Radiomics in cone-beam breast CT for the prediction of axillary lymph node metastasis in breast cancer: a multi-center multi-device study.

OBJECTIVES: To develop a radiomics model in contrast-enhanced cone-beam breast CT (CE-CBBCT) for preoperative prediction of axillary lymph node (ALN) status and metastatic burden of breast cancer. METHODS: Two hundred and seventy-four patients who underwent CE-CBBCT examination with two scanners between 2012 and 2021 from two institutions were enrolled. The primary tumor was annotated in each patient image, from which 1781 radiomics features were extracted with PyRadiomics. After feature selection, support vector machine models were developed to predict ALN status and metastatic burden. To avoid overfitting on a specific patient subset, 100 randomly stratified splits were made to assign the patients to either training/fine-tuning or test set. Area under the receiver operating characteristic curve (AUC) of these radiomics models was compared to those obtained when training the models only with clinical features and combined clinical-radiomics descriptors. Ground truth was established by histopathology. RESULTS: One hundred and eighteen patients had ALN metastasis (N + (≥ 1)). Of these, 74 had low burden (N + (1~2)) and 44 high burden (N + (≥ 3)). The remaining 156 patients had none (N0). AUC values across the 100 test repeats in predicting ALN status (N0/N + (≥ 1)) were 0.75 ± 0.05 (0.67~0.93, radiomics model), 0.68 ± 0.07 (0.53~0.85, clinical model), and 0.74 ± 0.05 (0.67~0.88, combined model). For metastatic burden prediction (N + (1~2)/N + (≥ 3)), AUC values were 0.65 ± 0.10 (0.50~0.88, radiomics model), 0.55 ± 0.10 (0.40~0.80, clinical model), and 0.64 ± 0.09 (0.50~0.90, combined model), with all the ranges spanning 0.5. In both cases, the radiomics model was significantly better than the clinical model (both p < 0.01) and comparable with the combined model (p = 0.56 and 0.64). CONCLUSIONS: Radiomics features of primary tumors could have potential in predicting ALN metastasis in CE-CBBCT imaging. CLINICAL RELEVANCE STATEMENT: The findings support potential clinical use of radiomics for predicting axillary lymph node metastasis in breast cancer patients and addressing the limited axilla coverage of cone-beam breast CT. KEY POINTS: • Contrast-enhanced cone-beam breast CT-based radiomics could have potential to predict N0 vs. N + (≥ 1) and, to a limited extent, N + (1~2) vs. N + (≥ 3) from primary tumor, and this could help address the limited axilla coverage, pending future verifications on larger cohorts. • The average AUC of radiomics and combined models was significantly higher than that of clinical models but showed no significant difference between themselves. • Radiomics features descriptive of tumor texture were found informative on axillary lymph node status, highlighting a higher heterogeneity for tumor with positive axillary lymph node.

Horizontal transfer and driving factors of extended-spectrum ß-lactamase-producing resistance genes in mice intestine after the ingestion of contaminated water.

Extended-spectrum ß-lactamase (ESBL)-producing Escherichia coli (E. coli) has been identified in various water environments, posing a serious risk to public health. However, whether and how ESBL-producing genes in water-derived E. coli can spread among mammalian gut microbiota via drinking water is largely unclear. To address this problem, horizontal transfer characterization of ESBL-producing genes in mice gut microbiota was determined after the oral ingestion of contaminated water by ESBL-producing E. coli, and then the driving factors were comprehensively examined from multiple different perspectives. The results showed that water-borne ESBL-producing E. coli can colonize in the mice intestine, the ESBL-producing genes can horizontally spread among gut microbiota, and the recipient bacteria include opportunistic pathogens Klebsiella pneumoniae and Salmonella enterica. This horizontal spread may be attributed to the intestinal micro-environment changes caused by the ingestion of contaminated water by ESBL-producing E. coli. These changes, including gut microbiota diversity, increased levels of inflammatory response and reactive oxygen species, cell membrane permeability, and expression levels of conjugative transfer-related genes, are all major driving factors for horizontal transfer of ESBL-producing genes in mice gut microbiota. Our findings highlight the potential for ESBL-producing E. coli to spread resistance genes to mammalian gut microbiota during ingestion of contaminated water.

Clinical Research Progress of Small Molecule Compounds Targeting Nrf2 for Treating Inflammation-Related Diseases.

Studies have found that inflammation is a symptom of various diseases, such as coronavirus disease 2019 (COVID-19) and rheumatoid arthritis (RA); it is also the source of other diseases, such as Alzheimer's disease (AD), Parkinson's disease (PD), lupus erythematosus (LE), and liver damage. Nrf2 (nuclear factor erythroid 2-related factor 2) is an important multifunctional transcription factor in cells and plays a central regulatory role in cellular defense mechanisms. In recent years, several studies have found a strong association between the activation of Nrf2 and the fight against inflammation-related diseases. A number of small molecule compounds targeting Nrf2 have entered clinical research. This article reviews the research status of small molecule compounds that are in clinical trials for the treatment of COVID-19, rheumatoid arthritis, Alzheimer's disease, Parkinson's disease, lupus erythematosus, and liver injury.

An infrared image-enhancement algorithm in simulated prosthetic vision: Enlarging working environment of future retinal prostheses.

BACKGROUND: Most existing retinal prostheses contain a built-in visible-light camera module that captures images of the surrounding environment. Thus, in case of insufficient or lack of visible light, the camera fails to work, and the retinal prostheses enter a dormant or "OFF" state. A simple and effective solution is replacing the visible-light camera with a dual-mode camera. The present research aimed to achieve two main purposes: (1) to explore whether the dual-mode camera in prosthesis recipients works under no visible-light conditions and (2) to assess its performance. METHODS: To accomplish these aims, we enrolled subjects in a psychophysical experiment under simulated prosthetic vision conditions. We found that the subjects could complete some simple visual tasks, but the recognition performance under the infrared mode was significantly inferior to that under the visible-light mode. These results inspired us to develop and propose a feasible infrared image-enhancement processing algorithm. Another psychophysical experiment was performed to verify the feasibility of the algorithm. RESULTS: The obtained results showed that the average efficiency of the subjects completing visual tasks using our enhancement algorithm (0.014 ± 0.001) was significantly higher (p < 0.001) than that of subjects using direct pixelization (0.007 ± 0.001). CONCLUSIONS: We concluded that a dual-mode camera could be a feasible solution to improving the performance of retinal prostheses as the camera adapted better to the specific existing ambient light conditions. Dual-mode cameras combined with this infrared image-enhancement algorithm could provide a promising direction for the design of future retinal prostheses.

The visual cortical responses to sinusoidal transcorneal electrical stimulation.

Retinal stimulation has become a widely utilized approach to restore visual function for individuals with retinal degenerative diseases. Although the rectangular electrical pulse is the primary stimulus waveform used in retinal neuromodulation, it remains unclear whether alternate waveforms may be more effective. Here, we used the optical intrinsic signal imaging system to assess the responses of cats' visual cortex to sinusoidal electrical stimulation through contact lens electrode, analyzing the response to various stimulus parameters (frequency, intensity, pulse width). A comparison between sinusoidal and rectangular stimulus waveform was also investigated. The results indicated that the optimal stimulation frequency for sinusoidal electrical stimulation was approximately 20 Hz, supporting the hypothesis that low-frequency electrostimulation induces more responsiveness in retinal neurons than high-frequency electrostimulation in case of sinusoidal stimulation. We also demonstrated that for low-frequency retinal neuromodulation, sinusoidal pulses are more effective than rectangular ones. In addition, we found that compared to current intensity, the effect of the sinusoidal pulse width on cortical responses was more prominent. These results suggested that sinusoidal electrical stimulation may provide a promising strategy for improved retinal neuromodulation in clinical settings.

Bacillus amyloliquefaciens BLCC1-0238 Alone or in Combination with Mannan-Oligosaccharides Alleviates Subclinical Necrotic Enteritis in Broilers.

Subclinical necrotic enteritis (SNE) is a severe intestinal disease in broilers which brings huge economic losses to poultry industry. Herein, the effects of Bacillus amyloliquefaciens BLCC1-0238 (B. amyloliquefaciens BLCC1-0238) alone or in combination with mannan-oligosaccharides (MOS) on the SNE challenge model in broilers were comprehensively explored. A total of 360 broilers were randomly divided into 4 groups, including an SNE infection control (IC), an antibiotic pretreatment control (AC), a B. amyloliquefaciens BLCC1-0238 pretreatment (BP), and a B. amyloliquefaciens BLCC1-0238 + MOS pretreatment (BMP). The results showed that compared with the IC, three pretreatment groups significantly improved the growth performance, lowered the overall mortality, and reduced intestinal mucosal lesions in broilers. Additionally, the expression levels of claudin-3 and peroxisome proliferator-activated receptor-gamma coactivator-1α in the BP and BMP groups and the levels of mucin-2 and mechanistic target of rapamycin in the BMP group were significantly upregulated compared with the IC. By contrast, the expression levels of interferon-γ, interleukin-10, and secretory immunoglobulin A in the BP and BMP groups were significantly downregulated. In conclusion, these findings show that B. amyloliquefaciens BLCC1-0238 in combination with MOS can exert synergetic effects by the interplay between them on improving growth performance and combating the SNE infection in broilers.

[Animal Manure Fertilization Promotes Antibiotic Resistance Gene Dissemination Among Manure, Soil, and Vegetables].

Increasing antibiotic resistant genes (ARG) are being detected in animal manure-amended soil, causing public concerns. However, the effects of animal manure fertilization on the ARG of vegetable endophytes remain unknown. In this study, high-throughput quantitative PCR (HT-qPCR) was used to explore the effects of poultry manure fertilization on the bacterial community and resistome in the soil, root endophytes, and leaf endophytes from Cichorium endivia L.. The results indicated that poultry manure fertilization not only enhanced the number of ARG in soil and root endophytes, but also increased the abundance of ARG in the soil, root endophytes, and leaf endophytes. The relationship analysis demonstrated that the ARG profile in the soil and vegetable endophytes was significantly correlated with the bacterial community, and the ARG abundance was highly correlated with Proteobacteria, Acidobacteria, Actinobacteria, and Cyanobacteria. In addition, the overlapping ARG between the Cichorium endivia L. endophyte and soil indicated the internal spread route of the ARG from the manure to Cichorium endivia L through the soil. In conclusion, poultry manure fertilization increased the diversity and abundance of the ARG in vegetables via the manure-soil-vegetable pathway.

Occurrence and characteristics of Escherichia coli mcr-1-like in rabbits in Shandong, China.

Polymyxin is regarded as the last retort to fight against multidrug-resistant (MDR) Enterobacteriaceae. The emergency and spread of polymyxin-associated resistance gene mcr-1 evoked great panic of no medicine to cure the bacterial infection in society. mcr-1 is widespread in domestic and wild animals. Therefore, continuous monitoring of its prevalence and characteristics is required. In this study, we used a polymerase chain reaction (PCR)-based method to detect the mcr-1 of Escherichia coli isolated from rabbits of Tai'an, China, and determined the characteristics of mcr-1-bearing plasmids. A total of 55 non-duplicated E. coli was recovered from the swabs of rabbit faeces. Plasmid profiling, plasmid and chromosome PCR, complete genome sequencing, a conjugation experiment, lactose fermentation experiment, multilocus sequence typing and polymyxin resistance tests were performed to determine the characteristics of mcr-1-bearing plasmids. 14.6% (8/55) of the specimens were mcr-1 positive. The mcr-1-positive E. coli harboured more drug-resistant genes compared with the mcr-1-negative specimens, and results showed four sequence types. Overall, these findings suggested the possible threat of the transmission of mcr-1 from rabbits to humans, especially since the gene is located on transferable plasmids making horizontal transfer relatively easy. Since food-producing animals are necessary for our daily diet, worldwide cooperation is needed in fighting the spread of this drug resistance gene to avoid human infections with MDR pathogenic bacteria.

Glyphosate escalates horizontal transfer of conjugative plasmid harboring antibiotic resistance genes.

Glyphosate has been frequently detected in water environments because of the wide use for controlling weed in farm lands and urban areas. Presently, the focus of the majority of studies is placed on the toxicity of glyphosate on humans and animals. However, the effects of glyphosate on horizontal transfer of conjugative plasmid carrying antibiotic resistance gene (ARG) are largely unknown. Here, we explored the ability and potential mechanism of glyphosate for accelerating horizontal transfer of conjugative plasmid-mediated ARG. The results showed that glyphosate can effectively boost horizontal transfer rate of conjugative plasmid carrying ARG. The possible mechanism analysis demonstrated that over-production of reactive oxygen species and reactive nitrogen species effectively regulated expression levels of bacterial outer membrane protein and conjugative transfer-related genes, thereby resulting into elevated horizontal transfer rate of plasmid-mediated ARG. In conclusion, this study casts new understanding into the biological effects of glyphosate on ARG.

Glyphosate-induced lipid metabolism disorder contributes to hepatotoxicity in juvenile common carp.

Residues of glyphosate (GLY) are widely detected in aquatic systems, raising potential environmental threats and public health concerns, but the mechanism underlying GLY-induced hepatotoxicity in fish has not been fully elucidated yet. This study was designed to explore the hepatotoxic mechanism using juvenile common carp exposed to GLY for 45 d, and plasma and liver samples were collected at 15 d, 30 d, and 45 d to analyze the assays. First, GLY-induced hepatic damage was confirmed by serum liver damage biomarker and hepatic histopathological analysis. Next, changes in oxidative stress biomarkers, gene expression levels of pro- and anti-inflammatory cytokines, and lipid metabolism-related parameters in collected samples were analyzed to clarify their roles in GLY-induced hepatic damage. Data showed that oxidative stress was an early event during GLY exposure, followed by hepatic inflammatory response. Lipid metabolism disorder was a late event during GLY exposure, as evidenced by overproduced hepatic free fatty acids, enhanced lipogenesis-related gene expression levels, reduced lipolysis-related gene expression levels, and resultant hepatic lipid accumulation. Collectively, these findings demonstrate that GLY induces hepatotoxicity in fish through involvement of oxidative stress, inflammatory response, and lipid metabolism disorder, which are intimately interrelated with each other during GLY exposure.

High fat diet-triggered non-alcoholic fatty liver disease: A review of proposed mechanisms.

Obesity is characterized by the deposition of excessive body fat, and is caused by energy imbalance, especially when consuming fat-rich diets. High fat diet (HFD)-associated obesity is greatly common in patients with non-alcoholic fatty liver disease (NAFLD) that is emerging as one of the most universal causes of liver disease worldwide, especially in Western countries. In spite of its high prevalence, only a small proportion of affected individuals will become inflamed, followed by fibrosis and chronic liver diseases, and most patients only show simple steatosis. In this case, the full comprehension of the mechanisms underlying the progression of NAFLD is of extreme significance; in spite of progress in this field, awareness on the development of NAFLD is still incomplete. Traditionally, liver steatosis is commonly connected with HFD, obesity, and insulin resistance (IR). Recently, various possible mechanisms have been put forward for liver damage, including endoplasmic reticulum stress, perturbation of autophagy, mitochondrial dysfunction, hepatocellular apoptosis, gut microbiota imbalance, dysregulation of microRNAs, and genetic/epigenetic risk factors, as well as an increase in inflammatory responses, among many others. Collectively, these proposed mechanisms allow for a variety of hits acting together on subjects to mediated NAFLD and will offer a more accurate explanation for progression of NAFLD. Therefore, this review summarizes the present information concerning NAFLD after HFD exposure, as well as discusses possible mechanisms through which it may arise.

mTORC1 activation contributes to autophagy inhibition via its recruitment to lysosomes and consequent lysosomal dysfunction in cadmium-exposed rat proximal tubular cells.

Autophagy dysregulation is implicated in cadmium (Cd)-induced nephrotoxicity. The mammalian target of rapamycin complex 1 (mTORC1) is a negative regulator of autophagy, but its role in Cd-induced autophagy inhibition and possible regulatory mechanisms remains poorly understood. In the present study, Cd exposure activated mTORC1 in primary rat proximal tubular (rPT) cells, and two mTORC1 inhibitors (rapamycin and torin 1) were separately utilized to inhibit Cd-induced mTORC1 activation. Data showed that Cd-inhibited autophagic flux was markedly restored by two mTORC1 inhibitors, respectively, as evidenced by immunoblot analysis of autophagy marker proteins and tandem red fluorescent protein-green fluorescent protein-microtubule associated protein light chain 3 (RFP-GFP-LC3) fluorescence microscopy assay. Importantly, Cd exposure triggered the recruitment of mTORC1 onto lysosome membrane assessed by immunofluorescence co-localization analysis, which was obviously inhibited by rapamycin or torin 1. Moreover, Cd-induced lysosomal alkalization, suppressed vacuolar ATPases (V-ATPases) protein levels and impaired lysosomal degradation capacity were markedly reversed by rapamycin or torin 1. In summary, these findings demonstrate that Cd recruits mTORC1 to lysosome membrane to induce its activation, which results in lysosomal dysfunction and resultant autophagy inhibition in rPT cells.

Protective effect and mechanism of ginsenoside Rg1 in cerebral ischaemia-reperfusion injury in mice.

BACKGROUND: Ginsenoside Rg1 is regarded as one of main bioactive compounds responsible for pharmaceutical actions of ginseng with little toxicity and has been shown to have possibly neuroprotective effects. However, the mechanism of its neuroprotection for acute ischemic stroke is still elusive. The purpose of present study is thus to assess the neuroprotective effects of the ginsenoside Rg1 against neurological injury in a mice model of cerebral ischemia/reperfusion (I/R), and then to explore the mechanisms for these neuroprotective effects. METHODS: Mices were pretreated with ginsenoside Rg1 20,40?mg?kg?1?d?1, ig, for 7d, respectively, then subjected to cerebral ischenmia (middle cerebral artery occlusion) for 2?h and reperfusion for 22?h. The infarct volume and the neurological deficit were determined by TTC staining and Longa?s scoring, respectively. The protein expression of brain-derived neurotrophic factor (BDNF) was analyzed by Immunohistochemistry and Western blot, respectively. Interleukin-1? (IL-1?), tumor necrosis factor alpha (TNF-?) and interleukin-6 (IL-6) expression in serum was measured by ELISA kit. High-performance liquid chromatography (HPLC) was used to explore the contents of Glu and Asp. RESULTS: Compared with the ischemia/reperfusion group, ginsenoside Rg1 40?mg/kg group has significantly reduced infarct volume, neurological deficit scores (P?

Antimicrobial resistance and integrons of ESBL-producing thermotolerant coliforms from a water reservoir in Tai'an, China.

INTRODUCTION: The contamination of water environments by extended-spectrum ß-lactamases-producing thermotolerant coliforms (ESBL-TC) has aroused public concern. However, little epidemiological data on ESBL-TC isolates from water reservoirs is available in China. METHODOLOGY: This study was designed to investigate antibiotic resistance, bla gene types, and the presence of integrons (class 1, 2, and 3) and gene cassettes in ESBL-TC isolated from the Huangqian Reservoir of Tai'an, China. RESULTS: A total of 96 non-duplicate ESBL-TC were obtained in this study and the ESBL genes included blaCTX-M-14 (n = 47), blaCTX-M-15 (n = 27), blaCTX-M-55 (n = 18), blaSHV-12 (n = 4), blaCTX-M-3 (n = 3), and blaCTX-M-123 (n = 1). Eighty-three of the ninety-six ESBL-TC contained class 1 integrons (86.5%), and 2 isolates harbored class 2 integrons. The sizes of gene cassette regions within integrons were ranged from 0.2 kb to 3.2 kb. CONCLUSIONS: The findings of this study indicated the widespread presence of ESBL-TC strains in the Huangqian Reservoir and spotlighted the potential role of water bodies as reservoirs for antibiotic resistant genes.

A lower dose of fluorescein sodium is more suitable for confocal laser endomicroscopy: a feasibility study.

BACKGROUND AND AIMS: Image quality can be guaranteed with the conventional dosage of fluorescein sodium in probe-based confocal laser endomicroscopy (pCLE). However, yellow discoloration of the skin seriously affects daily life and simultaneously increases the risk of adverse events such as allergic reactions. The aim of this study was to test whether a lower dosage of fluorescein sodium can provide satisfactory image quality and to compare the diagnostic accuracy of gastric intestinal metaplasia (GIM) through a randomized blind controlled trial. METHODS: Consecutive patients were randomly assigned to different doses of fluorescein sodium. Image quality was determined by the endoscopists' subjective assessments and signal-to-noise ratio (SNR) assessment systems. Skin discoloration was tested using a neonatal transcutaneous jaundice detector. In addition, consecutive patients with a known or suspected diagnosis of GIM were examined by pCLE with the lower dose and the traditional dose. RESULTS: Only 0.01 mL/kg dose of 10% fluorescein sodium led to a significant decrease in image quality (P < .05), and a dose of 0.02 mL/kg had the highest SNR value (P < .05). There were no significant differences in skin discoloration between the 0.01 mL/kg and 0.02 mL/kg doses (P = .148) and no statistical difference in the diagnostic accuracy of pCLE for GIM between the 0.02 mL/kg and 0.10 mL/kg doses (P > .05). The kappa values for the correlation between pCLE and histopathology were 0.867 (95% confidence interval, 0.782-0.952) and 0.891 (95% confidence interval, 0.811-0.971). CONCLUSIONS: The 0.02 mL/kg dose of 10% fluorescein sodium seems to be the best dose for pCLE in the upper GI tract, with comparable image quality with the conventional dose and insignificant skin discoloration. This dose is also very efficient for the diagnosis of GIM.

Characterization of Extended-Spectrum ß-Lactamase-Producing Escherichia coli Isolates from Pigs and Farm Workers.

Food-producing animals can serve as reservoirs for extended-spectrum ß-lactamase (ESBL)-producing Escherichia coli . The present study aimed to characterize and compare ESBL-carrying E. coli isolates from both pigs and farm workers. Rectal swabs were obtained from 60 pigs on four pig-fattening farms (15 samples per farm), and rectal swabs were taken from 40 farm workers on these farms (10 samples per farm). ESBL-carrying E. coli isolates from the workers and pigs were characterized by ESBL genotype, antibiotic susceptibility, enterobacterial repetitive intergenic consensus type, and multilocus sequence type. ESBL-producing E. coli was detected in 34 (56.7%) of 60 pigs, and 20.0% (8 of 40) of the farm workers were positive for ESBL-producing E. coli . More importantly, ESBL-producing E. coli isolates with the same ß-lactamase genes, antibiotic resistance profiles, enterobacterial repetitive intergenic consensus types, and multilocus sequence types were detected in both pigs and workers on the same pig farm. These findings were suggestive for transfer of ESBL-producing E. coli between animals and humans.

FvBck1, a component of cell wall integrity MAP kinase pathway, is required for virulence and oxidative stress response in sugarcane Pokkah Boeng pathogen.

Fusarium verticillioides (formerly F. moniliforme) is suggested as one of the causal agents of Pokkah Boeng, a serious disease of sugarcane worldwide. Currently, detailed molecular and physiological mechanism of pathogenesis is unknown. In this study, we focused on cell wall integrity MAPK pathway as one of the potential signaling mechanisms associated with Pokkah Boeng pathogenesis. We identified FvBCK1 gene that encodes a MAP kinase kinase kinase homolog and determined that it is not only required for growth, micro- and macro-conidia production, and cell wall integrity but also for response to osmotic and oxidative stresses. The deletion of FvBCK1 caused a significant reduction in virulence and FB1 production, a possibly carcinogenic mycotoxin produced by the fungus. Moreover, we found the expression levels of three genes, which are known to be involved in superoxide scavenging, were down regulated in the mutant. We hypothesized that the loss of superoxide scavenging capacity was one of the reasons for reduced virulence, but overexpression of catalase or peroxidase gene failed to restore the virulence defect in the deletion mutant. When we introduced Magnaporthe oryzae MCK1 into the FvBck1 deletion mutant, while certain phenotypes were restored, the complemented strain failed to gain full virulence. In summary, FvBck1 plays a diverse role in F. verticillioides, and detailed investigation of downstream signaling pathways will lead to a better understanding of how this MAPK pathway regulates Pokkah Boeng on sugarcane.

Functional characterization of Rho family small GTPases in Fusarium graminearum.

Rho GTPases have multiple cellular and metabolic functions, including vesicular trafficking and pathogenesis, as signaling molecules in fungi. Wheat scab, caused by Fusarium graminearum, is one of the most important wheat diseases worldwide, yet the mechanisms associated with making this fungus such a devastating pathogen remain largely ambiguous. In an effort to better understand F. graminearum virulence, we functionally characterized all six Rho GTPases in F. graminearum. FgRHO1 was determined to be essential for fungal survival, while FgRho3 demonstrated functions only in vegetative growth and conidiation. Other four Rho GTPases, FgRho2, FgRho4, FgCdc42 and FgRac1, were multifunctional and were involved in sexual development and pathogenesis. While FgRho2 and FgRho4 were both involved in cell wall integrity, only FgRho4 showed a role in nuclear division and septum formation. FgRho4, FgCdc42 and FgRac1 were also important for hyphal growth and conidiation. All single deletion mutants showed impaired growth, particularly in conidial morphology, when compared to the wild-type progenitor. FgRac1 deletion mutants displayed a precocious, multi-site germ tube formation as well as hyperbranching of hyphae. Significantly we learned that FgRac1 negatively controls DON production whereas FgRho4 plays a positive role. FgCla4 was identified as a downstream target of FgRac1, but was dispensable for sexual development. We determined that FgRho GTPases contribute diversely to growth, conidiogenesis, sexual reproduction, DON production and pathogenesis in F. graminearum.