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Computational and Structural... 2023Performing lifespan assays with () nematodes manually is a time consuming and laborious task. Therefore, automation is necessary to increase productivity. In this...
Performing lifespan assays with () nematodes manually is a time consuming and laborious task. Therefore, automation is necessary to increase productivity. In this paper, we propose a method to automate the counting of live using deep learning. The survival curves of the experiment are obtained using a sequence formed by an image taken on each day of the assay. Solving this problem would require a very large labeled dataset; thus, to facilitate its generation, we propose a simplified image-based strategy. This simplification consists of transforming the real images of the nematodes in the Petri dish to a synthetic image, in which circular blobs are drawn on a constant background to mark the position of the . To apply this simplification method, it is divided into two steps. First, a Faster R-CNN network detects the , allowing its transformation into a synthetic image. Second, using the simplified image sequence as input, a regression neural network is in charge of predicting the count of live nematodes on each day of the experiment. In this way, the counting network was trained using a simple simulator, avoiding labeling a very large real dataset or developing a realistic simulator. Results showed that the differences between the curves obtained by the proposed method and the manual curves are not statistically significant for either short-lived N2 (p-value log rank test 0.45) or long-lived (p-value log rank test 0.83) strains.
PubMed: 37867965
DOI: 10.1016/j.csbj.2023.10.007 -
Cureus Sep 2023Aim The present study aims to compare the antibacterial efficacy of a novel automated endodontic irrigation device with that of a manual irrigation technique. Materials...
Aim The present study aims to compare the antibacterial efficacy of a novel automated endodontic irrigation device with that of a manual irrigation technique. Materials and methods The present study considered 45 extracted single-rooted teeth. After sectioning the teeth, the specimens were inoculated with () bacteria for three weeks. Instrumentation was done using ProTaper Gold rotary files to size 50 and 5% taper. Based on the irrigation protocol, the experimental samples were divided into Group I: control (normal saline irrigation) (n = 15); Group II: conventional needle irrigation (n = 15); and Group III: automated irrigation (n = 15). The irrigation in Group III was done using the automated irrigation device. After the experimental irrigation, the pre-weighted dentinal shaving was collected in Eppendorf tubes, diluted 10 times, and incubated in the Petri dish with tryptone soy agar (TSA) for 48 hours. Finally, the colony-forming unit (CFU) counts were assessed. IBM SPSS Statistics for Windows version 23.0 (Armonk, NY, USA, IBM Corp.) was used for data analysis. Intergroup comparisons were made using the non-parametric Kruskal-Wallis test. Results The mean CFU count (CFU/ml) for the groups was: Group 1 (normal saline irrigation): 3.67x10; Group 2 (manual irrigation): 2.69× 10; Group 3 (automated irrigation): 1.57× 10. A statistically significant reduction in CFU count was seen among the groups assessed (p<0.01). The automated irrigation group exhibited the most substantial reduction in CFU count. Group 2 showed a significant difference compared to Group 1 (p<0.05). Conclusions The novel automated endodontic irrigation device was superior to manual irrigation in reducing CFU counts. This indicates that the automated irrigation device has the potential to enhance bacterial elimination efficacy during endodontic treatment.
PubMed: 37842411
DOI: 10.7759/cureus.45200 -
Sensors (Basel, Switzerland) Oct 2023Colony-Forming Unit (CFU) counting is a complex problem without a universal solution in biomedical and food safety domains. A multitude of sophisticated heuristics and...
Colony-Forming Unit (CFU) counting is a complex problem without a universal solution in biomedical and food safety domains. A multitude of sophisticated heuristics and segmentation-driven approaches have been proposed by researchers. However, U-Net remains the most frequently cited and used deep learning method in these domains. The latter approach provides a segmentation output map and requires an additional counting procedure to calculate unique segmented regions and detect microbial colonies. However, due to pixel-based targets, it tends to generate irrelevant artifacts or errant pixels, leading to inaccurate and mixed post-processing results. In response to these challenges, this paper proposes a novel hybrid counting approach, incorporating a multi-loss U-Net reformulation and a post-processing Petri dish localization algorithm. Firstly, a unique innovation lies in the multi-loss U-Net reformulation. An additional loss term is introduced in the bottleneck U-Net layer, focusing on the delivery of an auxiliary signal that indicates where to look for distinct CFUs. Secondly, the novel localization algorithm automatically incorporates an agar plate and its bezel into the CFU counting techniques. Finally, the proposition is further enhanced by the integration of a fully automated solution, which comprises a specially designed uniform Petri dish illumination system and a counting web application. The latter application directly receives images from the camera, processes them, and sends the segmentation results to the user. This feature provides an opportunity to correct the CFU counts, offering a feedback loop that contributes to the continued development of the deep learning model. Through extensive experimentation, the authors of this paper have found that all probed multi-loss U-Net architectures incorporated into the proposed hybrid approach consistently outperformed their single-loss counterparts, as well as other comparable models such as self-normalized density maps and YOLOv6, by at least 1% to 3% in mean absolute and symmetric mean absolute percentage errors. Further significant improvements were also reported through the means of the novel localization algorithm. This reaffirms the effectiveness of the proposed hybrid solution in addressing contemporary challenges of precise in vitro CFU counting.
PubMed: 37837169
DOI: 10.3390/s23198337 -
PloS One 2023The 'Astrocyte Network' and the understanding of its communication has been posed as a new grand challenge to be investigated by contemporary science. However,...
The 'Astrocyte Network' and the understanding of its communication has been posed as a new grand challenge to be investigated by contemporary science. However, communication studies in astrocyte networks have investigated traditional petri-dish in vitro culture models where cells are closely packed and can deviate from the stellate form observed in the brain. Using novel cell patterning approaches, highly organised, regular grid networks of astrocytes on chip, to single-cell fidelity are constructed, permitting a stellate-like in vitro network model to be realised. By stimulating the central cell with a single UV nanosecond laser pulse, the initiation/propagation pathways of stellate-like networks are re-explored. The authors investigate the mechanisms of intercellular Ca2+ communication and discover that stellate-like networks of adult human astrocytes in vitro actually exploit extracellular ATP release as their dominant propagation pathway to cells in the network locally; being observed even down to the nearest neighbour and next nearest neighbouring cells-contrary to the reported gap junction. This discovery has significant ramifications to many neurological conditions such as epilepsy, stroke and aggressive astrocytomas where gap junctions can be targeted. In cases where such gap junction targeting has failed, this new finding suggests that these conditions should be re-visited and the ATP transmission pathway targeted instead.
Topics: Humans; Adult; Astrocytes; Calcium; Calcium Signaling; Gap Junctions; Cell Communication; Communication; Adenosine Triphosphate; Cells, Cultured
PubMed: 37788259
DOI: 10.1371/journal.pone.0289350 -
Polymers Sep 2023The diverse applications of metal oxide-biopolymer matrix as a nanocomposite heterogenous catalyst have caused many researches to scrutinize the potential of this...
The diverse applications of metal oxide-biopolymer matrix as a nanocomposite heterogenous catalyst have caused many researches to scrutinize the potential of this framework. In this study, a novel hybrid barium oxide-chitosan nanocomposite was synthesized through a facile and cost-effective co-precipitation method by doping barium oxide nanoparticles within the chitosan matrix at a weight percentage of 20 wt.% BaO-chitosan. A thin film of the novel hybrid material was produced by casting the nanocomposite solution in a petri dish. Several instrumental methods, including Fourier-transform infrared (FTIR), scanning electron microscope (SEM), energy dispersive spectroscopy (EDS), and X-ray diffraction (XRD), were used to analyze and characterize the structure of the BaO-CS nanocomposite. The chemical interaction with barium oxide molecules resulted in a noticeable displacement of the most significant chitosan-specific peaks in the FTIR spectra. When the surface morphology of SEM graphs was analyzed, a dramatic morphological change in the chitosan surface was also discovered; this morphological change can be attributed to the surface adsorption of BaO molecules. Additionally, the patterns of the XRD demonstrated that the crystallinity of the material, chitosan, appears to be enhanced upon interaction with barium oxide molecules with the active sites, OH and NH groups, along the chitosan backbone. The prepared BaO-CS nanocomposite can be used successfully as an effective heterogenous recyclable catalyst for the reaction of ,-(alkane-diyl)bis(2-chloroacetamide) with 2-(arylidinehydrazine)-1-carbothioamide as a novel synthetic approach to prepare 2-hydrazonothiazol-4(5)-ones. This new method provides a number of benefits, including quick and permissive reaction conditions, better reaction yields, and sustainable catalysts for multiple uses.
PubMed: 37765671
DOI: 10.3390/polym15183817 -
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 -
Plant Disease Sep 2023Pecan (Carya illinoinensis) is an important economic forest crops widely cultivated in China. From June to September in both 2021 and 2022, severe leaf disease...
Pecan (Carya illinoinensis) is an important economic forest crops widely cultivated in China. From June to September in both 2021 and 2022, severe leaf disease resembling anthracnose was observed in 6.6-ha pecan orchard in Jintan (31°42'23.84″ N, 119°21'22.90″ E), Jiangsu Province. The disease severity was about 15 to 25% with 5 to 12% incidence on 100 surveyed trees of the orchard in 2022. Symptoms initially appeared as small gray-bark sunken lesions, which gradually developed to big sunken lesions with brown edges and irregular-shaped. Small fragments (4 × 4 mm) from the necrotic borders of infected leaves were surfaced sterilized, plated on potato dextrose agar (PDA) and then incubated in darkness at 25°C for 3 days. Pure cultures were obtained by monosporic isolation. Twenty-one isolates with similar characteristics were obtained from the infected leaves (isolation frequency about 90%). The upper side of colonies on the PDA plates was milky, and the reverse side was pale yellow at the center and pale white at the margin. After 10 days of growth on the PDA medium, these isolates produced spores separately. . Through electron microscopic observation, conidia were smooth walled, hyaline, aseptate, guttulate, cylindrical with rounded ends with 15 to 20.5 × 5.3 to 6.7 μm (mean 18.5 × 5.8 μm, n = 50) in size. These morphological characteristics were similar to those of the species of Colletotrichumspp (Weir et al. 2012, Fu et al. 2019). To further identify the isolates, the regions of internal transcribed spacer (ITS), actin (ACT), calmodulin (CAL), chitin synthase (CHSI), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), and beta-tubulin 2 (TUB2) loci of the three representative isolates (JSJT-1, JSJT-2, and JSJT-3) were amplified and sequenced with the primer pairs ITS-1F/ITS-4, ACT-512F/ACT-783R, CL1/CL2A, CHS-79F/CHS-345R, GDF/GDR and T1/T2 primers, respectively (Weir et al. 2012). Sequences of them were deposited in GenBank under nos. OR214960 to OR214962 (ITS), OR228543 to OR228545 (ACT),OR228546 to OR228548 (CAL), OR228549 to OR228551 (CHSI), OR228552 to OR228554 (GAPDH), and OR228555 to OR228557 (TUB2). Multilocus phylogenetic analysis revealed that the three isolates and C. aenigma were clustered in the same clade. Based on the results of morphological and molecular analysis, these isolates were identified as C. aenigma. The pathogenicity of three isolates was tested on leaves of pecan seedlings. Suspensions of conidia were obtained by scraping the surface of a 10-day-old sporulated petri dish PDA cultures into sterile water. Suspensions were adjusted to a density of 2 × 106 conidia/ml with a hemocytometer.The conidial suspension of each isolate was sprayed evenly on the surface of leaves from three healthy pecan seedlings. Sterilized distilled water was used for negative controls. The pathogenicity experiment was repeated three times. Finally, all inoculated plants were kept in a light-incubator at 28°C under 100% relative humidity and 12 h photoperiod. Two weeks after inoculation, the inoculated plants developed symptoms similar to those of the original diseased plants, while controls remained asymptomatic. C. aenigma were re-isolated from from inoculated leaves. C. aenigma has been reported as the causal agent of anthracnose on several economically important plants, such as grape ( Kim et al. 2021), tree peonies (Wang et al.2023), chili (Diao et al. 2017), and pear (Fu et al. 2019), but this is the first report of C. aenigma causing anthracnose on pecan in China. Identification of C. aenigma as a pathogen of pecan is important for implementing control management strategies for pecan disease. References: Diao, Y. Z., et al. 2017. Persoonia. 38:20. Fu, M., et al. 2019. Persoonia. 42:1. Kim, J. S., et al. 2021. Plant Dis. 105:2729. Weir, B. S., et al. 2012. Stud. Mycol.. 73:115. Wang, Y. L., et al. 2023. Plant Dis. 107(4):1242. The author(s) declare no conflict of interest. Keywords: Colletotrichum aenigma, Anthracnose, Carya illinoinensis, Pathogenicity.
PubMed: 37721520
DOI: 10.1094/PDIS-07-23-1378-PDN -
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 -
Cureus Jul 2023This investigation was carried out to evaluate the color stability of a nanocomposite restorative material and the erosive potential of carbonated soft drinks...
AIM
This investigation was carried out to evaluate the color stability of a nanocomposite restorative material and the erosive potential of carbonated soft drinks (Coca-Cola; The Coca-Cola Company, Atlanta, Georgia, United States) and packaged orange juice (Real Fruit Power Orange; Dabur Ltd, Ghaziabad, Uttar Pradesh, India) on its surface micromorphology.
MATERIALS AND METHODS
Sixty discs (2mm thick and 10mm diameter) of nanocomposite material (Herculite Précis; KaVo Kerr, Brea, California, United States) were prepared using a silicon cylindrical mold. Initially, all the specimens were stored in artificial saliva in five Petri dishes; 12 specimens in each dish. In the Petri dishes, the specimens were immersed in the respective beverages once or twice a day. Before and after each immersion, the specimens were stored in artificial saliva at room temperature. Artificial saliva was changed each day, i.e., every 24 hours. The whole procedure was carried out for three months and then evaluated for color stability using a spectrophotometer and surface micromorphology using a scanning electron microscope. Now, the exposure of specimens to aerated drinks (Coca-Cola) and packaged orange juice (Real Orange) was put to a halt, and specimens were kept continuously in artificial saliva. This procedure was carried out for one month and then evaluated for color stability. The information was analyzed using PASW Statistics for Windows, Version 18.0 (Released 2009; SPSS Inc., Chicago, United States). A p-value of 0.05 was considered significant.
RESULTS
The p-value after three months, which is < 0.001 (p<0.05) indicates that the mean color difference values for groups I, II, III, IV, and V show a statistically significant change between the five groups, and similarly, the p-value after one month, which is < 0.001 (p<0.05) indicates that the mean color difference values for groups I, II, III, IV, and V show a statistically significant change between the five groups. Specimens immersed in the carbonated drink twice a day showed clinically more color change than packaged orange juice and artificial saliva on the composite restorative material. Coca-Cola, an aerated drink, was shown to have a higher erosive potential on the composite restorative material than Real Fruit Power Orange and fake saliva.
CONCLUSION
The findings are consistent with the hypothesis that repeated exposure to carbonated beverages (such as Coca-Cola and packaged juice) degrades the surface qualities of dental restorations.
PubMed: 37583717
DOI: 10.7759/cureus.41905