Early Stages Of Infection Research Articles

Construction and validation of a novel multi-epitope in silico vaccine design against the paramyosin protein of Opisthorchis viverrini using immunoinformatics analyses

Liver fluke infection caused by Opisthorchis viverrini (O. viverrini) remains a significant but neglected health threat across Southeastern Asia. The early infective anabolic growth stage of O. viverrini expresses and exposes proteins integral for the growth and maturation of immature worms to the adult catabolic stage. Among these proteins, paramyosin emerged as a distinct immunogenic protein during opisthorchiasis. The functional region of the paramyosin protein known as myosin tail was selected to design a multi-epitope vaccine (MEV) to elicit T and B cell immune responses in susceptible human hosts utilizing various immunoinformatics and in silico vaccinology tools. The vaccine candidate had several B- and T-cell epitopes that stimulate both humoral and cellular immune responses. Moreover, in silico structural, docking, and dynamic analyses showed that the construct interacted with target immune receptors effectively, which may result in sufficient immunological stimulation. Analysis of simulated coverage efficacy also supports vaccine application in the field. Cloning and expression of the vaccine candidate were determined to be viable based on physicochemical and in silico assessments. These results reveal that the vaccine candidate developed herein is stable and potentially useful in addressing opisthorchiasis. The promising result of this study establishes a strong platform for initiating laboratory and efficacy trials for the vaccine candidate.

Low potential of fish as a source of infection with Angiostrongylus cantonensis

Abstract Objectives Fish are hypothesized to act as paratenic hosts for the zoonotic nematode Angiostrongylus cantonensis, cause of human eosinophilic meningitis. There is a lack of data confirming the relevance of fish in A. cantonensis life cycle and their contribution to human infection. Materials and methods We conducted a series of experiments to investigate survival and infectivity of A. cantonensis larvae in Clarias gariepinus (catfish; n=30) and Oreochromis niloticus (tilapia; n=24). Each fish was inoculated with 10,000 third-stage larvae (L3). Larval survival was assessed through artificial digestion of fish tissues one, two, and three weeks post-infection. To investigate early stages of infection, four catfish were inoculated with 10,000 L3 each and sacrificed 3 days post-infection. qPCR and histopathological examinations were performed to evaluate larval distribution and tissue reactions. Two infected catfish, sacrificed one week post-infection, were used to feed Wistar rats. Results After 45 days, the rats were not shedding L1, indicating the absence of infection. One week post-infection, dead larvae were present in digested tissues of both fish species, and the same was observed two and three weeks after exposure. qPCR analysis revealed that intestine was the most heavily infected organ. Histopathology identified dead larvae within granulomas in intestines and liver. Early-stage infection experiment showed that fish sacrificed three days post-inoculation contained viable L3 which were infective to Wistar rats. Conclusions While A. cantonensis L3 can survive and remain infective in fish for a short period, they typically die within first few days post-infection. This suggests that fish may not be significant long-term paratenic hosts for A. cantonensis but may play a temporary role in its transmission to mammals (including humans) and birds. These results are consistent with previous studies on freshwater shrimps and highlight the importance of understanding aquatic host interactions in the epidemiology of this zoonotic food-borne pathogen.

Trypanosoma cruzi reprograms mitochondrial metabolism within the anterior midgut of its vector Rhodnius prolixus during the early stages of infection

BackgroundTrypanosoma cruzi is transmitted to humans by hematophagous bugs belonging to the Triatominae subfamily. Its intra-vectorial cycle is complex and occurs exclusively in the insect's midgut. Dissecting the elements involved in the cross-talk between the parasite and its vector within the digestive tract should provide novel targets for interrupting the parasitic life cycle and affecting vectorial competence. These interactions are shaped by the strategies that parasites use to infect and exploit their hosts, and the host's responses that are designed to detect and eliminate parasites. The objective of the current study is to characterize the impact of T. cruzi establishment within its vector on the dynamics of its midgut.MethodsIn this study, we evaluated the impact of T. cruzi infection on protein expression within the anterior midgut of the model insect Rhodnius prolixus at 6 and 24 h post-infection (hpi) using high-throughput quantitative proteomics.ResultsShortly after its ingestion, the parasite modulates the proteome of the digestive epithelium by upregulating 218 proteins and negatively affecting the expression of 11 proteins involved in a wide array of cellular functions, many of which are pivotal due to their instrumental roles in cellular metabolism and homeostasis. This swift response underscores the intricate manipulation of the vector's cellular machinery by the parasite. Moreover, a more in-depth analysis of proteins immediately induced by the parasite reveals a pronounced predominance of mitochondrial proteins, thereby altering the sub-proteomic landscape of this organelle. This includes various complexes of the respiratory chain involved in ATP generation. In addition to mitochondrial metabolic dysregulation, a significant number of detoxifying proteins, such as antioxidant enzymes and P450 cytochromes, were immediately induced by the parasite, highlighting a stress response.ConclusionsThis study is the first to illustrate the response of the digestive epithelium upon contact with T. cruzi, as well as the alteration of mitochondrial sub-proteome by the parasite. This manipulation of the vector's physiology is attributable to the cascade activation of a signaling pathway by the parasite. Understanding the elements of this response, as well as its triggers, could be the foundation for innovative strategies to control the transmission of American trypanosomiasis, such as the development of targeted interventions aimed at disrupting parasite proliferation and transmission within the triatomine vector.Graphical

Virulent Effector Hasp155 of Puccinia striiformis f. sp. tritici Suppresses Plant Immunity and Promotes Fungus Infection.

As a kind of obligate biotrophic fungus, Puccinia striiformis f. sp. tritici (Pst) secretes vast effectors via haustoria to host cells during the infection to inhibit host defense responses and promote fungal invasion. In this study, based on the completion of genome sequencing and haustorial transcriptome sequencing of Pst, we identified a Pst effector (Hasp155) that is significantly induced in the early stage of Pst infection to wheat. The 18 N-terminal amino acids of Hasp155 encoded a signal peptide with a secretory function. Transient expression of Hasp155 in Nicotiana benthamiana inhibited Bax-induced cell death as well as chitin-triggered callose deposition and defense-related gene expression. Moreover, delivery of the Hasp155 protein into wheat cells via type three secretion systems (TTSS) led to reduced plant immunity to nonpathogenic bacteria and to the avirulent Pst race with decreased H2O2 accumulation and promoted Pst development. Furthermore, transgenic overexpression of Hasp155 significantly renders wheat resistance susceptible, resulting in a decreased defense response and increased Pst pathogenicity. Overall, these results indicate that Hasp155 is an important effector of Pst pathogenicity by suppressing plant immunity.

Computational identification and analysis of CNP0269688 as a natural product inhibitor disrupting the interaction between the HIV matrix domain and tRNA.

Our research is dedicated to combating HIV by targeting its Matrix (MA) domain, which is crucial for viral assembly and replication. This strategy specifically aims to interrupt early-stage infection and deter drug resistance by focusing on this essential domain. Due to the MA domain's conservation across different HIV strains, our approach promises broad-spectrum efficacy, which is particularly crucial in regions marked by significant genetic diversity and resistance issues. In our study, we introduce CNP0269688, a natural product that exhibits high affinity for the HIV-1 Matrix. Through detailed molecular dynamics simulations, we have assessed the compound's structural stability and interaction dynamics, particularly its potential to hinder Protein-tRNA interactions. This analysis lays the groundwork for future experimental investigations. Our efforts are steps toward enhancing HIV treatment, reducing viral transmission, and curbing drug resistance, with the ultimate aim of controlling and eradicating the pandemic, thereby contributing significantly to public health and scientific advancement.

Murine C3 of the complement system affects infection by Leptospira interrogans

Leptospirosis is an infectious neglected disease estimated to affect more than one million people worldwide each year. The Complement System plays a vital role in eliminating infectious agents. However, its precise role in leptospirosis remains to be fully understood. We investigated the importance of C3 in L. interrogans serovar Kennewicki strain Pomona Fromm (LPF) infection. Lack of C3 leads to decreased leukocyte number, impaired inflammatory response and failure to eliminate bacteria during the early stages of infection, which may cause interstitial nephritis later. These findings could be explained, at least in part, by the lower presence of local opsonins. Furthermore, antibody production against Leptospira was compromised in the absence of C3, highlighting the importance of CR2 in B lymphocyte proliferation and the adjuvant role of C3d in humoral immunity. Leptospires can be eliminated through the urine, and according to our study, the lack of C3 delays the elimination of LPF through urine during the early stages of the infection. These results strongly suggest the crucial role of C3 protein in orchestrating an appropriate inflammatory response against LPF infection and in effectively eliminating the bacteria from the body during the acute phase of leptospirosis.

Vascular network-mediated systemic spread of Pseudomonas syringae pv. actinidiae causes the bacterial canker of kiwifruit

Pseudomonas syringae pv. actinidiae (Psa) causes destructive kiwifruit bacterial canker by invading vascular tissues across multiple plant organs. However, the cellular mechanism underlying its systemic transmission and cell-to-cell movement within these specialized vascular conduits remains unclear. In this study, a Psa-GFP strain and various microscopic techniques were used to investigate the interaction between kiwifruit and Psa. Our results reveal that Psa strategically exploits host vascular conduits for systemic movement, with the xylem vessel being the predominant avenue. In the phloem, Psa exhibits adaptive alteration in bacterial shape to traverse sieve pores, facilitating its systemic spread along sieve tubes and inducing phloem necrosis. Within the xylem, Psa breaches pit membranes to migrate between adjacent vessels. Furthermore, phloem fibers act as an initial barrier at the early stages of infection, delaying Psa's entry into vascular tissues during its journey to the xylem. Additionally, at the junctions of stem–stem or stem-leaf, branch trace or leaf trace mediates the bacterial organ-to-organ translocation, thus facilitating the systemic progression of disease. In conclusion, our findings shed light on the cellular mechanism employed by Psa to exploit the woody plant's vascular network for infection, thereby enhancing a better understanding of the biology of this poorly defined bacterium. These insights carry implications for the pathogenesis of Psa and other vascular pathogens, offering theoretical guidance for effective control strategies.

Root-knot nematodes exploit the catalase-like effector to manipulate plant reactive oxygen species levels by directly degrading H2O2.

Plants produce reactive oxygen species (ROS) upon infection, which typically trigger defence mechanisms and impede pathogen proliferation. Root-knot nematodes (RKNs, Meloidogyne spp.) represent highly detrimental pathogens capable of parasitizing a broad spectrum of crops, resulting in substantial annual agricultural losses. The involvement of ROS in RKN parasitism is well acknowledged. In this study, we identified a novel effector from Meloidogyne incognita, named CATLe, that contains a conserved catalase domain, exhibiting potential functions in regulating host ROS levels. Phylogenetic analysis revealed that CATLe is conserved across RKNs. Temporal and spatial expression assays showed that the CATLe gene was specifically up-regulated at the early infection stages and accumulated in the subventral oesophageal gland cells of M. incognita. Immunolocalization demonstrated that CATLe was secreted into the giant cells of the host plant during M. incognita parasitism. Transient expression of CATLe significantly dampened the flg22-induced ROS production in Nicotiana benthamiana. In planta assays confirmed that M. incognita can exploit CATLe to manipulate host ROS levels by directly degrading H2O2. Additionally, interfering with expression of the CATLe gene through double-stranded RNA soaking and host-induced gene silencing significantly attenuated M. incognita parasitism, highlighting the important role of CATLe. Taken together, our results suggest that RKNs can directly degrade ROS products using a functional catalase, thereby manipulating host ROS levels and facilitating parasitism.

A Putative Effector Pst-18220, from Puccinia striiformis f. sp. tritici, Participates in Rust Pathogenicity and Plant Defense Suppression.

Stripe rust, caused by Puccinia striiformis f. sp. tritici (Pst), stands out as one of the most devastating epidemics impacting wheat production worldwide. Resistant wheat varieties had swiftly been overcome due to the emergence of new virulent Pst strains. Effectors secreted by Pst interfere with plant immunity, and verification of their biological function is extremely important for controlling wheat stripe rust. In this study, we identified an effector, Pst-18220, from Puccinia striiformis f. sp. tritici (Pst), which was induced during the early infection stage of Pst. Silencing the expression of Pst-18220 through virus-mediated host-induced gene silencing (HIGS) resulted in a decreased number of rust pustules. In Nicotiana benthamiana, it significantly suppressed cell death induced by Pseudomonas syringae pv. tomato (Pto) DC3000. In Arabidopsis, plants with stable overexpression of Pst-18220 showed increased susceptibility to Pto DC3000, accompanied by a decrease in the expression level of pattern-triggered immunity (PTI)/effector-triggered immunity (ETI)-related genes, namely, AtPCRK1, AtPCRK2, and AtBIK1. These results emphasize the significant role of the Pst candidate effector, Pst-18220, in rust pathogenicity and the suppression of plant defense mechanisms. This broadens our understanding of effectors without any known motif.

A review on Paratuberculosis (John’s disease) current diagnostic approaches and future prospects

The objective of the present study is to review the current diagnostic tools used for detection of paratuberculosis infection in animals. It also showed the sensitivity and specificity of each test as well as the difficulty of fecal culture as gold standard technique for paratuberculosis infection diagnosis. It also reviewed the limitation of commercial Elisa tests as they cannot be used for diagnosis of animals at early stages of infection. Lateral flow test has been recently developed. We suggest it could be used as rapid, accurate useful tool for diagnosis of MAP infection.

Expression Pattern and Functional Analysis of MebHLH149 Gene in Response to Cassava Bacterial Blight.

The significant reduction in cassava (Manihot esculenta Crantz) yields attributed to cassava bacterial blight (CBB) constitutes an urgent matter demanding prompt attention. The current study centered on the MebHLH149 transcription factor, which is acknowledged to be reactive to CBB and exhibits augmented expression levels, as indicated by laboratory transcriptome data. Our exploration, encompassing Xanthomonas phaseoli pv. manihotis strain CHN01 (Xpm CHN01) and hormone stress, disclosed that the MebHLH149 gene interacts with the pathogen at the early stage of infection. Furthermore, the MebHLH149 gene has been discovered to be responsive to the plant hormones abscisic acid (ABA), methyl jasmonate (MeJA), and salicylic acid (SA), intimating a potential role in the signaling pathways mediated by these hormones. An analysis of the protein's subcellular localization suggested that MebHLH149 is predominantly located within the nucleus. Through virus-induced gene silencing (VIGS) in cassava, we discovered that MebHLH149-silenced plants manifested higher disease susceptibility, less ROS accumulation, and significantly larger leaf spot areas compared to control plants. The proteins MePRE5 and MePRE6, which are predicted to interact with MebHLH149, demonstrated complementary downregulation and upregulation patterns in response to silencing and overexpression of the MebHLH149 gene. This implies a potential interaction between MebHLH149 and these proteins. Both MePRE5 and MePRE6 genes are involved in the initial immune response to CBB. Notably, MebHLH149 was identified as a protein that physically interacts with MePRE5 and MePRE6. Based on these findings, it is hypothesized that the MebHLH149 gene likely functions as a positive regulator in the defense mechanisms of cassava against CBB.

Extrahepatic intraabdominal hydatid cyst: a case report

BackgroundHydatid disease, also known as echinococcosis, is a zoonotic parasitic infection caused by the larvae of the Echinococcus tapeworm. It is endemic in various regions worldwide, particularly in rural areas of countries in southern South America, Central Asia, China, parts of Africa, the Mediterranean, and parts of the Middle East. The disease primarily affects the liver (60–70% of cases) and the lungs (10–25% of cases), but it can involve any organ, including the brain, bones, and rarely the pelvic region, as seen in our case report. Hydatid disease typically follows an asymptomatic course in the early stages of the primary infection and may remain so potentially for years or even permanently. If symptoms occur, they depend on various factors, such as the number, size, and location among other factors. Typically, hydatid disease presents with nonspecific symptoms. Common symptoms include abdominal pain, hepatomegaly, as well as anaphylaxis in case of cyst rupture. Extrahepatic intra-abdominal isolated hydatic cyst is a rare finding (6–11%).Case presentationIn our case, a 70 year-old Asian white male presented with right thigh pain radiating to the lower leg, which is an atypical presentation for an extrahepatic intraabdominal hydatid cyst. Primary intraabdominal hydatid cysts involving the pelvic region are relatively rare, and such cases pose diagnostic and management challenges.ConclusionThis case report underscores the challenges in diagnosing and managing extrahepatic intraabdominal hydatid cysts, particularly in atypical presentations. A combination of clinical evaluation, serological studies, and imaging techniques facilitates accurate diagnosis.

Acute Chikungunya Virus Infection Triggers a Diverse Range of T Helper Lymphocyte Profiles.

Chikungunya virus (CHIKV) is an arbovirus causing acute febrile illness with severe joint pain, often leading to chronic arthralgia. This study investigated the adaptive immune responses during the early stages of symptomatic acute CHIKV infection, focusing on the transcription factors and cytokines linked to Th1, Th2, Th17, and Treg cells. Thirty-six individuals were enrolled: nine healthy controls and 27 CHIKV-positive patients confirmed by qRT-PCR. Blood samples were analyzed for the mRNA expression of transcription factors (Tbet, GATA3, FoxP3, STAT3, RORγt) and cytokines (IFN-γ, IL-4, IL-17, IL-22, TGF-β, IL-10). The results showed the significant upregulation of Tbet, GATA3, FoxP3, STAT3, and RORγt in CHIKV-positive patients, with RORγt displaying the highest increase. Correspondingly, cytokines IFN-γ, IL-4, IL-17, and IL-22 were upregulated, while TGF-β was downregulated. Principal component analysis (PCA) confirmed the distinct immune profiles between CHIKV-positive and healthy individuals. A correlation analysis indicated that higher Tbet expression correlated with a lower viral load, whereas FoxP3 and TGF-β were associated with higher viral loads. Our study sheds light on the intricate immune responses during acute CHIKV infection, characterized by a mixed Th1, Th2, Th17, and Treg response profile. These results emphasize the complex interplay between different adaptive immune responses and how they may contribute to the pathogenesis of Chikungunya fever.

Early detection of sugarcane smut and mosaic diseases via hyperspectral imaging and spectral-spatial attention deep neural networks

Early detection of sugarcane diseases is important for effective management strategy. However, it is a challenging task as many diseases can hardly be detected in the early stage of infection due to a lack of human-eye-detectable symptoms. For enhanced detection, hyperspectral imaging technologies can capture light from a broad spectral range which often includes part of the invisible spectral range (e.g., near-infrared), and thereby can potentially capture the features for early detection of sugarcane diseases before the development of visible symptoms. Moreover, deep neural networks are well-developed frameworks from the field of deep learning that can automatically extract features for association with specific disease-related spectra. In this research, early detection of sugarcane smut and sugarcane mosaic diseases was achieved using a combined hyperspectral imaging and deep learning approach that was subsequently applied to track disease development over a five-month trial period. Additionally, the first large-scale high-resolution hyperspectral image dataset of inoculated and healthy (control) sugarcane plants was created and released. Subsequently, experimental results indicated that hyperspectral images containing features that can be used for the early detection of the two target sugarcane diseases were extracted by deep neural network models efficiently. The detection accuracy for both diseases was above 90 % before the appearance of visible symptoms.

Temporal dynamics of N-hydroxypipecolic acid and salicylic acid pathways in the disease response to powdery mildew in wheat

Wheat (Triticum aestivum) is a major staple crop worldwide, and its yields are significantly threatened by wheat powdery mildew (Blumeria graminis f. sp. tritici). Enhancing disease resistance in wheat is crucial for meeting global food demand. This study investigated the disease response in wheat, focusing on the bioactive small molecules salicylic acid (SA), pipecolic acid (Pip), and N-hydroxypipecolic acid (NHP), to provide new insights for molecular breeding. We found that endogenous levels of SA, Pip, and NHP significantly increased in infected plants, with Pip and NHP levels rising earlier than those of SA. Notably, the rate of increase of NHP was substantially higher than that of SA. The gene expression levels of SARD1 and CBP60g, which are transcription factors for SA, Pip, and NHP biosynthesis, increased significantly during the early stages of infection. We also found that during the later stages of infection, the expression of ALD1, SARD4, and FMO1, which encode enzymes for Pip and NHP biosynthesis, dramatically increased. Additionally, ICS1, which encodes a key enzyme involved in SA biosynthesis, also showed increased expression during the later stages of infection. The temporal changes in ICS1 transcription closely mirrored the behavior of endogenous SA levels, suggesting that the ICS pathway is the primary route for SA biosynthesis in wheat. In conclusion, our results suggest that the early accumulation of Pip and NHP cooperates with SA in the disease response against wheat powdery mildew infection.

Getting insights into chemical composition and antiherpetic capability of jujube (Ziziphus jujuba mill.) drupes

Food plant diversity in bioactive compounds makes them an exploitable resource in the search for effective natural products to prevent or treat viral infections. Therefore, in the framework aimed at studying the antiviral properties of extractive mixtures from fruits (and their waste) grown in the Campania Region (Italy), jujube drupes (Zizyphus jujuba Mill.) were our focus. The drupes were dissected into their peel, pulp and seed parts, each of which was extracted by ultrasound-assisted maceration and further fractionated, thus obtaining, beyond the sugar fraction, a polyphenolic fraction and a lipid fraction. UHPLC-HR MS/MS tools highlighted that the polyphenolic component of the seed was strongly dissimilar from that of the edible parts, being constituted by swertisin and its derivatives. Moreover, the peel mostly accounted for triglycosylated flavonols, whereas the pulp was rich in volatile aromatic glycosides. Among lipids, p-coumaroyl triterpenes mainly characterized the peel. All fractions were screened for their cytotoxicity, and non-toxic concentrations of each extract were tested against herpes simplex virus type 1 (HSV-1) by plaque assays. Molecular tests and Western blot analyses were also carried out. The jujube mixtures, in detail the peel and pulp polyphenolic fractions, and peel lipophilic fraction (the latter enriched mainly in ursane-type triterpenes), showed a marked inhibitory activity against HSV-1 acting in the early stages of viral infection and preventing attachment of the virus to the host cell. The acquired data suggest jujube active mixtures as promising candidates for the prevention and treatment of herpetic lesions.

Effects of the oxoaporphine alkaloid hernandonine on dengue virus. Evidence for its mechanisms of action

BackgroundDengue, caused by the dengue virus (Orthoflavivirus dengue, encompassing DENV types 1-4), is a member of the Flaviviridae family. The symptoms of dengue range from subclinical or mild manifestations to potentially fatal complications. The management of severe dengue is exceptionally challenging due to the absence of effective antiviral medications. In this context, natural products, whether in the form of pure compounds or standardized plant extracts, have emerged as a promising source for the development of novel antiviral therapeutics. Hernandonine, an oxoaporphine alkaloid found in Hernandia nymphaeifolia (C. Presl) Kubitzki. serves both as a metabolite and an inhibitor of human immunodeficiency virus type 1 (HIV-1) integrase. PurposeThis study investigated the ability of hernandonine to inhibit DENV infection and explored its potential mechanisms. Study designTo assess the in vitro anti-DENV activity, cells or induced pluripotent stem cell (iPSC)-derived cerebral organoids were exposed to hernandonine before or after infection with DENV. Along with hernandonine, the endocytosis modulators, genistein, wortmannin, Methyl-β-cyclodextrin (MβCD) and lovastatin, were used in the assays. MethodsThe DENV infectivity and virion production in cells or cerebral organoids treated with compounds were determined. Various methods, including cell and cerebral organoids imaging, protein and gene detection were conducted to explore their antiviral mechanisms. ResultsThe results revealed notable antiviral properties of hernandonine, particularly in inhibiting DENV during the early stages of infection. Mechanistic analysis demonstrated that, akin to genistein, wortmannin, methyl-β-cyclodextrin (MβCD), and lovastatin, hernandonine exerted an influence on cholesterol-rich lipid rafts. It also restrained the pseudopodial movement ability of cells, potentially through the downregulation of cytoskeleton and endocytosis regulatory genes or protein expression. Moreover, hernandonine's virucidal activity was demonstrated. Hernandonine's inhibition of DENV infection was further validated in a disease-relevant iPSC-derived cerebral organoids model, a novel DENV-2 infection system worthy of further application. ConclusionThis study evidenced the potential of hernandonine as a novel candidate in the fight against DENV infection.

Deciphering early responsive signature genes in rice blast disease: an integrated temporal transcriptomic study.

Rice blast disease, caused by Magnaporthe oryzae, reigns as the top-most cereal killer, jeopardizing global food security. This necessitates the timely scouting of pathogen stress-responsive genes during the early infection stages. Thus, we integrated time-series microarray (GSE95394) and RNA-Seq (GSE131641) datasets to decipher rice transcriptome responses at 12- and 24-h post-infection (Hpi). Our analysis revealed 1580 differentially expressed genes (DEGs) overlapped between datasets. We constructed a protein-protein interaction (PPI) network for these DEGs and identified significant subnetworks using the MCODE plugin. Further analysis with CytoHubba highlighted eight plausible hub genes for pathogenesis: RPL8 (upregulated) and RPL27, OsPRPL3, RPL21, RPL9, RPS5, OsRPS9, and RPL17 (downregulated). We validated the expression levels of these hub genes in response to infection, finding that RPL8 exhibited significantly higher expression compared with other downregulated genes. Remarkably, RPL8 formed a distinct cluster in the co-expression network, whereas other hub genes were interconnected, with RPL9 playing a central role, indicating its pivotal role in coordinating gene expression during infection. Gene Ontology highlighted the enrichment of hub genes in the ribosome and protein translation processes. Prior studies suggested that plant immune defence activation diminishes the energy pool by suppressing ribosomes. Intriguingly, our study aligns with this phenomenon, as the identified ribosomal proteins (RPs) were suppressed, while RPL8 expression was activated. We anticipate that these RPs could be targeted to develop new stress-resistant rice varieties, beyond their housekeeping role. Overall, integrating transcriptomic data revealed more common DEGs, enhancing the reliability of our analysis and providing deeper insights into rice blast disease mechanisms.

Inhibitors of the small membrane (M) protein viroporin prevent Zika virus infection.

Flaviviruses, including Zika virus (ZIKV), are a significant global health concern, yet no licensed antivirals exist to treat disease. The small Membrane (M) protein plays well-defined roles during viral egress and remains within virion membranes following release and maturation. However, it is unclear whether M plays a functional role in this setting. Here, we show that M forms oligomeric membrane-permeabilising channels in vitro, with increased activity at acidic pH and sensitivity to the prototypic channel-blocker, rimantadine. Accordingly, rimantadine blocked an early stage of ZIKV cell culture infection. Structure-based channel models, comprising hexameric arrangements of two trans-membrane domain protomers were shown to comprise more stable assemblages than other oligomers using molecular dynamics (MD) simulations. Models contained a predicted lumenal rimantadine binding site, as well as a second druggable target region on the membrane-exposed periphery. In silico screening enriched for repurposed drugs/compounds predicted to bind to either one site or the other. Hits displayed superior potency in vitro and in cell culture compared with rimantadine, with efficacy demonstrably linked to virion-resident channels. Finally, rimantadine effectively blocked ZIKV viraemia in preclinical models, supporting that M constitutes a physiologically relevant target. This could be explored by repurposing rimantadine, or development of new M-targeted-therapies.

Post-translational modifications on protein VII are important during the early stages of adenovirus infection.

Due to the importance of post-translational modification (PTM) in cellular function, viruses have evolved to both take advantage of and be susceptible to such modification. Adenovirus encodes a multifunctional protein called protein VII, which is packaged with the viral genome in the core of virions and disrupts host chromatin during infection. Protein VII has several PTMs whose addition contributes to the subnuclear localization of protein VII. Here, we used mutant viruses that abrogate or mimic these PTMs on protein VII to interrogate their impact on protein VII function during adenovirus infection. We discovered that acetylation of the lysine in positions 2 or 3 (K2 or K3) is deleterious during early infection as mutation to alanine led to greater intake of protein VII to the nucleus and enhanced early gene expression. Furthermore, we determined that protein VII is acetylated at alternative residues late during infection which may compensate for the mutated sites. Lastly, due to the role of the early viral protein E1A in viral gene activation, we investigated the interaction between protein VII and E1A and demonstrated that protein VII interacts with E1A through a chromatin-mediated interaction. Together, these results emphasize that the complexity of virus-host interactions is intimately tied to post-translational modification.