Search Results (29)

Nucleopolyhedroviruses of lepidopteran larvae (Alphabaculovirus, Baculoviridae) form the basis for effective and highly selective biological insecticides for the control of caterpillar pests of greenhouse and field crops and forests. Horizontal transmission is usually achieved following the release of large quantities of viral occlusion bodies (OBs) from virus-killed insects. In the present review, I examine the evidence for productive midgut infection in different host species and the resulting transmission through the release of OBs in the feces (frass) of the host. This has been a neglected aspect of virus transmission since it was initially studied over six decades ago. The different host–virus pathosystems vary markedly in the quantity of OBs released in feces and in their ability to contaminate the host’s food plant. The release of fecal OBs tends to increase over time as the infection progresses. Although based on a small number of studies, the prevalence of transmission of fecal inoculum is comparable with that of recognized alternative routes for transmission and dissemination, such as cannibalism and interactions with predators and parasitoids. Finally, I outline a series of predictions that would affect the importance of OBs in feces as a source of inoculum in the environment and which could form the basis for future lines of research. Full article

The open reading frame 103 (p48) of Autographa californica multiple nucleopolyhedrovirus (AcMNPV) is one of the 38 core baculovirus genes. p48 has been shown to be essential for the production of infectious budded virions (BVs), nuclear egress of nucleocapsids, envelopment of the nucleocapsid, and embedding of occlusion-derived virions (ODVs) into occlusion bodies (OBs). However, the structure–function relationship of P48 remains unclear. In this study, we showed that four conserved cysteines (C127, C130, C138, and C141) in P48 may form a zinc finger motif based on a predicted structure analysis, and we investigated the roles of these cysteines in P48 function. AcMNPV bacmids lacking p48 or containing mutated p48 were generated. Transfection/infection assays showed that C127, C130, C138, and C141 in P48 were crucial for infectious BV production. Electron microscopy analysis further confirmed that these four cysteines played critical roles in the transport of nucleocapsids out of the nucleus for BV production, and in ODV envelopment. These results demonstrate that the conserved cysteines C127, C130, C138, and C141, related to the putative zinc finger motif, are critical for P48 function in baculovirus infection. Full article

Weight information plays a pivotal role in sturgeon breeding and production management. However, manual measurement is time consuming and labor intensive due to the immense size of the sturgeon. Due to the unique body shape of the sturgeon, traditional image segmentation algorithms struggle to extract the necessary features from sturgeon images, which makes them unsuitable for this particular species. Moreover, accurately measuring weight in an occlusion environment is difficult. To address these challenges, an improved YOLOv5s model with a context augmentation module, focal-efficient intersection over union, and soft non-maximum suppression was proposed in this paper. To validate the model’s feasibility, the improved YOLOv5s model was first pre-trained using the sturgeon dataset, followed by further training on the occlusion dataset for segmentation tasks. Based on the phenotypic data obtained from the improved model, a multilayer perceptron method was used to estimate the sturgeon’s weight accurately. Experimental results demonstrated that the average precision of the improved YOLOv5s model reached 89.80% under occlusion conditions, and the correlation coefficient of noncontact weight measurement results reached 89.80%. The experimental results showed that the improved algorithm effectively performs segmentation of sturgeon in occlusion conditions and can accurately estimate the mass. Full article

Insecticides based on baculoviruses have become an alternative for pest control in different agricultural crops. The aim of this study was to assess the biological activity of the bioinsecticide Cartugen^™ (SfMNPV: Baculoviridae: Alphabaculovirus) on larvae of Spodoptera frugiperda J. E. Smith (Lepidoptera: Noctuidae) resistant to Bt corn expressing the insecticidal proteins Cry1A.105+Cry2Ab2+Cry1F. In addition, we assessed the efficiency of SfMNPV on S. frugiperda control in the field from natural infestation of the pest during two agricultural seasons. The results showed that no larvae survived 10 days after being inoculated with Bt and non-Bt corn leaves contaminated with 1.50 × 10¹⁰ occlusion bodies (OBs)/L (equivalent to the recommended dose of the product). However, when using doses equivalent to 50% (7.50 × 10⁹ OBs/L) and 25% (3.75 × 10⁹ OBs/L), the larval mortality ranged from 21.12% to 46.55%, respectively. Although larvae resistant to the Cry1A.105+Cry2Ab2+Cry1F proteins, when exposed to 50% of the SfMNPV dose (7.50 × 10⁹ OBs/L), showed reductions in larval weight (52 to 67% reduction), pupal weight (32 to 59% reduction), and total fecundity (67 to 86% reduction) compared to the control. Furthermore, doses above 25% (3.75 × 10⁹ OBs/L⁻¹) caused a population decrease in the growth of the species in both Bt and non-Bt corn according to the fertility life table. In the field, at 7 and 10 DAA (days after application), corn plants sprayed with SfMNPV (1.50 × 10¹⁰ OBs/L) showed reductions in leaf damage according to the Davis scale. However, from 14 to 21 DAA, there was an increase in leaf damage in corn leaves from both treatments, with or without the application of SfMNPV. This shows that SfMNPV may be an important strategy in the integrated management and resistance management of S. frugiperda. Full article

Viral infection can regulate the cell cycle, thereby promoting viral replication. Hijacking and altering the cell cycle are important for the virus to establish and maintain a latent infection. Previously, Spodoptera exigua multiple nucleopolyhedrovirus (SeMNPV)-latently infected P8-Se301-C1 cells, which grew more slowly than Se301 cells and interfered with homologous SeMNNPV superinfection, were established. However, the effects of latent and superinfection with baculoviruses on cell cycle progression remain unknown. In this study, the cell cycle profiles of P8-Se301-C1 cells and SeMNPV or Autographa californica multiple nucleopolyhedrovirus (AcMNPV)-infected P8-Se301-C1 cells were characterized by flow cytometry. The results showed that replication-related genes MCM4, PCNA, and BAF were down-regulated (p < 0.05) in P8-Se301-C1 cells, and the S phase of P8-Se301-C1 cells was longer than that of Se301 cells. P8-Se301-C1 cells infected with SeMNPV did not arrest in the G2/M phase or affect the expression of Cyclin B and cyclin-dependent kinase 1 (CDK1). Furthermore, when P8-Se301-C1 cells were infected with SeMNPV after synchronized treatment with hydroxyurea and nocodazole, light microscopy and qRT-PCR analysis showed that, compared with unsynchronized cells and S and G2/M phase cells, SeMNPV-infected P8-Se301-C1 cells in G1 phase induced G2/M phase arrest, and the amount of virus adsorption and intracellular viral DNA replication were significantly increased (p < 0.05). In addition, budded virus (BV) production and occlusion body (OB)-containing cells were both increased at 120 h post-infection (p < 0.05). The expression of Cyclin B and CDK1 was significantly down-regulated at 48 h post-infection (p < 0.05). Finally, the arrest of SeMNPV-infected G1 phase cells in the G2/M phase increased BV production (p < 0.05) and the number of OB-containing cells. In conclusion, G1 phase infection and G2/M arrest are favorable to SeMNPV proliferation in P8-Se301-C1 cells, thereby alleviating the homologous superinfection exclusion. The results contribute to a better understanding of the relationship between baculoviruses and insect cell cycle progression and regulation. Full article

Spodoptera frugiperda (fall armyworm) is one of the most important maize pests in the world and the baculovirus Spodoptera frugiperda multiple nucleopolyhedrovirus (SfMNPV), a natural pathogen of this pest, has been used as a biopesticide for its control. At present, in vivo strategies at the commercial scale are employed by multiplying the virus in the host insect in biofactory facilities; however, in vitro large-scale production is an interesting alternative to overcome the limitations of baculoviruses massal production. This study aimed to develop the process of the SfMNPV in vitro production by evaluating the effects of different multiplicities of infection (MOI) and nutritional supplements, morphological and molecular analysis of the infection on the growth of Sf9 cells and virus production. The Bioreactor Stirred Tank Reactor (STR) approach with glutamine-supplemented Sf-900 III serum free culture medium, combined with the MOI of 1.0, showed the best viral production performance, with a specific productivity above 300 occlusion bodies (OBs)/cell and volumetric productivity of 9.0 × 10¹¹ OBs/L. Full article

Outbreaks of Anticarsia gemmatalis (Hübner, 1818) (Lepidoptera: Erebidae), a major pest of soybean, can be controlled below economic thresholds with methods that do not involve the application of synthetic insecticides. Formulations based on natural isolates of the Anticarsia gemmatalis multiple nucleopolyhedrovirus (AgMNPV) (Baculoviridae: Alphabaculovirus) played a significant role in integrated pest management programs in the early 2000s, but a new generation of chemical insecticides and transgenic soybean have displaced AgMNPV-based products over the past decade. However, the marked genotypic variability present among and within alphabaculovirus isolates suggests that highly insecticidal genotypic variants can be isolated and used to reduce virus production costs or overcome isolate-dependent host resistance. This study aimed to select novel variants of AgMNPV with suitable insecticidal traits that could complement the existing AgMNPV active ingredients. Three distinct AgMNPV isolates were compared using their restriction endonuclease profile and in terms of their occlusion body (OB) pathogenicity. One isolate was selected (AgABB51) from which eighteen genotypic variants were plaque purified and characterized in terms of their insecticidal properties. The five most pathogenic variants varied in OB pathogenicity, although none of them was faster-killing or had higher OB production characteristics than the wild-type isolate. We conclude that the AgABB51 wild-type isolates appear to be genotypically structured for fast speed of kill and high OB production, both of which would favor horizontal transmission. Interactions among the component variants are likely to influence this insecticidal phenotype. Full article

Baculoviruses are entomopathogens that carry large, double-stranded circular DNA genomes and infect insect larvae of Lepidoptera, Hymenoptera and Diptera, with applications in the biological control of agricultural pests, in the production of recombinant proteins and as viral vectors for various purposes in mammals. These viruses have a variable genetic composition that differs between species, with some sequences shared by all known members, and others that are lineage-specific or unique to isolates. Based on the analysis of nearly 300 sequenced genomes, a thorough bioinformatic investigation was conducted on all the baculoviral protein coding sequences, characterizing their orthology and phylogeny. This analysis confirmed the 38 protein coding sequences currently considered as core genes, while also identifying novel coding sequences as candidates to join this set. Accordingly, homology was found among all the major occlusion body proteins, thus proposing that the polyhedrin, granulin and CUN085 genes be considered as the 39th core gene of Baculoviridae. Full article

The marked adoption of bioinsecticides in Brazilian agriculture in recent years is, at least partially, explained by the increasingly higher levels of insect pest resistance to synthetic insecticides. In particular, several baculovirus-based products have been registered in the last 5 years, including Helicoverpa armigera nucleopolyhedrovirus (HearNPV: Baculoviridae: Alphabaculovirus (Armigen^®)). Understanding the susceptibility of Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae) to HearNPV is an important step toward development of robust Integrated Pest Management (IPM) and Insect Resistance Management programs (IRM) aimed at managing this serious insect pest. In this study, droplet feeding bioassays were used to characterize the baseline susceptibility to HearNPV (Armigen^®) in H. armigera populations collected from major soybean and cotton-growing regions in Brazil. We defined and validated a diagnostic concentration for susceptibility monitoring of H. armigera populations to HearNPV. Additionally, cross-resistance between HearNPV and the insecticides flubendiamide and indoxacarb was evaluated by testing HearNPV in a susceptible strain and in resistant strains of H. armigera to these insecticides. A low interpopulation variation of H. armigera to HearNPV was detected. The LC₅₀ values ranged from 1.5 × 10⁵ to 1.1 × 10⁶ occlusion bodies (OBs) per mL (7.3-fold variation). The mortality rate at the identified diagnostic concentration of 6.3 × 10⁸ OBs/mL, based on the calculated LC₉₉, ranged from 98.6 to 100% in populations of H. armigera collected from 2018 to 2020. No cross-resistance was detected between HearNPV and flubendiamide or indoxacarb. These results suggest that HearNPV (Armigen^®) can be an effective tool in IPM and IRM programs to control H. armigera in Brazil. Full article

Helicoverpa armigera single nucleopolyhedrovirus (HearSNPV) is a virulent pathogen of lepidopterans in the genera Heliothis and Helicoverpa, whereas Helicoverpa armigera multiple nucleopolyhedrovirus (HearSNPV) is a different virus species with a broader host range. This study aimed to examine the consequences of coocclusion of HearSNPV and HearMNPV on the pathogenicity, stability and host range of mixed-virus occlusion bodies (OBs). HearSNPV OBs were approximately 6-fold more pathogenic than HearMNPV OBs, showed faster killing by approximately 13 h, and were approximately 45% more productive in terms of OB production per larva. For coocclusion, H. armigera larvae were first inoculated with HearMNPV OBs and subsequently inoculated with HearSNPV OBs at intervals of 0–72 h after the initial inoculation. When the interval between inoculations was 12–24 h, OBs collected from virus-killed insects were found to comprise 41–57% of HearSNPV genomes, but the prevalence of HearSNPV genomes was greatly reduced (3–4%) at later time points. Quantitative PCR (qPCR) analysis revealed the presence of HearSNPV genomes in a small fraction of multinucleocapsid ODVs representing 0.47–0.88% of the genomes quantified in ODV samples, indicating that both viruses had replicated in coinfected host cells. End-point dilution assays on ODVs from cooccluded mixed-virus OBs confirmed the presence of both viruses in 41.9–55.6% of wells that were predicted to have been infected by a single ODV. A control experiment indicated that this result was unlikely to be due to the adhesion of HearSNPV ODVs to HearMNPV ODVs or accidental contamination during ODV band extraction. Therefore, the disparity between the qPCR and end-point dilution estimates of the prevalence of mixed-virus ODVs likely reflected virus-specific differences in replication efficiency in cell culture and the higher infectivity of pseudotyped ODVs that were produced in coinfected parental cells. Bioassays on H. armigera, Spodoptera frugiperda and Mamestra brassicae larvae revealed that mixed-virus OBs were capable of infecting heterologous hosts, but relative potency values largely reflected the proportion of HearMNPV present in each mixed-virus preparation. The cooccluded mixtures were unstable in serial passage; HearSNPV rapidly dominated during passage in H. armigera whereas HearMNPV rapidly dominated during passage in the heterologous hosts. We conclude that mixed-virus coocclusion technology may be useful for producing precise mixtures of viruses with host range properties suitable for the control of complexes of lepidopteran pests in particular crops, although this requires validation by field testing. Full article

Hidradenitis suppurativa (HS) is a chronic, inflammatory skin disease usually occurring after puberty with painful, deep-seated, inflammatory lesions in the apocrine gland-bearing areas of the body. Although HS pathogenesis is still unproven, recent major research advantages have increased our knowledge of the mechanisms behind HS lesions. Particularly, follicular occlusion followed by follicular rupture has been shown to be crucial to HS development, leading to immune response activation, and resulting in typical clinical HS lesions. Moreover, an increased and imbalanced cytokine production, such as interleukin (IL) 17 and tumor necrosis factor (TNF) α, may play a role in HS. In recent years, paradoxical adverse events have been described during treatment. Since the recent increased use of biologic treatments in HS, an increased number of paradoxical HS occurrences have been reported. In this review, we analyzed all current data on paradoxical HS triggered by biological drugs. Full article

Over time, we have come to recognize a very complex network of physiological changes enabling wound healing. An immunological process enables the body to distinguish damaged cells and begin a cleaning mechanism by separating damaged proteins and cells with matrix metalloproteinases, a complement reaction, and free radicals. A wide variety of cell functions help to rebuild new tissue, dependent on energy provision and oxygen supply. Like in an optimized “bio-reactor,” disturbance can lead to prolonged healing. One of the earliest investigated local factors is the pH of wounds, studied in close relation to the local perfusion, oxygen tension, and lactate concentration. Granulation tissue with the wrong pH can hinder fibroblast and keratinocyte division and proliferation, as well as skin graft takes. Methods for influencing the pH have been tested, such as occlusion and acidification by the topical application of acidic media. In most trials, this has not changed the wound’s pH to an acidic one, but it has reduced the strong alkalinity of deeper or chronic wounds. Energy provision is essential for all repair processes. New insights into the metabolism of cells have changed the definition of lactate from a waste product to an indispensable energy provider in normoxic and hypoxic conditions. Neovascularization depends on oxygen provision and lactate, signaling hypoxic conditions even under normoxic conditions. An appropriate pH is necessary for successful skin grafting; hypoxia can change the pH of wounds. This review describes the close interconnections between the local lactate levels, metabolism, healing mechanisms, and pH. Furthermore, it analyzes and evaluates the different possible ways to support metabolism, such as lactate enhancement and pH adjustment. The aim of wound treatment must be the optimization of all these components. Therefore, the role of lactate and its influence on wound healing in acute and chronic wounds will be assessed. Full article

The multifunctional role of the human skin is well known. It acts as a sensory and immune organ that protects the human body from harmful environmental impacts such as chemical, mechanical, and physical threats, reduces UV radiation effects, prevents moisture loss, and helps thermoregulation. In this regard, skin disorders related to skin integrity require adequate treatment. Lipid nanoparticles (LN) are recognized as promising drug delivery systems (DDS) in treating skin disorders. Solid lipid nanoparticles (SLN) together with nanostructured lipid carriers (NLC) exhibit excellent tolerability as these are produced from physiological and biodegradable lipids. Moreover, LN applied to the skin can improve stability, drug targeting, occlusion, penetration enhancement, and increased skin hydration compared with other drug nanocarriers. Furthermore, the features of LN can be enhanced by inclusion in suitable bases such as creams, ointments, gels (i.e., hydrogel, emulgel, bigel), lotions, etc. This review focuses on recent developments in lipid nanoparticle systems and their application to treating skin diseases. We point out and consider the reasons for their creation, pay attention to their advantages and disadvantages, list the main production techniques for obtaining them, and examine the place assigned to them in solving the problems caused by skin disorders. Full article

The rise of deep learning technology has broadly promoted the practical application of artificial intelligence in production and daily life. In computer vision, many human-centered applications, such as video surveillance, human-computer interaction, digital entertainment, etc., rely heavily on accurate and efficient human pose estimation techniques. Inspired by the remarkable achievements in learning-based 2D human pose estimation, numerous research studies are devoted to the topic of 3D human pose estimation via deep learning methods. Against this backdrop, this paper provides an extensive literature survey of recent literature about deep learning methods for 3D human pose estimation to display the development process of these research studies, track the latest research trends, and analyze the characteristics of devised types of methods. The literature is reviewed, along with the general pipeline of 3D human pose estimation, which consists of human body modeling, learning-based pose estimation, and regularization for refinement. Different from existing reviews of the same topic, this paper focus on deep learning-based methods. The learning-based pose estimation is discussed from two categories: single-person and multi-person. Each one is further categorized by data type to the image-based methods and the video-based methods. Moreover, due to the significance of data for learning-based methods, this paper surveys the 3D human pose estimation methods according to the taxonomy of supervision form. At last, this paper also enlists the current and widely used datasets and compares performances of reviewed methods. Based on this literature survey, it can be concluded that each branch of 3D human pose estimation starts with fully-supervised methods, and there is still much room for multi-person pose estimation based on other supervision methods from both image and video. Besides the significant development of 3D human pose estimation via deep learning, the inherent ambiguity and occlusion problems remain challenging issues that need to be better addressed. Full article

The biomedical field is characterized by an ever-increasing production of sequential data, which often come in the form of biosignals capturing the time-evolution of physiological processes, such as blood pressure and brain activity. This has motivated a large body of research dealing with the development of machine learning techniques for the predictive analysis of such biosignals. Unfortunately, in high-stakes decision making, such as clinical diagnosis, the opacity of machine learning models becomes a crucial aspect to be addressed in order to increase the trust and adoption of AI technology. In this paper, we propose a model agnostic explanation method, based on occlusion, that enables the learning of the input’s influence on the model predictions. We specifically target problems involving the predictive analysis of time-series data and the models that are typically used to deal with data of such nature, i.e., recurrent neural networks. Our approach is able to provide two different kinds of explanations: one suitable for technical experts, who need to verify the quality and correctness of machine learning models, and one suited to physicians, who need to understand the rationale underlying the prediction to make aware decisions. A wide experimentation on different physiological data demonstrates the effectiveness of our approach both in classification and regression tasks. Full article