Target Spots Research Articles

MALDI target functionalization with deposited thin films of lanthanum stearate – An efficient tool for in situ enrichment of human globin adducts of chlorinated organic compounds

“Lab-on-a-plate” methodology encapsulates a number of different approaches aimed to shorten extensive sample processing for matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) analysis. Lately, we have proposed two approaches to the enrichment of chlorine-containing adducts of human globin (HHb) in a “lab-on-a-plate” format based on different techniques for the sorbent application to MALDI target spots: (i) droplet-free electrospraying of metal oxide nanoparticles and (ii) deposition of lanthanum stearate thin films (FLa). The latter approach has turned out to be more time- and cost-effective and, therefore, seems to be more promising. In this work, we assessed the quantitative characteristics (sensitivity, selectivity) of the “thin-film” procedure in order to estimate its analytical potential. The FLa-modified MALDI target was used for the on-spot enrichment of HHb adducts from the tryptic digests of the protein incubated with two model mono- and dichlorine-containing alkylating agents. The sensitivity of the technique is in the range of hundreds of fmol. Furthermore, the FLa-based procedure shows high selectivity (maximum molar ratio of modified/unmodified form of LLGNVLVC112VLAHHFGK is 1:500). As a case study, on-plate extraction of HHb adducts, formed due to the interaction of the protein with in vitro generated diclofenac metabolites, was performed on FLa-functionalized spots. This resulted in specific extraction of the HHb adducts with two diclofenac oxidation products from the tryptic digest of modified HHb. The developed approach appears to be effective for in situ enrichment of chlorine-containing adducts of HHb.

MR-PVHEE: A novel magnetic resonance-guided parallel-beam very high-energy electron radiotherapy system

PurposeMR-guided very high-energy electron (VHEE) radiotherapy can combine the imaging advantages of MR with the dosimetry advantages of VHEE radiotherapy, which is of significant clinical value. However, VHEEs are highly susceptible to the Lorentz force generated by the MR magnetic field, and coupling MR and VHEEs is challenging. Here, we present a novel MR-guided parallel-beam VHEE (MR-PVHEE) radiotherapy system and confirm its feasibility and effectiveness. MethodThe proposed MR-PVHEE radiotherapy system includes an ingenious coupling model of MR and VHEEs and an electromagnetic steering parameters commissioning (ESP-COM) method for accurate delivery. First, by generating parallel VHEE (PVHEE) beamlets that point in the same direction as the MR main magnetic field, the coupling model based on triple-stage electromagnetic steering can significantly lessen the twisting and deflecting of the VHEE beamlets' delivery trajectory caused by the MR magnetic field. The dosimetric advantage of VHEEs will be enhanced by delicately utilizing the MR magnetic field to constrain lateral scatter. Then, the ESP-COM method based on Bayesian black-box optimization for the PVHEE beamlets is designed by constructing a cost function using the position and direction information of particles in phase space and optimizing the parameters to deliver the beamlets accurately to the target spots. Finite element electromagnetic modeling and Monte Carlo particle transport simulation are used to validate the MR-PVHEE. ResultsMR-PVHEE can generate the PVHEE beamlets in the same direction as the MR magnetic field and accurately deliver them to the target spots. The average position error is within 0.5 mm, and the average direction error is about 0.5°, which can meet the requirements of clinical treatment accuracy. MR-PVHEE can reduce the VHEE lateral penumbra and increase the dose drop gradient. ConclusionFor the first time, MR and VHEE radiotherapy are successfully coupled in this study, and the innovative MR-PVHEE radiotherapy system can elicit new modalities for MR-guided charged particle radiotherapy and spatially fractionated radiotherapy.

B7-H3 and ICAM-1 are potentially therapeutic targets for thyroid carcinoma

Although most differentiated thyroid carcinoma has a clinically favorable prognosis, some of specific types of thyroid cancer (such as anaplastic thyroid carcinoma and advanced papillary thyroid carcinoma) show fatal outcomes and require novel treatments. Immunotherapy is a promising avenue for the treatment of advanced thyroid carcinoma. B7-H3 (B7 homolog 3 protein) and ICAM-1 (intercellular adhesion molecule 1), as two important immune checkpoints (ICPs), is becoming hopeful target spots for immunotherapy. A growing amount of evidence has suggested that B7-H3 and ICAM-1 are upregulated in papillary thyroid carcinoma. However, their expression level in specific types of thyroid cancer remains largely unclear. In the present study, we explored the expression level of B7-H3 and ICAM-1 in different types of thyroid carcinoma. In the groups of the TCGA cohort, both B7-H3 and ICAM-1 mRNA were highly expressed in thyroid carcinoma. Furthermore, the patients with Stage2, 61-80y, Follicular thyroid papillary carcinoma and N0 had lower B7-H3 and ICAM-1 mRNA expression. In the groups of our cohort, PTCs and ATCs showed frequently moderate to strong expression of B7-H3 and ICAM-1 protein expression. The significant relevance of B7-H3 staining score with ICAM-1 staining score was observed in TCGA database and our cohort, which might open avenues for the combination therapy in advanced thyroid cancer.

Hedyotis diffusa Willd and Astragalus membranaceus May Exert Anti-colon Cancer Effects by Affecting AKTI Expression, as Determined by Network Pharmacology and Molecular Docking.

Network pharmacology is a novel approach that uses bioinformatics to predict multitarget drugs and ingredient-target interactions in various diseases. A thorough search of previously published studies revealed that Hedyotis diffusa Willd (HDW) and Astragalus membranaceus (AM) possess anticancer activity. Colon cancer (CC) is one of the most common malignant tumors of the digestive tract and occurs in the colon. Herein, we explored the effect of two drugs in the treatment of CC. The present study aimed to predict and verify the effect of these two drugs in the treatment of CC. To explore the molecular mechanisms of the "HDW-AM" drug in the treatment of CC, we analyzed its principal efficiency in terms of ingredients, target spots, and pathways via network pharmacology, molecular docking, and experimental verification. The ingredients and their gene target sites were searched and screened through the TCMSP platform according to specific filtering conditions. Subsequently, components corresponding to the gene targets were chosen to construct the drug component-target network. The GEO (Gene Expression Omnibus) dataset was used to collect and screen for gene chips under CC and normal conditions, obtain differential genes, and construct a volcano map. The intersection genes between drug and disease targets were screened, the ".tsv" file was downloaded from the STRING platform and imported into Cytoscape 3.8.0 for visualization, a protein-protein interaction (PPI) network was constructed, the core targets were identified, and the common components with core targets were docked through Autodock Tools-1.5.6. Gene Ontology (GO) analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis were carried out through the Metascape platform to determine the major pathways. The CCK-8 (Cell Counting Kit-8) assay verified the effect of AKT1 on cell proliferation after treatment with quercetin. After the screening, 3658 DEGs (1841 downregulated and 1817 upregulated) were obtained from the GSE75970 gene chip; 21 active components and 220 targets were identified from the drugs. Subsequently, ten core genes (including AKT1, P53, and CASP3) and six major components were screened. GO functional analysis and KEGG analysis revealed that "HDWAM" regulates cell migration and motility through the combination of a transcription regulator complex, membrane rafts, vesicle lumen, and protein kinases via the MAPK, PI3K-Akt, and IL17 signaling pathways. The molecular docking results suggested that quercetin binds to AKT1, TP53, TNF, and CASP3. HDW-AM may exert a therapeutic effect on CC by modulating AKT1, TP53, TNF, and CASP3 and through signaling pathways. A CCK-8 cytotoxicity assay verified that quercetin affects cell viability through AKT1. The current study provides a theoretical basis for an in-depth investigation into the molecular mechanism of the "HDW-AM" drug in CC treatment via network pharmacology, molecular docking, and experimental verification.

Genome sequencing and comparative genome analysis of Rhizoctonia solani AG-3.

Rhizoctonia solani AG-3 is a plant pathogenic fungus that belongs to the group of multinucleate Rhizoctonia. According to its internal transcribed spacer (ITS) cluster analysis and host range, it is divided into TB, PT, and TM subgroups. AG-3 TB mainly causes tobacco target spots, AG-3 PT mainly causes potato black scurf, and AG-3 TM mainly causes tomato leaf blight. In our previous study, we found that all 36 tobacco target spot strains isolated from Yunnan (Southwest China) were classified into AG-3 TB subgroup, while only two of the six tobacco target spot strains isolated from Liaoning (Northeast China) were classified into AG-3 TB subgroup, and the remaining four strains were classified into AG-3 TM subgroup, which had a unique taxonomic status, and there was no previous report on the whole genome information of AG-3 TM subgroup. In this study, the whole genomes of R. solani AG-3 strains 3T-1 (AG-3 TM isolated from Liaoning) and MJ-102 (AG-3 TB isolated from Yunnan) isolated from tobacco target spot in Liaoning and Yunnan were sequenced by IIumina and PacBio sequencing platforms. Comparative genomic analysis was performed with the previously reported AG-3 PT strain Rhs1AP, revealing their differences in genomes and virulence factors. The results indicated that the genome size of 3T-1 was 42,103,597 bp with 11,290 coding genes and 49.74% GC content, and the genome size of MJ-102 was 41,908,281 bp with 10,592 coding genes and 48.91% GC content. Through comparative genomic analysis with the previously reported strain Rhs1AP (AG-3 PT), it was found that the GC content between the genomes was similar, but the strains 3T-1 and MJ-102 contained more repetitive sequences. Similarly, there are similarities between their virulence factors, but there are also some differences. In addition, the results of collinearity analysis showed that 3T-1 and MJ-102 had lower similarity and longer evolutionary distance with Rhs1AP, but the genetic relationship between 3T-1 and MJ-102 was closer. This study can lay a foundation for studying the molecular pathogenesis and virulence factors of R. solani AG-3, and revealing its genomic composition will also help to develop more effective disease control strategies.

199. Fast Detection of Ceftazidime-avibactam-resistant Enterobacterales with VITEK-MSTM Incorporating a Direct-on-target Microdroplet Growth Assay

Abstract Background The direct-on-target microdroplet growth assay (DOT-MGA) was evaluated for its ability to rapidly detect ceftazidime-avibactam resistant Enterobacterales. Methods In this study, 47 non-duplicate CRE isolates were used to determine unequivocally the performance of ceftazidime-avibatam resistance detection of 7 organisms found in clinical isolates. The isolates were incubated for 3 to 6 h in the presence and absence of 8 µg/mL ceftazidime-avibatam in the broth applied as microdroplets directly on to the matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) target spots. Following the incubation period, broth was separated from the microbes by pipette. MALDI-TOF MS was employed to evaluate the growth of isolates in each spot and to identify the microbes simultaneously. The micro-broth dilution method, recommended by EUCAST was used as a reference method. A PCR-based assay was carried out to evaluate the carbapenem resistance mechanisms by screening for the presence of various β-lactamase-encoding genes. Results It was found that a 3 h incubation time was insufficient for MS to identify isolates. At this time-point, the identification of valid results of the growth control was only 25.0%. Therefore, the study of 3 h incubation times was not continued. For ceftazidime-avibactam-resistant Enterobacter strains, 83.7% of growth controls were successfully detected after the 4 h-time-point of incubation when the sensitivity and specificity of drug resistance detection were both 100%. Following incubation for 5 h, the rate of growth controls, sensitivity and specificity were 98.0%, 100% and 100%. When the incubation was extended to 6 h, the rate of growth controls, sensitivity and specificity reached an ideal 100%. Conclusion The VITEK MS-based DOT-MGA is a fast, convenient and accurate in detecting Enterobacterales that is resistant to ceftazidime-avibatam within 5 to 6 h. The assay can also determine the resistance status of an isolate and the potential novel resistant mechanisms involved. Disclosures All Authors: No reported disclosures

Generating captions in English and Marathi language for describing health of cotton plant

<span>Humans’ basic needs include food, shelter, and clothing. Cotton is the foundation of the textile industry. It is also one of the most profitable non-food crops for farmers around the world. Different diseases have a significant impact on cotton yield. Cotton plant leaves are adversely affected by aphids, army worms, bacterial blight, powdery mildew, and target spots. This paper proposes an encoder decoder model for generating captions in English and Marathi language to describe health of cotton plant from aerial images. The cotton disease captions dataset (CDCD) was developed to assess the effectiveness of the proposed approach. Experiments were conducted using various convolutional neural network (CNN) models, such as VGG-19, InceptionResNetV2, and EfficientNetV2L. The quality of generated caption is evaluated on BiLingual evaluation understudy (BLEU) metrics and using subjective criteria. The results obtained for captions generated in English and Marathi language are comparable. The network combination of EfficientNetV2L and long short-term memory (LSTM) has outperformed the other combinations.</span>

Network pharmacology and molecular docking-based investigation on traditional Chinese medicine Astragalus membranaceus in oral ulcer treatment.

To analyze the mechanism of Astragalus membranaceus (AM) in molecular level in the oral ulcer (OU) treatment with reference to network pharmacology. Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform database was used in screening the AM active components and AM action targets; GeneCards database was used to screen OU targets; the common target were screened by Venny online tool; Cytoscape software was applied to construct the target gene regulation map of AM active components; STRING database was used to construct the protein-protein interaction network and the key targets were screened as per degree value; gene ontology enrichment and KEGG pathway enrichment of interactive genes were calculated through David database. There were 17 active ingredients and 429 target spots in Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform database. There are 606 target genes for OU in GeneCards database. There are 67 common targets, including 10 key targets: IL10, IL6, TNF, IL1B, CXCL8, CCL2, TLR4, IL4, ICAM1, and IFNG. It involves 30 gene ontology terms and 20 KEGG signal channels. The molecular docking results showed that quercetin and kaempferol had a good binding activity with IL6, IL1B, TNF, and CCL2. Network pharmacological analysis shows that AM can regulate multiple signal pathways through multiple targets to treat OU.

Tumor-associated macrophages in nanomaterial-based anti-tumor therapy: as target spots or delivery platforms.

Targeting tumor-associated macrophages (TAMs) has emerged as a promising approach in cancer therapy. This article provides a comprehensive review of recent advancements in the field of nanomedicines targeting TAMs. According to the crucial role of TAMs in tumor progression, strategies to inhibit macrophage recruitment, suppress TAM survival, and transform TAM phenotypes are discussed as potential therapeutic avenues. To enhance the targeting capacity of nanomedicines, various approaches such as the use of ligands, immunoglobulins, and short peptides are explored. The utilization of live programmed macrophages, macrophage cell membrane-coated nanoparticles and macrophage-derived extracellular vesicles as drug delivery platforms is also highlighted, offering improved biocompatibility and prolonged circulation time. However, challenges remain in achieving precise targeting and controlled drug release. The heterogeneity of TAMs and the variability of surface markers pose hurdles in achieving specific recognition. Furthermore, the safety and clinical applicability of these nanomedicines requires further investigation. In conclusion, nanomedicines targeting TAMs hold great promise in cancer therapy, offering enhanced specificity and reduced side effects. Addressing the existing limitations and expanding our understanding of TAM biology will pave the way for the successful translation of these nano-therapies into clinical practice.

Cruise report CAGE15-3

The cruise had two main objectives: Deployment of the two CAGE ocean floor observatories (OS1 and OS2) at shallow PKF site and deeper PKF site; CAGE ocean floor observatories were designed and build as collaborative work of CAGE scientists with Kongsberg engineers. Observatories have identical set up except that only one of them have side looking multibeam. The set up is: Seabed Platform/seabed lander/mooring frame (x2) – OS1 has black mooring frame, OS2 has grey mooring frame, CTD (x2), Oxygen sensor (x2), CH4 sensor (x2), CO2 sensor (x2), pH sensor (x1), Fluorometer (x2), ADCP (x2), Current profiler (x1), Multibeam echosounder (x1) – grey lander, OS2, Broadband Hydrophone (x2), Flowmeter (x2) For specific description of each lander, please refer to ‘taking over’ documents (WP4 team leader). Landers and sensors arrived to Tromsø with a track from Hamburg, A. Silyakova was a reference person to receive goods and shipping documents. Time period between 24 and 26 of June was the assemblage of observatories and tests of telemetry/communication/camera on a launcher. Pär Jeanson (PhD student WP4) and Reidar Kaasa (substitute engineer instead of Anoop in WP4) from CAGE were assigned to receive training on observatory assemblage/communication. 26 of June – taking over procedure. Sites for the deployment were discussed during preparatory phase. Water depth at the sites could not exceed 500 meters due to restrictions in relation to the recovery rope, which is only 500 meters long. Photographs from the tow cam used during CAGE 15-2 cruise (chief scientist on the cruise G. Panieri) revealed sites with bacterial mats on the ocean floor. Prior each deployment we did echosounder and multibeam survey to know where flares are highly concentrated. Information from the survey was mapped instantly. Target spots for both observatories were chosen based on all this combined information. Oceanographic survey in the area of shallow PKF methane flares; From the “Testing seep fertilization hypothesis” proposal:‘ During cruise CAGE 14-1 the USGS-GAS system detected elevated methane fluxes near the coast and over the shelf seep site in ca. 90m water depth. Methane fluxes above the 240 and 400m site are much less, although slightly elevated with respect to the open ocean (e.g. Vestnesa). Unexpectedly, high methane concentrations (up to 20nM) are often accompanied by low CO2 concentrations. Initial estimates of the total CO2 budget show that under those conditions seep areas are CO2 sinks. What are the biological, geochemical and hydrographic conditions that made these seeps a CO2 sink? Are the observations from CAGE 14-1 repeatable? And ultimately, what are the processes causing the strong CO2 consumption.’It was decided to test seep fertilization hypothesis during CAGE 15-3 cruise by conducting comprehensive water sampling for biogeochemical environment in the entire water column above the area of methane flares. At the same time, USGS-GAS system was onboard allowing to simultaneously measuring surface water/lower atmosphere gas concentrations. This potentially allows calculating vertical gas flux from one realm to another. Depending on available time, collaborators equipment and human resources, water from 64 CTD stations was samples for following parameters: CH4 concentration; discrete sampling to introduce into CDRS system – 13C CH4, CO2; pH; DIC and 13C DIC; DOC; MOx; FISH; DNA; DMSP; CDOM; Nutrients (nitrate, silicate, phosphate). The cruise may be known as: CAGE15_3

CTNI-10. RANDOMIZED PIVOTAL STUDY OF BLOOD-BRAIN BARRIER (BBB) DISRUPTION USING EXABLATE MODEL 4000 WITH STANDARD OF CARE (SOC) THERAPY IN NON-SMALL CELL LUNG CANCER (NSCLC) BRAIN METASTASES: LIMITLESS TRIAL

Abstract BACKGROUND NSCLC is the most common cause of brain metastases. The Exablate focused ultrasound with circulating microbubble resonators uses the concept of real-time imaging to treat small target spots focusing energy through the skull in noninvasive manner, thereby disrupting the BBB. There is preclinical evidence of synergy of FUS-mediated BBB with immunotherapy beyond improved drug delivery due to BBB disruption (BBBD). METHODS LIMITLESS is a prospective, multi-center, randomized, two-arm, controlled, pivotal clinical trial to evaluate the safety and efficacy of additional targeted BBB disruption using Exablate Model 4000 for the treatment of NSCLC brain metastases in subjects who are undergoing planned pembrolizumab monotherapy compared to Pembrolizumab alone in 2:1 randomization. The inclusion criteria include age ≥ 18 years, KPS ≥ 70, NSCLC (EGFR and ALK negative) with up to 3 brain metastases, one of which meets the RANO-BM criteria for measurable Disease (1 cm x 1 cm) with normal organ function. The primary endpoint is to test superiority of Exablate BBBD targeted to their brain metastases over the standard of care without Exablate BBBD as determined by Objective Response Rate (ORR) based on RANO- BM by 6 months. The secondary endpoints include best objective response rate and time to response for brain metastases by treatment arm. Other end points include progression-free survival (PFS), overall survival (OS), Intracranial and extracranial PFS, quality of life assessments. The Bayesian design statistical analysis will assume superior ORR of 60% in Exablate group compared to 30% in SOC group for a total sample size of 96 subjects (64 subjects in Exablate and 32 in SOC) for 80% power using a two-sided Pearson's Chi-Square test with alpha = 0.05. For the upper-bound estimate ORR of 45% in Exablate group and 30% in SOC group will need 369 subjects (246 subjects in Exablate and 123 in SOC).

CTNI-46. PIVOTAL STUDY TO EVALUATE SAFETY AND EFFICACY OF EXABLATE MODEL 4000 USING MICROBUBBLE RESONATORS TO TEMPORARILY MEDIATE BLOOD-BRAIN BARRIER DISRUPTION FOR LIQUID BIOPSY IN GLIOBLASTOMA (LIBERATE)

Abstract BACKGROUND Glioblastoma (GBM) has dismal outcome of 14-16 months. One of the challenges of drug development in GBM is lack of requisite circulating-free (cfDNA) in blood samples to develop biomarker driven targeted therapy trials. The Exablate focused ultrasound with circulating microbubble resonators uses the concept of real-time imaging to treat small target spots focusing energy in a non-invasive manner, thereby disrupting the Blood-Brain Barrier (BBBD) with potential to increase cfDNA in blood. METHODS LIBERATE is a prospective, multi-center, self-controlled pivotal ongoing clinical trial in subjects with suspected GBM evaluating the safety and effectiveness of Exablate Model 4000 to disrupt BBB to achieve a greater proportion of cfDNA in samples taken post-BBB disruption as compared to the sample taken before BBBD. Fifty eligible subjects with suspected GBM who are scheduled to undergo brain tumor resection or biopsy at 10 centers in the U.S. will participate in this study. The primary endpoint is defined for each subject as ratio between his/her cfDNA amount found in the blood sample 1-hour post-BBBD procedure compared to the cfDNA amount found in the blood sample collected pre-BBBD procedure. This study proposes to demonstrate that on average there is at least 2-fold increase in cfDNA (measured by central laboratory) at 1 hour post BBBD. Confirmatory secondary analysis will evaluate correlation between patterns obtained in panel of biomarkers evaluated in the resected tumor tissue and/or biopsy sample and blood sample collected 1-hour post-BBBD. Exploratory endpoints will include: (1) sensitivity of detection of known somatic mutations in the circulating blood samples (circulating tumor DNA) before and after BBBD, (2) time of greatest yield of cfDNA in the samples collected post BBBD blood samples (at 30 min, 1-hour, 2-hour and 3-hour) to determine time of greatest yield, (3) correlation of biomarkers between the imaging and the post-BBBD biomarker samples. (NCT05383872)

Focal Microscopy Observe the Mechanism of Gegen Qinlian Decoction Treating Type 2 Diabetic Nephropathy in Nude Mice.

The incidence of diabetes is increasing year by year; among them, the rising trend of T2D is particularly obvious, and because it has many complications and poor prognosis, it has become one of the diseases that seriously endanger human health. Gegen Qinlian Decoction (GQD) has achieved good results in the treatment of type 2 diabetes (T2D). It can not only improve the anti-insulin effect of nude mice with type 2 diabetic, but also has the characteristics of small side effects and remarkable curative effect; it can improve the function of tissues and organs by multiple target spots and multiple ways. The method was to raise 30 nude mice with high fat and high sugar and inject small doses of streptozotocin (STZ). The treated nude mice were randomly divided into rosiglitazone group, Gegen Qinlian group, and model group, normally rearing 10 nude mice as a control group. The model group and the blank group were treated with 0.9% sodium chloride solution at 10 mL/kg, and the rosiglitazone group was intragastric administration with 3 mg/kg rosiglitazone solution. The GQD group was treated with 18.2 g/kg GQD, once a day, and lasting for 4 weeks. Use blood glucose meter to detect the FBG content in nude mice, use the radioimmunoassay to detect the amount of FINS, and calculate HOMA-IR. Use immunoenzyme linked assay method to detect the level of serum inflammatory factors. The experimental results showed that compared with the model group, the FBG of the rosiglitazone group mice was significantly decreased; the levels of the rosiglitazone group and the Gegen Qinlian group were significantly lower. Therefore, it is concluded that GQD can treat T2D in nude mice.

Identification of the types of disease for tomato plants using a modified gray wolf optimization optimized MobileNetV2 convolutional neural network architecture driven computer vision framework

SUMMARYTomato is a widely consumed fruit across the world due to its high nutritional values. Leaf diseases in tomato are very common which incurs huge damages but early detection of leaf diseases can help in avoiding that. The existing practices for detecting different diseases by the human experts are costly, time consuming and subjective in nature. Computer vision plays important role toward early detection of tomato leaf detection. However, implementation of computationally less expensive model and improvement of detection performance is still open. This article reports a computer vision based system to classify seven different categories of diseases, namely, bacterial spot, early blight, late blight, leaf mold, septoria leaf spot, spider mites, and target spots using optimized MobileNetV2 architecture. A modified gray wolf optimization approach has been adopted for optimization of MobileNetV2 hyperparameters for improved performance. The model has been validated using standard internal and external validation methods and found to provide the classification accuracy in the tune of 98%. The results reflect the promising potential of the presented framework for early detection of tomato leaf diseases which can help to avoid substantial agricultural loss.

Speed and safety of mass spectrometry for identification of Burkholderia pseudomallei directly from spiked blood cultures

Burkholderia pseudomallei is a bipolar Gram-negative bacillus and the causative agent of melioidosis; an infectious disease which commonly presents with bacteraemia. Data regarding direct from blood culture identification of B. pseudomallei using the Vitek mass spectrometer (MS) are limited. The authors aim to assess the safety and sensitivity of the Vitek MS for identification of B. pseudomallei from spiked positive blood culture samples. Safety was assessed by determining the ability of the standard MS α-cyano-4-hydroxycinnamic acid (CHCA) matrix solution to inactivate B. pseudomallei. Organism identification using the manufacturer's blood culture extraction method was compared to an in-house technique. Additionally, identification following abbreviated agar incubation of blood culture broth was performed. All 70 MS target spots were inactivated by the matrix solution. The manufacturer's blood culture extraction method identified 0/26 (0%) B. pseudomallei samples. An in-house method using the spun deposit from blood culture broth samples identified 38/38 (100%) B. pseudomallei samples. MS analysis of a blood culture broth drop on Chocolate agar following a 6 h incubation identified 30/32 (94%) samples. Decreased time to diagnosis of melioidosis bacteraemia is likely to improve patient outcomes. This study adds to the literature with regards to the utility of MALDI-TOF MS identification of B. pseudomallei both directly from positive blood culture broth and a subsequent 6 h plate incubation. The use of a standard matrix solution inactivates the organism, and use of the spun deposit from a positive blood culture broth is most effective for early identification of B. pseudomallei.

Mechanisms of Vitamin C Regulating Immune and Inflammation Associated with Neonatal Hypoxic-Ischemic Encephalopathy Based on Network Pharmacology and Molecular Simulation Technology.

Background There are still controversies about the curative effect of vitamin C in treating HIE, and its mechanism of action is not entirely clear. This study is designed to explore the potential molecular mechanism of vitamin C in treating neonatal hypoxic ischemic encephalopathy (HIE). Methods The effect targets of vitamin C and the pathogenic targets of neonatal HIE were obtained via retrieval of public databases to screen out the molecular targets of vitamin C acting on neonatal HIE. Gene Ontology (GO) functional annotations and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis were performed on the main targets. Vitamin C and the optimum target structural components are subjected to molecular docking and molecular dynamics simulation analysis via computer software so as to verify their binding activity and stability. Result Based on 16 overlapping targets of vitamin C and HIE, seven main targets were identified in this study. According to GO and KEGG analysis, molecular functions (top 25 items) and signal pathways (21 items) related to inflammatory reaction, immune response, and cell transcriptional control may be potential pathways for vitamin C to treat neonatal HIE. Molecular docking and molecular dynamics simulation were adopted to definitively determine the 4 optimum core target spots. Conclusion The efficacy of vitamin C on HIE is involved in the immunoregulation and inflammation-related functional processes and signal pathways. These molecular mechanisms, including core targets, will contribute to the clinical practice of neonatal HIE in the future.

Phenotypic and molecular characterization of the resistance to azoxystrobin and pyraclostrobin inFusarium graminearumpopulations from Brazil

AbstractWheat farmers rely on fungicides to protect fields against several foliar and flowering diseases, including Fusarium head blight (FHB). A range of active ingredients is used in isolation or in dual premixes that include a dimethylation inhibitor (DMI) or a quinone outside inhibitor (QoI) fungicide. Comprehensive information about fungicide resistance inF.graminearumis available for DMIs, while for QoIs the data are scarce. We characterized 225 strains obtained from two states in southern Brazil, Rio Grande do Sul (RS) and Paraná (PR), in relation to their response to two QoIs. The median EC50(effective concentration leading to 50% inhibition of conidial germination) value for azoxystrobin (n = 25 isolates) was 2.20 μg/ml in the PR population and 4.04 μg/ml in the RS population. For pyraclostrobin (n = 50), the median EC50was 0.28 μg/ml in the PR population and 0.24 μg/ml in the RS population. Evidence of cross‐resistance could not be detected. Screening using a discriminatory dose (DD) for azoxystrobin in a larger number of isolates from PR (n = 75) and RS (n = 100) states allowed the detection of 50% and 28% sensitive strains, respectively. Using the DD for pyraclostrobin, 33% and 18.8% were classified as less sensitive in the PR and RS isolates, respectively. In RS, the frequency of less‐sensitive isolates increased over time (2007–2011). No point mutation at any of the target spots (F129L, G137R, G143A) was detected. Our results represent an important step towards the establishment of a sensitivity profile for two of the most commonly used QoIs in commercial premixes targeting FHB control.

ВЫЯВЛЕНИЕ ГЕНОВ ФЕРМЕНТОВ У БАКТЕРИЙ ВИДА BACILLUS SUBTILIS МЕТОДОМ REAL-TIME PCR

Soil is a vital and valuable natural resource that sustains life on earth. Appropriate soil functioning depends on the balance of its structure and composition, as well as physical, chemical and biological properties. Often, this balance is disturbed under the influence of various abiotic, biotic and anthropogenic factors. Therefore, soil restoration is of great importance in order to prevent possible adverse effects on living systems and to preserve the environment for future generations. Various studies confirmed the effectiveness of introduction of rhizobacteria to improve soil fertility, increase growth and productivity of agricultural crops. Bacillus subtilis is one of the most common rhizobacteria used in agriculture. Many B. subtilis strains are capable of fixing atmospheric nitrogen, solubilizing phosphorus and potassium, thereby contributing to an increase in the amount of macroelements necessary for plant nutrition in soil. The aim of this work was to search for genes responsible for synthesis of phytase, nitrogenase, and alkaline phosphatase enzymes in strains of bacteria of Bacillus subtilis species by real-time PCR. To determine the presence of genes encoding the synthesis of the desired enzymes in Bacillus subtilis, an in-silico analysis of the annotated genomes of this bacterial species presented in the NCBI information database was carried out. Then the selection of primers for screening the target spots was made. According to the results of the study, 10 out of 19 isolated Bacillus subtilis strains contained all three required DNA regions responsible for synthesis of phytase, nitrogenase, and alkaline phosphatase enzymes.

A classification technique of civil objects by artificial neural networks using estimation of entropy on synthetic aperture radar images

Abstract. The article discusses the method for the classification of non-moving civil objects for information received from unmanned aerial vehicles (UAVs) by synthetic aperture radar (SAR). A theoretical approach to analysis of civil objects can be estimated by cross-entropy using a naive Bayesian classifier. The entropy of target spots on SAR images revaluates depending on the altitude and aspect angle of a UAV. The paper shows that classification of the target for three classes able to predict with fair accuracy P=0,964 based on an artificial neural network. The study of results reveals an advantage compared with other radar recognition methods for a criterion of the constant false-alarm rate (PCFAR<0.01). The reliability was confirmed by checking the initial data using principal component analysis.

Association Between the Degree of Inclusion of Components Identified on Fluorescein or Indocyanine Green Angiography in Target Spots and Relapse of Exudate in Eyes with Polypoidal Choroidal Vasculopathy and Typical Age-Related Macular Degeneration After Photodynamic Therapy.

PurposeTo investigate the association between the inclusion of components identified on images in target spots of photodynamic therapy (PDT) and exudate relapse in eyes with age-related macular degeneration (AMD).MethodsForty-one eyes (39 patients) with polypoidal choroidal vasculopathy (PCV) and 32 eyes (31 patients) with typical AMD (tAMD) who underwent PDT were retrospectively investigated. Each component identified on fluorescein (FA) or indocyanine angiography (IA), optical coherence tomography (OCT), or color photography was graded as not depicted, covered with a margin ≥500 µm or <500 µm, and protruding. Associations between these grades and the dry rate (proportion of subjects with continuous absence of exudate over following 12-month period) and the relapse index (2 × number of injections administered + accumulation of exudate for 12 months post-PDT) were investigated.ResultsIn PCV, the association between worse coverage and decreasing dry rates for feeder vessels and polyps approached statistical significance. With coverage margins ≥500 µm, dry rate tended to be greater than with coverage margins <500 µm for feeder vessels, classic lesions, and occult lesions on FA. In the tAMD group, coverage with margins ≥500 µm tended to yield a higher dry rate than coverage with margins <500 µm for CNV on IA. Coverage with margins ≥500 µm for occult and classic lesions on FA yielded no dry subjects, and all subjects with classic lesions or staining had recurrence (P = 0.009 and 0.050). Worse coverage and worse dry rate in PCV and worse relapse index in tAMD were related to pigment epithelial detachment on OCT (P = 0.040 and 0.006).ConclusionPolyps in PCV and pigment epithelial detachment (PED) in tAMD were verified as appropriate targets, corresponding to the existing guidelines, and feeder vessels, classic lesion, occult lesion, and PED in PCV and CNV on IA in tAMD were suggested as further targets. OCT was superior to FA for evaluating PED.