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S-Adenosyl-l-methionine (SAM)-mediated methylation of biomolecules controls their function and regulates numerous vital intracellular processes. Analogs of SAM with a reporter group in place of the S-methyl group are widely used to study these processes. However, many of these analogs are chemically unstable that largely limits their practical application. We have developed a new compound, SAM-PH, which contains an H-phosphinic group (-P(O)(H)OH) instead of the SAM carboxylic group. SAM-PH is significantly more stable than SAM, retains functional activity in catechol-O-methyltransferase and methyltransferase WBSCR27 reactions. The last is associated with Williams–Beuren syndrome. Rac-SAM-PH was synthesized chemically, while (R,S)-SAM-PH and its analogs were prepared enzymatically either from H-phosphinic analogs of methionine (Met-PH) or H-phosphinic analog of S-adenosyl-l-homocysteine (SAH-PH) using methionine adenosyltransferase 2A or halide methyltransferases, respectively. SAH-PH undergoes glycoside bond cleavage in the presence of methylthioadenosine nucleosidase like natural SAH. Thus, SAM-PH and its analogs are promising new tools for investigating methyltransferases and incorporating reporter groups into their substrates.

In Indonesia, there is a rich diversity of cultures, one of which is traditional houses. Traditional houses essentially have the potential to represent the way of life, culture, and local economy. Traditional houses in Indonesia, particularly in the Malaka region, are important cultural symbols that can be regarded as cultural icons in Malaka and Indonesia. They provide a historical perspective, heritage, and reflect the progress of society in a civilization. The Convolutional Neural Network (CNN) method is used in this research. In this study, the CNN algorithm is applied to classify traditional house objects. These traditional house objects are divided into two categories: Kolibein Traditional House and Laleik Traditional House. The objective of this research is to classify traditional houses in Malaka, namely Kolibein Traditional House and Laleik Traditional House, and also to determine the accuracy level of CNN classification results. The previously created model is tested using test data to assess its accuracy. The testing is conducted on 20 data points, with 10 data points in each respective class. The testing results show that the classification of Kolibein and Laleik traditional houses is error- free or very accurate. Based on the model developed for classifying Kolibein and Laleik traditional houses using the Convolutional Neural Network method, it is evident that this method is capable of producing accurate results. The obtained results indicate that the accuracy, based on the classification report using images of Kolibein and Laleik traditional houses, reaches 100%. Therefore, it can be concluded that the constructed CNN model has a high level of accuracy.

Aim. To study overall drug resistance genes (resistome) in the human gut microbiome and the changes in these genes during COVID-19 in-hospital therapy. Materials and methods. A single-center retrospective cohort study was conducted. Only cases with laboratory-confirmed SARS-CoV-2 RNA using polymerase chain reaction in oro-/nasopharyngeal swab samples were subject to analysis. The patients with a documented history of or current comorbidities of the hepatobiliary system, malignant neoplasms of any localization, systemic and autoimmune diseases, as well as pregnant women were excluded. Feces were collected from all study subjects for subsequent metagenomic sequencing. The final cohort was divided into two groups depending on the disease severity: mild (group 1) and severe (group 2). Within group 2, five subgroups were formed, depending on the use of antibacterial drugs (ABD): group 2A (receiving ABD), group 2AC (receiving ABD before hospitalization), group 2AD (receiving ABD during hospitalization), group 2AE (receiving ABD during and before hospitalization), group 2B (not receiving ABD). Results. The median number of antibiotic resistance (ABR) genes (cumulative at all time points) was significantly higher in the group of patients treated with ABD: 81.0 (95% CI 73.8–84.5) vs. 51.0 (95% CI 31.1–68.4). In the group of patients treated with ABD (2A), the average number of multidrug resistance genes (efflux systems) was significantly higher than in controls (group 2B): 47.0 (95% CI 46.0–51.2) vs. 21.5 (95% CI 7.0–43.9). Patients with severe coronavirus infection tended to have a higher median number of ABR genes but without statistical significance. Patients in the severe COVID-19 group who did not receive ABD before and during hospitalization also had more resistance genes than the patients in the comparison group. Conclusion. This study demonstrated that fewer ABR genes were identified in the group with a milder disease than in the group with a more severe disease associated with more ABR genes, with the following five being the most common: SULI, MSRC, ACRE, EFMA, SAT.

Abstract Neurological side effects arising from chemotherapy, such as severe pain and cognitive impairment, are a major concern for cancer patients. These major side effects can lead to reduction or termination of chemotherapy medication in patients, negatively impacting their prognoses. With cancer survival rates improving dramatically, addressing side effects of cancer treatment has become pressing. Here, we use iPSC-derived human neurons to investigate the molecular mechanisms that lead to neurotoxicity induced by vincristine, a common chemotherapeutic used to treat solid tumors. Our results uncover a novel mechanism by which vincristine causes a local increase in mitochondrial proteins that produce reactive oxygen species (ROS) in the axon. Vincristine triggers a cascade of axon pathology, causing mitochondrial dysfunction that leads to elevated axonal ROS levels and SARM1-dependent axon degeneration. Importantly, we show that the neurotoxic effect of increased axonal ROS can be mitigated by the small molecule mitochondrial division inhibitor 1 (mdivi-1) and antioxidants glutathione and mitoquinone, identifying a novel therapeutic avenue to treat the neurological effects of chemotherapy.

The article describes the existential category of the Other and the Otherness in V. Sorokin's screenplay for the film called 4, which was directed by I. Hrzhanovsky in 2004. The Otherness helped V. Sorokin to realize his artistic idea of post-Soviet Russia. According to Jean Paul Sartre, the Other conveys a feeling of the last days: without a single scene of physical violence, the reader develops a feeling of nausea. The essence of the Other is based on the principles of rejection, fear, and inevitability; the Other is the only verifier of one’s existence. In 4, the frightening atmosphere of the Other World is created by such artistic means as coding, dying, masking, and deconstruction of symmetry. They highlight the artistic intent to portray the chaotic nature of being and the absence of time, where human fate is meaningless, and people are reduced to functions in the game of life. The Otherness of the chronotype gives the script and the film a new code: the loss of the holy number four desacralizes existence, thus condemning the world and people in it to a life of hopeless dismay.

The mechanisms of action of pyrimidine nucleoside derivatives on model lipid membranes of various compositions were studied. A systematic analysis of the tested agents’ effects on the membrane physicochemical properties was performed. Differential scanning microcalorimetry data indicated that the ability of nucleoside derivatives to disorder membrane lipids depended on the types of nucleoside bases and membrane-forming lipids. The 5′-norcarbocyclic uracil derivatives were found to be ineffective, while N4-alkylcytidines demonstrated the most pronounced effects, significantly decreasing the dipalmitoylphosphocholine melting temperature and cooperativity of phase transition. The elongation of hydrophobic acyl radicals potentiated the disordering action of N4-alkylcytidines, while an increase in hydrophilicity due to replacing deoxyribose with ribose inhibited this effect. The ability of compounds to form transmembrane pores was also tested. It was found that 5-alkyluridines produced single, ion-permeable pores in phosphatidylglycerol membranes, and that methoxy-mycolic acid and trehalose monooleate potentiated the pore-forming activity of alkyloxymethyldeoxyuridines. The results obtained open up perspectives for the development of innovative highly selective anti-tuberculosis agents, which may be characterized by a low risk of developing drug resistance due to the direct action on the membranes of the pathogen.

Enzymatic transglycosylation of the fleximer base 4-(4-aminopyridine-3-yl)-1H-pyrazole using recombinant E. coli purine nucleoside phosphorylase (PNP) resulted in the formation of “non-typical” minor products of the reaction. In addition to “typical” N1-pyrazole nucleosides, a 4-imino-pyridinium riboside and a N1-pyridinium-N1-pyrazole bis-ribose derivative were formed. N1-Pyrazole 2′-deoxyribonucleosides and a N1-pyridinium-N1-pyrazole bis-2′-deoxyriboside were formed. But 4-imino-pyridinium deoxyriboside was not formed in the reaction mixture. The role of thermodynamic parameters of key intermediates in the formation of reaction products was elucidated. To determine the mechanism of binding and activation of heterocyclic substrates in the E. coli PNP active site, molecular modeling of the fleximer base and reaction products in the enzyme active site was carried out. As for N1-pyridinium riboside, there are two possible locations for it in the PNP active site. The presence of a relatively large space in the area of amino acid residues Phe159, Val178, and Asp204 allows the ribose residue to fit into that space, and the heterocyclic base can occupy a position that is suitable for subsequent glycosylation. Perhaps it is this “upside down” arrangement that promotes secondary glycosylation and the formation of minor bis-riboside products.

Abstract Background In light of the growing appreciation for the role of collective cell motility in metastasis, a deeper understanding of the underlying signaling pathways will be critical to translating these observations to the treatment of advanced cancers. Here, we examine the contribution of Wnt/planar cell polarity (Wnt/PCP), one of the non-canonical Wnt signaling pathways and defined by the involvement of the tetraspanin-like proteins Vangl1 and Vangl2, to breast tumor cell motility, collective cell invasiveness and mammary tumor metastasis. Methods Vangl1 and Vangl2 knockdown and overexpression and Wnt5a stimulation were employed to manipulate Wnt/PCP signaling in a battery of breast cancer cell lines representing all breast cancer subtypes, and in tumor organoids from MMTV-PyMT mice. Cell migration was assessed by scratch and organoid invasion assays, Vangl protein subcellular localization was assessed by confocal fluorescence microscopy, and RhoA activation was assessed in real time by fluorescence imaging with an advanced FRET biosensor. The impact of Wnt/PCP suppression on mammary tumor growth and metastasis was assessed by determining the effect of conditional Vangl2 knockout on the MMTV-NDL mouse mammary tumor model. Results We observed that Vangl2 knockdown suppresses the motility of all breast cancer cell lines examined, and overexpression drives the invasiveness of collectively migrating MMTV-PyMT organoids. Vangl2-dependent RhoA activity is localized in real time to a subpopulation of motile leader cells displaying a hyper-protrusive leading edge, Vangl protein is localized to leader cell protrusions within leader cells, and actin cytoskeletal regulator RhoA is preferentially activated in the leader cells of a migrating collective. Mammary gland-specific knockout of Vangl2 results in a striking decrease in lung metastases in MMTV-NDL mice, but does not impact primary tumor growth characteristics. Conclusions We conclude that Vangl-dependent Wnt/PCP signaling promotes breast cancer collective cell migration independent of breast tumor subtype and facilitates distant metastasis in a genetically engineered mouse model of breast cancer. Our observations are consistent with a model whereby Vangl proteins localized at the leading edge of leader cells in a migrating collective act through RhoA to mediate the cytoskeletal rearrangements required for pro-migratory protrusion formation.

Abstract Background Therapeutic interventions that optimize angiogenic activities may reduce rates of end-stage kidney disease, critical limb ischemia, and lower extremity amputations in individuals with diabetic kidney disease (DKD). Infusion of autologous mesenchymal stromal cells (MSC) is a promising novel therapy to rejuvenate vascular integrity. However, DKD-related factors, including hyperglycemia and uremia, might alter MSC angiogenic repair capacity in an autologous treatment approach. Methods To explore the angiogenic activity of MSC in DKD, the transcriptome of adipose tissue-derived MSC obtained from DKD subjects was compared to age-matched controls without diabetes or kidney impairment. Next-generation RNA sequencing (RNA-seq) was performed on MSC (DKD n = 29; Controls n = 9) to identify differentially expressed (DE; adjusted p 1) messenger RNA (mRNA) and microRNA (miRNA) involved in angiogenesis (GeneCards). Paracrine-mediated angiogenic repair capacity of MSC conditioned medium (MSCcm) was assessed in vitro using human umbilical vein endothelial cells incubated in high glucose and indoxyl sulfate for a hyperglycemic, uremic state. Results RNA-seq analyses revealed 133 DE mRNAs (77 upregulated and 56 down-regulated) and 208 DE miRNAs (119 up- and 89 down-regulated) in DKD-MSC versus Control-MSC. Interestingly, miRNA let-7a-5p, which regulates angiogenesis and participates in DKD pathogenesis, interacted with 5 angiogenesis-associated mRNAs (transgelin/TAGLN, thrombospondin 1/THBS1, lysyl oxidase-like 4/LOXL4, collagen 4A1/COL4A1 and collagen 8A1/COL8A1). DKD-MSCcm incubation with injured endothelial cells improved tube formation capacity, enhanced migration, reduced adhesion molecules E-selectin, vascular cell adhesion molecule 1 and intercellular adhesion molecule 1 mRNA expression in endothelial cells. Moreover, angiogenic repair effects did not differ between treatment groups (DKD-MSCcm vs. Control-MSCcm). Conclusions MSC from individuals with DKD show angiogenic transcriptome alterations compared to age-matched controls. However, angiogenic repair potential may be preserved, supporting autologous MSC interventions to treat conditions requiring enhanced angiogenic activities such as DKD, diabetic foot ulcers, and critical limb ischemia.

Between 2018 and 2021, several mastic trees (Pistacia lentiscus var. Chia) sampled in the field and the nursery of the Chios Mastiha Growers Association (CMGA) were analyzed to determine the cause of dominant diseases. Symptoms included defoliation, leaf, and twig blight, wilting and/or apoplexy of trees and apoplexy of young hardwood cuttings. Moreover, brown discoloration had also been observed on older woody parts of the trees such as branches and tree trunks. Several pathogens have been isolated and identified as the causing agents. Neopestalotiopsis and Alternaria species were isolated consistently from necrotic tissues of mastic trees (branches, twigs, and leaves) in the field and the nursery. All fungal isolates’ pathogenicity was confirmed by applying Koch’s postulates on young mastic trees under glasshouse conditions. Fungal pathogens were identified by sequence analyses of the ITS, β-tubulin, and histone gene regions. Alternaria species were analyzed further by sequencing the endopolygalacturonase (endoPG) and the Alternaria major allergen (Alta1) genes. More specifically, the isolates were identified as Neopestalotiopsis clavispora, Alternaria arborescens, and A. alternata based on morphological features and sequence analyses. This is the first report of N. clavispora, A. arborescens, and A. alternata on P. lentiscus var. Chia.

The author deals with the refraction of the category of “tolerance” in O. Zhanaydarov's play “Aldar” through the interpretation of the image of the Migrant Other. The analysis of the work is built from the standpoint of the new drama and communicative violence as one of its main principles. The depiction of a migrant in the play is subject to a number of characteristics that contribute to the overall idea - to reflect acute inequality, global chaos, and the deconstruction of moral attitudes. Hence, the concept of tolerance undergoes complete decomposition, degradation - destruction. The discourse of the Other allows the playwright to narrate on the basis of a performative principle using a certain cultural code, which is also considered as the Other. By constructing a character's conflict with a system that is based on the foundation of violence, the author implements the principle of hyperrealism, which in turn exposes previously hidden problems that corrode society. The Other in this case turns out to be a signal of the need to reshape the “sick,” destructive world order, which is interpreted by society as something self-evident, unshakable. However, whether the Other can become an effective lever for understanding tolerance in its “healthy” meanings by changing the violent, perverted principles of reality - this question has not yet been answered.

Abstract Background Loss of control eating (LOC-E) in youth predicts the later development of full-syndrome binge-eating disorder (BED), and therefore, could be a relevant target for prevention treatments. To develop these treatments, it is important to understand the underlying disease processes and mechanisms. Based on the putative role of neurocognitive impairments in the pathogenesis of LOC-E, treatments that modulate these neurocognitive factors warrant further exploration. For instance, stimulants are an effective treatment for impulsivity in youth with attention deficit/hyperactivity disorder (ADHD) and have been shown to improve symptoms of BED in adults. Notably, stimulants have not been examined as a treatment for LOC-E in youth. To explore this gap, we aim to measure change in LOC-E episodes and secondary outcomes in youth with comorbid ADHD and LOC-E who are being started on stimulants. Methods We will collect prospective observational data on forty 8-to-13-year-old youth diagnosed with comorbid ADHD and LOC-E who are initiating a stimulant for ADHD. Prior to stimulant initiation, participants will complete baseline measures including LOC-E episode frequency in the last 3 months (primary outcome), and secondary outcomes including disordered eating cognitions, emotions and behaviors, ADHD symptom severity, parental LOC-E, impulsivity and reward sensitivity, and anxiety/mood severity. Outcome measurements will be gathered again at 3-months after initiating the stimulant. Within-patient standardized effect sizes with 95% confidence intervals will be calculated from baseline to 3-month follow-up for all outcomes. Discussion Many individuals with LOC-E or binge eating do not fully remit over the course of psychotherapy. Whereas psychotherapy may address psychological and sociocultural domains associated with LOC-E, some individuals with neurocognitive impairments (e.g., ADHD) and neurobiological deficits (e.g., low intrasynaptic dopamine or norepinephrine) may benefit from adjunctive treatment that targets those factors. This will be the first study to provide pilot data for future studies that could examine both the effect of stimulants on LOC-E in youth and underlying mechanisms. Trial registration Trial registration number: NCT05592119

The regulation of plasma glucose levels is effected by insulin. Here, the authors reveal atomic detail of how peptides distinct from insulin bind to and activate the insulin receptor, with implications for design of small-molecule insulin mimetics.

Salak is a potential horticultural sector that is a leading commodity in Banjarnegara. Salak fruit varieties have fruit categories that have their advantages. Variants of salak fruit include ivory salak, granulated sugar salak, pondoh salak, and honey salak. Based on data released from the relevant government agencies, further research was carried out related to analyzing and conducting research to predict salak fruit varieties. This variety is suitable for land in every area in Banjarnegara with predictive analysis using the C4.5 algorithm. This method has been widely developed to classify and predict a case with a fairly high degree of accuracy. From this study, researchers hope that it can contribute farmers to determining the type of salak fruit that is most suitable for the land they own so that later the harvest obtained by farmers can be maximized

Menentukan sumber pembelajaran pendukung yang efektif dalam E-Learning merupakan suatu permasalahan yang kompleks, karena keputusan yang diberikan untuk berbagai kriteria yang ditujukan untuk memberi prioritas kepada alternatif yang digunakan. Pendekatan Fuzzy-Analytic Hierarchy Process (Fuzzy AHP) termasuk salah satu prosedur yang bisa menjadi solusi dari permasalahan ini. Metode ini dapat digunakan untuk menentukan keputusan dalam penaksiran kepada kriteria yang sudah ada sebelumnya. Dari perhitungan pengujian yang dilakukan, YouTube mendapat peringkat tertinggi yang lalu dibawahnya diikuti oleh Google, Brainly, dan yang terakhir Slide Player. Dari perhitungan tersebut, dapat disimpulkan bahwa YouTube bisa dijadikan sebagai sumber pembelajaran pendukung dalam sistem pembelajaran E-Learning.

Human FACT (FACT) is a multifunctional histone chaperone involved in transcription, replication and DNA repair. Curaxins are anticancer compounds that induce FACT-dependent nucleosome unfolding and trapping of FACT in the chromatin of cancer cells (c-trapping) through an unknown molecular mechanism. Here, we analyzed the effects of curaxin CBL0137 on nucleosome unfolding by FACT using spFRET and electron microscopy. By itself, FACT adopted multiple conformations, including a novel, compact, four-domain state in which the previously unresolved NTD of the SPT16 subunit of FACT was localized, apparently stabilizing a compact configuration. Multiple, primarily open conformations of FACT-nucleosome complexes were observed during curaxin-supported nucleosome unfolding. The obtained models of intermediates suggest “decision points” in the unfolding/folding pathway where FACT can either promote disassembly or assembly of nucleosomes, with the outcome possibly being influenced by additional factors. The data suggest novel mechanisms of nucleosome unfolding by FACT and c-trapping by curaxins.

Saat ini seluruh penduduk dunia berjuang melawan wabah covid-19. Cara yang efektif melawan covid adalah melawannya dengan meningkatkan imut tubuh. Salah satu cara meningkatkan imun tubuh adalah menjaga hidup sehat dengan kebiasaan berolahraga. Namun faktanya sebagian besar orang cenderung enggan berolahraga dengan berbagai alasan. Maka penelitian ini menganalisis bagaimana motivasi masyarakat dalam berolahraga selama pandemic Covid-19 dengan analisis core drive Octalysis sehingga dapat digali lebih jauh doronga-dorongan apa saja yang mungkin lemah dan yang kuat sehingga bisa dijadikan referensi bagi pihak-pihak yang berkepentingan untuk menyelenggarakan kegiatan olahraga yang lebih diminati. Hasil analisis menunjukkan bahwa kecenderungan masyarakat berolahraga semasa pandemic Covid-19 adalah termotivasi ingin menjaga kesehatan diri sendiri dan menginspirasi orang lain untuk hidup sehat. Sedangkan suasana kompetisi maupun kerjasama dalam olah raga tidak terlalu dibutuhkan karena masyarakat tidak terlalu termotivasi dari dua hal tersebut.

There are many queuing systems that accept single arrivals, accumulate them and service only as a group. Examples of such systems exist in various areas of human life, from traffic of transport to processing requests on a computer network. Therefore, our study is actual. In this paper some class of finite Markovian queueing models with single arrivals and group services are studied. We considered the forward Kolmogorov system for corresponding class of Markov chains. The method of obtaining bounds of convergence on the rate via the notion of the logarithmic norm of a linear operator function is not applicable here. This approach gives sharp bounds for the situation of essentially non-negative matrix of the corresponding system, but in our case it does not hold. Here we use the method of differential inequalities to obtaining bounds on the rate of convergence to the limiting characteristics for the class of finite Markovian queueing models. We obtain bounds on the rate of convergence and compute the limiting characteristics for a specific non-stationary model too. Note the results can be successfully applied for modeling complex biological systems with possible single births and deaths of a group of particles.

BackgroundSpecific phobias represent the largest category of anxiety disorders. Previous work demonstrated that stimulating the ventromedial prefrontal cortex (vmPFC) with repetitive Transcranial Magnetic Stimulation (rTMS) may improve response to exposure therapy for acrophobia.ObjectiveTo examine feasibility of accelerating extinction learning in subjects with spider phobia using intermittent Theta Burst Stimulation (iTBS) rTMS of vmPFC.MethodsIn total, 17 subjects with spider phobia determined by spider phobia questionnaires [Spider Phobia Questionnaire (SPQ) and Fear of Spiders questionnaire (FSQ)] underwent ratings of fear of spiders as well as behavioral and skin conductance data during a behavioral avoidance test (BAT). Subjects then received a sequential protocol of in vivo spider exposure followed by iTBS for three sessions administered to either active or control treatment sites (vmPFC [n = 8] or vertex [n = 9], respectively), followed 1 week later by repetition of questionnaires and BAT.ResultsAll subjects improved significantly regardless of group across both questionnaires (FSQ η2 = 0.43, p = 0.004; SPQ η2 = 0.39, p = 0.008) and skin conductance levels during BAT (Wald χ2 = 30.9, p < 0.001). Subjects in the vmPFC group tolerated lower treatment intensity than in the control group, and there was a significant correlation between treatment intensity, BAT subjective distress improvement, and physiologic measures (all ρ > 0.5).ConclusionThis proof-of-concept study provides preliminary evidence that a sequential exposure and iTBS over vmPFC is feasible and may have rTMS intensity-dependent effects on treatment outcomes, providing evidence for future areas of study in the use of rTMS for phobias.

This paper aimed to characterize urbostratozems (Urbic Technosol, WRB) of Saint Petersburg located in industrial (“Electrodepo” railway station) and residential (region Polish Garden) zones. These soils were also compared with background (natural) soddy podzol soil (Umbric Albic Gleic Podzol, WRB) sampled in recreational zone (suburban park “Oranienbaum”). Soil samples were collected from soil horizons for chemical analysis and from top of soils for microbialogical analysis in June of 2012. Chemical properties (pH, total organic carbon, mobile forms of K and P) and content of heavy metals (Pb, Cu, Zn, Ni) in soils were determined. Culturable forms of microorganisms (bacteria and fungi) were studied. Assessment of the enzymatic activity of the soil was carried out by culturing of microorganisms-producers of protease, amylase, cellulase and lipase on special media. Biotesting using cress (Lepidium sativum L.) seeds had been carried out for assessment of soil phytotoxicity. It was found that chemical properties of urban and natural soils differ greatly. Heavy metal pollution was evident in both urban soils, but maximum concentrations of heavy metals were found in the soil of the industrial zone. Phytotoxocity had been also most pronouncend in the soil of the industrial zone. The natural soil exhibited significantly higher respiration activity than urbostratozems. The greatest difference in the structure of the bacterial and fungal communities was observed between the natural soil of the recreational zone and the urbostratozem of the industrial zone. Algae had been present in the urban soils of the residential zone that was not observed in the natural podzol. The minimum number of producers of all enzymes, except for cellulase, was observed in the soddy podzol in the recreational zone. The maximum number of protease and amylase producers was found in the soil of the industrial zone. Lipolytic activity was almost the same in all samples. It was found that more sensitive biological methods are needed for environmental assessment of urban soils. The results of the article can be used by soil scientists and environmental engineers for a comprehensive environmental assessment of the condition of urban soils and for creating new urban green spaces.

Background Sacubitril/valsartan has been incorporated into guidelines based on the results of the PARADIGM-HF trial, which demonstrated reduced mortality in stable patients with heart failure with reduced ejection fraction (HFrEF). Sacubitril/valsartan is recommended in addition to other HF therapies in place of an angiotensin-converting-enzyme inhibitor or angiotensin-receptor-blocker. Objectives To evaluate the safety and tolerability of sacubitril/valsartan initiation in a community hospital. Design/methods This single-center, retrospective review evaluated patients that received ≥24 hours of sacubitril/valsartan therapy August 2015-March 2018. The primary outcome included the incidence of hypotensive events during hospitalization. Secondary outcomes included: incidence of inpatient acute kidney injury (AKI) and hyperkalemia, rates of inpatient discontinuation, and change in ejection fraction (EF) ≥30 days after initiation. Results Of the 59 patients included, 21 (35.6%) experienced a hypotensive event. A total of 6 patients (10.2%) discontinued therapy while inpatient, which was more likely in patients that developed AKI (n = 3; p = 0.005) or those who experienced a hypotensive event (n = 5; p = 0.018). There was a significant difference in mean EF from baseline to ≥ 30 days post-initiation (24.8% vs. 33.2%; p = 0.018). Conclusion Careful patient selection and monitoring for hypotension, AKI, and hyperkalemia can help increase successful outcomes and improve patient safety.

The aim of this study was to develop three supervised self-organizing map (SOM) models for the automatic recognition of a systemic resistance state in plants after application of a resistance inducer. The pathosystem Fusarium oxysporum f. sp. radicis-lycopersici (FORL) + tomato was used. The inorganic, defense inducer, Acibenzolar-S-methyl (benzo-[1,2,3]-thiadiazole-7-carbothioic acid-S-methyl ester, ASM), reported to induce expression of defense genes in tomato, was applied to activate the defense mechanisms in the plant. A handheld fluorometer, FluorPen FP 100-MAX-LM by SCI, was used to assess the fluorescence kinetics response of the induced resistance in tomato plants. To achieve recognition of resistance induction, three models of supervised SOMs, namely SKN, XY-F, and CPANN, were used to classify fluorescence kinetics data, in order to determine the induced resistance condition in tomato plants. To achieve this, a parameterization of fluorescence kinetics curves was developed corresponding to fluorometer variables of the Kautsky Curves. SKN was the best supervised SOM, achieving 97.22% to 100% accuracy. Gene expression data were used to confirm the accuracy of the supervised SOMs.

Tuberculosis (TB) is the oldest human infection disease. Mortality from TB significantly decreased in the 20th century, because of vaccination and the widespread use of antibiotics. However, about a third of the world’s population is currently infected with Mycobacterium tuberculosis (Mtb) and the death rate from TB is about 1.4–2 million people per year. In the second half of the 20th century, new extensively multidrug-resistant strains of Mtb were identified, which are steadily increasing among TB patients. Therefore, there is an urgent need to develop new anti-TB drugs, which remains one of the priorities of pharmacology and medicinal chemistry. The antimycobacterial activity of nucleoside derivatives and analogues was revealed not so long ago, and a lot of studies on their antibacterial properties have been published. Despite the fact that there are no clinically used drugs based on nucleoside analogues, some progress has been made in this area. This review summarizes current research in the field of the design and study of inhibitors of mycobacteria, primarily Mtb.

Nitrogen-functionalization is an effective means of improving the catalytic performances of nanozymes. In the present work, plasma-assisted nitrogen modification of nanocolumnar Ni GLAD films was performed using an ammonia plasma, resulting in an improvement in the peroxidase-like catalytic performance of the porous, nanostructured Ni films. The plasma-treated nanozymes were characterized by TEM, SEM, XRD, and XPS, revealing a nitrogen-rich surface composition. Increased surface wettability was observed after ammonia plasma treatment, and the resulting nitrogen-functionalized Ni GLAD films presented dramatically enhanced peroxidase-like catalytic activity. The optimal time for plasma treatment was determined to be 120 s; when used to catalyze the oxidation of the colorimetric substrate TMB in the presence of H2O2, Ni films subjected to 120 s of plasma treatment yielded a much higher maximum reaction velocity (3.7⊆10-8 M/s vs. 2.3⊆10-8 M/s) and lower Michaelis-Menten coefficient (0.17 mM vs. 0.23 mM) than pristine Ni films with the same morphology. Additionally, we demonstrate the application of the nanozyme in a gravity-driven, continuous catalytic reaction device. Such a controllable plasma treatment strategy may open a new door toward surface-functionalized nanozymes with improved catalytic performance and potential applications in flow-driven point-of-care devices.

Pyrimidine nucleoside analogues are widely used to treat infections caused by the human immunodeficiency virus (HIV) and DNA viruses from the herpes family. It has been shown that 5-substituted uracil derivatives can inhibit HIV-1, herpes family viruses, mycobacteria and other pathogens through various mechanisms. Among the 5-substituted pyrimidine nucleosides, there are not only the classical nucleoside inhibitors of the herpes family viruses, 2′-deoxy-5-iodocytidine and 5-bromovinyl-2′-deoxyuridine, but also derivatives of 1-(benzyl)-5-(phenylamino)uracil, which proved to be non-nucleoside inhibitors of HIV-1 and EBV. It made this modification of nucleoside analogues very promising in connection with the emergence of new viruses and the crisis of drug resistance when the task of creating effective antiviral agents of new types that act on other targets or exhibit activity by other mechanisms is very urgent. In this paper, we present the design, synthesis and primary screening of the biological activity of new nucleoside analogues, namely, 5′-norcarbocyclic derivatives of substituted 5-arylamino- and 5-aryloxyuracils, against RNA viruses.

Introduction: Current atrial fibrillation (AF) guidelines recommend flecainide as a first-line rhythm control option in patients without structural heart disease. While there is proven efficacy in clinical trials and guideline support, it is hypothesized that flecainide may be underutilized due to negative outcomes in the CAST trial and that adverse effects are less common than previously perceived. Methods: This retrospective chart review evaluated patients ⩾18 years initiated on flecainide for AF from August 2011 to October 2016 by a cardiology provider at the study site. Exclusion criteria included:

Abstract Heterologous production of extracellular polyhydroxybutyrate (PHB) depolymerases (PhaZs) has been of interest for over 30 years, but implementation is sometimes difficult and can limit the scope of research. With the constant development of tools to improve recombinant protein production in Escherichia coli, we propose a method that takes characteristics of PhaZs from different bacterial strains into account. Recombinant His‐tagged versions of PhaZs (rPhaZ) from Comamonas testosteroni 31A, Cupriavidus sp. T1, Marinobacter algicola DG893, Pseudomonas stutzeri, and Ralstonia sp. were successfully produced with varying expression, solubility, and purity levels. PhaZs from C. testosteroni and P. stutzeri were more amenable to heterologous expression in all aspects; however, using the E. coli Rosetta‐gami B(DE3) expression strain and establishing optimal conditions for expression and purification (variation of IPTG concentration and use of size exclusion columns) helped circumvent low expression and purity for the other PhaZs. Degradation activity of the rPhaZs was compared using a simple PHB plate‐based method, adapted to test for various pH and temperatures. rPhaZ from M. algicola presented the highest activity at 15°C, and rPhaZs from Cupriavidus sp. T1 and Ralstonia sp. had the highest activity at pH 5.4. The methods proposed herein can be used to test the production of soluble recombinant PhaZs and to perform preliminary evaluation for applications that require PHB degradation.

Two novel, origami-inspired, metamaterials were designed, mechanically tested, and modelled. One novel origami model was folded using a triangular based crease pattern and the other was folded using a rectangular based crease pattern. The origami-inspired metamaterial sheets were fabricated from polylactic acid using fused deposition additive manufacturing. Several configurations, parameterized by varying the fold angle, were mechanically tested under compression and impact loads. It was found that the specific elastic compression modulus of these novel designs was higher, ranging from 594 MPa/kg to 926 MPa/kg, than existing origami-inspired structures made based on the popular Ron-Resch design, which had specific elastic compression moduli between 15 MPa/kg to 365 MPa/kg. A finite element model further analysed the stress distribution of the core structures under compression loads. The impact testing results showed that the pattern of the tessellated cores affected the amount of impact force transferred through the samples, whereas the fold angle of the origami-inspired design had little impact on the results. The rectangular structure was shown to transfer approximately 50–75% of the force transferred by the triangular structure under impact loads. Keywords: Origami, Metamaterial, Fused deposition modelling, Compression, Impact, Finite element modelling

Assessing and managing nitrogen sustainably is imperative for achieving the 17 UN Sustainable Development Goals (SDGs) targeted for 2030. South Asian countries, aware of the environmental and health impacts of nitrogen pollution, regionally as well as globally, piloted the 2019 UN resolution on sustainable nitrogen management, calling for urgent policy action. This paper assists South Asia policy development by providing new insights into nitrogen-related policies in the region; it makes a step-change advance on an existing global analysis and database. We built on available methods to better identify, classify and analyse 966 nitrogen-related policies for the region. We compared the global and regional nitrogen policy landscapes to explain the benefits of a deeper policy assessment. The policies we classified as having ‘higher’ relevance—those with direct reference to nitrogen and/or its potential impacts—represent the current nitrogen policy landscape for South Asia. We show that a small proportion of policies (9%) consider multiple pollution sources, sectors, nitrogen threats and impacts, with integrative policy instruments. A 5% of policies also consider both non-point and point sources of pollution, representing standout policies. More such policies with an integrated approach are vital in addressing the complexities of nitrogen pollution. Adapting existing and drafting new policies are both required to deal with other current and emerging nitrogen issues. Our analysis provides evidence for a roadmap for sustainable nitrogen policy in South Asia and beyond and supports efforts to reduce the threats posed by nitrogen pollution to achieve the SDGs.

Abstract Background The majority of undiagnosed diseases manifest with objective findings that warrant further investigation. The Undiagnosed Diseases Network (UDN) receives applications from patients whose symptoms and signs have been intractable to diagnosis; however, many UDN applicants are affected primarily by subjective symptoms such as pain and fatigue. We sought to characterize presenting symptoms, referral sources, and demographic factors of applicants to the UDN to identify factors that may determine application outcome and potentially differentiate between those with undiagnosed diseases (with more objective findings) and those who are less likely to have an undiagnosed disease (more subjective symptoms). Methods We used a systematic retrospective review of 151 consecutive Not Accepted and 50 randomly selected Accepted UDN applications. The primary outcome was whether an applicant was Accepted, or Not Accepted, and, if accepted, whether or not a diagnosis was made. Objective and subjective symptoms and information on prior specialty consultations were collected from provider referral letters. Demographic data and decision data on network acceptance were gathered from the UDN online portal. Results Fewer objective findings and more subjective symptoms were found in the Not Accepted applications. Not Accepted referrals also were from older individuals, reported a shorter period of illness, and were referred to the UDN by their primary care physicians. All of these differences reached statistical significance in comparison with Accepted applications. The frequency of subspecialty consults for diagnostic purposes prior to UDN application was similar in both groups. Conclusions The preponderance of subjective and lack of objective findings in the Not Accepted applications distinguish these from applicants that are accepted for evaluation and diagnostic efforts through the UDN. Not Accepted applicants are referred primarily by their primary care providers after multiple specialist consultations fail to yield answers. Distinguishing between patients with undiagnosed diseases with objective findings and those with primarily subjective findings can delineate patients who would benefit from further diagnostic processes from those who may have functional disorders and need alternative pathways for management of their symptoms. Trial registration clinicaltrials.gov NCT02450851, posted May 21st 2015.

Biosolid application is an effective strategy, alternative to synthetic chemicals, for enhancing plant growth and performance and improving soil properties. In previous research, biosolid application has shown promising results with respect to tomato resistance against Fusarium oxysporum f. sp. radicis-lycopersici (Forl). Herein, we aimed at elucidating the effect of biosolid application on the plant–microbiome response mechanisms for tomato resistance against Forl at a molecular level. More specifically, plant–microbiome interactions in the presence of biosolid application and the biocontrol mechanism against Forl in tomato were investigated. We examined whether biosolids application in vitro could act as an inhibitor of growth and sporulation of Forl. The effect of biosolid application on the biocontrol of Forl was investigated based on the enhanced plant resistance, measured as expression of pathogen-response genes, and pathogen suppression in the context of soil microbiome diversity, abundance, and predicted functions. The expression of the pathogen-response genes was variably induced in tomato plants in different time points between 12 and 72 h post inoculation in the biosolid-enriched treatments, in the presence or absence of pathogens, indicating activation of defense responses in the plant. This further suggests that biosolid application resulted in a successful priming of tomato plants inducing resistance mechanisms against Forl. Our results have also demonstrated that biosolid application alters microbial diversity and the predicted soil functioning, along with the relative abundance of specific phyla and classes, as a proxy for disease suppression. Overall, the use of biosolid as a sustainable soil amendment had positive effects not only on plant health and protection, but also on growth of non-pathogenic antagonistic microorganisms against Forl in the tomato rhizosphere and thus, on plant–soil microbiome interactions, toward biocontrol of Forl.

This paper dwells on exposures' analysis of open-environment operated swarm robotic systems. Flocking algorithms for swarm robotics, which use alignment and averaging of direction in local communication radius (after C. Reynolds' model) are considered. The problem where a small group of informed robots has to guide the swarm along a desired direction is also considered. Couzin I. D. et al. (2005) showed that even a minority of informed individuals is able to move the group toward the target Hereafter great number of swarm robotics' flocking algorithms were developed in order to increase swarm system's handiness. Several implicit leadership algorithms for distributed robotic systems were proposed. The idea of this paper is to study and simulate the operation of such kind of above mentioned algorithms in the situation when there are saboteurs which destructively impact swarm system. Specially developed by authors simulation software allows to vary the parameters of system operation, which can be rather the number of agents or their interaction features or etc. On the basis of the experimental data the authors produce the analysis of features in order to determine their relevance to the algorithm.

Swarm robotic systems are actively developed and widely studied in the world scientific practice. It is expected that multiagent, distributed approach to creating artificial intelligence of autonomous systems will allow to solve a great number of complex problems in the areas of environmental protection, medicine, cleaning, patrolling, etc. Thereby the research of these systems (design and testing) in terms of information security becomes relevant. The key to a wide practical use of swarm robotic systems is the development of specific guidelines and algorithms for the organization of group actions. This research proposes the use of trust and reputation model for information security of swarm system. Swarm's agents generate trust levels to each other basing on the analysis of situation on the kth iteration step of the algorithm and using their sensor devices. On the calculated trust levels the collective recognition of saboteurs is carried out. To perform experiment software simulator was designed. It allows to vaiy the basic parameters of swarm robotic system (number of agents, number of targets, range of communication, number of saboteurs).

Widespread latent herpes viral infections within a population can lead to the development of co-infections in HIV-infected patients. These infections are not particularly dangerous for healthy individuals and often occur with minimal symptoms, but for those who are immunocompromised, these infections can accelerate the acute phase of HIV infection and AIDS. Thus, the idea of designing compounds that could combine activity against HIV and co-infections would seem promising. In that regard, eleven compounds were synthesized that represent conjugates of non-nucleoside HIV reverse transcriptase inhibitors and nucleoside inhibitors of the herpes family viruses with the hope that these novel heterodimers will result in dual activity against HIV and concomitant herpes virus infections.

Nowadays, Inventory management has become a major concern for company because it can bring value added. Inventory management focused on maintaining and managing the inventory level in order to fulfill the customer demand with the lowest cost. In retailing and wholesaling, multi items from the same supplier are jointly ordered. This mean, the order interval of each item must be the same. But, there are several manufacturing companies that have not achieving the optimum solution in inventory management for multi items. It leads the company to increase the total inventory cost. Most of them are caused by the improper order interval. Thus, appropriate of inventory management may reduce the total inventory cost. This research tried to identify this problem. The first step that should be done is by identifying the problems, scopes, and objectives of the research which aimed to minimize the inventory control management. Then collecting data and analyzing the supported data in inventory management. By using the total inventory cost analysis as a parameter in comparing the actual system and the proposal system. This research also used forecasting methodology in order to predicting the future demand. In the final section is by implementing the Economic Order Interval in ordering process for determining the optimum order interval. The result in this thesis show that the proposal system can reduce the total inventory cost over 21% from the current condition.

Carbocyclic nucleosides have long played a role in antiviral, antiparasitic, and antibacterial therapies. Recent results from our laboratories from two structurally related scaffolds have shown promising activity against both Mycobacterium tuberculosis and several parasitic strains. As a result, a small structure activity relationship study was designed to further probe their activity and potential. Their synthesis and the results of the subsequent biological activity are reported herein.

High activity levels of a transgene can be very useful, making a transgene easier to evaluate for safety and efficacy. High activity levels can also increase the economic benefit of the production of high value proteins in transgenic plants. The goal of this research is to determine if recurrent selection for activity of a transgene will result in higher activity, and if selection for activity of a transgene controlled by a native promoter will also increase protein levels of the native gene with the same promoter. To accomplish this goal we used transgenic maize containing a construct encoding green fluorescent protein controlled by the promoter for the maize endosperm-specific 27 kDa gamma zein seed storage protein. We carried out recurrent selection for fluorescence intensity in two breeding populations. After three generations of selection, both selected populations were significantly more fluorescent and had significantly higher levels of 27 kDa gamma zein than the unselected control populations. These higher levels of the 27 kDa gamma zein occurred independently of the presence of the transgene. The results show that recurrent selection can be used to increase activity of a transgene and that selection for a transgene controlled by a native promoter can increase protein levels of the native gene with the same promoter via proxy selection. Moreover, the increase in native gene protein level is maintained in the absence of the transgene, demonstrating that proxy selection can be used to produce non-transgenic plants with desired changes in gene expression.

A series of novel 5′-norcarbocyclic derivatives of 5-alkoxymethyl or 5-alkyltriazolyl-methyl uracil were synthesized and the activity of the compounds evaluated against both Gram-positive and Gram-negative bacteria. The growth of Mycobacterium smegmatis was completely inhibited by the most active compounds at a MIC99 of 67 μg/mL (mc2155) and a MIC99 of 6.7⁻67 μg/mL (VKPM Ac 1339). Several compounds also showed the ability to inhibit the growth of attenuated strains of Mycobacterium tuberculosis ATCC 25177 (MIC99 28⁻61 μg/mL) and Mycobacterium bovis ATCC 35737 (MIC99 50⁻60 μg/mL), as well as two virulent strains of M. tuberculosis; a laboratory strain H37Rv (MIC99 20⁻50 μg/mL) and a clinical strain with multiple drug resistance MS-115 (MIC99 20⁻50 μg/mL). Transmission electron microscopy (TEM) evaluation of M. tuberculosis H37Rv bacterial cells treated with one of the compounds demonstrated destruction of the bacterial cell wall, suggesting that the mechanism of action for these compounds may be related to their interactions with bacteria cell walls.

Here we report the synthesis and biological activity of new 5′-norcarbocyclic derivatives of bicyclic pyrrolo- and furano[2,3-d]pyrimidines with different substituents in the heterocyclic ring. Lead compound 3i, containing 6-pentylphenyl substituent, displays inhibitory activity with respect to a number of tumor cells with a moderate selectivity index value. Compound 3i induces cell death by the apoptosis pathway with the dissipation of mitochondrial potential.

Bis(monoacylglycero)phosphates (BMPs), a class of lipids highly enriched within endolysosomal organelles, are key components of the lysosomal intraluminal vesicles responsible for activating sphingolipid catabolic enzymes. While BMPs are understudied relative to other phospholipids, recent reports associate BMP dysregulation with a variety of pathological states including neurodegenerative diseases and lysosomal storage disorders. Since the dramatic lysosomal remodeling characteristic of cellular transformation could impact BMP abundance and function, we employed untargeted lipidomics approaches to identify and quantify BMP species in several in vitro and in vivo models of breast cancer and comparative non-transformed cells and tissues. We observed lower BMP levels within transformed cells relative to normal cells, and consistent enrichment of docosahexaenoic acid (22:6) fatty acyl chain-containing BMP species in both human- and mouse-derived mammary tumorigenesis models. Our functional analysis points to a working model whereby 22:6 BMPs serve as reactive oxygen species scavengers in tumor cells, protecting lysosomes from oxidant-induced lysosomal membrane permeabilization. Our findings suggest that breast tumor cells might divert polyunsaturated fatty acids into BMP lipids as part of an adaptive response to protect their lysosomes from elevated reactive oxygen species levels, and raise the possibility that BMP-mediated lysosomal protection is a tumor-specific vulnerability that may be exploited therapeutically.

The soilborne fungus Fusarium oxysporum f. sp. radicislycopersici causes tomato foot and root rot (TFRR), which can be controlled by the addition of the nonpathogenic fungus F. oxysporum Fo47 to the soil. To improve our understanding of the interactions between the two Fusarium strains on tomato roots during biocontrol, the fungi were labeled using different autofluorescent proteins as markers and subsequently visualized using confocal laser scanning microscopy. The results were as follows. i) An at least 50- fold excess of Fo47over F. oxysporum f. sp. radicis-lycopersici was required to obtain control of TFRR. ii) When seedlings were planted in sand infested with spores of a single fungus, Fo47 hyphae attached to the root earlier than those of F. oxysporum f. sp. radicis-lycopersici. iii) Subsequent root colonization by F. oxysporum f. sp. radicis-lycopersici was faster and to a larger extent than that by Fo47. iv) Under disease-controlling conditions, colonization of tomato roots by the pathogenic fungus was significantly reduced. v) When the inoculum concentration of Fo47 was increased, root colonization by the pathogen was arrested at the stage of initial attachment to the root. vi) The percentage of spores of Fo47 that germinates in tomato root exudate in vitro is higher than that of the pathogen F. oxysporum f. sp. radicis-lycopersici. Based on these results, the mechanisms by which Fo47 controls TFRR are discussed in terms of i) rate of spore germination and competition for nutrients before the two fungi reach the rhizoplane; ii) competition for initial sites of attachment, intercellular junctions, and nutrients on the tomato root surface; and iii) inducing systemic resistance.

Ultrasound attenuates when propagating through aqueous dispersions of carbon nanotubes. We established a significant difference in the ultrasound attenuation frequency spectra for the nanotubes oriented either along or across the ultrasound beam. This effect can be used for calculating geometrical and rheological properties of nanofiber materials dispersed in liquids. Keywords: Ultrasound attenuation, Carbon nanotubes, Nanofibers

BackgroundIn light of the growing appreciation for the role of collective cell motility in metastasis, a deeper understanding of the underlying signaling pathways will be critical to translating these observations to the treatment of advanced cancers. Here, we examine the contribution of Wnt/planar cell polarity (Wnt/PCP), one of the non-canonical Wnt signaling pathways and defined by the involvement of the tetraspanin-like proteins Vangl1 and Vangl2, to breast tumor cell motility, collective cell invasiveness and mammary tumor metastasis.MethodsVangl1 and Vangl2 knockdown and overexpression and Wnt5a stimulation were employed to manipulate Wnt/PCP signaling in a battery of breast cancer cell lines representing all breast cancer subtypes, and in tumor organoids from MMTV-PyMT mice. Cell migration was assessed by scratch and organoid invasion assays, Vangl protein subcellular localization was assessed by confocal fluorescence microscopy, and RhoA activation was assessed in real time by fluorescence imaging with an advanced FRET biosensor. The impact of Wnt/PCP suppression on mammary tumor growth and metastasis was assessed by determining the effect of conditional Vangl2 knockout on the MMTV-NDL mouse mammary tumor model.ResultsWe observed that Vangl2 knockdown suppresses the motility of all breast cancer cell lines examined, and overexpression drives the invasiveness of collectively migrating MMTV-PyMT organoids. Vangl2-dependent RhoA activity is localized in real time to a subpopulation of motile leader cells displaying a hyper-protrusive leading edge, Vangl protein is localized to leader cell protrusions within leader cells, and actin cytoskeletal regulator RhoA is preferentially activated in the leader cells of a migrating collective. Mammary gland-specific knockout of Vangl2 results in a striking decrease in lung metastases in MMTV-NDL mice, but does not impact primary tumor growth characteristics.ConclusionsWe conclude that Vangl-dependent Wnt/PCP signaling promotes breast cancer collective cell migration independent of breast tumor subtype and facilitates distant metastasis in a genetically engineered mouse model of breast cancer. Our observations are consistent with a model whereby Vangl proteins localized at the leading edge of leader cells in a migrating collective act through RhoA to mediate the cytoskeletal rearrangements required for pro-migratory protrusion formation.

The fungus Fusarium oxysporum f. sp. radicis-lycopersici causes foot and root rot of tomato plants, which can be controlled by the bacteria Pseudomonas fluorescens WCS365 and P. chlororaphis PCL1391. Induced systemic resistance is thought to be involved in biocontrol by P. fluorescens WCS365. The antifungal metabolite phenazine-1-carbox-amide (PCN), as well as efficient root colonization, are essential in the mechanism of biocontrol by P. chlororaphis PCL1391. To understand the effects of bacterial strains WCS365 and PCL1391 on the fungus in the tomato rhizosphere, microscopic analyses were performed using different autofluorescent proteins as markers. Tomato seedlings were inoculated with biocontrol bacteria and planted in an F. oxysporum f. sp. radicis-lycopersici-infested gnotobiotic sand system. Confocal laser scanning microscope analyses of the interactions in the tomato rhizosphere revealed that i) the microbes effectively compete for the same niche, and presumably also for root exudate nutrients; ii) the presence of either of the two bacteria negatively affects infection of the tomato root by the fungus; iii) both biocontrol bacteria colonize the hyphae extensively, which may represent a new mechanism in biocontrol by these pseudomonads; and iv) the production of PCN by P. chlororaphis PCL1391 negatively affects hyphal growth and branching, which presumably affects the colonization and infecting ability of the fungus.

The fungus Fusarium oxysporum f. sp. radicis-lycopersici is the causal agent of tomato foot and root rot disease. The green fluorescent protein (GFP) was used to mark this fungus in order to visualize and analyze the colonization and infection processes in vivo. Transformation of F. oxysporum f. sp. radicis-lycopersici was very efficient and gfp expression was stable for at least nine subcultures. Microscopic analysis of the transformants revealed homogeneity of the fluorescent signal, which was clearly visible in the hyphae as well as in the chlamydospores and conidia. To our knowledge, this is the first report in which this is shown. The transformation did not affect the pathogenicity. Using confocal laser scanning microscopy, colonization, infection, and disease development on tomato roots were visualized in detail and several new aspects of these processes were observed, such as (i) the complete colonization pattern of the tomato root system; (ii) the very first steps of contact between the fungus and the host, which takes place at the root hair zone by mingling and by the attachment of hyphae to the root hairs; (iii) the preferential colonization sites on the root surface, which are the grooves along the junctions of the epidermal cells; and (iv) the absence of specific infection sites, such as sites of emergence of secondary roots, root tips, or wounded tissue, and the absence of specific infection structures, such as appressoria. The results of this work prove that the use of GFP as a marker for F. oxysporum f. sp. radicis-lycopersici is a convenient, fast, and effective approach for studying plant-fungus interactions.

Remote sensing techniques are routinely used in plant species discrimination and of weed mapping. In the presented work, successful Silybum marianum detection and mapping using multilayer neural networks is demonstrated. A multispectral camera (green-red-near infrared) attached on a fixed wing unmanned aerial vehicle (UAV) was utilized for the acquisition of high-resolution images (0.1 m resolution). The Multilayer Perceptron with Automatic Relevance Determination (MLP-ARD) was used to identify the S. marianum among other vegetation, mostly Avena sterilis L. The three spectral bands of Red, Green, Near Infrared (NIR) and the texture layer resulting from local variance were used as input. The S. marianum identification rates using MLP-ARD reached an accuracy of 99.54%. Τhe study had an one year duration, meaning that the results are specific, although the accuracy shows the interesting potential of S. marianum mapping with MLP-ARD on multispectral UAV imagery.

In the present study, the detection and mapping of Silybum marianum (L.) Gaertn. weed using novelty detection classifiers is reported. A multispectral camera (green-red-NIR) on board a fixed wing unmanned aerial vehicle (UAV) was employed for obtaining high-resolution images. Four novelty detection classifiers were used to identify S. marianum between other vegetation in a field. The classifiers were One Class Support Vector Machine (OC-SVM), One Class Self-Organizing Maps (OC-SOM), Autoencoders and One Class Principal Component Analysis (OC-PCA). As input features to the novelty detection classifiers, the three spectral bands and texture were used. The S. marianum identification accuracy using OC-SVM reached an overall accuracy of 96%. The results show the feasibility of effective S. marianum mapping by means of novelty detection classifiers acting on multispectral UAV imagery.