Innate Mechanisms Research Articles

Effect of silymarin on the parameters of oxidative stress in hearts in the course of diabetes mellitus in Wistar rats

Chronic hyperglycemia is one of the most important causes of the formation of oxygen free radicals in diabetes, which damage various organs. Damage to proteins and lipids in the heart can lead to the development of diabetic cardiomyopathy, which in turn may cause cardiovascular failure. In order to counteract oxidative stress, the innate antioxidant mechanisms of the system should be supported by supplementation with exogenous antioxidants, for instance of plant origin. In this experiment, the role of silymarin, a plant-derived flavonolignan on the oxidative stress parameters and oxidative damage markers in the hearts was determined in an experimental model of diabetes. Male Wistar rats were injected with streptozotocin at a dose of 60 mg/kg (ip). Then, silymarin was administered at doses of 50 and 100 mg/kg for four weeks via a gastric tube. In diabetic animals an increase in malondialdehyde, advanced oxidation protein products, as well as increased activity of antioxidant enzymes in heart tissues was noted. Administration of silymarin inhibited oxidative stress by increasing the total antioxidant response, decreasing the concentration of malondialdehyde and advanced oxidation protein products and a slight increase in the level of glutathione. It can be concluded that the redox state in the examined tissue improved. Based on the results in can be concluded that silymarin demonstrates a potential in preventing the development of oxidative damage in the heart tissue in the course of experimental diabetes.

A thalamic-primary auditory cortex circuit mediates resilience to stress

Resilience enables mental elasticity in individuals when rebounding from adversity. In this study, we identified a microcircuit and relevant molecular adaptations that play a role in natural resilience. We found that activation of parvalbumin (PV) interneurons in the primary auditory cortex (A1) by thalamic inputs from the ipsilateral medial geniculate body (MG) is essential for resilience in mice exposed to chronic social defeat stress. Early attacks during chronic social defeat stress induced short-term hyperpolarizations of MG neurons projecting to the A1 (MGA1 neurons) in resilient mice. In addition, this temporal neural plasticity of MGA1 neurons initiated synaptogenesis onto thalamic PV neurons via presynaptic BDNF-TrkB signaling in subsequent stress responses. Moreover, optogenetic mimicking of the short-term hyperpolarization of MGA1 neurons, rather than merely activating MGA1 neurons, elicited innate resilience mechanisms in response to stress and achieved sustained antidepressant-like effects in multiple animal models, representing a new strategy for targeted neuromodulation.

B cells in the tumor microenvironment: Multi-faceted organizers, regulators, and effectors of anti-tumor immunity.

Our understanding of tumor-infiltrating lymphocytes (TILs) is rapidly expanding beyond Tcell-centric perspectives to include B cells and plasma cells, collectively referred to as TIL-Bs. In many cancers, TIL-Bs carry strong prognostic significance and are emerging as key predictors of response to immune checkpoint inhibitors. TIL-Bs can perform multiple functions, including antigen presentation and antibody production, which allow them to focus immune responses on cognate antigen to support both Tcell responses and innate mechanisms involving complement, macrophages, and natural killer cells. In the stroma of the most immunologically "hot" tumors, TIL-Bs are prominent components of tertiary lymphoid structures, which resemble lymph nodes structurally and functionally. Additionally, TIL-Bs participate in a variety of other lympho-myeloid aggregates and engage in dynamic interactions with the tumor stroma. Here, we summarize our current understanding of TIL-Bs in human cancer, highlighting the compelling therapeutic opportunities offered by their unique tumor recognition and effector mechanisms.

Response and Resistance to Trametinib in MAP2K1-Mutant Triple-Negative Melanoma

The development of targeted therapies for non-BRAF p.Val600-mutant melanomas remains a challenge. Triple wildtype (TWT) melanomas that lack mutations in BRAF, NRAS, or NF1 form 10% of human melanomas and are heterogeneous in their genomic drivers. MAP2K1 mutations are enriched in BRAF-mutant melanoma and function as an innate or adaptive resistance mechanism to BRAF inhibition. Here we report the case of a patient with TWT melanoma with a bona fide MAP2K1 mutation without any BRAF mutations. We performed a structural analysis to validate that the MEK inhibitor trametinib could block this mutation. Although the patient initially responded to trametinib, he eventually progressed. The presence of a CDKN2A deletion prompted us to combine a CDK4/6 inhibitor, palbociclib, with trametinib but without clinical benefit. Genomic analysis at progression showed multiple novel copy number alterations. Our case illustrates the challenges of combining MEK1 and CDK4/6 inhibitors in case of resistance to MEK inhibitor monotherapy.

Advances in immune checkpoint inhibitor combination strategies for microsatellite stable colorectal cancer.

Immune checkpoint inhibitors have reshaped the prognostic of several tumor types, including metastatic colorectal tumors with microsatellite instability (MSI). However, 90-95% of metastatic colorectal tumors are microsatellite stable (MSS) in which immunotherapy has failed to demonstrate meaningful clinical results. MSS colorectal tumors are considered immune-cold tumors. Several factors have been proposed to account for this lack of response to immune checkpoint blockade including low levels of tumor infiltrating lymphocytes, low tumor mutational burden, a high rate of WNT/β-catenin pathway mutations, and liver metastases which have been associated with immunosuppression. However, studies with novel combinations based on immune checkpoint inhibitors are showing promising activity in MSS colorectal cancer. Here, we review the underlying biological facts that preclude immunotherapy activity, and detail the different immune checkpoint inhibitor combinations evaluated, along with novel immune-based therapies, to overcome innate mechanisms of resistance in MSS colorectal cancer.

Chemical Basis of Combination Therapy to Combat Antibiotic Resistance.

The antimicrobial resistance crisis is a global health issue requiring discovery and development of novel therapeutics. However, conventional screening of natural products or synthetic chemical libraries is uncertain. Combination therapy using approved antibiotics with inhibitors targeting innate resistance mechanisms provides an alternative strategy to develop potent therapeutics. This review discusses the chemical structures of effective β-lactamase inhibitors, outer membrane permeabilizers, and efflux pump inhibitors that act as adjuvant molecules of classical antibiotics. Rational design of the chemical structures of adjuvants will provide methods to impart or restore efficacy to classical antibiotics for inherently antibiotic-resistant bacteria. As many bacteria have multiple resistance pathways, adjuvant molecules simultaneously targeting multiple pathways are promising approaches to combat multidrug-resistant bacterial infections.

Anti-Inflammatory Actions of G-Protein-Coupled Estrogen Receptor 1 (GPER) and Brain-Derived Estrogen Following Cerebral Ischemia in Ovariectomized Rats.

Global cerebral ischemia can elicit rapid innate neuroprotective mechanisms that protect against delayed neuronal death. Brain-derived 17β-estradiol (BDE2), an endogenous neuroprotectant, is synthesized from testosterone by the enzyme aromatase (Aro) and is upregulated by brain ischemia and inflammation. Our recent study revealed that G1, a specific G-protein-coupled estrogen receptor 1 (GPER) agonist, exerts anti-inflammatory and anti-apoptotic roles after global cerebral ischemia (GCI). Herein, we aimed to elucidate whether G1 modulates the early inflammatory process and the potential underlying mechanisms in the ovariectomized rat hippocampal CA1 region. G1 was found to markedly reduce pro-inflammatory (iNOS, MHCII, and CD68) and to enhance anti-inflammatory (CD206, Arginase 1, IL1RA, PPARγ, and BDNF) markers after 1 and 3 days of reperfusion after GCI. Intriguingly, the neuroprotection of G1 was blocked by the Aro inhibitor, letrozole. Conversely, the GPER antagonist, G36, inhibited Aro-BDE2 signaling and exacerbated neuronal damage. As a whole, this work demonstrates a novel anti-inflammatory role of GPER, involving a synergistic mediation with BDE2 during the early stage of GCI.

A Model for Changes in Germination Synchrony and Its Implements to Study Weed Population Dynamics: A Case Study of Brassicaceae

In every agricultural system, weed seeds can be found in every cubic centimeter of soil. Weed seeds, as a valuable trait underlying the fate of weed populations, exhibit differing levels of seed dormancy, ensuring their survival under uncertain conditions. Seed dormancy is considered as an innate mechanism that constrains germination under suitable conditions that would otherwise stimulate germination of nondormant seeds. This work provides new insight into changes in germination patterns along the dormant to nondormancy continuum in seeds with physiological dormancy. Notable findings are: (1) germination synchrony can act as a new parameter that quantitatively describes dormancy patterns and, subsequently, weed population dynamics, (2) germination synchrony is dynamic, suggesting that the more dormancy decreases, the more synchrony is obtainable, (3) after-ripening and stratification can function as a synchronizing agent that regulates germination behavior. Freshly harvested seeds of Brassica napus with type 3 of non-deep physiological dormancy showed the most synchronous germination, with a value of 3.14, while a lower level of germination asynchrony was found for newly harvested seeds of Sinapis arvensis with type 1 of non-deep physiological dormancy, with an asynchrony value of 2.25. After-ripening and stratification can act as a synchronizing factor through decreasing the asynchrony level and increasing synchrony. There is a firm relationship between seed dormancy cycling and germination synchrony patterns, ensuring their survival and reproductive strategies. By germinating in synchrony, which is accompanied by cycling mechanisms, weeds have more opportunities to persist. The synchrony model used in the present study predicts germination behavior and synchrony along the dormant to nondormancy continuum in weed seeds with physiological dormancy, suggesting a useful method for the quantification of germination strategies and weed population dynamics.

Order of statistical learning depends on perceptive uncertainty.

Statistical learning (SL) is an innate mechanism by which the brain automatically encodes the n-th order transition probability (TP) of a sequence and grasps the uncertainty of the TP distribution. Through SL, the brain predicts a subsequent event (e n+1 ) based on the preceding events (e n ) that have a length of "n". It is now known that uncertainty modulates prediction in top-down processing by the human predictive brain. However, the manner in which the human brain modulates the order of SL strategies based on the degree of uncertainty remains an open question. The present study examined how uncertainty modulates the neural effects of SL and whether differences in uncertainty alter the order of SL strategies. It used auditory sequences in which the uncertainty of sequential information is manipulated based on the conditional entropy. Three sequences with different TP ratios of 90:10, 80:20, and 67:33 were prepared as low-, intermediate, and high-uncertainty sequences, respectively (conditional entropy: 0.47, 0.72, and 0.92 bit, respectively). Neural responses were recorded when the participants listened to the three sequences. The results showed that stimuli with lower TPs elicited a stronger neural response than those with higher TPs, as demonstrated by a number of previous studies. Furthermore, we found that participants adopted higher-order SL strategies in the high uncertainty sequence. These results may indicate that the human brain has an ability to flexibly alter the order based on the uncertainty. This uncertainty may be an important factor that determines the order of SL strategies. Particularly, considering that a higher-order SL strategy mathematically allows the reduction of uncertainty in information, we assumed that the brain may take higher-order SL strategies when encountering high uncertain information in order to reduce the uncertainty. The present study may shed new light on understanding individual differences in SL performance across different uncertain situations.

Immune Biology and Persistence of Helicobacter pylori in Gastric Diseases.

Helicobacter pylori is a prevalent pathogen, which affects more than 40% of the global population. It colonizes the human stomach and persists in its host for several decades or even a lifetime, if left untreated. The persistent infection has been linked to various gastric diseases, including gastritis, peptic ulcers, and an increased risk for gastric cancer. H. pylori infection triggers a strong immune response directed against the bacterium associated with the infiltration of innate phagocytotic immune cells and the induction of a Th1/Th17 response. Even though certain immune cells seem to be capable of controlling the infection, the host is unable to eliminate the bacteria as H. pylori has developed remarkable immune evasion strategies. The bacterium avoids its killing through innate recognition mechanisms and manipulates gastric epithelial cells and immune cells to support its persistence. This chapter focuses on the innate and adaptive immune response induced by H. pylori infection, and immune evasion strategies employed by the bacterium to enable persistent infection.

Associating multiple vision transformer layers for fine-grained image representation

- Accurate discriminative region proposal has an important effect for fine-grained image recognition. The vision transformer (ViT) brings about a striking effect in computer vision due to its innate multi-head self-attention mechanism. However, the attention maps are gradually similar after certain layers, and since ViT used a classification token to achieve classification, it is unable to effectively select discriminative image patches for fine-grained image classification. To accurately detect discriminative regions, we propose a novel network AMTrans, which efficiently increases layers to learn diverse features and utilizes integrated raw attention maps to capture more salient features. Specifically, we employ DeepViT as backbone to solve the attention collapse issue. Then, we fuse each head attention weight within each layer to produce an attention weight map. After that, we alternatively use recurrent residual refinement blocks to promote salient feature and then utilize the semantic grouping method to propose the discriminative feature region. A lot of experiments prove that AMTrans acquires the SOTA performance on four widely used fine-grained datasets under the same settings, involving Stanford-Cars, Stanford-Dogs, CUB-200-2011, and ImageNet.

Dhatusarata and its utility in the assessment of innate immunity: A review

Acharya Sushruta says that the supreme essence of all the Dhatus beginning with Rasa and ending with Shukra is known as Ojas and that is the Bala (strenth) of the body. And strength is not depending only on physical build up but it depends on the Sara means the excellence quality of Dhatus. In the Ayurvedic text, this Dhatusarata is described with respect to Sapta Dhatu and Sattva (mind) i.e. Ashtavidhsarata which is helpful in determining the degree of strength. Examination of Dhatusarata is done at physical and psychological level. Thus biological immunity can refer to constitutive physical innate mechanisms, such as the physical protection afforded against infection by skin, the activity of natural killer (NK) cells against virus-infected cells, or the natural resistance power of the body. It is of two types innate and adoptive. Innate immunity, also known as native immunity. It is defined as the first line of defense against pathogens, representing a critical systemic response to prevent infection and maintain homeostasis, contributing to the activation of an adaptive immune response. Aim: To interpret the utility of Dhatusarata in the assessment of Innate immunity. Objectives: To take a review on Innate immunity as per Modern science to interpret the utility of Dhatusarata in the assessment of innate immunity.

Antiaging properties of antioxidant photoprotective polymeric nanoparticles loaded with coenzyme-Q10

Skin is the most extensive organ within our body. It is continually subjected to stress factors, among which ultraviolet irradiation, a key factor responsible in skin aging since it leads to reactive oxygen species production. In order to fight against these oxidative species, the human body has an innate robust antioxidant mechanism composed of several different substances, one of which is coenzyme Q10. Its capacity to increase cellular energy production and excellent antioxidant properties have been proved, as well as its antiaging properties being able to attenuate cellular damage induced by ultraviolet irradiation in human dermal fibroblasts. However, its high hydrophobicity and photolability hampers its therapeutic potential. In this context, the objective of this work consists of the preparation of chitosan-rosmarinic acid conjugate-based nanoparticles to encapsulate coenzyme Q10 with high encapsulation efficiencies in order to improve its bioavailability and broaden its therapeutic use in skin applications. Hyaluronic acid coating was performed giving stable nanoparticles at physiological pH with 382 ± 3 nm of hydrodynamic diameter (0.04 ± 0.02 polydispersity) and − 18 ± 3 mV of surface charge. Release kinetics studies showed a maximum of 82 % mass release of coenzyme Q10 after 40 min, and radical scavenger activity assay confirmed the antioxidant character of chitosan-rosmarinic acid nanoparticles. Hyaluronic acid-coated chitosan-rosmarinic acid nanoparticles loaded with coenzyme Q10 were biocompatible in human dermal fibroblasts and exhibited interesting photoprotective properties in ultraviolet irradiated cells. In addition, nanoparticles hindered the production of reactive oxygen species, interleukin-6 and metalloproteinase-1, as well as caspase-9 activation maintaining high viability values upon irradiation of dermal fibroblasts. Overall results envision a great potential of these nanovehicles for application in skin disorders or antiaging treatments.

Heat and drought induced transcriptomic changes in barley varieties with contrasting stress response phenotypes.

Drought and heat stress substantially impact plant growth and productivity. When subjected to drought or heat stress, plants exhibit reduction in growth resulting in yield losses. The occurrence of these two stresses together intensifies their negative effects. Unraveling the molecular changes in response to combined abiotic stress is essential to breed climate-resilient crops. In this study, transcriptome profiles were compared between stress-tolerant (Otis), and stress-sensitive (Golden Promise) barley genotypes subjected to drought, heat, and combined heat and drought stress for five days during heading stage. The major differences that emerged from the transcriptome analysis were the overall number of differentially expressed genes was relatively higher in Golden Promise (GP) compared to Otis. The differential expression of more than 900 transcription factors in GP and Otis may aid this transcriptional reprogramming in response to abiotic stress. Secondly, combined heat and water deficit stress results in a unique and massive transcriptomic response that cannot be predicted from individual stress responses. Enrichment analyses of gene ontology terms revealed unique and stress type-specific adjustments of gene expression. Weighted Gene Co-expression Network Analysis identified genes associated with RNA metabolism and Hsp70 chaperone components as hub genes that can be useful for engineering tolerance to multiple abiotic stresses. Comparison of the transcriptomes of unstressed Otis and GP plants identified several genes associated with biosynthesis of antioxidants and osmolytes were higher in the former that maybe providing innate tolerance capabilities to effectively combat hostile conditions. Lines with different repertoire of innate tolerance mechanisms can be effectively leveraged in breeding programs for developing climate-resilient barley varieties with superior end-use traits.

Cold exposure impairs extracellular vesicle swarm–mediated nasal antiviral immunity

The human upper respiratory tract is the first site of contact for inhaled respiratory viruses and elaborates an array of innate immune responses. Seasonal variation in respiratory viral infections and the importance of ambient temperature in modulating immune responses to infections have been well recognized; however, the underlying biological mechanisms remain understudied. We investigated the role of nasal epithelium-derived extracellular vesicles (EVs) in innate Toll-like receptor 3 (TLR3)-dependent antiviral immunity. We evaluated the secretion and composition of nasal epithelial EVs after TLR3 stimulation in human autologous cells and fresh human nasal mucosal surgical specimens. We also explored the antiviral activity and mechanisms of TLR3-stimulated EVs against respiratory viruses as well as the effect ofcool ambient temperature on TLR3-dependent antiviral immunity. We found that polyinosinic:polycytidylic acid, aka poly(I:C), exposure induced a swarm-like increase in the secretion of nasal epithelial EVs via the TLR3 signaling. EVs participated in TLR3-dependent antiviral immunity, protecting the host from viral infections through both EV-mediated functional delivery of miR-17 and direct virion neutralization after binding to virus ligands via surface receptors, including LDLR and ICAM-1. These potent antiviral immune defense functions mediated by TLR3-stimulated EVs were impaired by cold exposure via a decrease in total EV secretion as well as diminished microRNA packaging and antiviral binding affinity of individual EV. TLR3-dependent nasal epithelial EVs exhibit multiple innate antiviral mechanisms to suppress respiratory viral infections. Furthermore, our study provides a direct quantitative mechanistic explanation for seasonal variation in upper respiratory tract infection prevalence.

41P Single-cell signalling analysis of engineered γδ T cell biotherapeutics for the treatment of colorectal cancer

Colorectal cancer (CRC) is a devastating disease that kills ∼700,000 people worldwide annually, with immunotherapies struggling against the immunosuppressive CRC tumour microenvironment (TME). Tumour infiltrating γδ T cells confer a prognostic benefit to CRC patients and can kill cancer via multiple innate and adaptive mechanisms, including antibody-dependent cellular cytotoxicity (ADCC) against tumour antigens (e.g., B7-H3). We hypothesise that γδ T cells can be exploited as an anti-CRC biotherapeutic but their complex interactions within the CRC TME need to be elucidated.

Parental bonding in retrospect and adult attachment style: A comparative study between Spanish, Italian and Japanese cultures.

Attachment is an innate human relational mechanism that develops progressively from early childhood, influences individuals' representations and behaviors, shapes relationships, and affects the social and cultural environment. Parental bonding refers to the ability of parents to be emotionally and behaviorally available to the child during infancy. Attachment style refers to the individual's relational attitude in close relationships that influences adult love, bonding, handling relationships, and social exploration. The role of intergenerational, cultural and developmental factors influencing the relationship between the attachment style in adulthood and the parental bonding style recalled during childhood has been debated. This study explores the relationships between recalled parental bonding, adult attachment style, and cultural background in a sample of Spanish, Italian, and Japanese adults using a cross-sectional and cross-cultural design. For this purpose, the validated versions of the Experience in Close Relationship Scale and the Parental Bonding Instrument were administered to a non-clinical population of three hundred and five participants in the three countries. Results show that the most frequent adult attachment style is the secure style, followed by the dismissing-avoidant, the preoccupied, and the fearful-avoidant style. The dismissing-avoidant style was the most frequent insecure attachment style in the Japanese sample whereas the preoccupied style was the most frequent insecure attachment style in the Italians and Spaniards. Japanese are more anchored to the memory of maternal and paternal overprotection, which is related to more avoidance in actual close relationships. Spaniard's current relationships are mildly independent of recalled parental bonding, showing an association between lower current avoidance to primary parental care. In the Italian sample, there is no significant relationship between current adult close relationships and recalled parental bonding. These results suggest that different cultural models influence adult attachment representations differently, in terms of the weight placed on attachment-related avoidance, attachment-related anxiety, care, and overprotection in infant and adult relationships.

Transcriptomic Signature of Horseshoe Crab Carcinoscorpius rotundicauda Hemocytes' Response to Lipopolysaccharides.

Carcinoscorpius rotundicauda (C. rotundicauda) is one of the four species of horseshoe crabs (HSCs). The HSC hemocytes store defense molecules that are released upon encountering invading pathogens. The HSCs rely on this innate immunity to continue its existence as a living fossil for more than 480 million years. To gain insight into the innate mechanisms involved, transcriptomic analysis was performed on isolated C. rotundicauda hemocytes challenged with lipopolysaccharides (LPS), the main components of the outer cell membrane of gram-negative bacteria. RNA-sequencing with Illumina HiSeq platform resulted in 232,628,086 and 245,448,176 raw reads corresponding to 190,326,253 and 201,180,020 high-quality mappable reads from control and LPS-stimulated hemocytes, respectively. Following LPS-stimulation, 79 genes were significantly upregulated and 265 genes were downregulated. The differentially expressed genes (DEGs) were related to multiple immune functional categories and pathways such as those of the cytoskeleton, Toll and Imd, apoptosis, MAP kinase (MAPK), inositol phosphate metabolism, phagosome, leucocyte endothelial migration, and gram-negative bacterial infection, among others. This study provides important information about the mechanisms of response to LPS, which is relevant for the understanding the HSCs' immune response.

Genome-Wide Analysis of Innate Susceptibility Mechanisms of Escherichia coli to Colistin.

Colistin is an antibiotic that has seen increasing clinical use for the treatment of human infections caused by Gram-negative pathogens, particularly due to the emergence of multidrug-resistant pathogens. Colistin resistance is also a growing problem and typically results from alterations to lipopolysaccharides mediated by phosphoethanolamine (pETn) transferase enzymes which can be encoded on the chromosome, or plasmids. In this study, we used 'TraDIS-Xpress' (Transposon Directed Insertion site Sequencing with expression), where a high-density transposon mutant library including outward facing promoters in Escherichia coli BW25113 identified genes involved in colistin susceptibility. We examined the genome-wide response of E. coli following exposure to a range of concentrations of colistin. Our TraDIS-Xpress screen confirmed the importance of overexpression of the two-component system basSR (which regulates pETn transferases) but also identified a wider range of genes important for survival in the presence of colistin, including genes encoding membrane associated proteins, DNA repair machinery, various transporters, RNA helicases, general stress response genes, fimbriae and phosphonate metabolism. Validation experiments supported a role in colistin susceptibility for novel candidate genes tested. TraDIS-Xpress is a powerful tool that expands our understanding of the wider landscape of genes involved in response to colistin susceptibility mechanisms.

Similar Characteristics of siRNAs of Plant Viruses Which Replicate in Plant and Fungal Hosts

RNA silencing is a host innate antiviral mechanism which acts via the synthesis of viral-derived small interfering RNAs (vsiRNAs). We have previously reported the infection of phytopathogenic fungi by plant viruses such as cucumber mosaic virus (CMV) and tobacco mosaic virus (TMV). Furthermore, fungal RNA silencing was shown to suppress plant virus accumulation, but the characteristics of plant vsiRNAs associated with the antiviral response in this nonconventional host remain unknown. Using high-throughput sequencing, we characterized vsiRNA profiles in two plant RNA virus-fungal host pathosystems: CMV infection in phytopathogenic fungus Rhizoctonia solani and TMV infection in phytopathogenic fungus Fusarium graminearum. The relative abundances of CMV and TMV siRNAs in the respective fungal hosts were much lower than those in the respective experimental plant hosts, Nicotiana benthamiana and Nicotiana tabacum. However, CMV and TMV siRNAs in fungi had similar characteristics to those in plants, particularly in their size distributions, proportion of plus and minus senses, and nucleotide preference for the 5' termini of vsiRNAs. The abundance of TMV siRNAs largely decreased in F. graminearum mutants with a deletion in either dicer-like 1 (dcl1) or dcl2 genes which encode key proteins for the production of siRNAs and antiviral responses. However, deletion of both dcl1 and dcl2 restored TMV siRNA accumulation in F. graminearum, indicating the production of dcl-independent siRNAs with no antiviral function in the absence of the dcl1 and dcl2 genes. Our results suggest that fungal RNA silencing recognizes and processes the invading plant RNA virus genome in a similar way as in plants.