Search Results (52)

Aspergillus flavus contaminates food commodities and produces carcinogenic aflatoxins. Pamamycin, a macrodiolide antibiotic from Streptomyces alboflavus TD-1, shows potent antifungal activity, yet its action against A. flavus and efficacy in complex food matrices largely remains unknown. Here, pamamycin was purified and evaluated using in vitro assays together with a peanut kernel model. Pamamycin reduced colony formation of A. flavus on PDA in a concentration-dependent manner, with near-complete inhibition at 4.0 mg/L on surface-treated PDA plates. Microscopy revealed progressive deformation and collapse of conidia and hyphae. Pamamycin increased membrane permeability, as indicated by elevated extracellular nucleic acid leakage, and impaired cell envelope integrity, as reflected by alkaline phosphatase release. In addition, pamamycin reduced Rh123-associated fluorescence, indicating an apparent dissipation of mitochondrial membrane potential under the tested conditions. Notably, at pamamycin concentrations of ≥0.5 mg/L, AFB1 accumulation was markedly reduced and fell below the limit of detection (LOD). This suppression was accompanied by distinct transcriptional changes in the aflatoxin regulatory network. RT–qPCR showed concentration-dependent repression of the pathway-specific regulators aflR and aflS, whereas the global regulator veA displayed a biphasic response with transient upregulation at lower concentrations. Notably, at 0.5 mg/L, multiple structural genes (aflC, aflD, aflK, aflP, and aflQ) were reduced to near-background transcript levels, coinciding with the loss of detectable AFB1. In inoculated peanut kernels incubated under high-humidity conditions, pamamycin significantly reduced fungal colonization and decreased AFB1 accumulation by >99%. Transcriptomic analysis of cultures treated with 0.5 mg/L pamamycin further revealed extensive transcriptional reprogramming, with enrichment of pathways related to branched-chain amino acid biosynthesis, central carbon metabolism, and ABC transporters. Collectively, pamamycin inhibits A. flavus through combined disruption of cell envelope integrity, apparent mitochondrial potential collapse, and broad suppression of the aflatoxin biosynthetic pathway, supporting its potential utility for mitigating aflatoxin contamination in peanut kernels, pending further safety evaluation. Full article

Background/Objectives: The measurement of body fat percentage (%BF) could alert us to potential respiratory problems; however, differences in %BF values have been reported depending on the method used. Therefore, the objectives of this study were to determine whether there are differences in %BF obtained through skinfold measurements (SF) and bioelectrical impedance analysis (BIA), and their correlation with airflow limitation (AFL), and secondly, to observe the relationship between leptin and AFL. Materials and Methods: A cross-sectional study was conducted with 80 participants (40 men and 40 women, aged 18–30 years). Assessments of %BF were made using SF and BIA. Spirometric parameters and pulmonary volumes were measured. Plasma leptin levels were determined using ELISA. Bivariate correlations and gender differences were analyzed. Results: When comparing %BF measured by SF and BIA, no significant differences were found between the two methods in either females or males. Furthermore, in both men and women, there was a direct and significant correlation between %BF obtained through BIA and SF (r = 0.936; r = 0.789, p < 0.001, respectively). Leptin showed a significant correlation with airway resistance (Raw) and specific airway resistance (sRaw) in men (r = 0.506; r = 0.553, p < 0.001, respectively) and women (r = 0.537, p = 0.001; r = 0.489, p = 0.003, respectively). Leptin also showed a significant correlation with %BF measured by both SF and BIA in men (r = 0.675; r = 0.687, p < 0.001, respectively) and women (r = 0.583; r = 0.682, p < 0.001, respectively). Conclusions: BIA and SF offer comparable results in estimating %BF. The significant correlation between leptin, %BF, and FEV1/FVC suggests a possible pathophysiological mechanism mediated by adiposity that could affect pulmonary function even in young and clinically healthy individuals. Full article

We introduce a lightweight 1D ConvNeXtV2–based neural network for the robust detection of atrial fibrillation (AFib) and atrial flutter (AFL) from single-lead ECG signals. Trained on multiple public datasets (Icentia11k, CPSC-2018/2021, LTAF, PTB-XL, PCC-2017) and evaluated on MIT-AFDB, MIT-ADB, and NST, our model attained a state-of-the-art F1-score of 0.986 on MIT-AFDB. With only 770 k parameters and 46 MFLOPs per 10 s window, the network remained computationally efficient. Guided Grad-CAM visualizations confirmed attention to clinically relevant P-wave morphology and R–R interval irregularities. This interpretable architecture is, therefore, well-suited for deployment in resource-constrained wearable or bedside monitors. Future work will extend this framework to multi-lead ECGs and a broader spectrum of arrhythmias. Full article

Hamstring strain injuries (HSIs) are the most common time loss injury sustained in male Australian Football League (AFL) athletes, causing significant financial cost, time cost, and impaired team and individual performance. In a squad of 42 players, HSIs accounted for 4.86 new injuries sustained by players per club per AFL season in 2020. This is consistent with injury reporting over the last decade in AFL, despite best efforts to reduce the rate. This scoping review sought to firstly identify the reported hamstring injury prevention risk factors in elite AFL, discern the impact of these factors, and map the gaps in the current literature using a biopsychosocial understanding of injury prevention. The scoping review process was based on the Askey and O’Malley framework. Five relevant online databases (MEDLINE, Proquest, CINAHL, SPORTdiscuss, and EMBASE) were systematically searched using a series of Boolean and operator terms following the PRISMA-ScR protocol using the criteria: (1) assessing male professional/elite athletes in AFL; (2) written in English and peer-reviewed; (3) full text available; and (4) published after 2006. Only manuscripts that fit the search terms and inclusion criteria were retained in the scoping review. Following an initial search, 246 potential studies were identified, with 12 studies meeting the inclusion criteria after full-text screening. The risk factors examined were subclassified into modifiable and non-modifiable categories. Modifiable factors include high-speed running exposure, gluteus medius activation, eccentric hamstring strength, shorter bicep femoris fascicle length, use of interchange, and hamstring stiffness. Non-modifiable factors include previous history of HSI and limb injury, age, and size of injury on MRI. This scoping review highlights the need for continued monitoring of high-speed running volumes as rapid increases in completed distances present as a substantial risk factor. The modifiable mechanistic risk factors of eccentric hamstring strength and hamstring stiffness were identified as important components of player screening to reduce the risk of future HSI. Risk factors identified throughout will help develop comprehensive injury profiling for athletes. Further research is warranted to develop a holistic approach to injury profiling. Full article

Poultry feeds with cereal grain-based constituents are vulnerable to fungal contamination during the processing and storage stages. A total of 100 samples of compound poultry feedstuffs were collected from the cities of Riyadh, Alhassa, Qassim, and Jeddah in Saudi Arabia. A quantitative enumeration of fungal colony-forming units (CFUs) was performed on Dichloran Rose Bengal Chloramphenicol Agar (DRBC) and Czapek Iprodione Dichloran Agar (CZID) media. Aspergillus flavus was the most predominant species, accounting for 18.714 × 10³ and 3.956 × 10³ CFU/g, with frequencies of 84 and 42% in the feed samples on DRBC and CZID media, respectively. The levels of different mycotoxins were estimated by the HPLC technique. One hundred percent of the compound poultry feedstuff samples were contaminated by mycotoxins such as AFB1, AFB2, AFG1, AFG2, FB1, DON, T2, OTA, and ZEN. Aflatoxins were recorded in 84% of the tested samples, of which 70 samples were contaminated by AFB1, ranging from 0.03 to 0.40 μg/kg. The aflatoxin analysis of the fungal species revealed that 89% and 100% of A. flavus and A. parasiticus isolates were aflatoxigenic, and all of them exhibited the presence of the aflR, omt-1, ver-1, and nor-1 genes. According to the PCR protocol based on FLA1, two primers were successful in directly and rapidly detecting A. flavus in the poultry feedstuff samples. Full article

Conidia are important for the dispersal of Aspergillus flavus, which usually generates aflatoxin B1 (AFB1) and poses a threat to the safety of agricultural food. The development of conidia is usually susceptible to changes in environmental conditions, such as nutritional status and light. However, how the light signal is involved in the conidiation in A. flavus is still unknown. In this study, LreA was identified to respond to blue light and mediate the promotion of conidiation in A. flavus, which is related to the central development pathway. At the same time, blue light inhibited the biosynthesis of AFB1, which was mediated by LreA and attributed to the transcriptional regulation of aflR and aflS expression. Our findings disclosed the function and mechanism of the blue light sensor LreA in regulating conidiation and AFB1 biosynthesis, which is beneficial for the prevention and control of A. flavus and mycotoxins. Full article

Aflatoxin B1 (AFB1) is the most potent naturally occurring carcinogen for humans and animals produced by the common fungus Aspergillus flavus (A. flavus). Aflatoxin (AF) contamination in commodities is a global concern related to the safety of food and feed, and it also impacts the agricultural economy. In this study, we investigated the AFB1-inhibiting activity of a new benzaldehyde derivative, 2-[(2-methylpyridin-3-yl)oxy]benzaldehyde (MPOBA), on A. flavus. It was found that MPOBA inhibited the production of AFB1 by A. flavus, with an IC₅₀ value of 0.55 mM. Moreover, the inhibition of conidiation was also observed at the same concentration. The addition of MPOBA resulted in decreased transcript levels of the aflR gene, which encodes a key regulatory protein for the biosynthesis of AF, and also decreased transcript levels of the global regulator genes veA and laeA. These results suggested that MPOBA has an effect on the regulatory mechanism of the development and differentiation of conidia, leading to the inhibition of AFB1 production. In addition, the cytotoxicity study showed that MPOBA had a very low cytotoxic effect on the Madin-Darby canine kidney (MDCK) cell line. Therefore, MPOBA may be a potential compound for developing practically effective agents to control AF contamination. Full article

Aflatoxins are immunosuppressive and carcinogenic secondary metabolites, produced by the filamentous ascomycete Aspergillus flavus, that are hazardous to animal and human health. In this study, we show that multiplexed host-induced gene silencing (HIGS) of Aspergillus flavus genes essential for fungal sporulation and aflatoxin production (nsdC, veA, aflR, and aflM) confers enhanced resistance to Aspergillus infection and aflatoxin contamination in groundnut (<20 ppb). Comparative proteomic analysis of contrasting groundnut genotypes (WT and near-isogenic HIGS lines) supported a better understanding of the molecular processes underlying the induced resistance and identified several groundnut metabolites that might play a significant role in resistance to Aspergillus infection and aflatoxin contamination. Fungal differentiation and pathogenicity proteins, including calmodulin, transcriptional activator-HacA, kynurenine 3-monooxygenase 2, VeA, VelC, and several aflatoxin pathway biosynthetic enzymes, were downregulated in Aspergillus infecting the HIGS lines. Additionally, in the resistant HIGS lines, a number of host resistance proteins associated with fatty acid metabolism were strongly induced, including phosphatidylinositol phosphate kinase, lysophosphatidic acyltransferase-5, palmitoyl-monogalactosyldiacylglycerol Δ-7 desaturase, ceramide kinase-related protein, sphingolipid Δ-8 desaturase, and phospholipase-D. Combined, this knowledge can be used for groundnut pre-breeding and breeding programs to provide a safe and secure food supply. Full article

The contamination of peanuts, with Aspergillus flavus and subsequent aflatoxins (AFs) is considered to be one of the most serious, safety problems in the world. Water activity (a_w) and temperature are limiting, factors for fungal growth and aflatoxin production during storage. The objectives of this study were to integrate data on the effects of temperature (34, 37, and 42 °C) and water activity (a_w; 0.85, 0.90, and 0.95) on growth rate aflatoxin B1 (AFB1) production and up- or-downregulation of the molecular expression of biosynthetic AFB1 genes divided into three types based on their A. flavus isolate composition and AFB1 capacity in vitro: A. flavus KSU114 (high producer), A. flavus KSU114 (low producer), and A. flavus KSU121 (non-producer). The A. flavus isolates were shown to be resilient in terms of growth on yeast extract sucrose agar media when exposed to temperature and water activity as pivotal environmental factors. The optimal conditions for the fungal growth of three isolates were a temperature of 34 °C and water activity of 0.95 a_w; there was very slow fungal growth at the highest temperature of 42 °C, with different a_w values causing inhibited fungal growth. The AFB1 production for the three isolates followed the same pattern with one exception: A. flavus KSU114 failed to produce any AFB1 at 42 °C with different a_w values. All tested genes of A. flavus were significantly up- or downregulated under three levels of interaction between temperature and a_w. The late structural genes of the pathway were significantly upregulated at 34 °C under a_w 0.95, although aflR, aflS and most of the early structural genes were upregulated. Compared to 34 °C with an a_w value of 0.95, most of the expressed genes were significantly downregulated at 37 and 42 °C with a_w values of 0.85 and 0.90. Additionally, two regulatory genes were downregulated under the same conditions. The expression level of laeA was also completely associated with AFB1 production, while the expression level of brlA was linked to A. flavus colonization. This information is required to forecast the actual effects of climate change on A. flavus. The findings can be applied to improve specific food technology processes and create prevention strategies to limit the concentrations of potential carcinogenic substances in peanuts and their derivatives. Full article

AF-X1 is a commercial aflatoxin biocontrol product containing the non-aflatoxigenic (AF-) strain of Aspergillus flavus MUCL54911 (VCG IT006), endemic to Italy, as an active ingredient. The present study aimed to evaluate the long-term persistence of VCG IT006 in the treated fields, and the multi-year influence of the biocontrol application on the A. flavus population. Soil samples were collected in 2020 and 2021 from 28 fields located in four provinces in north Italy. A vegetative compatibility analysis was conducted to monitor the occurrence of VCG IT006 on the total of the 399 isolates of A. flavus that were collected. IT006 was present in all the fields, mainly in the fields treated for 1 yr or 2 consecutive yrs (58% and 63%, respectively). The densities of the toxigenic isolates, detected using the aflR gene, were 45% vs. 22% in the untreated and treated fields, respectively. After displacement via the AF- deployment, a variability from 7% to 32% was noticed in the toxigenic isolates. The current findings support the long-term durability of the biocontrol application benefits without deleterious effects on each fungal population. Nevertheless, based on the current results, as well as on previous studies, the yearly applications of AF-X1 to Italian commercial maize fields should continue. Full article

Aflatoxin B1 is one of the most deleterious types of mycotoxins. The application of an endophytic fungus for biodegradation or biosuppression of AFB1 production by Aspergillus flavus was investigated. About 10 endophytic fungal species, isolated from healthy maize plants, were screened for their in vitro AFs-degrading activity using coumarin medium. The highest degradation potential was recorded for Trichoderma sp. (76.8%). This endophyte was identified using the rDNA-ITS sequence as Trichoderma harzianum AYM3 and assigned an accession no. of ON203053. It caused a 65% inhibition in the growth of A. flavus AYM2 in vitro. HPLC analysis revealed that T. harzianum AYM3 had a biodegradation potential against AFB1. Co-culturing of T. harazianum AYM3 and A. flavus AYM2 on maize grains led to a significant suppression (67%) in AFB1 production. GC-MS analysis identified two AFB1-suppressing compounds, acetic acid and n-propyl acetate. Investigating effect on the transcriptional expression of five AFB1 biosynthesis-related genes in A. flavus AYM2 revealed the downregulating effects of T. harzianum AYM3 metabolites on expression of aflP and aflS genes. Using HepaRG cell line, the cytotoxicity assay indicated that T. harazianum AYM3 metabolites were safe. Based on these results, it can be concluded that T. harzianum AYM3 may be used to suppress AFB1 production in maize grains. Full article

A method for accurately analyzing electrocardiograms (ECGs), which are obtained from electrical signals generated by cardiac activity, is essential in heart disease diagnosis. However, rhythms are typically obtained with relatively few data samples and similar characteristics, making them difficult to classify. To solve these issues, we proposed a novel method that distinguishes a given ECG rhythm using a beat score map (BSM) image. Through the proposed method, the associations between beats and previously used features, such as the R–R interval, were considered. Rhythm classification was implemented by training a convolutional neural network model and using transfer learning with the created BSM image. As a result, the proposed method for ECG rhythms with small data samples showed significant results. It also showed good performance in differentiating atrial fibrillation (AFIB) and atrial flutter (AFL) rhythms, which are difficult to distinguish due to their similar characteristics. The performance for rhythms with a small number of samples of the proposed method is 20% better than an existing method. In addition, the performance based on the F-1 score for classifying AFIB and AFL of the proposed method is 30% better than the existing method. This study solved the previous limitations caused by small sample numbers and similar rhythms. Full article

Screening for microorganisms that inhibit aflatoxin production from environments showed that Penicillium citrinum inhibited aflatoxin production by Aspergillus parasiticus. The inhibitory substance in the culture medium of P. citrinum was confirmed to be citrinin (CTN). RT-PCR analyses showed that CTN did not inhibit expressions of aflatoxin biosynthetic genes (aflR, pksL1, and fas-1) of A. parasiticus, whereas feeding experiments using A. parasiticus showed that CTN inhibited the in vivo conversion of dihydrosterigmatocystin to AFB₂·AFG₂. These results suggest that CTN inhibits a certain post-transcriptional step in aflatoxin biosynthesis. CTN in the culture medium of A. parasiticus was found to be decreased or lost with time, suggesting that a certain metabolite produced by A. parasiticus is the cause of the CTN decrease; we then purified, characterized, and then analyzed the substance. Physico-chemical analyses confirmed that the metabolite causing a decrease in CTN fluorescence was kojic acid (KA) and the resulting product was identified as a novel substance: (1R,3S,4R)-3,4-dihydro-6,8-dihydroxy-1-(3-hydroxy-6-(hydroxymethyl)-4-oxo-4H-pyran-2-yl)-3,4,5-trimethyl-1H-isochromene-7-carboxylic acid, which was named “CTN-KA adduct”. Our examination of the metabolites’ toxicities revealed that unlike CTN, the CTN-KA adduct did not inhibit aflatoxin production by A. parasiticus. These results indicate that CTN’s toxicity was alleviated with KA by converting CTN to the CTN-KA adduct. Full article

The basic biological function of glutamine synthetase (Gs) is to catalyze the conversion of ammonium and glutamate to glutamine. This synthetase also performs other biological functions. However, the roles of Gs in fungi, especially in filamentous fungi, are not fully understood. Here, we found that conditional disruption of glutamine synthetase (AflGsA) gene expression in Aspergillus flavus by using a xylose promoter leads to a complete glutamine deficiency. Supplementation of glutamine could restore the nutritional deficiency caused by AflGsA expression deficiency. Additionally, by using the xylose promoter for the downregulation of AflgsA expression, we found that AflGsA regulates spore and sclerotic development by regulating the transcriptional levels of sporulation genes abaA and brlA and the sclerotic generation genes nsdC and nsdD, respectively. In addition, AflGsA was found to maintain the balance of reactive oxygen species (ROS) and to aid in resisting oxidative stress. AflGsA is also involved in the regulation of light signals through the production of glutamine. The results also showed that the recombinant AflGsA had glutamine synthetase activity in vitro and required the assistance of metal ions. The inhibitor molecule L-α-aminoadipic acid suppressed the activity of rAflGsA in vitro and disrupted the morphogenesis of spores, sclerotia, and colonies in A. flavus. These results provide a mechanistic link between nutrition metabolism and glutamine synthetase in A. flavus and suggest a strategy for the prevention of fungal infection. Full article

Aflatoxins (AF) and ochratoxin A (OTA) are fungal metabolites that have carcinogenic, teratogenic, embryotoxic, genotoxic, neurotoxic, and immunosuppressive effects in humans and animals. The increased consumption of plant-based foods and environmental conditions associated with climate change have intensified the risk of mycotoxin intoxication. This study aimed to investigate the abilities of eleven selected LAB strains to reduce/inhibit the growth of Aspergillus flavus, Aspergillus parasiticus, Aspergillus carbonarius, Aspergillus niger, Aspergillus welwitschiae, Aspergillus steynii, Aspergillus westerdijkiae, and Penicillium verrucosum and AF and OTA production under different temperature regiments. Data were treated by ANOVA, and machine learning (ML) models able to predict the growth inhibition percentage were built, and their performance was compared. All factors LAB strain, fungal species, and temperature significantly affected fungal growth and mycotoxin production. The fungal growth inhibition range was 0–100%. Overall, the most sensitive fungi to LAB treatments were P. verrucosum and A. steynii, while the least sensitive were A. niger and A. welwitschiae. The LAB strains with the highest antifungal activity were Pediococcus pentosaceus (strains S11sMM and M9MM5b). The reduction range for AF was 19.0% (aflatoxin B1)-60.8% (aflatoxin B2) and for OTA, 7.3–100%, depending on the bacterial and fungal strains and temperatures. The LAB strains with the highest anti-AF activity were the three strains of P. pentosaceus and Leuconostoc mesenteroides ssp. dextranicum (T2MM3), and those with the highest anti-OTA activity were Leuconostoc paracasei ssp. paracasei (3T3R1) and L. mesenteroides ssp. dextranicum (T2MM3). The best ML methods in predicting fungal growth inhibition were multilayer perceptron neural networks, followed by random forest. Due to anti-fungal and anti-mycotoxin capacity, the LABs strains used in this study could be good candidates as biocontrol agents against aflatoxigenic and ochratoxigenic fungi and AFL and OTA accumulation. Full article