Stages Of Infection Research Articles

The glutamate receptor antagonist ifenprodil inhibits hepatitis E virus infection.

Infection with hepatitis E virus (HEV) represents a global problem, with over 20 million people infected annually. No specific antiviral drugs are available for treating HEV infection, necessitating the development of novel targeted therapeutics. Here, we report that the N-methyl-D-aspartate receptor (NMDAR) antagonist ifenprodil, a clinically approved drug used to treat idiopathic pulmonary fibrosis (IPF), is an HEV inhibitor in liver-derived cells. In vitro investigation demonstrates that ifenprodil suppresses viral protein expression in a dose-dependent manner in human hepatoma cells by inhibiting early stages of viral infection. We also found that ifenprodil modulates host cell intrinsic biological processes distinct from virus-induced innate immunity, inhibiting HEV RNA accumulation in primary human hepatocytes. Finally, the inhibitory effect of ifenprodil in vivo was also tested in rabbits challenged with the HEV-3ra CHN-BJ-R14 strain. Fecal virus shedding was below the limit of detection in two animals for both ribavirin-treated and ifenprodil-treated rabbits compared to vehicle-treated control animals. Our data demonstrate that ifenprodil is an effective anti-HEV compound with potential as a therapeutic candidate for the treatment of HEV infection.

B-048 Proposal of a new monitoring algorythm for patients with a HIV positive result with the Elecsys HIV DUO test

Abstract Background Worldwide HIV infections remain a public health problem, contributing significantly to morbidity. In Mexico we have had a progressive adaptation to the HIV epidemic providing free therapy and testing (4th generation tests) for the whole population in anbulatory centers for HIV/AIDS prevention and medical care. During 2022 Mexico reported 12,374 new cases. In the last decade there has been an increasing interest in the identification of patients HIV positives subgroups, based on disease progression and serological features. To date, 6 generations of tests for HIV detection have been developed, strongly reducing the window period for a positive result. Elecsys HIV Duo is a 5th generation immunological test that provides in a separate manner an antibody and antigen results for HIV detection. These results show a better insight of the serologycal status reducing the window period post infection. By using these tests we want to provide an early detection of new cases that gives a better prognosis for the patient and propose a new monitoring for patients with a positive antigen result but that have not yet produced antibodies. Such algorythm is currently lacking in the Mexican public health services. Methods A retrospective statystical analysis of HIV results of samples procesed with the Elecsys HIV DUO test during the january-april 2023 period in the National Reference Center of Salud Digna correlated with western blot results. Results The study population was 6405 patients, 6340 (98.98%) had a negative result in the test and 65 (1.02%) a positive test. Based in antibody/antigen results we categorized the positives results in three grupos: Group 1 Ab positive/Ag negative (16), group 2 Ab positive/Ag positive (32) and group 3 Ab negative/Ag positive (7). When correlated with the WB results all the patients from group 1 and 2 had a positive result while all patients in group 3 had a negative result. Conclusions Actually Mexican public health services do not have access to 5th generation HIV tests like the Elecsys HIV DUO test, therefore, public health profesional are not qualified to interprete these results and do not have a stated algorythm for patientes belonging to group 3. Therefore and based in our results we suggest 5th generation test (or superior if available) and propose a new patient monitoring algorythm with a molecular confirmatory test for viral detection. This proposal is strongly supported by the fact the we have a population that have not generated antibodys during the first weeks of post infection and will test negative in a serologycal confirmatory test like WB. Screening for patients in early infection stages is important because if we provide early therapy which may lead to infection reversal or a less impact in C4+ T cells numbers. Studies like this provide information for the generation of public politics in benefit of the population.

Proteomic analysis of global protein acetylation in response to WSSV infection in a crustacean Cherax quadricarinatus

Protein acetylation, a crucial post-translational modification, plays a significant role in antiviral immunity of host and viral replication processes after infection. Our previous research showed that an acetyltransferase KAT2A significantly enhanced white spot syndrome virus (WSSV) replication in hematopoietic tissue (Hpt) cells from Cherax quadricarinatus, suggesting that alterations in intracellular acetylation states affect viral infection. Nevertheless, the mechanisms by which WSSV invasion regulates intracellular protein acetylation remain unexplored. To explore the connection between protein acetylation and WSSV infection, differentially acetylated proteins (DAcPs) were identified in Hpt cells during WSSV infection by proteomic analysis. The results revealed that 25 DAcPs were identified with primarily involving in transporter activity and proton ATPase activity complex during the early stage of WSSV infection, i.e., the intracellular transport of the virions. Furthermore, that 36 DAcPs associated with chromatin assembly and acetyltransferase complex were identified in the replication stage of WSSV infection, i.e., the process of extensive viral replication. Additionally, acetylation at the K27 and K36 sites of histone H3 was strongly activated during WSSV replication; similarly, the inhibition of histone acetylation by acetyltransferase inhibiter C646 significantly suppressed WSSV replication, implying that WSSV replication requires high levels of histone acetylation, while the acetylation at the K27 and K36 sites of histone H3 probably serves as critical viral regulatory targets. This study deepens the understanding of WSSV-host interaction, and lays the groundwork for future research into the relationship between protein acetylation and viral infection in crustaceans.

Potential role of host autophagy in Clonorchis sinensis infection.

An in vivo mouse model of Clonorchis sinensis (C. sinensis) infection with or without the administration of autophagy inhibitor chloroquine (CQ) stimulation was established to assess the possible involvement of autophagic response during C. sinensis infection. Abnormal liver function was observed at 4, 6, and 8weeks post-infection, as indicated by elevated levels of ALT/GPT, AST/GOT, TBIL, and α-SMA in the infected groups. These findings indicated that C. sinensis infection activated autophagy, as shown by a decreased LC3II/I ratio and accumulated P62 expression in infected mice. Interestingly, CQ administration exhibited dual and opposing effects during the infection. In the early stage of infection, the engagement of CQ appeared to mitigate symptoms by reducing inflammation and fibrotic responses. However, in the later stage of infection, CQ might contribute to parasite survival by evading autophagic targeting, thereby exacerbating hepatic impairment and worsening liver fibrosis. Autophagy in liver was suppressed throughout the infection. These observations attested that C. sinensis infection triggered autophagy, and highlighted a complex role for CQ, with both protective and detrimental effects, in the in vivo process of C. sinensis infection.

Single cell transcriptomics reveals dysregulated immnue homeostasis in different stages in HPV-induced cutaneous squamous cell carcinoma.

In order to explore the huge impact of impaired immnue homeostasis on the occurrence and development of cutaneous squamous cell carcinoma (cSCC), and investigate characterization of the cellular components and their changes which is crucial to understanding the pathologic process of HPV-induced cSCC, we diagnosed and followed up on a very rare HPV-induced cSCC patient who progressed at a very fast rate and transferred to death quickly. We performed single-cell RNA sequencing (scRNA-seq) of 11 379 cells from the skin tissues of this patient with four different skin statuses after HPV infection. Immunofluorescence experiments were used for validation. scRNA-seq identified that CD52+ HLA-DOA- macrophages only existed in paracancerous cutaneous squamous cell carcinoma (pc-cSCC) and cSCC tissue. Besides, immune cells including CD8+ exhausted T cells and CD4+ regulatory T cells as well as matrix cells like MMP1+, and MMP11+ fibroblasts were gradually increased. Meanwhile, COMP+ ASPN+ fibroblasts gradually decreased. Cell interaction analysis revealed enhancement in interactions between monocytes/macrophages, fibroblasts and tumour-specific keratinocytes. scRNA-seq was performed in HPV-induced cSCC for the first time, to explore the correlation between infection and tumour. It is the first time to study the development of tumours from different stages of infection in HPV-induced cSCC. In this study, the tumour itself and the tumour microenvironment were both analysed and explored. And it was validated in clinical samples from different patients. Our findings reveal the dynamic immnue homeostasis from normal skinto cSCC tissue, this alteration might drive HPV-induced cSCC.

Revolution in malaria detection: unveiling current breakthroughs and tomorrow's possibilities in biomarker innovation.

The ongoing battle against malaria has seen significant advancements in diagnostic methodologies, particularly through the discovery and application of novel biomarkers. Traditional diagnostic techniques, such as microscopy and rapid diagnostic tests, have their limitations in terms of sensitivity, specificity, and the ability to detect low-level infections. Recent breakthroughs in biomarker research promise to overcome these challenges, providing more accurate, rapid, and non-invasive detection methods. These advancements are critical in enhancing early detection, guiding effective treatment, and ultimately reducing the global malaria burden. Innovative approaches in biomarker detection are leveraging cutting-edge technologies like next-generation sequencing, proteomics, and metabolomics. These techniques have led to the identification of new biomarkers that can be detected in blood, saliva, or urine, offering less invasive and more scalable options for widespread screening. For instance, the discovery of specific volatile organic compounds in the breath of infected individuals presents a revolutionary non-invasive diagnostic tool. Additionally, the integration of machine learning algorithms with biomarker data is enhancing the precision and predictive power of malaria diagnostics, making it possible to distinguish between different stages of infection and identify drug-resistant strains. Looking ahead, the future of malaria detection lies in the continued exploration of multi-biomarker panels and the development of portable, point-of-care diagnostic devices. The incorporation of smartphone-based technologies and wearable biosensors promises to bring real-time monitoring and remote diagnostics to even the most resource-limited settings.

Assessment of enteric helminth parasites in bushmeat in Ghana

Anthropogenic activities, such as hunting wild animals for food, increase the risk of zoonotic transmission of infective stages of parasites to humans. The handling, processing and consumption of wild animal meat, popularly known as ‘bushmeat’, as well as exposure to wildlife habitats, can pose a significant risk to human health through the transfer of parasitic infective stages. This study sought to assess the enteric helminth parasite burden and potentially zoonotic helminths in fresh, wild animal carcasses being processed for food. Parasitological analysis of samples of rectal and intestinal contents collected from a total of fifty (50) wild animal carcasses belonging to eight (8) different species at the Atwemonom Bushmeat Market in Kumasi showed nine (9) genera of enteric helminth parasites with an overall prevalence of 71.0%. Individual parasite species prevalence was assessed, with Ascaris sp. showing 25% and 87.5% by coprological and molecular assessment, respectively. Molecular analysis showed a higher parasite species prevalence in all samples analyzed. Species-specific analysis indicated the presence of two potentially zoonotic parasites, Strongyloides stercoralis and Trichuris trichiura, in wild animals, indicating the need to intensify one health approach in wild animal parasitic infections. Data from this study suggest that wild animals in addition to being natural hosts, may also serve as reservoirs for numerous parasites of medical and veterinary importance.

A Narrative Review on the Potential of Black Seeds (Nigella Sativa) in the Management of West Nile Virus Infection

Abstract: West Nile virus (WNV) infection is a mosquito-borne illness that is endemic to Africa, Asia, the Middle East, Australia, and Europe. WNV-infected individuals can only be managed with symptomatic therapy and supportive care at this point of time. Thus, we have analyzed the possible beneficial effects of black seeds (N. sativa) in this literature review. To find relevant articles, various online databases, such as the Web of Science, Medline, PMC, PubMed, Embase, EBSCO, Google Scholar, Science Direct, and reference lists were searched. The antiviral, analgesic, anti-inflammatory, antioxidant, and immunomodulatory effects of black seeds (N. sativa) have been established by numerous clinical, animal, in vivo, in vitro, and in silico studies. It has been found that black seeds (N. sativa) have the potential to be included as adjuvant therapy in addition to symptomatic treatment and supportive care in the early stages of WNV infection once randomized controlled clinical trials (RCTs) affirm the safety and efficacy of black seeds (N. sativa) in WNV infection management as they have been considered the gold standard of treatment in clinical studies.

Lithospermic acid inhibits dengue virus infection through binding with envelope proteins

The dengue virus has emerged as a global pandemic, highlighting the urgent need for the immediate development of antiviral therapeutics. Lithospermum erythrorhizon, a medicinal plant commonly used in China for various ailments including viral infections, inflammation, rheumatism, and cancer, showed promising antiviral properties in our research. Specifically, both the ethanol extract of Lithospermum erythrorhizon and its active component, lithospermic acid, demonstrated significant inhibitory effects on Dengue virus (DENV) replication in Vero cells, with EC50 values of 6.50 μg/mL(95% CI: 2.25 to 18.98)and 15.00 μM(95% CI: 12.13 to 18.07), respectively. Notably, lithospermic acid exhibited potent antiviral activity across multiple cell lines against DENV, impeding virus replication and specifically impeding the expression of viral E and NS3 proteins during the early stages of DENV infection. Experimental assays involving RNase digestion and sucrose density gradient analysis confirmed that lithospermic acid did not directly inactivate DENV but rather interfered with viral processes. Furthermore, the compound was found to bind to the E protein of DENV, effectively inhibiting viral infection and mitigating the cytopathic effects induced by DENV. Collectively, these findings underscore the potential of lithospermic acid as a promising candidate for the development of therapeutic strategies targeting DENV infection.

A virulent milRNA inhibits host immunity by silencing a host receptor-like kinase MaLYK3 and facilitates infection by Fusarium oxysporum f. sp. cubense.

MicroRNA-like RNAs (milRNAs) play a significant role in the infection process by plant-pathogenic fungi. However, the specific functions and regulatory mechanisms of fungal milRNAs remain insufficiently elucidated. This study investigated the function of Foc-milR138, an infection-induced milRNA secreted by Fusarium oxysporum f. sp. cubense (Foc), which is the causal agent of Fusarium wilt of banana. Initially, through precursor gene knockout and phenotypic assessments, we confirmed that Foc-milR138 acts as a virulent milRNA prominently upregulated during the early stages of Foc infection. Subsequent bioinformatic analyses and transient expression assays in Nicotiana benthamiana leaves identified a host receptor-like kinase gene, MaLYK3, as the direct target of Foc-milR138. Functional investigations of MaLYK3 revealed its pivotal role in triggering immune responses of N. benthamiana by upregulating a suite of resistance genes, bolstering reactive oxygen species (ROS) accumulation and callose deposition, thereby fortifying disease resistance. This response was markedly subdued upon co-expression with Foc-milR138. Expression pattern analysis further verified the specific suppression of MaLYK3 by Foc-milR138 during the early root infection by Foc. In conclusion, Foc secretes a virulent milRNA (Foc-milR138) to enter the host banana cells and inhibit the expression of the plant surface receptor-like kinase MaLYK3, subverting the disease resistance activated by MaLYK3, and ultimately facilitating pathogen invasion. These findings shed light on the roles of fungal milRNAs and their targets in resistance and pathogenicity, offering promising avenues for the development of disease-resistant banana cultivars.

Dual effects of ipecac alkaloids with potent antiviral activity against foot-and-mouth disease virus as replicase inhibitors and direct virucides

ABSTRACT Foot-and-Mouth Disease (FMD) is a contagious, blistering disease caused by the Foot-and-Mouth Disease virus (FMDV), which affects livestock globally. Currently, no commercial antiviral agent is available for effective disease control. This study investigated the antiviral potential of natural-derived alkaloids against FMDV in BHK-21 cells. Twelve alkaloids were assessed for their antiviral activities at various stages of FMDV infection, including pre-viral entry, post-viral entry, and prophylactic assays, as well as attachment and penetration assays by evaluating cytopathic effect reduction and directed-virucidal effects. The results showed that ipecac alkaloids, cephaeline (CPL) and emetine (EMT), exhibited dual effects with robust antiviral efficacy by reducing cytopathic effect and inhibiting FMDV replication in a dose-dependent manner. Evaluation through immunoperoxidase monolayer assay and RT-PCR indicated effectiveness at post-viral entry stage, with sub-micromolar EC50 values for CPL and EMT at 0.05 and 0.24 µM, respectively, and high selective indices. Prophylactic effects prevented infection with EC50 values of 0.23 and 0.64 µM, respectively. Directed-virucidal effects demonstrated significant reduction of extracellular FMDV, with CPL exhibiting a dose-dependent effect. Furthermore, the replicase (3Dpol) inhibition activity was identified using the FMDV minigenome assay, which revealed strong inhibition with IC50 values of 0.15 µM for CPL and 4.20 µM for EMT, consistent with the decreased negative-stranded RNA production. Molecular docking confirmed the interaction of CPL and EMT with residues in the active site of FMDV 3Dpol. In conclusion, CPL and EMT exhibited promising efficacy through their dual effects and provide an alternative approach for controlling FMD in livestock.

Redox Status and Protein Glutathionylation in Binase-Treated HPV16-Positive SiHa Carcinoma Cells

Human papillomavirus type 16 (HPV16) belongs to viruses of the high-risk type and is associated by overexpression of E6 and E7 oncoproteins, which determine the oncogenic properties of the virus, such as immortalization and malignant transformation of proliferating epithelial cells. The biogenesis of redox-sensitive proteins E6 and E7 at the early stages of viral infection leads to blocking of the cell antioxidant defense system and ubiquintin-dependent degradation of p53 and Rb tumor suppressors. Maintaining high rates of tumor cell proliferation contributes to an increase in the level of reactive oxygen species (ROS) and a shift in the redox balance towards oxidative processes. Reduced glutathione (GSH) provides antioxidant protection to tumor cells through S-glutathionylation of thiol groups of redox-sensitive proteins, which leads to the appearance of multidrug-resistant forms of cancer. In this regard, drugs restoring redox balance and increasing susceptibility to antitumor therapy are of particular importance. We have established that, Bacillus pumilus RNase (binase) modulates the redox-dependent regulatory mechanisms that ensure tumor cell resistance to apoptosis in HPV-16-positive SiHa cells of cervical squamous cell carcinoma,. Binase in nontoxic concentrations initiates a number of pre-apoptogenic changes, i.e., decreases ROS and reduced glutathione (GSH) levels, suppresses the expression of the E6 oncoprotein, activates the expression of the p53 tumor suppressor, and reduces the mitochondrial potential of tumor cells. Binase-induced disruption of the integrity of the mitochondrial membrane is a signal for activation of the mitochondrial apoptosis pathway.

Single-cell transcriptional profiling reveals cell type-specific responses to duck reovirus infection in the Bursa of Fabricius of Cairna moschata

Duck reovirus (DRV) is a universal waterfowl virus that causes significant economic losses in the duck industry. However, the role of the host innate immune response of the Bursa of Fabricius to DRV infection is largely unknown. In the present study, we constructed a single-cell resolution transcriptomic atlas of the Bursa of Fabricius of Cairna moschata after infection with HN10 (a novel DRV). Ten cell-type marker genes were used to annotate the cell type, indicating a high degree of cell heterogeneity in the Bursa of Fabricius. Most of the innate and adaptive immune system-related genes were highly expressed in T cells, B cells, neutrophils, macrophages, and DCs. In the Bursa of Fabricius, the proportions of DCs and macrophages were largely increased by HN10 infection at 14 d, suggesting that DCs and macrophages play important roles in the long-term viral response. Notably, a number of innate and adaptive immune system-related genes were highly expressed at 24 h after HN10 infection, indicating that the Bursa of Fabricius has a very strong immune function even in the early developmental stage. In the immune system, the NOD-like receptor signaling pathway and RIG-I-like receptor signaling pathway were significantly activated at the early stage of HN10 infection, while the Toll-like receptor signaling pathway was significantly activated at the late stage. Enrichment analysis suggested that different immune signaling pathways play roles in specific developmental stages. Our data provide an opportunity to reveal the immune response to DRV infection at the single-cell level.

RipE expression correlates with high ATP levels in Ehrlichia, which confers resistance during the extracellular stage to facilitate a new cycle of infection.

Ehrlichiosis is a potentially life-threatening disease caused by infection with the obligatory intracellular bacteria Ehrlichia species. Ehrlichia japonica infection of mice provides an animal model of ehrlichiosis as it recapitulates full-spectrum and lethal ehrlichiosis in humans. The E. japonica transposon mutant of EHF0962, which encodes a previously uncharacterized hypothetical protein, is attenuated in both infection and virulence in mice. EHF0962 was hence named here as resistance-inducing protein of Ehrlichia (RipE). Using this ΔripE mutant, we studied how RipE protein contributes to Ehrlichia pathogenesis. Ehrlichia species have an intracellular developmental cycle and a brief extracellular stage to initiate a new cycle of infection. Majority of RipE proteins were expressed on the surface of the smaller infectious dense-core stage of bacteria. Extracellular ΔripE E. japonica contained significantly less adenosine triphosphate (ATP) and lost infectivity more rapidly in culture compared with wild-type (WT) E. japonica. Genetic complementation in the ΔripE mutant or overexpression of ripE in WT E. japonica significantly increased bacterial ATP levels, and RipE-overexpressing E. japonica was more virulent in mice than WT E. japonica. RipE is conserved among Ehrlichia species. Immunization of mice with recombinant RipE induced an in vitro infection-neutralizing antibody, significantly prolonged survival time after a lethal dose of E. japonica challenge, and cross-protected mice from infection by Ehrlichia chaffeensis, the agent of human monocytic ehrlichiosis. Our findings shed light on the extracellular stage of Ehrlichia, highlighting the importance of RipE and ATP levels in Ehrlichia for extracellular resistance and the next cycle of infection. Thus, RipE is a critical Ehrlichia protein for infection as such can be a potential vaccine target for ehrlichiosis.

Outcomes after Endovascular Revascularization for Chronic Limb Threatening Ischemia from the only Vascular Quality Initiative (VQI) Center in Asia

IntroductionThis study compares chronic limb-threatening ischemia (CLTI) disease characteristics and endovascular revascularization outcomes in a multi-ethnic Asian cohort versus their North American counterparts. MethodsThe Society for Vascular Surgery Vascular Quality Initiative (SVS VQI) registry database from the first and currently the only VQI center in Asia was reviewed to identify patients with CLTI who underwent endovascular revascularization between July 2019 and April 2024. Standardized VQI reporting variables were compared against benchmarks derived from all participating centers in North America. Results2862 endovascular revascularization procedures from our center were benchmarked against 129347 procedures from 406 North American centers. Our cohort had a higher burden of comorbidities (diabetes mellitus, end-stage renal disease, cardiac disease) and presented with more advanced Wound, Ischemia, and Foot Infection (WIfI) stages. Our patients had more heavily calcified and longer (14.8 cm vs. 6.0 cm) diseased vessels with higher prevalence of multi-level (87% vs 54.6%), infrapopliteal (52.6% vs 38.9%) and inframalleolar (9.6% vs 2.4%) disease. Rates of technical success (92.7% vs 93%) and symptom improvement (39.1% vs 40.4%) were comparable between cohorts. However, 1-year mortality rates (28.9% vs 25.1%) and major amputation rates (13.3% vs 7.8%) were significantly higher. ConclusionShort term outcomes of technical success and symptom relief in our center were comparable to benchmark North American outcomes despite having a cohort with more diseased vessels, higher WIfI stages and comorbidities. However, this cohort fared worse in longer term outcomes of 1-year mortality and major amputation rates. Further studies are required to elucidate the causes to improve these outcomes.

First Report of Botryosphaeria dothidea Causing Leaf Blight on Parrotia subaequalis in China.

Parrotia subaequalis is of great ornamental value due to its unique bark, featuring interesting textures and colors, and its large, striking galls. These characteristics make it a popular choice for bonsai cultivation. (Yan et al. 2022) . In July 2023, an outbreak of leaf blight was observed on 40, six-month-old P. subaequalis seedlings in Anqing, Anhui, China, with an incidence rate of 80%. In the early stages of infection, small brown spots appear on the leaf surface, which gradually become round or irregular and darken to a deep brown color. As the disease progresses, the affected areas expand from the leaf margins towards the center, causing the leaf surface to become concave, wilt, and necrotize. This resulted in restricted plant growth, and in severe cases, partial or complete plant death. For isolation, 30 tissue slices (5 × 5 mm) were taken from the leaves of 10 symptomatic seedlings and surface sterilized with 75% ethanol for 5 seconds, followed by five rinses with sterilized distilled water. After two days of dark incubation at 28°C, hyphal tips of fungi were transferred onto new potato dextrose agar (PDA) plates and incubated until conidia production. Six unidentified isolates with similar morphological characteristics were obtained. The colonies, initially white, darken to black after 7 to 10 days of incubation. They produced colorless, aseptate conidia that were oblong or fusiform, measuring 18-26 μm in length and 5-8 μm in width (n=50). The morphological characteristics of the isolates resembled those of Botryosphaeria (Udayanga et al. 2015) . Isolate IS2116-1 was further confirmed through molecular methods. The rDNA internal transcribed spacer (ITS) region, translation elongation factor 1-α (TEF1-α), and beta-tubulin (TUB2) genes were amplified and sequenced using the primers ITS1/ITS4 (White et al., 1990), EF1-728F/EF1-986R, and Bt2a/Bt2b (Ferreira et al., 2021; Carbone et al., 1999), respectively. BLAST analysis revealed that the ITS (OR958722) sequence was 100% similar to the B. dothidea isolate HZ5(MH329650.1), TEF1-a (PP214058) sequence was 100% similar to the B. dothidea strain JZB310220(ON890458.1), and strain TUB2 (PP214057) sequence was 99.78% similar to the B. dothidea strain L14 (KR260833.1). Maximum likelihood analyses were performed for the combined ITS、TUB2、TEF datasets using PhyloSuite v1.2.2, the resulting phylogenetic tree indicated that isolate IS2116-1 clustered together with Botryosphaeria dothidea in a clade with 97% bootstrap support(Zheng et al. 2020) . Pathogenicity tests were conducted on 3-6 month-old P. subaequalis seedlings (n = 5) grown in a greenhouse. A conidial suspension (106 spores/ml) collected from the isolates was sprayed onto P. subaequalis seedlings, while the control was treated with distilled water. All plants were maintained in a growth chamber at 28°C with a 12-h photoperiod. The experiment was conducted twice independently . After 20 days of inoculation, brownish lesions similar to those observed in the field appeared on the treated plants, while the noninoculated control plants remained symptomless. The pathogen was reisolated from the leaves of the obviously diseased seedlings and confirmed as B. dothidea through morphological and sequence analysis. No isolates were obtained from uninoculated control plants, thus fulfilling Koch's hypothesis. This report marks the first record of B. dothidea causing leaf blight in P. subaequalis. In light of the rarity of natural P. subaequalis populations, it is imperative to assess both the extent of disease spread and its economic impact. These insights are crucial for devising strategies to protect this endangered species from disease threats and to preserve its ecological significance.

Correlation between CRP and ferritin in dengue patients: A hospital-based prospective observational study

ABSTRACT Background: In India, dengue is one of the health challenges and a leading cause of hospitalization, especially during the monsoon. C-reactive protein (CRP) is an independent marker for dengue severity. The present study aimed to analyze the correlation between CRP and ferritin and with other hematological markers in dengue fever (DF). Methods: A hospital-based prospective study was conducted over one year. With the patient’s consent, A total of 189 dengue-diagnosed hospitalized patients aged more than 18 years were included in the study group. Demographic features, total leukocytes, total platelet counts, packed cell volume, CRP, and ferritin were monitored in both severe and non-severe cases. Significant univariate parameters were correlated and significant associations were analyzed from the correlation coefficient and P value respectively. Result: A total of 189 dengue-diagnosed, hospitalized patients were recruited, among them 30 patients were diagnosed as severe and 159 patients were diagnosed as non-severe. The mean age ranged between 41 ± 16.7 and 42 ± 15.9 years. In univariate analysis, total leucocytes, CRP, and ferritin were significantly higher in dengue severe cases in comparison to non-severe dengue cases. The association between CRP and ferritin revealed a positive correlation whereas, the association between ferritin and total platelet count showed a negative correlation. Conclusion: A significant positive correlation between CRP and ferritin indicates that ferritin can be an additional biomarker along with CRP for the hospitalization of dengue patients and may be useful for severity prediction at an early stage of infection.

Host genetic diversity and body condition influence parasite resistance and clearance in a wild marine mammal population.

Host genetic variability can modulate infection resistance, although its role in infection clearance remains unclear. Hookworm disease (Uncinaria sp.) is the leading cause of pup mortality in several otariid species, although the parasite can be cleared through immune-mediated processes. We evaluated the association of host genetic diversity, body condition and immune response with hookworm resistance and/or clearance in the South American fur seal (Arctocephalus australis). Uninfected pups had higher heterozygosity than parasitized individuals, indicating a negative relationship between heterozygosity and the chances of infection. Likewise, pups that died of hookworm infection had lower heterozygosity than those that died of non-infectious causes. Interestingly, once infected, pups that survived hookworm infection had heterozygosities similar to pups that died of hookworm disease. However, pups that cleared the infection had a higher body mass and parasite-specific immunoglobulin G levels than those that did not recover or died of hookworm disease. Thus, although heterozygosity predicted resistance to and mortality from hookworm infections, it did not affect parasite clearance, which was facilitated by better body condition and adaptive immune responses. This demonstrates that host genetic variability and host-environment interactions influence disease dynamics, acting at different, well-defined stages of infection.

Classifying early apple scab infections in multispectral imagery using convolutional neural networks

Multispectral imaging systems combined with deep learning classification models can be cost-effective tools for the early detection of apple scab (Venturia inaequalis) disease in commercial orchards. Near-infrared (NIR) imagery can display apple scab symptoms earlier and at a greater severity than visible-spectrum (RGB) imagery. Early apple scab diagnosis based on NIR imagery may be automated using deep learning convolutional neural networks (CNNs). CNN models have previously been used to classify a range of apple diseases accurately but have primarily focused on identifying late-stage rather than early-stage detection. This study fine-tunes CNN models to classify apple scab symptoms as they progress from the early to late stages of infection using a novel multispectral (RGB-NIR) time series created especially for this purpose.This novel multispectral dataset was used in conjunction with a large Apple Disease Identification (ADID) dataset created from publicly available, pre-existing disease datasets. This ADID dataset contained 29,000 images of infection symptoms across six disease classes. Two CNN models, the lightweight MobileNetV2 and heavyweight EfficientNetV2L, were fine-tuned and used to classify each disease class in a testing dataset, with performance assessed through metrics derived from confusion matrices. The models achieved scab-prediction accuracies of 97.13 % and 97.57 % for MobileNetV2 and EfficientNetV2L, respectively, on the secondary data but only achieved accuracies of 74.12 % and 78.91 % when applied to the multispectral dataset in isolation. These lower performance scores were attributed to a higher proportion of false-positive scab predictions in the multispectral dataset. Time series analyses revealed that both models could classify apple scab infections earlier than the manual classification techniques, leading to more false-positive assessments, and could accurately distinguish between healthy and infected samples up to 7 days post-inoculation in NIR imagery.

Feasibility and potential of terahertz spectral and imaging technology for Apple Valsa canker detection: A preliminary investigation

Apple Valsa canker (AVC) caused by the Ascomycete Valsa mali, seriously constrains the production and quality of apple fruits. The symptomless incubation characteristics of Valsa mali make it highly challenging to detect AVC at an early infection stage. After infecting the wound of apple bark, the pathogenic hyphae of AVC will expand and colonize the phloem tissue. Meanwhile, various enzymes and toxic substances released by hyphae cause the decomposition of cellulose and lignin, and the generation of poisonous secondary metabolites in bark tissue. However, these early symptoms of AVC are invisible from the bark’s appearance. Fortunately, Terahertz Spectral Imaging (ThzSI) technology with the advantage of penetrating, and fingerprinting is promising for detecting hidden or slight symptoms of the fungal infection. This study is a preliminary investigation of terahertz frequency-domain spectra for AVC in the early stage of infection. Healthy and two-week-infected apple tree branches were prepared for capturing ThzS images, and the spectral data were preprocessed by Multivariate scattering correction (MSC), Savitzky-Golay convolution smoothing (SG), and standard normal variate (SNV) respectively to remove data noise and improve data quality. Principal component analysis (PCA), competitive adaptive reweighted sampling (CARS), and random frog (RFROG) were employed to extract the spectral feature bands to eliminate redundant data and improve computational efficiency. Machine learning models were established based on the spectral features to detect AVC at an early infection stage, where 11 of them exhibited the best performance with F1-score of 99.72%. To further explore disease information in spatial spectra, imaging data were acquired using terahertz imaging technology. Based on imaging data, pseudo-color imaging, histogram equalization, and Otsu segmentation were employed to visualize early infection areas in apple barks. Furthermore, histogram feature (HF), shape feature (SF), and local binary pattern (LBP) extracted from terahertz spectral images were utilized to establish the SVM, RF, and KNN models. HF-SF-KNN and HF-SF-LBP-KNN with the best performance achieved F1-score of 98.82%. This study presents a preliminary application of terahertz spectral and imaging technology for early-stage AVC detection and demonstrates its feasibility. Additionally, it provides a new way to detect AVC, which expands the application of ThzSI technology in tree disease detection in orchards and lays the foundation for further research.