collection (Additional file 1: Figure S9; strains per OTU were randomly chosen): a Flavobacterium (F; Bacteroidetes, #8 isolates for this genus in the reference stock; KHB002), a Pseudomonas (P; Proteobacteria, #83; KHB004), and a Janthinobacterium (J; Proteobacteria, #19, KHB023; Table 2). We also included a Microbacterium (M; Actinobacteria, #7; strain KHB073) because this genus was well-represented in the reference stock (numerous isolates could indicate that these bacteria were abundant on roots; Fig. 4b) and because we wanted the inoculated community to broadly reflect the abundant bacterial phyla of plant root microbiomes (Actinobacteria, Bacteroidetes, and Proteobacteria; [2, 25]). We inoculated these bacteria alone or in combination to the autoclaved microcosms (Fig. 1, step V) at densities of 106 cells mL−1 and planted surface-sterilized Trifolium seeds. We then monitored the community dynamics of the inoculated simplified community and scored effects of the bacteria inoculation on plant growth in three replicate experiments.

Output: contrast


Input: Claim: Some studies suggest that olfactory receptors may function through quantum effects. [SEP] Text: As it stands, therefore, the current 3-group type of comparison does not allow conclusions about associative learning. A second major weakness is apparent when considering the sketch in Figure 2g and the equation defining the response index (R. I. ) (line 480). The point is that the larvae that are located in the middle zone are not included in the denominator. This can inflate scores and is not appropriate. That is, suppose from a group of 30 animals (line 471) only 1 chooses the odour side and 29, bedazzled after 30-min quinine or sucrose exposure or otherwise confused by a given opto- or thermogenetic treatment, stay in the middle zone. . . a P. I. of 1. 0 would result. Unless experimentally demonstrated, claims that the thermogenetic effector shibire/ts reduces dopamine release from DANs are questionable. This is because firstly, there might be shibire/ts-insensitive ways of dopamine release, and secondly because shibire/ts may affect co-transmitter release from DANs. To implicate a role of dopamine in DANs, previous work used e. g. RNAi against the dopamine-synthesizing TH enzyme (Rohwedder et al, cited). It is not clear whether the genetic controls when using the Gal4/ UAS system are the homozygous, parental strains (XY-Gal4/ XY-Gal4 and UAS-effector/ UAS-effector), or as is standard in the field the heterozygous driver (XY-Gal4/ wildtype) and effector controls (UAS-effector/ wildtype) (in some cases effector controls appear to be missing, e. g. Figure 4d, Figure S4e, Figure S5c). As recently suggested by Yamada et al 2024, bioRxiv, high cAMP can lead to synaptic depression (sic).

Output: pass


Input: Claim: 3D printing is used to create custom prosthetics and implants tailored to individual patients. [SEP] Text: For all these patients, and in all these situations, subperiosteal implants may be an alternative, due to the digital revolution and the advent of modern digital technologies in dentistry [34,35,36,37, 46]. CBCT allows acquisition of 3D data of the patient’s residual bone volume, with considerable accuracy, and a low dose of radiation directed exclusively at the area of interest [27, 47]. The data acquired with the CBCT is then re-elaborated with appropriate reconstruction software, which allows generation of a virtual bone model of the area of interest [34,35,36,37]. Capturing an optical impression of the dentate arches allows modelling of a virtual diagnostic wax-up. Finally, the superimposition of bone and dental models, together with the virtual wax-up, makes it possible to design and model custom-made subperiosteal implants, designed and conceived specifically for the patient’s needs, both as a bone-supported structure and as a prosthetic emergency [34,35,36,37]. These 3D-printed implants represent a possible alternative solution for the rehabilitation of the atrophic posterior mandible of elderly patients who do not want or cannot undergo traditional regenerative techniques, preparatory to insertion of classical endosseous implants [34,35,36,37]. In particular, increasingly advanced DMLS techniques allow revisiting the old concept of subperiosteal implants. DMLS is a technique arose in the late 1990s in Germany, which has since been gaining importance in dentistry [29]. It is an additive manufacturing (AM) technique that uses a high-power laser to melt metallic powders together [29]. The procedure involves construction of a stratified 3D model, layer by layer. The CAD file is sliced into thin layers, creating a two-dimensional images series that, laced together, create the three-dimensionality of the product [29]. The powder of each layer is then selectively melted by the laser beam, and the process is continuously repeated layer by layer, until completion of the device. With DMLS, there is almost no limitation in the fabrication of complex objects, such as porous, hollow objects with interconnections, tunnels and crevices.

Output: corroborate


Input: Claim: Everyone's microbiome composition is identical. [SEP] Text: Table 2 Bacterial strains used in the microcosm experimentsFull size table After 25 days, we harvested the experiments and counted ≥106 bacterial colony forming units of the inoculated strains on the roots (Table 2). This confirmed that the chosen strains are also able to successfully colonize roots under the artificial growth conditions in the microcosms. We noted a lower biomass in one experiment compared to the two others, and this experiment-to-experiment variation indicated to us that numerous replicates are also needed when highly controlled conditions are used. With regard to the effects of individual bacteria inoculation on the plants, we found that the Flavobacterium negatively affected the growth of Trifolium, while the other bacteria did not have an effect on shoot biomass production (Fig. 6a). The combined application of the bacteria (FJMP) also did not have an apparent effect on biomass production but alleviated the negative impact of the Flavobacterium when grown alone. We measured the composition of the simplified community upon inoculation and after 25 days on the roots (Additional file 1: Supplementary methods for details). could not be captured with the community quantification method, and we noted a small proportion of additional OTU sequences possibly representing sequencing errors or contamination, or in root samples, being derived from seed endophytes. Despite these limitations, the analysis revealed that the three other inoculated members retained similar proportions on the roots during 25 days of incubation as compared to when they were inoculated (Fig. 6b). This observation indicated that the alleviation of the negative impact of the Flavobacterium was not due to out-competition of this community member, but rather that its negative activities may have been “buffered” by the other bacteria in the simplified community. Fig. 6Functional analysis of a simplified Trifolium root microbiota in microcosms. a Trifolium growth in microcosms in the absence of inoculated bacteria (nbc no-bacteria control), with specific strains (F Flavobacterium KHB002, J Janthinobacterium KHB023, M Microbacterium strain KHB073, P Pseudomonas KHB004) or the simplified community (FJMP). The graph reports the mean shoot fresh weight (n = 12; ± s. e. m. ) and the individual data points from the three independent experiments with four replicates each.

Output: contrast


Input: Claim: Some studies suggest that olfactory receptors may function through quantum effects. [SEP] Text: That would call into question the interpretation of low-Dop2R leading to high-cAMP, leading to high-dopamine release, and thus the authors interpretation of the matching effects of low-Dop2R and driving DANs. response: Reviewer #1 (Public Review): Weakness #1: The authors claim to have identified drivers that label single DANs in Figure 1, but their confocal images in Figure S1 suggest that many of those drivers label additional neurons in the larval brain. It is also not clear why only some of the 57 drivers are displayed in Figure S1. As introduced in the results section, we screened 57 driver strains based on previous studies, either they were reported identifying a single (a pair of) dopaminergic neuron (DAN) in larvae or identifying only several DANs in the adult brain indicating the potential of identifying single dopaminergic neuron in larvae. 1, TH-GAL4 was used to cover all neurons in the DL1 cluster, while R58E02 and R30G08 were well known drivers for pPAM. Fly strains in Figure 1h, k, l, and m were reported as single DAN strains in larvae4, while strains in Figure 1e, f, g were reported identifying only several DANs in adult brains5,6. We examined these strains and only some of them labeled single DANs in 3rd instar larval brains (Figure 1f, g, h, l and m). Among them, only strains in Figure 1f and h labeled single DAN in the brain hemisphere, without labeling other non-DANs. Other strains labeled non-DANs in addition to single DANs (Figure 1g, l and m). Taking ventral nerve cord (VNC) into consideration, strain in Figure 1h also labeled neurons in VNC (Figure S1e), while strain in Figure 1f did not (Figure S1c). In summary, the strain in Figure 1f (R76F02AD;R55C10DBD, labeling DAN-c1) is a strain we screened labeling only a single DAN in the 3rd instar larval brains.

Output: pass


Input: Claim: 3D printing is used to create custom prosthetics and implants tailored to individual patients. [SEP] Text: It is possible to manufacture titanium or titanium alloy implants, whether standard endosseous [29] or custom-made subperiosteal [34,35,36,37]. Surovas [35] demonstrated the feasibility and economic sustainability of the design and manufacture via DMLS of a custom-made subperiosteal implant in titanium alloy, able to adapt perfectly to the 3D-printed bone model. The steps for the design and fabrication of the custom-made implants were computerized tomography (CT) scan, CT data processing, 3D virtual model creation, modelling technique for custom implant, and data file preparation for 3D printing [35]. The custom-made subperiosteal implant was then fabricated in Ti6Al4V (a type 5 titanium alloy) using DMLS [35]. Cohen et al. [36] developed custom-made subperiosteal Ti6Al4V devices produced by AM and post-fabrication osteogenic micro- and nano-scale surface texture modification. The porous surface of these implants had the potential to stimulate human osteoblasts to produce osteogenic factors, and a high bone-to-implant contact was found for DMLS disks implanted in the rat calvaria and in the rabbit tibia [36]. When implanted in the human posterior mandible, three- and eight-month postoperative images showed new bone formation and osseointegration. The implants were stable and successful under function. These data are not surprising, given that the highly porous surface of titanium laser sintering implants has amply demonstrated in histological and histomorphometric human studies high levels of bone contact and osseointegration, with bone incorporation into the pores, in the posterior maxilla 2 months after insertion [48,49,50]. The possibility of obtaining a porous surface capable of stimulating bone formation represents one of the possible advantages of manufacturing subperiosteal implants with DMLS, and a major difference from traditional subperiosteal implants, which were fused and therefore presented a smooth surface. In a retrospective clinical study, Cerea et al. [34] presented an analogue–digital technique for fabricating custom-made subperiosteal implants, and reported on the survival and complication rates encountered when using these fixtures.

Output: corroborate


Input: Claim: Everyone's microbiome composition is identical. [SEP] Text: The letters indicate statistical significance at P < 0. 05 (Tukey’s HSD; analysis over the three experiments). Note, the Microbacterium (M, panel a) was not captured with the community quantification method. b Community composition of the simplified community (FJMP) at inoculation (input) and after 25 days on the roots. Sequences of other OTUs are indicated in gray Full size image DiscussionRoot microbiome compositionHere, we have characterized the bacterial communities on roots of T. pratense with respect to their composition and reported first steps towards experimentally testing their functions. Trifolium harbors a diverse root microbiome that differs qualitatively and quantitatively from that of the surrounding bulk soil (Fig. 2), confirming studies with other plant species [3–5, 26]. We found that OTU1, matching R. leguminosarum, accounted for a median 73. 5% of the root microbiome (Fig. 3b). We separately inspected root nodules and confirmed that Trifolium nodules were primarily inhabited by R. leguminosarum (Additional file 1: Figure S6) but also contained other bacteria taxa. This is in agreement with earlier work revealing within-nodule diversity in Trifolium repens and Trifolium fragiferum, which consisted of the dominant R. leguminosarum and the less-frequent rhizobia species Bradyrhizobium japonicum, Sinorhizobium sp. , and Mesorhizobium [27, 28]. For the purpose of the microcosm experiments, we described the root microbiome of Trifolium at a whole-root scale, sampling the entire root system including nodules. For a broader description of legume microbiomes, future work investigating the variation in multiple soil types and comparisons with non-legume plants is needed. Additionally, an in-depth spatial assessment of legume root microbiomes would be insightful, e. g. , by profiling the bacteria communities of root tissues with the nodules removed as well as inside the root nodules. The large number of DNA sequences allowed us to thoroughly characterize the Trifolium root microbiome beyond the dominant rhizobia members.

Output: contrast


Input: Claim: Some studies suggest that olfactory receptors may function through quantum effects. [SEP] Text: Others (Figure 1g, h, l, and m) we still describe them as strains labeling single DANs, but they also label one to several non-DANs. 1, we mainly showed the strains labeling single DANs. The labeling patterns of other screened driver strains were summarized in Table1. Since all brain images of the rest 47 strains are available, we will state in Fig S1 that additional brain images can be provided upon request. Weakness #2: Critically, R76F02-AD; R55C10-DBD labels more than one neuron per hemisphere in Figure S1c, and the authors cite Xie et al. (2018) to note that this driver labels two DANs in adult brains. Therefore, the authors cannot argue that the experiments throughout their paper using this driver exclusively target DAN-c1. Figure S1c shows single DA neuron in each brain hemisphere. Additional GFP (+) signals were often observed, but not from cell bodies of DANs because they were not stained by a TH antibody. These additional GFP (+) signals were mainly neurites, including axonal terminals, but could be false positive signals or weakly stained non-neuronal cell bodies. This conclusion was based on analysis of a total of 22 larval brains. We will add this in the text or Fig S1 caption. Enlarged insert of GFP (+) signals will be added also to Figure S1c. Weakness #3: Missing from the screen of 57 drivers is the driver MB320C, which typically labels only PPL1-γ1pedc in the adult and should label DAN-c1 in the larva. If MB320C labels DAN-c1 exclusively in the larva, then the authors should repeat their key experiments with MB320C to provide more evidence for DAN-c1 involvement specifically. We thank the reviewer for the suggestion. MB320C mainly labels PPL1-y1pedc in the adult brain, with one or two other weakly labeled cells. It will be interesting to investigate the pattern of this driver in 3rd instar larval brains. If it only covers DAN-c1, we can try to knock-down D2R in this strain to check whether it can repeat our results.

Output: pass


Input: Claim: 3D printing is used to create custom prosthetics and implants tailored to individual patients. [SEP] Text: In total, 70 partially or completely edentulous patients were included in the study and treated with custom-made DMLS subperiosteal implants, in both the maxilla and the mandible [34]. These implants were designed in an analogue way by the surgeon, directly on 3D models, then digitally designed and fabricated via DMLS. At two-year follow-up, three implants were lost due to recurrent, untreatable infections; the implant survival rate was 95. 8% [34]. With regard to complications, four patients had pain, discomfort or swelling after implant placement, giving an incidence of immediate postoperative complications of 5. 7% [34]. During the follow-up period, one patient experienced recurrent infections, representing an incidence of biological complications of 1. 4% [34]. The rate of prosthetic complications amounted to 8. 9% [34]. The authors concluded that application of custom-made DMLS titanium subperiosteal implants can represent a successful strategy for the prosthetic restoration of patients with severe bone deficiencies, and an alternative to conventional bone regenerative techniques. The clinical study of Cerea et al. [34] has the highest number of enrolled patients and the longest follow-up, though it should be noted that the process for fabrication of the implants was hybrid (analogue–digital) and all implants underwent an electropolishing treatment, capable of transforming the porous surface of the laser-sintered implants into a smooth one. This can radically change the response of hard and soft tissues to the implant. Moreover, the study included different categories of patients (including completely edentulous), with different fixation screws and in the maxillary area. In our present study we have focused our attention only on partially edentulous patients, and in particular on the rehabilitation of the posterior mandible. Our case series showed positive clinical outcomes with the use of subperiosteal custom-made DMLS implants. First, the overall accuracy of the implants was excellent. Only two implants had an insufficient fit (with values of 4 and 5 rated by the surgeon), because of the presence of scattering from neighbouring crowns or teeth in the original CBCT that interfered with the thresholding process. However, these implants were adapted to the surgical sites during the interventions. In our present study, the whole fabrication processes (from the intraoral scans and CBCT to the surgery) took approximately 2–3 weeks.

Output: corroborate


Input: Claim: Everyone's microbiome composition is identical. [SEP] Text: In addition to Rhizobium, Trifolium supports enriched OTUs from the genera Pantoea, Sphingomonas, Novosphingobium, and Pelomonas, among others, in its root microbiome (Additional file 1: Table S2). A review of relevant literature reveals that bacteria isolates of some of these genera have been found to be antagonistic to pathogens (Additional file 1: Table S2). This could possibly suggest a partitioning of complementary host services in the Trifolium root microbiome with “disease protection” and “nutrient provision” provided by the mentioned root-enriched genera and the nodule-inhabiting Rhizobia, respectively. However, because it is notoriously problematic to infer bacteria function from a taxonomy assignment [29], approaches other than 16S community sequencing are required for the functional understanding of the root microbiome. As a next step, such an indicative observation from cultivation independent microbiome analysis could be examined by testing reference stock bacteria belonging to these OTUs for their ability to suppress pathogens. Reference stocks and microcosms to study functions of the root microbiomeWith the isolation of root microbiome members (Fig. 5), setting up an experimental microcosm system (Additional file 1: Figure S1a–d) and testing for microbiota effects on plant growth (Fig. 6a), we delineate a possible approach to advance the functional understanding of the root microbiome. We built our reference stock (Fig. 4b) using one bacteria isolation medium, and at a sampling depth of 200 bacteria strains, we captured close to a quarter of the abundant members of the Trifolium root microbiome. Therefore, we believe that our work presents an encouraging example especially for smaller laboratories with limited resources. For future work, additional isolation media and growth conditions would likely permit us to broaden the reference stock and contribute to a targeted cultivation of “missing” Trifolium root microbiome members. Experimentation with inoculated plantsWe conducted multi-strain inoculation experiments with members of the Trifolium root microbiome to evaluate the suitability of microcosm growth system for plant-microbiota inoculation experiments. However, we first conducted the soil extract experiment (Additional file 1: Figure S7a, b, Figure S8; Supplementary methods and results) as a proof-of-concept to pre-screen microcosm-adapted bacteria strains.

Output: contrast


Input: Claim: Some studies suggest that olfactory receptors may function through quantum effects. [SEP] Text: This will be an interesting fly strain to test, but we believe that it will not be necessary for our current manuscript as DAN-c1 driver is very specific (for details, refer to our response to Reviewer#3). However, this line will be very useful for future experiments. Weakness #4: The authors claim that the SS02160 driver used by Eschbach et al. (2020) labels other neurons in addition to DAN-c1. Could the authors use confocal imaging to show how many other neurons SS02160 labels? Given that both Eschbach et al. and Weber et al. (2023) found no evidence that DAN-c1 plays a role in larval aversive learning, it would be informative to see how SS02160 expression compares with the driver the authors use to label DAN-c1. We did not have our own images showing DANs in brains of SS02160 driver cross line. However, Figure 1 in the paper of Eschbach et al. (2020) shows strongly labeled four neurons on each brain hemisphere9, indicating that this driver is not a strain only labeling one neuron, DAN-c1. Weakness #5: The claim that DAN-c1 is both necessary and sufficient in larval aversive learning should be reworded. Such a claim would logically exclude any other neuron or even the training stimuli from being involved in aversive learning (see Yoshihara and Yoshihara (2018) for a detailed discussion of the logic), which is presumably not what the authors intended because they describe the possible roles of other DANs during aversive learning in the discussion. We agree that the words ‘necessary’ and ‘sufficient’ are too exclusive for other neurons. As mentioned in the Discussion part, we do think other dopaminergic neurons may also be involved in larval aversive learning. We are going to re-phrase these words by replacing them with more logically appropriate words, such as ‘important’, ‘essential’, or ‘mediating’. Weakness #6: Moreover, if DAN-c1 artificial activation conveyed an aversive teaching signal irrespective of the gustatory stimulus, then it should not impair aversive learning after quinine training (Figure 2k).

Output: pass


Input: Claim: 3D printing is used to create custom prosthetics and implants tailored to individual patients. [SEP] Text: The mean duration of the surgery was 44. 3 min. Notably, this result was influenced by the two aforementioned cases in which adaptation was not fully satisfactory (those cases required respectively 85 and 67 min, from anaesthesia to sutures). Finally, in our study, all implants survived at 1 year after placement. Immediate postoperative complications had a low incidence (10%), were minor in nature, and resolved in a few days with pain-relieving and antibiotic therapies. The low incidence of complications was a direct result of the perfect fit of the custom-made implants to the patient’s residual anatomy; the surgical procedures were simplified and sped up, inducing a more comfortable result. The main advantages of our fully digital technique, compared to the conventional analogue technique used in the past for fabrication of subperiosteal implants, relies on the accuracy of the implants. This increases considerably the correspondence between the implant structure and the underlying bone, eliminating the need for a surgical session to capture a physical impression of the bone, reducing non-fitting problems and distributing the load more evenly. The excellent accuracy speeds up and simplifies the surgery, reducing the risk of bacterial contamination or infection. In addition, compared to the previously published study of Cerea et al. [34], in our present study the design and fabrication of the implants was entirely digital, and the implants did not undergo electropolishing, so they had a porous surface. In fact, the manufacturing of implants by DMLS, together with the treatment with organic acids, determine the formation of a porous surface, with concavities that continue in a structure with interconnections between the pores [29, 48,49,50,51]. This porosity, although usually lower than that generated by other AM procedures such as the electron beam melting (EBM) [52] has the potential to stimulate bone ingrowth [29, 48, 49], as well as soft tissue adhesion [50, 51], for better healing and long-term tissue stability. Naturally, the present full digital technique has some problems. For the full digital design of these implants, experience and knowledge of CAD software is required. In particular, the thresholding is delicate for creating the 3D bone model on which the implant will be designed. In fact, if the thresholding is wrong, problems will inevitably arise in the fit and adaptation of the implant during the operation.

Output: corroborate


Input: Claim: Everyone's microbiome composition is identical. [SEP] Text: We subsequently tested four bacteria strains, three of which were culturable members of the abundant root community (Fig. 5) and were also abundant members of the root microbiome in the soil extract experiment (Additional file 1: Figure S8). We chose to include a Microbacterium isolate because of its abundance in our reference stock (seven isolates, Fig. 4b) and its classification in the Actinobacteria, a phylum shown to be abundant in plant root microbiomes [25]. We confirmed that these strains successfully colonized plant roots as suggested by the higher abundances on roots compared to their initial inoculated density to the microcosms (Table 2). We could not capture the Microbacterium strain with the community quantification method (Fig. 6b), and similarly, none of the seven isolates from the reference stock clustered to any OTU in the entire dataset. A first possible explanation is that the Microbacterium is a rare but easily culturable microbiome member. Alternatively, the Microbacterium could be an abundant microbiome member, as indicated by the numerous isolates in the reference stock, but absent in the community profiles because of an observed mismatch in the priming site of the PCR primer 799F. A third possible explanation for the microcosms is that although the titer quantification revealed that the Microbacterium strain successfully colonized the plant roots in mono-associations, this strain was outcompeted in the simplified community by the other tested strains. Future experiments need to clarify these possibilities, but nevertheless, this is an example where cultivation and DNA-based approaches do not overlap, and a reminder that both methods have inherent limitations. While it is often discussed that PCR primers are biased towards certain bacterial taxa [30], the same is also true for isolation media, which have a specificity by favoring growth of certain bacterial groups [31]. We quantified the fresh weight of the shoot biomass in response to the bacteria in mono-associations or when the four bacteria were combined to a simplified community. We found that plants grew smaller when inoculated with the Flavobacterium strain in a mono-association, but that this negative plant growth response was alleviated when the Flavobacterium was inoculated in a community with the other strains (Fig. 6a). Since we measured that the Flavobacterium comprised roughly a third of the community (Fig.

Output: contrast


Input: Claim: Some studies suggest that olfactory receptors may function through quantum effects. [SEP] Text: While the authors interpret Figure 2k (and Figure 5) to indicate that artificial activation causes excessive DAN-c1 dopamine release, an alternative explanation is that artificial activation compromises aversive learning by overriding DAN-c1 activity that could be evoked by quinine. This is a great point! Yes, we cannot rule out the possibility that artificial activation compromises aversive learning by overriding DAN-c1 activity that could be evoked by quinine. The experimental results with TRPA1 could be caused by depletion of dopamine, or DA inactivation due to prolonged depolarization or adaptation. However, we still think that our hypothesis on the over-excitation of DAN-c1 is more consistent with our experimental results and other published data. Our justification is as follows: (1) Associative learning occurs only when the CS and US are paired. In wild type larvae, a specific odor (conditioned stimulus, CS, such as pentyl acetate) depolarizes a subset of Kenyon cells in the mushroom body, while gustatory unconditioned stimulus (US, quinine) induces dopamine release from DAN-c1 to the lower peduncle (LP) compartment in the mushroom body (Figure 7a). Only when the CS and US are paired, calcium influx caused by CS and Gas activated by D1R binding to dopamine will turn on a mushroom body specific version of adenylyl cyclase, rutabaga, which is the co-incidence detector in associative learning (Figure 7d). (2) Rutabaga transforms ATP into cAMP, activating PKA signaling pathway and modifying the synaptic strength from mushroom body neurons (MBN, also called Kenyan cells) to the mushroom body output neurons (MBON, Figure 7d). This change in synaptic strength will lead to learned responses when the same odor appears again. (3) In our work, we found D2R is expressed in DAN-c1, and knockdown D2R in DAN-c1 impairs larval aversive learning. As D2R reduces cAMP level and neuronal excitability3, we hypothesized that knockdown of D2R in DAN-c1 would remove the inhibition of D2R auto-receptor, and lead to more dopamine (DA) release when US (quinine) was delivered compared to the wild type larvae.

Output: pass


Input: Claim: 3D printing is used to create custom prosthetics and implants tailored to individual patients. [SEP] Text: It is therefore clear how, in the presence of CBCT with artefacts or substantial scattering in the area to be reconstructed, criticalities can emerge in the design of the implant, which in any case is not simple and requires adequate CAD knowledge. Some company services now offer help for the surgeon, at relatively low costs; these companies own the software and machines necessary for manufacturing the implants, and in this sense they represent the best solution for the clinician. In any case, surgery for placement of a subperiosteal implant is technically more complex than the classic positioning of endosseous implants ad modum Brånemark. The clinician’s surgical skills play a fundamental role in the insertion and in the management of any biological and technical complications. This study has limits. Firstly, it is just a case series; a larger sample of patient would be advisable to draw more specific conclusions about the reliability of these implants. Secondly, this study has a short follow-up of 1 year, and therefore does not report on the possible mid- or long-term complications that may affect these custom-made subperiosteal implants. For example, the progressive atrophy of the bone in elderly patients could determine the mobilization in the implant, in the medium or long period. Extending the follow-up is therefore mandatory, as it could lead to more solid conclusions. ConclusionsIn the past, subperiosteal implants represented a possible solution for prosthetic restoration of the atrophic posterior mandible. However, their use presented several technical and surgical issues (including the need for two surgical sessions, and often poor adaptation to the surgical site) that were in part responsible for the high percentage of complications in the short and long term. Today, each patient must be examined in their complexity, evaluating the best solution considering every aspect, physiological, pathological, aesthetic and economic. Digital technologies such as DMLS allow revisiting the old concept of subperiosteal implants in a modern way, for treatment of the posterior atrophic mandible of elderly patients who do not want to undergo to bone regenerative procedures. In the present study, ten elderly patients were treated with custom-made 3D-printed subperiosteal titanium implants. During surgery, the fit of the implants was satisfactory. Only two implants had insufficient fit, because of the presence of scattering in the CBCT; however, these were adapted to the surgical site during the intervention.

Output: corroborate


Input: Claim: Everyone's microbiome composition is identical. [SEP] Text: 6b), we excluded the possibility that the loss of the negative growth effect was due to the bacterium being outcompeted by the other inoculated strains. Instead, the growth compromising activities of the Flavobacterium were possibly counteracted by one or more of the co-inoculated isolates, or alternatively, it did not reach a sufficient cell density in the simplified community treatment. The reference stock bacteria and microcosms present valuable resources for future microbiota manipulation experiments in which the contribution of the plant root microbiome to plant growth can be investigated. One next step would be to identify the functional traits, e. g. , related to bio-control or plant growth promotion, of the reference stock bacteria using bioassays and/or genome sequencing. We expect that different strains that mapping to the same OTU would interact differently with the host plant, and thus the testing of the functional range among bacteria within an OTU presents another next step. In summary, there are countless opportunities for microcosm experiments. For example, the microbiota of Trifolium can be manipulated with regard to its taxonomic or trait composition or with regard to its diversity and tested for effects on plant growth. Furthermore, the interplay among community members or the dynamics of community assembly can be examined in more detail. Finally, microbiota induced effects on plant growth under stress conditions such as high salinity, reduced nutrient availability, or pathogens can be investigated. ConclusionsWe have reported a multi-step approach (Fig. 1) combining cultivation-dependent and independent methods to describe and functionally examine the root microbiome of Trifolium. The need to experimentally manipulate a microbiota requires reference stocks of isolates, and we believe that reductionist plant-microbiota systems will permit a systematic examination of the root microbiome functions. Further studies employing targeted manipulations of the root microbiome can help in the development of new tools to increase the sustainability of other agricultural plant species [17] and investigate the relationship between microbiome diversity and plant performance [16]. MethodsPreparation of experimental soil, plant cultivation, and harvestExperimental soilAll experiments of this study were conducted with a natural experimental soil collected from the area outside the experimental plots of the long-term and Tillage (FAST) experiment (47° 26′ 20″ N 8° 31′ 40″ E). The experimental soil is a loamy sand with the following physicochemical characteristics: pH 6.

Output: contrast


Input: Claim: Some studies suggest that olfactory receptors may function through quantum effects. [SEP] Text: The elevated DA release along with calcium influx caused by CS increases the cAMP level in MBN, which leads to the learning deficit (over-excitation, Figure 7b). Mutant larvae with excessive cAMP, dunce, showed aversive learning deficiency, supporting our hypothesis2. (4) Our results of TRPA1 can be explained by this over-excitation hypothesis. When DAN-c1 is activated (34C) in distilled water group, the artificial activation mimicked the gustatory activation of quinine. The larvae showed the aversive learning responses towards the odor (Figure 2k DW group). When DAN-c1 is activated (34C) in sucrose group, the artificial activation mimicked the gustatory activation of quinine, so the larvae showed a learning response combining both appetitive and aversive learning (Figure 2k SUC group). (5) When DAN-c1 is activated (34C) in quinine group, the artificial activation and the gustatory activation of quinine lead to elevated DA release from DAN-c1. During training, this elevated DA caused over-excitation of MBN, leading to failure of aversive learning (Figure 2k QUI group), which had a similar phenotype compared to larvae with D2R knockdown in DAN-c1. (6) Similarly, optogenetic activation of DAN-c1 during aversive training, leads to elevated DA release from DAN-c1 (both gustatory activation of quinine and artificial activation). This would also cause over-excitation of MBN, and lead to failure of aversive learning. Artificial activation in other stages (resting or testing) won’t cause elevated DA release during training, so the aversive learning was not affected (Figure 5b). (7) However, when optogenetic activation was applied during training, we did not observe aversive learning responses in the distilled water group, or a reduction in the sucrose group (Figure 5c, Figure 5d). Our explanation is that the optogenetic stimulus we applied is too strong, DAN-c1 has already released elevated DA in both groups. So, the aversive learning in these groups has already been impaired, they just showed the corresponding learning responses to distilled water or sucrose.

Output: pass


Input: Claim: 3D printing is used to create custom prosthetics and implants tailored to individual patients. [SEP] Text: A digital workflow for modeling of custom dental implants Download PDF Download eeaccess Published: 06 June 2019 A digital workflow for modeling of custom dental implants : orcid. org/0000-0002-7822-81311 3D Printing in Medicine 1985. The first documented case of an orthopedic device made using 3D digital technology was a mandibular subperiosteal implant made by James RA [7]. At that time, direct metal printing was not available; therefore, CT data were used to create anatomical models of patients’ mandibles, which were then actualized as implants using traditional metal casting technology. After direc metal laser sintering (DMLS) technology became available from EOS (Krailling, Germany) in 1995 [6], it became possible to make three-dimensional objects in metal with virtually any complexity. This opened up possibilities for completely digital manufacturing processes—a route from patient anatomy virtualization to digital design and direct printing in biologically compatible metal or alloy (such as steel, CoCr (cobalt-chromium) alloy, Ti64 alloy, or pure titanium). Introduced into the dental field, these techniques expanded the ways in which custom subperiosteal implants could be made. With currently available technologies (as of 2018), it is possible to make subperiosteal dental implants using any of the following four methods: 1. Conventional: bone impression and lost wax casting metal 2. Transitional: CT, 3D jaw model printing, lost wax casting in metal 3. Digital: CT, modeling on virtual jaw model, printing in metal using DMLS or similar technology 4. Hybrid: bone impression, silicone impression or stone model optical scanning, modeling of implant on virtual jaw model, printing in metal using DMLS or similar technology The first method is the known classic, long-established approach that requires a second surgery for implant placement. During the first surgery, an impression is taken with silicone material. Then, the implant is modeled on a refractory model, cast in metal, and finished between surgeries—usually 2–3 weeks. The implant is put into place in a second surgical procedure. The second approach avoids the second surgery by using CT data obtained in advance and a refractory model for casting, duplicated from a 3D printed model.

Output: corroborate