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Plant Disease Aug 2023Bletilla striata (Thunb.) is a perennial herb plant of the orchidaceous family and is used as an ornamental plant in Europe and the United States. Furthermore, it is...
Bletilla striata (Thunb.) is a perennial herb plant of the orchidaceous family and is used as an ornamental plant in Europe and the United States. Furthermore, it is important as traditional Chinese medicine (TCM) in East Asian countries, such as China, Japan, Korea, Mongolia, and Myanmar (Gou et al. 2022). In April 2023, a severe disease similar to gray mold occurred in a B. striata plantation in Anqing, Anhui province, China (N30°27'15″, E116°18'32″), causing disease on about 20% of the plants in the field. Early symptoms were characterized by brown spots or stripes on the leaves of B. striata, and as the disease progressed, large brown irregular spots appeared. Eventually disease spots coalesced, covering the entire leaf surface and causing leaf death. A gray mildew layer was observed on the senescent leaves. To investigate the causal agent, 10 plants with typical symptoms were collected from the field. Leaf pieces (5 × 5 mm) from the border of infected areas were soaked in 75% ethanol for 10 seconds, and then transferred into 0.1% mercury bichloride for three min, rinsed three times with sterile water, and transferred to PDA at 25 °C for three days. Pure cultures were obtained by single spore isolation, and the resulting colonies were morphologically similar, indicating a single pathogen, of which the representative BSFC-7 was selected for further study. BSFC-7 colonies were initially white to gray-brown, and cottony aerial hyphae grew over the entire petri dish after five days of incubation. Grayish, branched conidiophores and their terminal unicellular conidia were observed under a microscope after additional two days at 25 °C. Conidia were colorless or gray, elliptical or oval, and 7.06-12.54 × 8.33-13.55 μm (n=30). Sclerotia appeared in BSFC-7 culture up to about two weeks and were black, hard, and round or irregularly shaped (0.81-4.32 × 0.97-5.68 mm, n=20). The morphological characteristics fit the description of Botrytis cinerea (Li et al. 2016). To further identify the species, genomic DNA of BSFC-7 was extracted. PCR analysis was performed with species-specific primer pairs C729+/C729- and two nuclear genes G3PDH and RPB2 with their corresponding primer pairs G3PDH-F/G3PDH-R and RPB2-F/RPB2-R (Rigotti et al. 2002; Aktaruzzaman et al. 2018). The sequences for all three PCR products of C729, G3PDH, and RPB2 (GenBank accession nos. OR287069, OR255923, and OR255924 respectively) exhibited 99 to 100% similarity with other B. cinerea isolates. In the pathogenicity test, detached leaves of B. striata were inoculated with the BSFC-7 isolate. The leaves were soaked in sodium hypochlorite (1%) for two min, washed with sterile distilled water, and then inoculated with 10 µl of conidial suspension (106 conidia/ml). Sterile water was used as control and samples were incubated at 25 °C. After three days, all leaves inoculated with conidia showed dark brown water-soaked lesions similar to those observed in the field, while the control leaves remained healthy. The pathogen was re-isolated from the affected leaves, fulfilling Koch's postulates. B. cinerea is a common pathogen on a wide range of host plant species worldwide and has been reported to infect B. striata in Yunnan province, China (Romanazzi and Feliziani 2014; Zhang et al. 2020). To our knowledge, this is the first report of B. cinerea causing leaf spots on B. striata in Anhui province, China. This study will provide a basis for controlling the prevalence and economic losses of gray mold on B. striata.
PubMed: 37642545
DOI: 10.1094/PDIS-07-23-1384-PDN -
Potential of Plant-Based Extracts to Alleviate Sorbitol-Induced Osmotic Stress in Cabbage Seedlings.Plants (Basel, Switzerland) Mar 2024In light of expected climate change, it is important to seek nature-based solutions that can contribute to the protection of our planet as well as to help overcome the...
In light of expected climate change, it is important to seek nature-based solutions that can contribute to the protection of our planet as well as to help overcome the emerging adverse changes. In an agricultural context, increasing plant resistance to abiotic stress seems to be crucial. Therefore, the scope of the presented research was focused on the application of botanical extracts that exerted positive effects on model plants growing under controlled laboratory conditions, as well as plants subjected to sorbitol-induced osmotic stress. Foliar spraying increased the length and fresh mass of the shoots (e.g., extracts from , , and ) and the roots (e.g., , , and ) of cabbage seedlings grown under stressful conditions, as well as their content of photosynthetic pigments (, , and ) along with total phenolic compounds (, , and ). The antioxidant activity of the shoots measured with the use of DDPH (, , , and ), ABTS (, , , , and ), and FRAP (, , , , and ) assays was also enhanced in plants exposed to osmotic stress. Based on these findings, the most promising formulation based on was selected and subjected to transcriptomic analysis. The modification of the expression of the following genes was noted: (glutathione S-transferase), (chlorophyll A-B binding protein), (S-adenosylmethionine-dependent methyltransferases), (chlorophyll A-B binding protein), (GDSL lipase/esterase), (heat shock protein 70 family), (DnaJ Chaperone), (pre-mRNA splicing Prp18-interacting factor), (heat shock protein Hsp90 family), (gibberellin regulated protein), (B-box-type zinc finger), (RmlC-like cupin domain superfamily), (myb_SHAQKYF: myb-like DNA-binding domain, SHAQKYF class), (DA1-like Protein). Gene Ontology functional analysis indicated that the application of the extract led to a decrease in the expression of many genes related to the response to stress and photosynthetic systems, which may confirm a reduction in the level of oxidative stress in plants treated with biostimulants. The conducted studies showed that the use of innovative plant-based products exerted positive effects on crops and can be used to supplement current cultivation practices.
PubMed: 38592867
DOI: 10.3390/plants13060843 -
BioRxiv : the Preprint Server For... Mar 2024The small amount of human tissue available for testing is a paramount challenge in cancer drug development, cancer disease models, and personalized oncology....
The small amount of human tissue available for testing is a paramount challenge in cancer drug development, cancer disease models, and personalized oncology. Technologies that combine the microscale manipulation of tissues with fluid handling offer the exciting possibility of miniaturizing and automating drug evaluation workflows. This approach minimizes animal testing and enables inexpensive, more efficient testing of samples with high clinical biomimicry using scarce materials. We have developed an inexpensive platform based on an off-the-shelf robot that can manipulate microdissected tissues (µDTs) into user-programmed positions without using intricate microfluidic designs nor any other accessories such as a microscope or a pneumatic controller. The robot integrates complex functions such as vision and fluid actuation by incorporating simple items including a USB camera and a rotary pump. Through the robot's camera, the platform software optically recognizes randomly-seeded µDTs on the surface of a petri dish and positions a mechanical arm above the µDTs. Then, a custom rotary pump actuated by one of the robot's motors generates enough microfluidic lift to hydrodynamically pick and place µDTs with a pipette at a safe distance from the substrate without requiring a proximity sensor. The platform's simple, integrated construction is cost-effective and compact, allowing placement inside a tissue culture hood for sterile workflows. The platform enables users to select µDTs based on their size, place them in user-programmed arrays, such as multi-well plates, and control various robot motion parameters. As a case application, we use the robotic system to conduct semi-automated drug testing of mouse and human µDTs in 384-well plates. Our user-friendly platform promises to democratize microscale tissue research to clinical and biological laboratories worldwide.
PubMed: 38586030
DOI: 10.1101/2024.03.21.586169 -
Stem Cell Research & Therapy Oct 2023Thymic epithelial cells (TECs) are responsible for shaping the repertoires of T cells, where their postnatal regeneration depends on a subset of clonogenic TECs. Despite...
BACKGROUND
Thymic epithelial cells (TECs) are responsible for shaping the repertoires of T cells, where their postnatal regeneration depends on a subset of clonogenic TECs. Despite the implications for regenerative medicine, their cultivation and expansion remain challenging. Primary explant cell culture is a technique that allows the seeding and expansion of difficult-to-culture cells. Here, we report a reliable and simple culture system to obtain functional TECs and thymic interstitial cells (TICs).
METHODS
To establish primary thymic explants, we harvested 1 mm cleaned fragments of thymus from 5-week-old C57/BL6 mice. Tissue fragments of a complete thymic lobe were placed in the center of a Petri dish with 1 mL of DMEM/F-12 medium supplemented with 20% fetal bovine serum (FBS) and 1% penicillin‒streptomycin. To compare, thymic explants were also cultivated by using serum-free DMEM/F-12 medium supplemented with 10% KnockOut™.
RESULTS
We obtained high numbers of functional clonogenic TECs and TICs from primary thymic explants cultivated with DMEM/F-12 with 20% FBS. These cells exhibited a highly proliferative and migration profile and were able to constitute thymospheres. Furthermore, all the subtypes of medullary TECs were identified in this system. They express functional markers to shape T-cell and type 2 innate lymphoid cells repertoires, such as Aire, IL25, CCL21 and CD80. Finally, we also found that ≥ 70% of lineage negative TICs expressed high amounts of Aire and IL25.
CONCLUSION
Thymic explants are an efficient method to obtain functional clonogenic TECs, all mTEC subsets and different TICs AireIL25 with high regenerative capacity.
Topics: Mice; Animals; Immunity, Innate; Lymphocytes; Thymus Gland; Epithelial Cells; T-Lymphocytes; Cell Differentiation
PubMed: 37904232
DOI: 10.1186/s13287-023-03529-8 -
Mikrochimica Acta Apr 2024The therapeutic effect of gefitinib on colorectal cancer (CRC) is unclear, but it has been reported that stromal cells in the tumor microenvironment may have an impact...
The therapeutic effect of gefitinib on colorectal cancer (CRC) is unclear, but it has been reported that stromal cells in the tumor microenvironment may have an impact on drug sensitivity. Herein, we established a microfluidic co-culture system and explored the sensitivity of CRC cells co-cultured with cancer-associated fibroblasts (CAFs) to gefitinib. The system consisted of a multichannel chip and a Petri dish. The chambers in the chip and dish were designed to continuously supply nutrients for long-term cell survival and create chemokine gradients for driving cell invasion without any external equipment. Using this system, the proliferation and invasiveness of cells were simultaneously evaluated by quantifying the area of cells and the migration distance of cells. In addition, the system combined with live cell workstation could evaluate the dynamic drug response of co-cultured cells and track individual cell trajectories in real-time. When CRC cells were co-cultured with CAFs, CAFs promoted CRC cell proliferation and invasion and reduced the sensitivity of cells to gefitinib through the exosomes secreted by CAFs. Furthermore, the cells that migrated out of the chip were collected, and EMT-related markers were determined by immunofluorescent and western blot assays. The results demonstrated that CAFs affected the response of CRC cells to gefitinib by inducing EMT, providing new ideas for further research on the resistance mechanism of gefitinib. This suggests that targeting CAFs or exosomes might be a new approach to enhance CRC sensitivity to gefitinib, and our system could be a novel platform for investigating the crosstalk between tumor cells and CAFs and understanding multiple biological changes of the tumor cells in the tumor microenvironment.
Topics: Gefitinib; Humans; Coculture Techniques; Colorectal Neoplasms; Cell Proliferation; Antineoplastic Agents; Cancer-Associated Fibroblasts; Cell Movement; Epithelial-Mesenchymal Transition; Cell Line, Tumor; Lab-On-A-Chip Devices; Microfluidic Analytical Techniques; Exosomes; Tumor Microenvironment; Drug Resistance, Neoplasm
PubMed: 38647729
DOI: 10.1007/s00604-024-06362-9 -
Pest Management Science Mar 2024Myzus persicae, a serious sap-sucking pest of a large variety of host plants in agriculture, is traditionally controlled using chemical insecticides but there...
BACKGROUND
Myzus persicae, a serious sap-sucking pest of a large variety of host plants in agriculture, is traditionally controlled using chemical insecticides but there is interest in using biopesticides as restrictions are increasingly placed on the use of broad-spectrum pesticides.
RESULTS
Here, we show that in Petri dish experiments, high concentrations of the fungal entomopathogen Beauveria bassiana led to rapid mortality of M. persicae, although at a low concentration (1 × 10 conidia mL) there is a hormetic effect in which survival and fecundity are enhanced. Hormetic effects persisted across a generation with reduced development time and increased fecundity in the offspring of M. persicae exposed to B. bassiana. The whole-plant experiment points to a hormetic effect being detected in two out of three tested lines. The impact of these effects might also depend on whether M. persicae was transinfected with the endosymbiont Rickettsiella viridis, which decreases fecundity and survival compared with aphids lacking this endosymbiont. This fecundity cost was ameliorated in the generation following exposure to the entomopathogen.
CONCLUSION
Although B. bassiana is effective in controlling M. persicae especially at higher spore concentrations, utilization of this entomopathogen requires careful consideration of hormetic effects at lower spore concentrations, and further research to optimize its application for sustainable agriculture is recommended. © 2024 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.
PubMed: 38469952
DOI: 10.1002/ps.8075 -
Journal of Infection and Chemotherapy :... Feb 2024In daycare centers, infants come in close contact with each other, and contact, droplet, and mouth-to-mouth infections may occur owing to sharing of toys. Additional...
INTRODUCTION
In daycare centers, infants come in close contact with each other, and contact, droplet, and mouth-to-mouth infections may occur owing to sharing of toys. Additional effective disinfection methods should be considered aside from wiping with disinfectants-including alcohol or sodium hypochlorite solution-for environmental disinfection of daycare centers. We aimed to examine the usefulness of hypochlorous acid water atomization in the effective disinfection of the classroom environment and toys at a nursery school.
METHODS
Environmental cultures of the nursery and toys were prepared to evaluate the species and bacterial load and to assess the contaminated areas. Staphylococcus aureus petri dishes were placed at high-frequency contact sites, and hypochlorous acid water was atomized to achieve a 0.03-ppm atmospheric chlorine concentration. After the atomization, the amount of S. aureus bacteria on the Petri dish and the changes in bacterial count isolated from the environment and toys were evaluated.
RESULTS
Hypochlorous acid water atomization was performed for 5 h to avoid condensation. After a 3-h atomization, ≥99.99% of S. aureus was eliminated on petri dishes; furthermore, a significant disinfection effect was observed on environmental bacteria at least 1 h after atomization. For rubber and textile toys, the significant disinfection effect was observed 1 h after atomization, and for plastic toys, the effect was observed 3 h after atomization.
CONCLUSIONS
Hypochlorous acid water atomization is a useful strategy to disinfect nursery school classrooms.
Topics: Infant; Humans; Hypochlorous Acid; Schools, Nursery; Staphylococcus aureus; Water; Disinfectants; Bacteria; Anti-Bacterial Agents; Ethanol
PubMed: 37758000
DOI: 10.1016/j.jiac.2023.09.024 -
Nanomaterials (Basel, Switzerland) Feb 2024Among nanoparticles (NPs), titanium dioxide is one of the most highly manufactured worldwide and widely used in multiple products for both industrial use and personal...
Among nanoparticles (NPs), titanium dioxide is one of the most highly manufactured worldwide and widely used in multiple products for both industrial use and personal care products. This increases the probability of release into aquatic environments, potentially affecting these ecosystems. The present study aimed to evaluate TiO P25 NP toxicity in zebrafish embryos and eleutheroembryos by evaluating LC, hatching rate, embryo development, and chemical analysis of the TiO concentration accumulated in eleutheroembryo tissues. Zebrafish embryos ~2 h post-fertilization (hpf) were exposed to 75, 100, 150, 200, and 250 mg/L TiO P25 NPs for 48 and 96 h. A total of 40-60 embryos were placed in each Petri dish for the respective treatments. Three replicates were used for each treatment group. Ti concentrations were determined by inductively coupled plasma optical emission spectrometry (ICP-OES), and a conversion factor was used to calculate the TiO concentrations in the tissues. The highest calculated concentrations of TiO in zebrafish larvae were 1.0199 mg/L after 48 h and 1.2679 mg/L after 96 h of exposure. The toxicological results indicated that these NPs did not have a significant effect on the mortality and hatching of zebrafish embryos but did have an effect on their development. LC and LC were determined experimentally, and LC and LC were estimated using four different methods. Up to 11% of embryos also presented physical malformations. These effects can be detrimental to a species and affect ecosystems. Physical malformations were observed in all treatments, indicating teratogenic effects.
PubMed: 38392747
DOI: 10.3390/nano14040373 -
Cytokine Oct 2023Emerging evidence links interleukin-17A (IL-17A) to anxiety and stress. Circulating levels of IL-17A are elevated in patients with anxiety disorders, and pharmacological...
Emerging evidence links interleukin-17A (IL-17A) to anxiety and stress. Circulating levels of IL-17A are elevated in patients with anxiety disorders, and pharmacological blockade of IL-17 signaling or genetic deletion of IL-17 reduces anxiety-like behaviors in mice. Given that IL-17 is one of the most conserved cytokines among animal phyla, we tested the hypothesis that anti-IL-17 treatments reduce defensive responding in planarians, the simplest animal with bilateral symmetry and a CNS with cephalization. The endpoint selected was light avoidance, which is a common phenotype of planarians and rodents and an index of defensive responding that is reduced by anxiolytic compounds in both species. Planarians were placed at the midline of a Petri dish containing water or test solution that was equally split into light and dark halves. Planarians exposed to a selective IL-17A antibody (0.1, 1, 10 pM) over a 5-min interval spent more time in the light than water-exposed planarians. Cyanidin (0.01, 0.1 1, 10 µM), an anti-inflammatory flavonoid and non-selective IL-17A inhibitor, also increased time spent in the light. Motility was not affected by IL-17A antibody or cyanidin at concentrations that reduced light avoidance, although higher concentrations reduced motility (>10 µM). Our results show that IL-17A antagonists reduce defensive responding in planarians and suggest conservation of IL-17A effects on aspects of anxiety-related behaviors.
Topics: Animals; Mice; Antibodies; Anxiety; Interleukin-17; Planarians; Water; Stress, Psychological
PubMed: 37625214
DOI: 10.1016/j.cyto.2023.156345 -
Frontiers in Plant Science 2023Seed germination rate is one of the important indicators in measuring seed quality and seed germination ability, and it is also an important basis for evaluating the...
Seed germination rate is one of the important indicators in measuring seed quality and seed germination ability, and it is also an important basis for evaluating the growth potential and planting effect of seeds. In order to detect seed germination rates more efficiently and achieve automated detection, this study focuses on wild rice as the research subject. A novel method for detecting wild rice germination rates is introduced, leveraging the SGR-YOLO model through deep learning techniques. The SGR-YOLO model incorporates the convolutional block attention module (efficient channel attention (ECA)) in the Backbone, adopts the structure of bi-directional feature pyramid network (BiFPN) in the Neck part, adopts the generalized intersection over union (GIOU) function as the loss function in the Prediction part, and adopts the GIOU function as the loss function by setting the weighting coefficient to accelerate the detection of the seed germination rate. In the Prediction part, the GIOU function is used as the loss function to accelerate the learning of high-confidence targets by setting the weight coefficients to further improve the detection accuracy of seed germination rate. The results showed that the accuracy of the SGR-YOLO model for wild rice seed germination discrimination was 94% for the hydroponic box and 98.2% for the Petri dish. The errors of germination potential, germination index, and average germination days detected by SGR-YOLO using the manual statistics were 0.4%, 2.2, and 0.9 days, respectively, in the hydroponic box and 0.5%, 0.5, and 0.24 days, respectively, in the Petri dish. The above results showed that the SGR-YOLO model can realize the rapid detection of germination rate, germination potential, germination index, and average germination days of wild rice seeds, which can provide a reference for the rapid detection of crop seed germination rate.
PubMed: 38322421
DOI: 10.3389/fpls.2023.1305081