α-Ketoglutarate links p53 to cell fate during tumour suppression.

The tumour suppressor TP53 is mutated in the majority of human cancers, and in over 70% of pancreatic ductal adenocarcinoma (PDAC)1,2. Wild-type p53 accumulates in response to cellular stress, and regulates gene expression to alter cell fate and prevent tumour development2. Wild-type p53 is also known to modulate cellular metabolic pathways3, although p53-dependent metabolic alterations that constrain cancer progression remain poorly understood. Here we find that p53 remodels cancer-cell metabolism to enforce changes in chromatin and gene expression that favour a premalignant cell fate. Restoring p53 function in cancer cells derived from KRAS-mutant mouse models of PDAC leads to the accumulation of α-ketoglutarate (αKG, also known as 2-oxoglutarate), a metabolite that also serves as an obligate substrate for a subset of chromatin-modifying enzymes. p53 induces transcriptional programs that are characteristic of premalignant differentiation, and this effect can be partially recapitulated by the addition of cell-permeable αKG. Increased levels of the αKG-dependent chromatin modification 5-hydroxymethylcytosine (5hmC) accompany the tumour-cell differentiation that is triggered by p53, whereas decreased 5hmC characterizes the transition from premalignant to de-differentiated malignant lesions that is associated with mutations in Trp53. Enforcing the accumulation of αKG in p53-deficient PDAC cells through the inhibition of oxoglutarate dehydrogenase-an enzyme of the tricarboxylic acid cycle-specifically results in increased 5hmC, tumour-cell differentiation and decreased tumour-cell fitness. Conversely, increasing the intracellular levels of succinate (a competitive inhibitor of αKG-dependent dioxygenases) blunts p53-driven tumour suppression. These data suggest that αKG is an effector of p53-mediated tumour suppression, and that the accumulation of αKG in p53-deficient tumours can drive tumour-cell differentiation and antagonize malignant progression.

Large Lipoma of the Lateral Space of the Neck.

Giant lipoma is characterized by asymptomatic growth and compared with other parts of the body rarely occurs on the neck. If the tumor is localized in the lateral segment of the neck, it can show symptoms in the form of dysphagia and dyspnea. Preoperatively, it is important to perform computed tomography (CT) diagnostic to determine the size of the lesion and makes the plan for operative treatment. The paper presents the case of a 66-year-old patient with a tumor in the neck area as well as swallowing disorders and suffocation during sleep. Palpation revealed a tumor of soft consistency, and based on a CT scan of the neck, the differential diagnosis confirmed giant lipoma. Clinical picture and CT findings of giant neck lipoma are clear in most cases. Due to the atypical localization and size, the tumor should be removed to prevent possible functional disturbances. The treatment is operative, and a histopathological examination should exclude malignancy.

Upregulation of the Oct3/4 Network in Basal Breast Cancer Is Associated with Its Metastatic Potential and Shows Tissue Dependent Variability.

Adaptive plasticity of Breast Cancer stem cells (BCSCs) is strongly correlated with cancer progression and resistance, leading to a poor prognosis. In this study, we report the expression profile of several pioneer transcription factors of the Oct3/4 network associated with tumor initiation and metastasis. In the triple negative breast cancer cell line (MDA-MB-231) stably transfected with human Oct3/4-GFP, differentially expressed genes (DEGs) were identified using qPCR and microarray, and the resistance to paclitaxel was assessed using an MTS assay. The tumor-seeding potential in immunocompromised (NOD-SCID) mice and DEGs in the tumors were also assessed along with the intra-tumor (CD44+/CD24-) expression using flow cytometry. Unlike 2-D cultures, the Oct3/4-GFP expression was homogenous and stable in 3-D mammospheres developed from BCSCs. A total of 25 DEGs including Gata6, FoxA2, Sall4, Zic2, H2afJ, Stc1 and Bmi1 were identified in Oct3/4 activated cells coupled with a significantly increased resistance to paclitaxel. In mice, the higher Oct3/4 expression in tumors correlated with enhanced tumorigenic potential and aggressive growth, with metastatic lesions showing a >5-fold upregulation of DEGs compared to orthotopic tumors and variability in different tissues with the highest modulation in the brain. Serially re-implanting tumors in mice as a model of recurrence and metastasis highlighted the sustained upregulation of Sall4, c-Myc, Mmp1, Mmp9 and Dkk1 genes in metastatic lesions with a 2-fold higher expression of stem cell markers (CD44+/CD24-). Thus, Oct3/4 transcriptome may drive the differentiation and maintenance of BCSCs, promoting their tumorigenic potential, metastasis and resistance to drugs such as paclitaxel with tissue-specific heterogeneity.

Pharmaceutical excipients with potential to cause adverse effects in paediatric nasal medicines.

Some pharmaceutical excipients may cause adverse reactions, excipient-related interactions and/or contraindications. Due to the unique characteristics of the paediatric population, adverse effects may occur to substances generally thought safe. The proportion of topical nasal medicines approved for paediatric use and the prevalence and labelling of excipients with known effect (EKE) in these products were compared in Serbia as a non-EU country and Croatia and Slovenia as EU countries. The study was designed as a post-authorization safety study and safety of excipients was considered in accordance with recommendations of the European Medicines Agency (EMA). More than 90% of topical nasal medicines registered in the three countries were approved for paediatric use and more than half of these paediatric medicines contained EKE that may cause adverse effects. Benzalkonium chloride was found in 52.38%, 55.81% and 59.09% of these products in Serbia, Croatia and Slovenia, respectively. Propylene glycol, benzyl alcohol, ethanol, methyl paraben, propyl paraben and boric acid were also present in a few analysed preparations. A significant number of EKE labelling deficiencies were detected in all three countries, hindering healthcare professionals' access to information needed for adequate patient counselling. A revision of the nasal paediatric medicines' PLs and SmPCs is recommended.

Radical Scavenging Activity and Pharmacokinetic Properties of Coumarin-Hydroxybenzohydrazide Hybrids.

Free radicals often interact with vital proteins, violating their structure and inhibiting their activity. In previous studies, synthesis, characterisation, and the antioxidative properties of the five different coumarin derivatives have been investigated. In the tests of potential toxicity, all compounds exhibited low toxicity with significant antioxidative potential at the same time. In this paper, the radical scavenging activity of the abovementioned coumarin derivatives towards ten different radical species was investigated. It was found that all investigated compounds show good radical scavenging ability, with results that are in correlation with the results published in the previous study. Three additional mechanisms of radical scavenging activity were investigated. It was found that all three mechanisms are thermodynamically plausible and in competition. Interestingly, it was found that products of the Double Hydrogen Atom Transfer (DHAT) mechanism, a biradical species in triplet spin state, are in some cases more stable than singlet spin state analogues. This unexpected trend can be explained by spin delocalisation over the hydrazide bridge and phenolic part of the molecule with a low probability of spin pairing. Besides radical-scavenging activity, the pharmacokinetic and drug-likeness of the coumarin hybrids were investigated. It was found that they exhibit good membrane and skin permeability and potential interactions with P-450 enzymes. Furthermore, it was found that investigated compounds satisfy all criteria of the drug-likeness tests, suggesting they possess a good preference for being used as potential drugs.

Synthesis, Crystallographic, Quantum Chemical, Antitumor, and Molecular Docking/Dynamic Studies of 4-Hydroxycoumarin-Neurotransmitter Derivatives.

In this contribution, four new compounds synthesized from 4-hydroxycoumarin and tyramine/octopamine/norepinephrine/3-methoxytyramine are characterized spectroscopically (IR and NMR), chromatographically (UHPLC-DAD), and structurally at the B3LYP/6-311++G*(d,p) level of theory. The crystal structure of the 4-hydroxycoumarin-octopamine derivative was solved and used as a starting geometry for structural optimization. Along with the previously obtained 4-hydroxycoumarin-dopamine derivative, the intramolecular interactions governing the stability of these compounds were quantified by NBO and QTAIM analyses. Condensed Fukui functions and the HOMO-LUMO gap were calculated and correlated with the number and position of OH groups in the structures. In vitro cytotoxicity experiments were performed to elucidate the possible antitumor activity of the tested substances. For this purpose, four cell lines were selected, namely human colon cancer (HCT-116), human adenocarcinoma (HeLa), human breast cancer (MDA-MB-231), and healthy human lung fibroblast (MRC-5) lines. A significant selectivity towards colorectal carcinoma cells was observed. Molecular docking and molecular dynamics studies with carbonic anhydrase, a prognostic factor in several cancers, complemented the experimental results. The calculated MD binding energies coincided well with the experimental activity, and indicated 4-hydroxycoumarin-dopamine and 4-hydroxycoumarin-3-methoxytyramine as the most active compounds. The ecotoxicology assessment proved that the obtained compounds have a low impact on the daphnia, fish, and green algae population.

Comparison of Seniors in the Community: Risk Evaluation for Eating and Nutrition, version II and Mini Nutritional Assessment - Short Form in detecting nutritional risk among community-dwelling seniors in Bosnia and Herzegovina.

OBJECTIVE: The aim of the current study is to compare Seniors in the Community: Risk Evaluation for Eating and Nutrition, version II (SCREEN II) and Mini Nutritional Assessment - Short Form (MNA-SF), where each is used to identify nutritional risk prevalence among community-dwelling people aged 65 years and above in Bosnia and Herzegovina. DESIGN: A cross-sectional study. Nutritional risk assessed using the nutritionist's risk rating, anthropometric measurements, functional indicators, cognitive parameters, SCREEN II and MNA-SF. SETTING: The municipalities of Foca, East Sarajevo and Bijeljina, Bosnia and Herzegovina. PARTICIPANTS: Eight hundred twenty-one community-dwelling individuals aged ≥65 years. RESULTS: The prevalence of high nutritional risk per nutritionist's risk rating, SCREEN II and MNA-SF was 26, 60, and 7 %, respectively. With the nutritionist's rating score ≥5 as the criterion, the MNA-SF cut-off point of ≤11 (indicating any possible risk) had poor sensitivity (55·7 %), specificity (46·6 %) and AUC (0·563; P = 0·024). When the criterion of >7 was applied, good sensitivity (95·3 %) and specificity (88·9 %) were obtained for the MNA-SF cut-off score of ≤7. AUC for this comparison was 0·742 (considered fair). Cut-off points of <54 (AUC = 0·816) and <50 (AUC = 0·881) for SCREEN II (indicating moderate to high risk) corresponded with good sensitivity (82·2 %; 80·9 %) and fair specificity (72·1 %; 75·0 %). CONCLUSION: MNA-SF may have a limited role in nutritional risk screening among community-dwelling seniors in Bosnia and Herzegovina. SCREEN II has promising results in regard to validity, but further studies are warranted.

Effect of shading and grafting on yield and quality of tomato.

BACKGROUND: The need to increase marketable tomato yields and decrease losses due to sunburn and disease during the summer motivates farmers to adopt additional cultural practices such as shading and grafting. To investigate complex interactions among grafting, shading, and tomato cultivar, grafted and ungrafted tomatoes (cv. 'Optima' F1 and cv. 'Big Beef' F1 ) were grown in the soil under net-house cover, using pearl and red nets, and in unshaded conditions (open fields). Tomato fruit at the red stage of maturity were used for the analysis of quality traits, and total and marketable yields were recorded during the whole production season. RESULTS: Grafting and shading in tomato production might be considered as cultivation practices to increase the marketable tomato yield. A decrease in sugar content increased the uptake of some micro elements (Fe and Zn) and macro elements (Ca). In some cases, firmer and less elastic skin may be expected due to grafting. Shading with pearl net might result in fruit with lower firmness and higher total, and particularly malic, acid content. CONCLUSION: Shading with colored nets and grafting provide alternative strategies for achieving higher fruit yields and avoiding or reducing a decrease in tomato quality caused by environmental stresses such as excessive radiation and temperature in the summer cropping season. © 2019 Society of Chemical Industry.

Arabidopsis leucine-rich repeat receptor-like kinase NILR1 is required for induction of innate immunity to parasitic nematodes.

Plant-parasitic nematodes are destructive pests causing losses of billions of dollars annually. An effective plant defence against pathogens relies on the recognition of pathogen-associated molecular patterns (PAMPs) by surface-localised receptors leading to the activation of PAMP-triggered immunity (PTI). Extensive studies have been conducted to characterise the role of PTI in various models of plant-pathogen interactions. However, far less is known about the role of PTI in roots in general and in plant-nematode interactions in particular. Here we show that nematode-derived proteinaceous elicitor/s is/are capable of inducing PTI in Arabidopsis in a manner dependent on the common immune co-receptor BAK1. Consistent with the role played by BAK1, we identified a leucine-rich repeat receptor-like kinase, termed NILR1 that is specifically regulated upon infection by nematodes. We show that NILR1 is essential for PTI responses initiated by nematodes and nilr1 loss-of-function mutants are hypersusceptible to a broad category of nematodes. To our knowledge, NILR1 is the first example of an immune receptor that is involved in induction of basal immunity (PTI) in plants or in animals in response to nematodes. Manipulation of NILR1 will provide new options for nematode control in crop plants in future.

Divergent expression of cytokinin biosynthesis, signaling and catabolism genes underlying differences in feeding sites induced by cyst and root-knot nematodes.

Cyst and root-knot nematodes are obligate parasites of economic importance with a remarkable ability to reprogram root cells into unique metabolically active feeding sites. Previous studies have suggested a role for cytokinin in feeding site formation induced by these two types of nematodes, but the mechanistic details have not yet been described. Using Arabidopsis as a host plant species, we conducted a comparative analysis of cytokinin genes in response to the beet cyst nematode (BCN), Heterodera schachtii, and the root-knot nematode (RKN), Meloidogyne incognita. We identified distinct differences in the expression of cytokinin biosynthesis, catabolism and signaling genes in response to infection by BCN and RKN, suggesting differential manipulation of the cytokinin pathway by these two nematode species. Furthermore, we evaluated Arabidopsis histidine kinase receptor mutant lines ahk2/3, ahk2/4 and ahk3/4 in response to RKN infection. Similar to our previous studies with BCN, these lines were significantly less susceptible to RKN without compromising nematode penetration, suggesting a requirement of cytokinin signaling in RKN feeding site formation. Moreover, an analysis of ahk double mutants using CycB1;1:GUS/ahk introgressed lines revealed contrasting differences in the cytokinin receptors mediating cell cycle activation in feeding sites induced by BCN and RKN.

Coronary Chronic Total Occlusion (CTO): A Review.

Coronary artery chronic total occlusions (CTO) are frequently encountered during coronary angiography; however percutaneous recanalization has historically been technically challenging and an important determinant for referral to coronary artery bypass surgery or for medical therapy alone. Recent advances in interventional equipment and innovative approaches to crossing CTO have significantly increased the success rate of percutaneous treatment. Although there is only one relevant randomized control trial (RCT) performed to date, several large, nonrandomized studies have consistently reported improvement in clinical outcomes, including improved survival and relief of angina, when successful percutaneous treatment of CTO was compared with unsuccessful revascularization. These positive observational results have encouraged the initiation of several RCTs which will provide more robust evidence on clinical outcomes of CTO-PCI compared with guideline-directed medical therapy (GDMT) alone.

A parasitic nematode releases cytokinin that controls cell division and orchestrates feeding site formation in host plants.

Sedentary plant-parasitic cyst nematodes are biotrophs that cause significant losses in agriculture. Parasitism is based on modifications of host root cells that lead to the formation of a hypermetabolic feeding site (a syncytium) from which nematodes withdraw nutrients. The host cell cycle is activated in an initial cell selected by the nematode for feeding, followed by activation of neighboring cells and subsequent expansion of feeding site through fusion of hundreds of cells. It is generally assumed that nematodes manipulate production and signaling of the plant hormone cytokinin to activate cell division. In fact, nematodes have been shown to produce cytokinin in vitro; however, whether the hormone is secreted into host plants and plays a role in parasitism remained unknown. Here, we analyzed the spatiotemporal activation of cytokinin signaling during interaction between the cyst nematode, Heterodera schachtii, and Arabidopsis using cytokinin-responsive promoter:reporter lines. Our results showed that cytokinin signaling is activated not only in the syncytium but also in neighboring cells to be incorporated into syncytium. An analysis of nematode infection on mutants that are deficient in cytokinin or cytokinin signaling revealed a significant decrease in susceptibility of these plants to nematodes. Further, we identified a cytokinin-synthesizing isopentenyltransferase gene in H. schachtii and show that silencing of this gene in nematodes leads to a significant decrease in virulence due to a reduced expansion of feeding sites. Our findings demonstrate the ability of a plant-parasitic nematode to synthesize a functional plant hormone to manipulate the host system and establish a long-term parasitic interaction.

Fast-Responding Bio-Based Shape Memory Thermoplastic Polyurethanes.

Novel fast response shape-memory polyurethanes were prepared from bio-based polyols, diphenyl methane diisocyanate and butane diol for the first time. The bio-based polyester polyols were synthesized from 9-hydroxynonanoic acid, a product obtained by ozonolysis of fatty acids extracted from soy oil and castor oil. The morphology of polyurethanes was investigated by synchrotron ultra-small angle X-ray scattering, which revealed the inter-domain spacing between the hard and soft phases, the degree of phase separation, and the level of intermixing between the hard and soft phases. We also conducted thorough investigations of the thermal, mechanical, and dielectric properties of the polyurethanes, and found that high crystallization rate of the soft segment gives these polyurethanes unique properties suitable for shape-memory applications, such as adjustable transition temperatures, high degree of elastic elongations, and good mechanical strength. These materials are also potentially biodegradable and biocompatible, therefore suitable for biomedical and environmental applications.

Secondary metabolites and metal content dynamics in Teucrium montanum L. and Teucrium chamaedrys L. from habitats with serpentine and calcareous substrate.

The purpose of this comparative analysis is the determination of the total quantity of metals (Mg, Ca, K, Ni, Fe, Mn, Zn, Cu, Cr and Pb) in soil samples, above ground plant parts and tea made of plants Teucrium montanum and T. chamaedrys from different serpentine and calcareous habitats as well as of the total quantity of phenolic compounds and antioxidant activity. The obtained results showed that the quantities of certain metals (Mg, Fe, Ni and Mn) in the soil from the serpentine habitats were greater in comparison with other metals (Ca, Zn and Pb) which were more frequently found in the soil from the calcareous habitats. The results demonstrated that the analysed plant samples from the serpentine habitats contained higher quantity of Fe, Ni and Cr as opposed to the plant samples from the calcareous habitats which contained greater quantity of Ca and Zn. Although the studied species accumulate analysed metals in different quantities, depending on the substrate type, they are not hyperaccumulators of these metals. The use of these species from serpentine habitats for tea preparation is safe to a great extent, because in spite of the determined metal absorption by plant organs, the tea does not contain dangerous quantity of heavy metals. The results showed greater total quantity of phenolic compounds and the higher level of antioxidant activity in the plant samples from serpentine habitats in comparison with the samples from calcareous habitats, which is an indicator of one of the mechanisms of adaptation to the serpentine habitat conditions.

Fourteen monogenic genes account for 15% of nephrolithiasis/nephrocalcinosis.

Nephrolithiasis is a prevalent condition with a high morbidity. Although dozens of monogenic causes have been identified, the fraction of single-gene disease has not been well studied. To determine the percentage of cases that can be molecularly explained by mutations in 1 of 30 known kidney stone genes, we conducted a high-throughput mutation analysis in a cohort of consecutively recruited patients from typical kidney stone clinics. The cohort comprised 272 genetically unresolved individuals (106 children and 166 adults) from 268 families with nephrolithiasis (n=256) or isolated nephrocalcinosis (n=16). We detected 50 likely causative mutations in 14 of 30 analyzed genes, leading to a molecular diagnosis in 14.9% (40 of 268) of all cases; 20 of 50 detected mutations were novel (40%). The cystinuria gene SLC7A9 (n=19) was most frequently mutated. The percentage of monogenic cases was notably high in both the adult (11.4%) and pediatric cohorts (20.8%). Recessive causes were more frequent among children, whereas dominant disease occurred more abundantly in adults. Our study provides an in-depth analysis of monogenic causes of kidney stone disease. We suggest that knowledge of the molecular cause of nephrolithiasis and nephrocalcinosis may have practical implications and might facilitate personalized treatment.

Autosomal dominant spondylocostal dysostosis is caused by mutation in TBX6.

In humans, congenital spinal defects occur with an incidence of 0.5-1 per 1000 live births. One of the most severe syndromes with such defects is spondylocostal dysostosis (SCD). Over the past decade, the genetic basis of several forms of autosomal recessive SCD cases has been solved with the identification of four causative genes (DLL3, MESP2, LFNG and HES7). Autosomal dominant forms of SCD have also been reported, but to date no genetic etiology has been described for these. Here, we have used exome capture and next-generation sequencing to identify a stoploss mutation in TBX6 that segregates with disease in two generations of one family. We show that this mutation has a deleterious effect on the transcriptional activation activity of the TBX6 protein, likely due to haploinsufficiency. In mouse, Tbx6 is essential for the patterning of the vertebral precursor tissues, somites; thus, mutation of TBX6 is likely to be causative of SCD in this family. This is the first identification of the genetic cause of an autosomal dominant form of SCD, and also demonstrates the potential of exome sequencing to identify genetic causes of dominant diseases even in small families with few affected individuals.

Effect of coloured shade-nets on plant leaf parameters and tomato fruit quality.

BACKGROUND: The concept of photo-selective netting using commercial cultivation practices was studied in a tomato (Solanum lycopersicum 'Vedetta') summer cultivation in south Serbia (under high solar radiation 910 W m(-2) , with a photosynthetic photon flux density of 1661 µmol m(-2) s(-1) ), under four different coloured shade-nets (pearl, red, blue and black) with 40% relative shading. The aim of the study was to determine how different environmental control technologies (coloured shade-nets as screen house or plastic-house integrated with coloured shade-nets) could influence plant parameters, production and quality traits in tomato fruits cultivated in south Serbia (Balkan region). RESULTS: The leaf area index (LAI) ranged from 4.6 to 5.8 in open field and plastic tunnels plants (control) with maximum LAI values of 7.9-8.2 in net houses with red colour nets. Shade-grown leaves generally have higher total chlorophyll and carotenoids content than do control leaves. Pericarp thickness was significantly higher tomatoes grown under pearl (7.215.82 µm), red (7099.00 µm) and blue nets (6802.29 µm) compared to other treatments and to control (6202.48 µm). The highest concentration of lycopene was detected in tomatoes grown in plastic houses integrated with red colour nets (64.9 µg g(-1) fresh weight). The plastic house and open field (control) tomato production had a taste index mean value of 1.09-1.10. This is significantly higher than the values determined for the treatments with different coloured shade-nets. CONCLUSION: These results show that red and pearl photo-selective nets create optimal growing conditions for the growth of the plant and produce fruits with thicker pericarp, the highest lycopene content, a satisfactory level of taste index and can be further implemented within protected cultivation practices.

Mechanism, kinetics and selectivity of selenocyclization of 5-alkenylhydantoins: an experimental and computational study.

The mechanism and selectivity of a bicyclic hydantoin formation by selenium-induced cyclization are investigated. The proposed mechanism involves the intermediates formed by an electrophilic addition of the selenium reagent on a double bond of the starting 5-alkenylhydantoin prior the cyclization. These intermediates are readily converted into the more stable cyclic seleniranium cations. A key step of the mechanism is an intramolecular cyclization which is realized through an anti-attack of the internal nucleophile, the amidic nitrogen, to the seleniranium cation yielding the intermediate imidazolinium cations. Their deprotonation is followed by the formation of the fused bicyclic reaction products. Important intermediates and key transition states are studied by using density functional theory (DFT) methods. The pathways of the reaction are investigated in detail. There are two regioselective pathways related to 5-exo and 6-endo products. Theoretical calculations and the monitoring of the cyclization reaction using (1)H NMR spectroscopy are in a good agreement with the proposed mechanism and are consistent with our experimental results. The preferred pathway for formation of 5-exo products is confirmed.

Myo-inositol oxygenase is important for the removal of excess myo-inositol from syncytia induced by Heterodera schachtii in Arabidopsis roots.

The enzyme myo-inositol oxygenase is the key enzyme of a pathway leading from myo-inositol to UDP-glucuronic acid. In Arabidopsis, myo-inositol oxygenase is encoded by four genes. All genes are strongly expressed in syncytia induced by the beet cyst nematode Heterodera schachtii in Arabidopsis roots. Here, we studied the effect of a quadruple myo-inositol oxygenase mutant on nematode development. We performed metabolite profiling of syncytia induced in roots of the myo-inositol oxygenase quadruple mutant. The role of galactinol in syncytia was studied using Arabidopsis lines with elevated galactinol levels and by supplying galactinol to wild-type seedlings. The quadruple myo-inositol oxygenase mutant showed a significant reduction in susceptibility to H. schachtii, and syncytia had elevated myo-inositol and galactinol levels and an elevated expression level of the antimicrobial thionin gene Thi2.1. This reduction in susceptibility could also be achieved by exogenous application of galactinol to wild-type seedlings. The primary function of myo-inositol oxygenase for syncytium development is probably not the production of UDP-glucuronic acid as a precursor for cell wall polysaccharides, but the reduction of myo-inositol levels and thereby a reduction in the galactinol level to avoid the induction of defence-related genes.

Investigations on Microbes Attached to the Cuticle of Phytonematodes.

Nematodes form various associations with soil microbiome. Experimental studies on nematode-attached microbes can improve mechanistic understanding of these associations and lead to new discoveries relevant for the field of nematode biocontrol. Microbial attachment to the surface of phytonematodes is very specific and influenced by a multitude of factors, including the designation of nematodes and microbes, environmental and biological factors in soil, time of incubation, and the ratio and evolutionary trajectories between nematodes and microbes. Here, we describe how the classical nematological and microbiological techniques can be coupled with the advanced molecular tools to study the microbial attachment to phytonematodes in soil. We focus on the characterization of nematode-attached microbes using classical microbiological approaches and high-throughput amplicon sequencing and on the effects of nematode-attached microbes on plant defense responses.