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  • Juhl Philipsen posted an update 1 week, 6 days ago

    Septation in filamentous fungi is a normal part of development, which involves the formation of cross-hyphal bulkheads, typically containing pores, allowing cytoplasmic streaming between compartments. Rosuvastatin Based on previous findings regarding septa and cell wall stress, we hypothesized that septa are critical for survival during cell wall stress. To test this hypothesis, we used known Aspergillus nidulans septation-deficient mutants (ΔsepH, Δbud3, Δbud4, and Δrho4) and six antifungal compounds. Three of these compounds (micafungin, Congo red, and calcofluor white) are known cell wall stressors which activate the cell wall integrity signaling pathway (CWIS), while the three others (cycloheximide, miconazole, and 2,3-butanedione monoxime) perturb specific cellular processes not explicitly related to the cell wall. Our results show that deficiencies in septation lead to fungi which are more susceptible to cell wall-perturbing compounds but are no more susceptible to other antifungal compounds than a control. This implies that septa play a critical role in surviving cell wall stress. IMPORTANCE The ability to compartmentalize potentially lethal damage via septation appears to provide filamentous fungi with a facile means to tolerate diverse forms of stress. However, it remains unknown whether this mechanism is deployed in response to all forms of stress or is limited to specific perturbations. Our results support the latter possibility by showing that presence of septa promotes survival in response to cell wall damage but plays no apparent role in coping with other unrelated forms of stress. Given that cell wall damage is a primary effect caused by exposure to the echinocandin class of antifungal agents, our results emphasize the important role that septa might play in enabling resistance to these drugs. Accordingly, the inhibition of septum formation could conceivably represent an attractive approach to potentiating the effects of echinocandins and mitigating resistance in human fungal pathogens.Titanium (Ti) is an element beneficial to plant growth. Application of titanium to roots or leaves at low concentrations can improve crop yield and performance. However, the effect of titanium ions on the bulk soil microbial community of planted crops remains unclear. This study aimed to explore the effects of titanium on soil bacterial and fungal communities. Field surveys were conducted to determine the effect of titanium ions on bulk soil microbial communities in pitaya and grape plantations of Panzhihua and Xichang areas, respectively. Full-length 16S rRNA and internal transcribed spacer (ITS) amplicon sequencing were performed using PacBio Sequel to further explore the composition and structure of soil microbiota. The application of titanium ions significantly altered the composition and structure of soil microbiota. Root irrigation with titanium ions in pitaya gardens reduced the diversity of soil fungi and bacteria. However, the decline in bacterial diversity was not statistically significant. Meanwhil cooccurrence relationships and improved the stability of the community. This study provides a basis for the importance of titanium ion application in crop cultivation.Methanocaldococcus sp. strain FS406-22, a hyperthermophilic methanogen, fixes nitrogen with a minimal set of known nif genes. Only four structural nif genes, nifH, nifD, nifK, and nifE, are present in a cluster, and a nifB homolog is present elsewhere in the genome. nifN, essential for the final synthesis of the iron-molybdenum cofactor of nitrogenase in well-characterized diazotrophs, is absent from FS406-22. In addition, FS406-22 encodes four novel hypothetical proteins, and a ferredoxin, in the nif cluster. Here, we develop a set of genetic tools for FS406-22 and test the functionality of genes in the nif cluster by making markerless in-frame deletion mutations. Deletion of the gene for one hypothetical protein, designated Hp4, delayed the initiation of diazotrophic growth and decreased the growth rate, an effect we confirmed by genetic complementation. NifE also appeared to play a role in diazotrophic growth, and the encoding of Hp4 and NifE in a single operon suggested they may work together in some way in the synthesis of the nitrogenase cofactor. No role could be discerned for any of the other hypothetical proteins, nor for the ferredoxin, despite the presence of these genes in a variety of related organisms. Possible pathways and evolutionary scenarios for the synthesis of the nitrogenase cofactor in an organism that lacks nifN are discussed. IMPORTANCE Methanocaldococcus has been considered a model genus, but genetic tools have not been forthcoming until recently. Here, we develop and illustrate the utility of positive selection with either of two selective agents (simvastatin and neomycin), negative selection, generation of markerless in-frame deletion mutations, and genetic complementation. These genetic tools should be useful for a variety of related species. We address the question of the minimal set of nif genes, which has implications for how nitrogen fixation evolved.Developing strategies to regulate the immune response poses significant challenges with respect to the clinical translation of tissue-engineered scaffolds. Prominent advancements have been made relating to macrophage-based therapies and biomaterials. Macrophages exhibit the potential to influence healing trajectory, and predominance of particular subtypes during early onset of healing influences repair outcomes. This study evaluated short- and long-term healing response and postoperative mechanical properties of genipin-cross-linked, electrochemically aligned collagen biotextiles with comparative administration of M0, M1, and M2 subtypes. Irrespective of macrophage subtype seeded, all the groups demonstrated existence of M2 macrophages at both time points as typified by arginase and Ym-1 expressions, and distinct absence of M1 macrophages, as indicated by lack of inducible nitric oxide synthase (iNOS) and interleukin-1β expression in all the groups for both time points. M2 macrophage-seeded collagen biotextilgineering remains limited. In this study, genipin-cross-linked collagen biotextiles were employed to deliver M0, M1, and M2 macrophages and evaluate tissue responses and postsurgical mechanical properties in vivo. M2-seeded collagen biotextiles showed reduced fibrous capsule and favorable healing response. These outcomes shed new light on designing tissue-engineered constructs that offer a novel cell-based therapeutic approach for applications requiring structural augmentation.We retrospectively reviewed 56 fingers with a type 1 flexor digitorum profundus avulsion (jersey finger) injury. Nineteen fingers were treated with a four-strand pull-out suture technique that did not include the distal palmar plate, and 37 were treated with a six-strand pull-out suture technique that included the distal palmar plate. An early active mobilization regimen was used. At follow-up (mean 13 and 14 months, respectively), total active interphalangeal joint motion was significantly better in the six-strand/palmar plate group, as were the results assessed with the Strickland-Glogovac grading system and patient satisfaction according to the Stark criteria. No repair ruptures occurred in this group versus two ruptures in the four-strand/no-palmar-plate group. We consider that the better results can be attributed to a stronger repair, which facilitated early active mobilization.Level of evidence III.1. This study evaluated and compared the oxidation status of meat from conventionally-farmed, organic and wild ducks during chilled storage in air or vacuum packaging. In total, 48 carcases from each group of ducks were investigated, and 24 carcases from each type of production were stored under vacuum or in air at 3°C ± 1°C. Sample analyses were conducted after 1, 3, 6 and 9 d of storage.2. The antioxidant capacity (2,2-diphenyl-1-picrylhydrazyl DPPH), thiobarbituric acid reactive substances (TBARS) and free fatty acids in the breast, thigh and wing muscles were determined.3. The antioxidant capacity of meat from conventional ducks was higher than that from organic and wild ducks. The muscles of organic and wild ducks were more susceptible to secondary oxidation (particularly when stored in air) than conventionally-raised birds. The free fatty acid levels in samples of breast and thigh meat from wild ducks on the last day of storage were higher than in conventional and organic ducks.4. The results showed that the oxidation status of meat from conventional ducks was more stable during refrigerated storage than the meat of organic and wild ducks.A variety of surgical techniques and tissue engineering strategies utilizing osteogenic potential of the periosteum have been developed for the repair of extended bone deficiencies. The aim of the present study was to assess the impact of an alternating protocol of periosteal distraction osteogenesis (PDO) on bone regeneration in an intraoral model. Eight adult, male Beagle dogs were used for the study. Two distraction devices were placed on each side of the mandible. After a 7-day latency period, distraction devices in all animals were manipulated at the rate of 0.5 mm for a total of 8 days. The pumping protocol in two test groups proceeded twice daily by alternating activation with relaxation. In the periosteal pumping/distraction (PPDO) group, the distraction screws were activated two times (at 12 and 24 h) and then turned back (at 36 h), and in the periosteal pumping (PP) group repeatedly activated and turned back (at 12 h). In the PDO group, only activation was performed once daily (positive control). Deration in the mandible. A periosteum-based regeneration approach may represent a valuable step toward creating a significant volume of hard and soft tissues, without need for autogenous bone harvesting or application of biomaterials.Assigning amplicon sequences to operational taxonomic units (OTUs) is an important step in characterizing microbial communities across large data sets. A notable difference between de novo clustering and database-dependent reference clustering methods is that OTU assignments from de novo methods may change when new sequences are added. However, one may wish to incorporate new samples to previously clustered data sets without clustering all sequences again, such as when comparing across data sets or deploying machine learning models. Existing reference-based methods produce consistent OTUs but only consider the similarity of each query sequence to a single reference sequence in an OTU, resulting in assignments that are worse than those generated by de novo methods. To provide an efficient method to fit sequences to existing OTUs, we developed the OptiFit algorithm. Inspired by the de novo OptiClust algorithm, OptiFit considers the similarity of all pairs of reference and query sequences to produce OTUs of the other, but the assignments can change when new sequences are added to a data set, making it difficult to compare different studies. Other approaches assign sequences to groups by comparing them to sequences in a reference database to produce consistent assignments, but the quality of the groups produced is reduced compared to that with OptiClust. We developed OptiFit, a new reference-based algorithm that produces consistent yet high-quality assignments like OptiClust. OptiFit allows researchers to compare microbial communities across different studies or add new data to existing studies without sacrificing the quality of the group assignments.