Search Results (90)

In November 2022, black-root and stem-rot symptoms were observed on the strawberry cultivar ‘Ssanta’ in Hohhot, Inner Mongolia Autonomous Region, China. In order to identify the causing agent of strawberry root rot, and select effective fungicides for controlling this disease, this study was carried out. The associated fungi were isolated from diseased strawberry plants, and the isolate that fulfilled Koch’s postulates was further identified based on morphological characteristics, together with the internal transcribed spacer regions and 28S rDNA sequences; then, the inhibitory activities of 11 commercial fungicides on the pathogenic strain were screened based on the mycelium growth method. Results showed that six candidate strains were isolated from diseased strawberry, and only the isolate CMGF-A caused typical root and stem rot on strawberry. The CMGF-A showed typical morphology of Acrostalagmus species, and sequence analyses revealed it as A. luteoalbus. The prochloraz was selected as the primary fungicide for effectively controlling CMGF-A, while tebuconazole, thiophanate-methyl, and difenoconazole·azoxystrobin could be used as alternatives. A. luteoalbus was previously reported to cause potato tuber disease, and red rust of needle mushroom, this work is the first report of A. luteoalbus causing strawberry root rot worldwide. This study provided helpful information for the diagnosis and management of strawberry root rot disease. Full article

Fruit rot seriously damages pomelo production. Given concerns regarding the safety of chemical agents, biological alternatives are becoming more preferable. Therefore, the experiment aimed to (i) identify the pathogens causing pomelo fruit rot disease and (ii) select Trichoderma spp. strains controlling the determined pathogens in Ben Tre, Vietnam. Three pathogenic fungal strains isolated from diseased pomelo fruits were selected. The three pathogenic fungal strains were randomly injected into 9 healthy pomelo fruits. The strain PCP-B02-A2 led to a completely rotten fruit on day 17 after infection, while strains PCP-B02-B2 and PCP-B03-A1 had infected spots whose lengths were 17.5 and 28.1 mm, became larger, and eventually led to the whole fruit rot. The pathogens were identified by the internal transcribed spacer (ITS) technique as Colletotrichum gloeosporioides PCP-B02-A2, Pseudopestalotiopsis camelliae sinensis PCP-B03-A1, and P. chinensis PCP-B02-B2. Twenty-five Trichoderma spp. strains were isolated. The ITS technique identified four strains, including Trichoderma asperellum TP-B01, T. harzianum TP-B08, T. harzianum TP-B09, and T. asperellum TP-C25. The PCP-B02-A2 strain had antagonism at 66.7–68.7%, while those of PCP-B02-B2 and PCP-B03-A1 were 64.2–71.1% and 55.7–57.4%, respectively. Full article

Due to the frequent use of fungicides in viticulture, resistant plant pathogens have emerged, necessitating environmentally friendly alternatives. This research aimed to determine the effect of ozonated water (OW) spraying on fungal colonies present on grapevine leaves and berries, as well as on the biochemical composition of the berries. ‘Regent’ grapevines were grown in a high plastic tunnel and sprayed with OW from post-flowering to harvest. The fungal population on the phyllosphere of grapevine leaves and berries was evaluated using the serial dilution plating method. The taxonomic composition of the predominant fungal colonies was characterized using internal transcribed spacer amplicon sequencing. OW treatment significantly decreased fungal colonies on grapes but had no significant effect on grapevine leaves. The fungal colonies were dominated by Botrytis cinerea, Penicillium brevicompactum, and Fusarium sp. OW treatment significantly reduced the total sugar content in grapes (from 160 to 154 g L⁻¹) and increased the total acid content (from 7.2 to 8.6 g L⁻¹). The fruit polyphenol content increased from 431 to 508 mg 100 g⁻¹, and antioxidant activity was significantly enhanced. It can be concluded that OW treatment is effective in reducing fungal colony forming units on grapes in vineyards. OW treatment affected the sugar, acid, and polyphenol content in grapes, but not to a degree that would present specific challenges for winemakers. Full article

Background: as the demand for total joint arthroplasty continues to grow each year, the healthcare burden is expected to increase due to periprosthetic joint infection (PJI). This review article aims to highlight the significance of biofilms in the pathogenesis of PJI and introduce alternative therapies that prevent bacterial adhesion to implants or enhance their eradication when infection occurs. Search strategy: we conducted a bibliographic search in PubMed using the following MeSH terms as follows: “no antibiotic treatment of PJI”, “bacterial biofilm eradication agents”, and “unconventional prevention of PJI”, among others. Most important results: after an initial analysis of the literature, we selected the most significant topics on novel PJI treatment methods and prevention strategies. A second PubMed search highlighted the following therapeutic modalities: the application of hydrogels on implant surfaces, the use of phage therapy, lysostaphin and antimicrobial peptides, the implementation of two-stage debridement, irrigation, implant retention and antibiotic therapy (DAIR), the intra-articular antibiotic infusion, and the use of methylene blue for biofilm eradication. Conclusions: the use of new cement spacers with xylitol, ammonium compounds, or silver nanoparticles is another promising technique to increase the eradication rate in two-stage revision. It is important for professionals to deeply understand the pathogenesis of PJI and the role of biofilms in its development in order to become familiar with these novel techniques that could reduce the burdens on healthcare systems. Full article

Introduction: Chronic osteomyelitis is a persistent infection of the bone that poses significant challenges, particularly when associated with pathological fractures and extensive bone defects. This case report highlights the application of Masquelet’s induced membrane technique (MIMT) in managing a complex distal femur defect in a 50-year-old male with a long-standing history of chronic osteomyelitis. The patient presented with a non-union fracture, severe osseous destruction, and infection, requiring a multidisciplinary approach. Case report: The first stage involved radical debridement of necrotic tissue and stabilization with a titanium plate and an antibiotic-impregnated polymethylmethacrylate spacer to induce a bioactive membrane. The second stage, performed 30 days later, after infection resolution, entailed removing the spacer, harvesting an autologous iliac crest bone graft, and filling the defect within the preserved membrane. Postoperative care included a tailored antibiotic regimen and gradual weight-bearing, leading to favorable clinical and radiological outcomes. Conclusions: This case demonstrates the utility of MIMT in reconstructing extensive bone defects while addressing infection. The technique provides a reliable and effective alternative to amputation, offering high success rates and functional restoration in complex cases. Full article

An HVDC-GIL system with a conical spacer in a radioactive environment is studied in this work using simulated data on COMSOL^® Multiphysics. Electromagnetic simulations on a 2D model were performed with varying ion-pair generation rates and potential applied to the system. This article explores machine learning methods to derive time to steady state, dark current, gas conductivity, and surface charge density expressions. The focus was on constructing symbolic representations, which could be interpretable and less prone to overfitting, using the symbolic regression (SR) and sparse identification of nonlinear dynamics (SINDy) algorithms. The study successfully derived the intended expressions, demonstrating the power of symbolic regression. Predictions of dark currents in the gas–ground electrode interface reported an absolute error and mean absolute percentage error (MAPE) of 1.04 × ${10}^{-4}$ pA and 0.01%, respectively. The solid–ground electrode interface reported an error of 8.99 × ${10}^{-5}$ pA and MAPE of 0.04%, showing strong agreement with simulation data. Expressions for time to steady state had a test error of approximately 110 h with MAPE of around 3%. Steady-state gas conductivity expression achieved an absolute error of 0.55 log(S/m) and MAPE of 1%. An interpretable equation was created with SINDy to model the time evolution of surface charge density, achieving a root mean squared error of 1.12 nC/m²/s across time-series data. These results demonstrate the capability of SR and SINDy to provide interpretable and computationally efficient alternatives to time-consuming numerical simulations of HVDC systems under radiation conditions. While the model provides useful insights, performance and practical applications of the expressions can improve with more diverse datasets, which might include experimental data in the future. Full article

The taxane molecules extracted from the bark of trees from the Taxus genus demonstrate anticancer activity and are the main components of the drug paclitaxel. Even though a great deal of research has been carried out to produce them synthetically, this manufacturing is still dependent on Taxus cell culture. Furthermore, these processes are not suitable for steady taxane production. Therefore, the search for alternative sources of taxane production has generated growing interest amongst the scientific community. The use of endophytic fungi for the obtention of taxane constitutes an attractive alternative. Here, we present an analysis of the taxane production of several microorganisms through LC–QTOF MS/MS. We isolated 21 endophytic fungus strains, identified through sequencing of their internal transcribed spacer region (ITS). The phylogenetic analysis showed the presence of 11 different Fungus genera, namely Aspergillus, Trichoderma, Neurospora, Penicillium, Curvularia, Arcopilus, Biscogniauxia, Hypoxylon, Sordaria, Xylariales, and Gelasinospora. These fungi have been previously described to synthetize distinct metabolites of medical interest, hence supporting the study of their potential taxane production. Here, we report the production of taxadiene by some of these fungi, namely Aspergillus sp. (1.35 μg/L), TgO (0.7 μg/L), Trichoderma harzianum (0.13 μg/L), and Hypoxylon sp. (0.3 μg/L). Furthermore, we also detected the production of baccatin III, a crucial precursor component of the drug paclitaxel. This is the first report of taxane production by fungi phylogenetically related to the Arcopilus and Endoxyla genera. Full article

Background: This comprehensive systematic review aims to explore and discuss existing treatment modalities for infections in total ankle arthroplasty (TAA), providing insights that may contribute to the establishment of a “standard of care” for these challenging cases. The study analyses the intricate landscape of infected TAA, addressing gaps in the current literature and emphasizing the need to refine treatment strategies. With the reported incidence of periprosthetic joint infection after TAA surpassing rates observed in total hip and knee replacements, the research navigates through various treatment modalities, underscoring the lack of a universally accepted standard of care. Methods: In this systematic review, following PRISMA guidelines, PubMed, Scopus, and Google Scholar, we identified 15 papers addressing the management strategies for infected TAA (162 infected ankle arthroplasty cases).These databases were chosen for their extensive coverage, strong relevance to the research topic, and ease of access, ensuring a thorough and focused retrieval of pertinent literature on the treatment of infected ankle prostheses. The review involved the identification and evaluation of articles providing insights into complications, treatment outcomes, and risk factors. Extracted data were summarized and reported. A descriptive analysis was performed, and when feasible, a statistical analysis was conducted. Results: Treatment modalities included irrigation and debridement (48.8%), revision total ankle arthroplasty (36.3%), primary arthrodesis (7.9%), spacer arthroplasty (4.5%), and primary amputation (3.9%). Complication rates varied, with 46.5% for irrigation and debridement, 20% for two-stage revision, 7.14% for primary arthrodesis, and 25% for spacer arthroplasty. Conclusions: The rising prevalence of TAA underscores the need for a definitive treatment protocol due to severe complications. This review emphasizes careful patient selection and accurate diagnosis. Irrigation and debridement are effective for acute infections, while two-stage revision is a valid alternative for chronic infections. High-quality randomized controlled trials are important for establishing an evidence-based treatment protocol. Full article

Hysterothylacium aduncum is one of six pathogens responsible for human anisakiasis. Infection with H. aduncum can cause acute abdominal symptoms and allergic reactions and is prone to misdiagnosis in clinical practice. This study aims to enhance the efficiency and accuracy of detecting H. aduncum in food ingredients. We targeted the internal transcribed spacer 1 (ITS 1) regions of Anisakis to develop a visual screening method for detecting H. aduncum using recombinase polymerase amplification (RPA) combined with the CRISPR/Cas12a system. By comparing the ITS 1 region sequences of eight nematode species, we designed specific primers and CRISPR RNA (crRNA). The specificity of RPA primers was screened and evaluated, and the CRISPR system was optimized. We assessed its specificity and sensitivity and performed testing on commercial samples. The results indicated that the alternative primer ADU 1 was the most effective. The final optimized concentrations were 250 nM for Cas12a, 500 nM for crRNA, and 500 nM for ssDNA. The complete test procedure was achievable within 45 min at 37 °C, with a limit of detection (LOD) of 1.27 pg/μL. The amplified product could be directly observed using a fluorescence microscope or ultraviolet lamp. Detection results for 15 Anisakis samples were entirely consistent with those obtained via Sanger sequencing, demonstrating the higher efficacy of this method for detecting and identifying H. aduncum. This visual detection method, characterized by simple operation, visual results, high sensitivity, and specificity, meets the requirements for food safety testing and enhances monitoring efficiency. Full article

Sanguinoderma (Ganodermataceae) is recognized as a valuable medicinal resource in Taiwan, China. Additionally, it serves as a traditional folk medicine for treating neurotic epilepsy in Malaysia. This study involved the collection of six specimens of Sanguinoderma from Yunnan Province, China. Employing multigene phylogenetic analysis of DNA sequences, including internal transcribed spacer (ITS), nuclear large subunit (LSU), RNA polymerase II second largest subunit (rpb2), translation elongation factor 1-alpha (tef1-α), mitochondrial small subunit (mtSSU), nuclear small subunit (nSSU) and morphological examinations, three new species, viz. Sanguinoderma concentricum, S. dehongense and S. ovisporum, are introduced. Sanguinoderma concentricum is characterized by a central stipe basidiomata, an orbicular to suborbicular pileus, a grayish-yellow surface with alternating concentric zones and wavy margin-like petals and regular pileipellis cells (4–8 × 17–28 μm). Sanguinoderma dehongense is characterized by a long stipe and flabelliform basidiomata, a dark-grayish yellow-to-dark-yellow pileus surface, irregular pileipellis cells and wavy margin and ellipsoid basidia (8–11 × 9–13 μm). Sanguinoderma ovisporum is characterized by a reniform basidiomata, a heterogeneous context and ovoid basidiospores (7.5–8.6 × 5.5–7.2 µm). A detailed description and illustrations of these new species are provided, as well as a morphological comparison with similar taxa. Full article

The Herpotrichiellaceae family is an important group of dematiaceous filamentous fungi, associated with a variety of pathogenic fungal species causing chromoblastomycosis (CBM) and phaeohyphomycosis (PHM), both with polymorphic clinical manifestations and worldwide incidence. Currently, the identification of this family and determination of the causative agent is challenging due to the subjectivity of morphological identification methods, necessitating the use of molecular techniques to complement diagnosis. In this context, genetic sequencing of the Internal Transcribed Spacer (ITS) has become the norm due to a lack of alternative molecular tools for identifying these agents. Therefore, this study aimed to develop PCR-Multiplex methodologies to address this gap. Sequences from the ITS and Large Subunit (LSU) of ribosomal DNA were used, and after manual curation and in vitro analyses, primers were synthesized for the identification of the targets. The primers were optimized and validated in vitro, resulting in two PCR-Multiplex methodologies: one for identifying the Herpotrichiellaceae family and the bantiana clade, and another for determining the species Fonsecaea pedrosoi and Fonsecaea monophora. Ultimately, the assays developed in this study aim to complement other identification approaches for these agents, reducing the need for sequencing, improving the management of these infections, and enhancing the accuracy of epidemiological information. Full article

Rifampicin is one of the mainstays in treating staphylococcal prosthetic joint infection (PJI). However, discontinuation due to intolerance, drug interactions, and adverse events is common. Two-stage revision surgery remains the gold standard, with the number of revision arthroplasties steadily increasing. This study aims to evaluate the effectiveness and safety of a novel two-stage revision protocol for staphylococcal prosthetic joint infection (PJI) utilizing bone cement spacers loaded with multiple high doses of antibiotics. Additionally, it seeks to analyze outcomes in patients ineligible for rifampicin treatment. A retrospective review of 43 cases of staphylococcal hip and knee prosthetic joint infections (PJIs) from 2012 to 2020 was conducted. In all instances, a commercial cement containing 1 g of gentamicin and 1 g of clindamycin, augmented with 4 g of vancomycin and 2 g of ceftazidime, was employed to cast a spacer manually after thorough surgical debridement. We report an eradication rate of 82%, with no significant differences observed (p = 0.673) between patients treated with (84%, n = 19) and without rifampicin (79%, n = 24). There were no disparities in positive culture rates (7%), spacer replacement (18%), or survival analysis (p = 0.514) after an average follow-up of 68 months (range 10–147) in the absence of systemic toxicity and surgical complications superimposable to those previously reported. In conclusion, two-stage revision with local high doses of ceftazidime, vancomycin, gentamicin, and clindamycin demonstrates high effectiveness in treating staphylococcal PJIs. Notably, systemic rifampicin does not influence the outcomes. This protocol, with multiple high doses of antibiotics loaded into the bone cement spacer, is presented as a viable and safe alternative for patients unsuitable for rifampicin treatment. Full article

In the present study, we compared the genetic variability of fragments from the internal transcribed spacer region (ITS) and the small subunit ribosomal DNA (SSUrDNA) as nuclear markers, in contrast with the ribosomal protein large two (rpl2) loci, placed in the mitochondrion-related organelles (MROs) within and among human fecal samples with Blastocystis. Samples were analyzed using polymerase chain reaction (PCR)-sequencing, phylogenies, and genetics of population structure analyses were performed. In total, 96 sequences were analyzed, i.e., 33 of SSUrDNA, 35 of rpl2, and 28 of ITS. Only three subtypes (STs) were identified, i.e., ST1 (11.4%), ST2 (28.6%), and ST3 (60%); in all cases, kappa indexes were 1, meaning a perfect agreement among ST assignations. The topologies of phylogenetic inferences were similar among them, clustering to each ST in its specific cluster; discrepancies between phylogeny and assignment of STs were not observed. The STRUCTURE v2.3.4 software assigned three subpopulations corresponding to the STs 1–3, respectively. The population indices were consistent with those previously reported by other groups. Our results suggest the potential use of the ITS and rpl2 genes as molecular markers for Blastocystis subtyping as an alternative approach for the study of the genetic diversity observed within and between human isolates of this microorganism. Full article

The demand for high-precision CRISPR/Cas9 systems in biomedicine is experiencing a notable upsurge. The editing system fdCas9 employs a dual-sgRNA strategy to enhance editing accuracy. However, the application of fdCas9 is constrained by the stringent requirement for two protospacer adjacent motifs (PAMs) of Cas9. Here, we devised an optimized editor, fRYdCas9, by merging FokI with the nearly PAM-less RYdCas9 variant, and two fRYdCas9 systems formed a dimer in a proper spacer length to accomplish DNA cleavage. In comparison to fdCas9, fRYdCas9 demonstrates a substantial increase in the number of editable genomic sites, approximately 330-fold, while maintaining a comparable level of editing efficiency. Through meticulous experimental validation, we determined that the optimal spacer length between two FokI guided by RYdCas9 is 16 base pairs. Moreover, fRYdCas9 exhibits a near PAM-less feature, along with no on-target motif preference via the library screening. Meanwhile, fRYdCas9 effectively addresses the potential risks of off-targets, as analyzed through whole genome sequencing (WGS). Mouse embryonic editing shows fRYdCas9 has robust editing capabilities. This study introduces a potentially beneficial alternative for accurate gene editing in therapeutic applications and fundamental research. Full article

Energy-level alignment is a crucial factor in the performance of thin-film devices, such as organic light-emitting diodes and photovoltaics. One way to adjust these energy levels is through chemical modification of the molecules involved. However, this approach may lead to unintended changes in the optical and/or electrical properties of the compound. An alternative method for energy-level adjustment at the interface is the use of self-assembling monolayers (SAMs). Initially, SAMs with passive spacers were employed, creating a surface dipole moment that altered the work function (WF) of the electrode. However, recent advancements have led to the synthesis of SAM molecules with active spacers. This development necessitates considering not only the modification of the electrode’s WF but also the ionization energy (IE) of the molecule itself. To measure both the IE of SAM molecules and their impact on the electrode’s WF, a relatively simple method is photo-electric emission spectroscopy. Solar cell performance parameters have a higher correlation coefficient with the ionization energy of SAM molecules with carbazole derivatives as spacers (up to 0.97) than the work function of the modified electrode (up to 0.88). Consequently, SAMs consisting of molecules with active spacers can be viewed as hole transport layers rather than interface layers. Full article