Zika Virus Research Articles

Formulation of Mosquito Repellant Freshener Gel with China Grass as Gel Base and Lemon Grass Oil as Mosquito -Repellant

Background: Mosquitoes transmit severe diseases, including malaria, dengue, and Zika virus, necessitating effective mosquito repellents. While traditional repellents are typically sprays or lotions, these can be inconvenient and leave residues. A more convenient alternative is mosquito repellent gel, offering ease of application, lasting effects, and a more enjoyable user experience. This study focuses on a gel-based mosquito-repelling air freshener containing lemongrass oil, known for its mosquito-repelling properties and fresh citrus-like scent. Methodology: The study developed a mosquito repellent gel infused with lemongrass oil, containing active constituents such as neral, isoneral, and limonene. Sensory assessments evaluated the gel’s color and odor using 150 individual’s comments, and a hedonic test was conducted with 180 participants aged between 25-30 to assess product acceptability when applied to clothing and inanimate objects. To determine the longevity of the gel, total liquid evaporation was measured by weight loss over a four-week period, and the repellent's effectiveness was tested in a closed room. Results: Sensory assessment confirmed an acceptable color and a citrus-like odor for the gel, while the hedonic test indicated a favorable user response. Weight loss calculations demonstrated the gel's sustained presence over time. The closed-room test confirmed the gel's effectiveness in repelling mosquitoes. Conclusion: The lemongrass oil-based mosquito repellent gel is a promising alternative to traditional repellents, offering both user convenience and effective mosquito protection. Its long-lasting formula and positive sensory characteristics make it a viable option for personal and environmental use in mosquito-prone areas.

Postnatal Zika and Dengue Infection and their Effects on Neurodevelopment Among Children Living in Rural Guatemala.

Prenatal Zika virus (ZIKV) infection leads to microcephaly and adverse neurodevelopment. The effects of postnatal ZIKV infection on the developing brain are unknown. We assessed the neurodevelopmental outcomes of children exposed postnatally during the ZIKV epidemic. A prospective study enrolled infants 0-3 months of age and their mothers, and children 1.5-3.5 years of age in rural Guatemala from 2017 and were followed for 12 months until 2019. Neurodevelopment was evaluated using the Mullen Scales of Early Learning (MSEL). ZIKV and dengue virus (DENV) infections were identified by polymerase chain reaction (PCR) using active surveillance. Serological analyses, stratified by age group flavivirus serostatus at enrollment, were conducted using a focus reduction neutralization test. Of 1371 enrolled participants, 1187 (86.6%) completed the study. No PCR-confirmed ZIKV infections were identified during the study period. One-third of 1.5-3.5-year-old children were ZIKV-seropositive at enrollment (likely postnatal infection). Twenty participants (5.8%) tested positive for DENV by PCR (11 infants, 5 children and 4 mothers); 15 (75%) were DENV-3 infections and 5 were DENV-2. The incidence of DENV infection in infants was 2.6%. No significant differences in MSEL scores were found between infants born seropositive versus seronegative for ZIKV or DENV. DENV seropositivity at enrollment in 1.5-5-year-old children was associated with lower MSEL scores for fine motor, visual reception and language, and microcephaly at 12 months versus seronegative children (all P < 0.05). Postnatal ZIKV infection in children from rural Guatemala was not associated with worse neurodevelopmental outcomes. DENV seropositivity was associated with a higher risk of microcephaly in infants and worse neurodevelopmental outcomes in children.

Platelet Reactivity to Zika and Dengue Non-Structural Protein 1 (NS1) Assessed by Flow Cytometry, Atomic Force Microscopy, and Quartz Crystal Microbalance.

Platelets, besides being traditionally associated with hemostasis, have been recently positioned as immune cells. Alterations in platelet number and function have been reported in some viral infections. Zika virus (ZIKV) and Dengue virus (DENV) are arboviruses that encode for a non-structural protein 1 (NS1). NS1 is mainly involved in the viral replication process and can also be secreted by infected cells and has been associated with immune response evasion. The assessment of platelet reactivity against these viral agents and their proteins, through the use of different innovative technologies such as flow cytometry (FC), atomic force microscopy (AFM), and quartz crystal microbalance (QCM), will allow further study of the pathophysiology of these emerging diseases. The aim of this study was to assess platelet reactivity to ZIKV and DENV NS1 protein through the use of FC, AFM, and QCM. Platelet-rich plasma (PRP) was stimulated with ZIKV and DENV NS1 protein in individual assays. The expression of P-selectin and the activity of the glycoprotein IIb-IIIa, platelet activation markers, were assessed by FC, morphological changes were assessed by AFM, and interaction between NS1 protein and platelet were evaluated by QCM. An increased expression of P-selectin and GP IIb-IIIa activity (p < 0.001) was observed when PRP was stimulated with ZIKV and DENV NS1 proteins. AFM images showed an increase in cell size and the appearance of pseudopods upon stimulation with the viral proteins. QCM results showed a significant increase in the oscillation frequency of the quartz precoated with ZIKV or DENV NS1 when PRP was injected (p < 0.001). FC, AFM, and QCM are techniques that can be used in the study of platelet response to viral structures such as NS1 protein, broadening the range of existing methodologies in the study of these cells. It is imperative to study platelets in arboviral infections to better understand their involvement in these diseases.

Repurposing Drugs for Infectious Diseases by Graph Convolutional Network with Sensitivity-Based Graph Reduction.

Computational systems biology employs computational algorithms and integrates diverse data sources, such as gene expression profiles, molecular interactions, and network modeling, to identify promising drug candidates through repurposing existing compounds in response to urgent healthcare needs. This study tackles the urgent need for rapid therapeutic development against emerging infectious diseases. We introduce a novel analytic expression for sensitivity analysis based on the Kronecker product and enhance model prediction performance using Graph Convolutional Networks (GCNs) with sensitivity-based graph reduction. Our algorithm refines prediction performance by leveraging sensitivity-based graph reduction. By integrating RNA-seq data, molecular interactions, and GCNs, we identify disease-related genes and pathways, construct heterogeneous graph models, and predict potential drugs. This approach involves novel analytical expressions that assess sensitivity to model loss, employing the Kronecker product approach. Subgraph analysis identifies nodes for removal, leading to a refined graph used for model retraining. This cost-effective pipeline focuses on computational methods for drug repurposing, targeting infectious diseases such as Zika virus and COVID-19 infection. Applied to these infections, our methodology integrates 659 proteins and 703 drugs for Zika virus, and 495 proteins and 468 drugs for COVID-19, along with their interactions derived from gene expression profiles. Top candidate drugs, such as Betamethasone phosphate and Bizelesin for Zika virus, and Chloroquine, Heparin Disaccharide, and Resveratrol for COVID-19, were validated through literature review or docking analysis. This scalable approach demonstrates promise in repurposing drugs for urgent healthcare challenges.

High glucose potentiates Zika virus induced-astroglial dysfunctions.

Zika virus (ZIKV) is a neurotropic flavivirus that induces congenital Zika syndrome and neurodevelopmental disorders. Given that ZIKV can infect and replicate in neural cells, neurological complications in adult brain are also observed. Glial cells may emerge to delay and/or prevent the development of ZIKV-induced neurodegeneration. These cells actively participate in metabolic, inflammatory and redox processes, and consequently, in the pathophysiology of neurodegenerative diseases, including diabetic encephalopathy. In this sense, changes in glucose metabolism can support the inflammatory activity of astroglial cells; however, the effects of increased glucose concentration during ZIKV infection have not yet been explored in astroglial cells. Here, we evaluated functional parameters of astroglial cells exposed to ZIKV upon normal and high glucose concentrations, focusing on inflammatory profile, oxidative stress, and expression of critical genes for astroglial functions. High glucose potentiated the pro-inflammatory and oxidative effects of ZIKV, as well as potentiated the downregulation of signaling pathways, such as Nrf-2 (nuclear factor erythroid derived 2 like 2), sirtuin 1 (SIRT1), peroxisome proliferator activated receptor gamma coactivator 1-alpha (PGC-1α), and poly (ADP-ribose) polymerase (PARP). In summary, our results suggest that high glucose can favor the activation of inflammatory signaling while impairing cytoprotective pathways in astroglial cells exposed to ZIKV and reinforce the hypothesis that this virus is highly neurotrophic, with significant impact in glial cells.

Evaluating the specificity of flavivirus proteases in Aedes aegypti cells for dengue virus 2-derived cleavage sites.

Flaviviruses are a diverse group of RNA viruses known for their significant impact on human health worldwide. We generated a series of reporters that included cleavage sequences from the dengue virus type 2 polyprotein and co-transfected with plasmids encoding various flavivirus proteases into Aedes aegypti cells, followed by fluorescent imaging and western blot analysis for the determination of proteolytic cleavage. Recombinant flavivirus NS2B3 proteases from medically significant and insect-specific flaviviruses were able to process reporters encoding cleavage sequences from the dengue virus type 2 polyprotein in vitro including proteases from dengue virus types 1-4, Zika virus, yellow fever virus, Aedes flavivirus, and cell-fusing agent virus. Reporters were not cleaved when transfected cells were infected with dengue virus type 2. Endoplasmic reticulum tethered reporters were also cleaved by protease alone but not by infectious virus. These results shed light on the ability of multiple flavivirus proteases to cleave sequences derived from outside of their genome and raise new questions concerning the requirements for effective cleavage by flavivirus proteases in trans.

A zebrafish-based in vivo model of Zika virus infection unveils alterations of the glutamatergic neuronal development and NS4A as a key viral determinant of neuropathogenesis.

Infection of pregnant women by Zika virus (ZIKV) is associated with severe neurodevelopmental defects in newborns through poorly defined mechanisms. Here, we established a zebrafish in vivo model of ZIKV infection to circumvent limitations of existing mammalian models. Leveraging the unique tractability of this system, we gained unprecedented access to the ZIKV-infected brain at early developmental stages. The infection of zebrafish larvae with ZIKV phenocopied the disease in mammals including a reduced head area and neural progenitor cells (NPC) infection and depletion. Moreover, transcriptomic analyses of NPCs isolated from ZIKV-infected embryos revealed a distinct dysregulation of genes involved in survival and neuronal differentiation, including downregulation of the expression of the glutamate transporter vglut1, resulting in an altered glutamatergic network in the brain. Mechanistically, ectopic expression of ZIKV protein NS4A in the larvae recapitulated the morphological defects observed in infected animals, identifying NS4A as a key determinant of neurovirulence and a promising antiviral target for developing therapies.

Zika Virus Model with the Caputo–Fabrizio Fractional Derivative

In this article, we examine a deterministic Zika virus model that takes into account the vector and sexual transmission route, in the absence of disease-induced deaths, symmetrically observing the impact of human knowledge and vector control. In order to construct the model, we suppose that the Zika virus is first spread to humans through mosquito bites, and then to their sexual partner. In this article, we conduct analytical studies which often begin by proving the existence and uniqueness of solutions for the Zika virus model using the fractional derivative from the Caputo–Fabrizio derivative. Then, the uniqueness of the solution is investigated. After that, we also identify under which circumstances and symmetry the model provides a unique solution.

Serotype-Specific Dengue Virus IgG Assay Using EDIII-Based Recombinant Proteins and Its Application in an Endemic Population in Northeast Brazil.

Dengue is the most prevalent arboviral disease globally, with Brazil currently experiencing a significant rise in cases. Dengue virus (DENV) typically co-circulates with other clinically and antigenically similar flaviviruses, such as Zika virus (ZIKV). The clinical diagnosis is difficult and accurate serological analysis represents an unmet challenge. Traditionally, serological analysis of DENV infection focuses on the acute phase via detection of NS1 or IgM. Developing IgG DENV-specific assays has been defiant due to the co-circulation of four antigenically distinct serotypes and a strong cross-reactivity with other arboviruses, particularly with ZIKV. The goal of this study was to produce recombinant domain III of the Envelope (EDIII) proteins of each DENV serotype and ZIKV and evaluate the ability to detect specific IgG antibodies. The antigens were tested on the ELISA platform to measure DENV-specific IgG in mice and patients infected with ZIKV and DENV. The assay differentiated serotype-specific IgG responses in susceptible mice (AG129) experimentally infected with DENV or ZIKV. In addition, the test demonstrated a robust performance achieving 87.8% sensitivity and 91.4% specificity when tested against 648 well-characterized sera collected from humans infected with DENV and/or ZIKV. The test was further applied to serum samples collected from 318 healthy individuals from an endemic region in Northwest/Central Brazil, without a previous diagnosis of DENV, and revealed that 65% of the samples reacted with at least one DENV serotype antigen, including 123 monotypic samples (88 for DENV-1) and 90 samples reacting with multiple antigens. Collectively, these results indicate that the IgG DENV-EDIII ELISA is a valuable tool for assessing the serological status of populations in endemic areas, particularly in regions where other flaviviruses, particularly ZIKV, co-circulate and offer support to establish public health policies against the disease.

Effect of Sexual Partnerships on Zika Virus Transmission in Virus-Endemic Region, Northeast Brazil.

The epidemiologic effects of Zika virus (ZIKV) sexual transmission in virus-endemic countries remain unclear. We conducted a 2-level, linear mixed-effects logistic regression analysis by using a recently acquired population-based ZIKV and chikungunya virus (CHIKV) serologic dataset obtained from persons residing in Northeast Brazil (n = 2,070 participants). We adjusted mathematical models for housing type and age of participants; the models indicated a significantly higher likelihood of ZIKV seropositivity among persons engaged in a sexual relationship within the same household (odds ratio 1.25 [95% CI 1.00-1.55]; p = 0.047), regardless of their partner's ZIKV serostatus, and among participants with a ZIKV-seropositive sex partner within the same household (odds ratio 1.54 [95% CI 1.18-2.01]; p = 0.002). CHIKV was also modeled as a control; no sex-associated effects were observed for CHIKV serology. Inclusion of ZIKV sexual transmission in prevention and control strategies is urgently needed, particularly in ZIKV-endemic regions.

A symbiotic mosquito-gut bacterium for flavivirus control.

Dengue virus (DENV) and Zika virus (ZIKV) have emerged as major global public health challenges, causing numerous infections and fatalities each year. However, current measures, including vaccines and treatments, are often limited or ineffective. This highlights the urgent need for novel preventive strategies to control the spread of key mosquito-borne viruses like DENV and ZIKV. In a recent study published in Science, Zhang et al. isolated a bacterium named Rosenbergiella_YN46 from the gut of field-caught Aedes albopictus mosquitoes in Yunnan Province, China. This commentary reviews their findings, published on April19,2024, which describe the symbiotic bacterium Rosenbergiella_YN46 and its ability to block flavivirus transmission, including both DENV and ZIKV. The bacterium shows promising potential for future dengue fever prevention and provides valuable insights into a novel biological approach for controlling mosquito-borne viral diseases.

Prenatal Zika Virus Exposure Alters the Interaction Between Affective Processing and Decision-Making in Juvenile Rhesus Macaques (Macaca mulatta).

Many challenges during pregnancy can disrupt fetal development and have varying consequences on the subsequent psychological development of infants. Notably, exposure to infectious pathogens during fetal development, such as those encountered in viral pandemics, has been associated with persistent developmental consequences on infants' brains and behavior. However, the underlying mechanisms and the degree to which neural plasticity over infancy may accommodate fetal insults remain unclear. To address this gap, we investigated the interaction between affective processing and decision-making in a cohort of rhesus monkey juveniles exposed to Zika virus (ZIKV) during fetal development, a pathogen known to profoundly disrupt central nervous system development. Ten juveniles exposed to ZIKV during their fetal development and nine procedure-matched controls (CONs) completed a judgment bias task with and without a negative mood induction. Although ZIKV exposure did not impact the monkeys' decision-making processes during the task or the magnitude of their behavioral responses to the mood induction procedure, it did alter the influence of mood induction on decision-making. Although CON monkeys exhibited significantly more conservative decision-making following negative mood induction, the decision-making of Zika-exposed monkeys remained consistent among conditions. These findings suggest that fetal exposure to ZIKV impacts the neural systems involved in integrating affective and cognitive information, with potential long-term implications for learning, memory, and emotion regulation.

Assessment of multifunctional activity of graphene oxide and guar gum-based nanomaterials against Aedes aegypti

Nanomaterials have been receiving much research attention in controlling insect pests and vectors. Properties, such as small surface-to-volume ratio, low dosages, durability, solubility, enhanced target activity, pore size and surface characteristics have enabled the design of precise and targeted insecticides through adsorption, encapsulation, and conjugation. The reported study aims to evaluate the efficacy of graphene oxide (GO) and guar gum (GG)-based nanomaterials against early fourth instar of Aedes aegypti, a mosquito responsible for transmitting diseases like dengue fever, Zika virus, and chikungunya, among many others. GO and a total of eight nanomaterials (SA-1 to SA-8) were formulated using GO, GG and their combinations with ferrous oxide and copper oxide. Each material was characterized using various biophysical techniques. The materials were evaluated for larvicidal potential by assessing their efficacy on the survival and morphology of Ae. aegypti, while the contact irritancy activity focused on determining their irritant properties upon direct exposure to the female adults. The larvicidal and irritant bioassays, conducted with these nanomaterials at different concentrations, demonstrated concentration-dependent mortality and significant behavioral responses. The exposures with nanomaterials also resulted in a deposition of black soot on the larval cuticle. The most effective nanomaterial was found as ferrous oxide nanomaterial which induced 100% larval mortality as well as significant contact irritancy. The results indicate the potential use of GO and GG-based nanomaterials against Ae. aegypti after concentration optimization.

Diagnostic Challenges of Arboviral Infections and Dengue Virus Serotype Distribution in Febrile Patients in East Java, Indonesia

ObjectivesArboviruses pose significant public health threats globally, with Southeast Asia being a hotspot for these infections. This study aimed to reassess the incidence of dengue, Zika, and chikungunya viruses in patients clinically diagnosed with dengue in East Java, Indonesia in 2023. MethodsThe study included 108 patients admitted to hospitals in Jember, with blood samples collected on admission. Multiplex RT-PCR was used to detect viral RNA for dengue, Zika, and chikungunya, while dengue serotypes were identified using real-time PCR. ResultsA total of n=67/108 patients (62%) tested positive for dengue virus (DENV), one patient tested positive for chikungunya and no cases of Zika were detected. Differences in laboratory parameters between DENV RNA-negative patients and confirmed dengue cases suggest possible misdiagnosis of dengue. Serotyping of DENV-positive samples revealed DENV-3 as predominant serotype in Jember, accounting for 34% of cases (n=23/67), followed by DENV-1 and DENV-2 at 19% each (n=13/67), and 6% for DENV-4 (n=4/67), while 21% (n=14/67) remained untyped. ConclusionsThis study highlights the nature of the dengue outbreak in Jember in 2023, where all four DENV serotypes were in circulation, and underlines the need for serological or nucleic acid-based methods to improve arbovirus diagnosis in the region.

Revolutionizing Antiviral Therapeutics: In silico Approaches for Emerging and Neglected RNA Viruses.

The 21st century has shown us how rapidly the pandemic can evolve and devastate the life of human beings without differentiating between the continents. Even after the global investment of billions of dollars into the healthcare sector, we are still lacking multiple therapeutics against emerging viruses. World Health Organization (WHO) has listed a number of viruses that could take the form of pandemics at anytime, depending upon their mutations. Among those listed, the SARS-CoV, Ebola, Zika, Nipah, and Chikungunya virus (CHIKV) are the most known viruses in terms of their number of outbreaks. The common feature among these viruses is their RNA-based genome. Developing a new therapeutic candidate for these RNA viruses in a short period of time is challenging. In silico drug designing techniques offer a simple solution to these problems by implementing supercomputers and complicated algorithms that can evaluate the inhibition activity of proposed synthetic compounds without actually doing the bioassays. A vast collection of protein crystal structures and the data on binding affinity are useful tools in this process. Taking this into account, we have summarized the in silico based therapeutic advances against SARS-CoV, Ebola, Zika, Nipah, and CHIKV viruses by encapsulating state-of-art research articles into different sections. Specifically, we have shown that computer- aided drug design (CADD) derived synthetic molecules are the pillars of upcoming therapeutic strategies against emerging and neglected viruses.

Improved Zika virus plaque assay using Vero/TMPRSS2 cell line.

While there are several methods of viral quantification, the plaque assay remains the gold standard for accurate quantification of replication-competent, cytopathic viruses including Zika virus (ZIKV). Vero cells are commonly used to titer ZIKV stock via the plaque assay. Prior to the initiation of this study, we observed that ZIKV-PRV strain plaque assays using Vero cells often yielded plaques that were very small, overly diffuse, and hard to count. We also observed that the ZIKV-CAM strain often did not produce plaques at all on Vero cells. This study shows that Vero cells expressing TMPRSS2 improve the morphology and visibility of plaques produced by ZIKV-PRV and ZIKV-CAM compared to regular Vero cells and may, therefore, be a better alternative to use for performing plaque assays for these strains and perhaps for other strains of ZIKV that are difficult to titer. However, Vero cells proved to be superior for generating high titer stock.

A novel pan-epitope based nanovaccine self-assembled with CpG enhances immune responses against flavivirus

BackgroundFlavivirus is a highly prevalent and outbreak-prone disease, affecting billions of individuals annually and posing substantial public health challenges. Vaccination is critical to reducing the global impact of flavivirus infections, making the development of a safe and effective vaccine a top priority. The self-assembled pan-epitope vaccine presents key advantages for improving immunogenicity and safety without relying on external vectors or adding immunomodulatory elements, both of which are essential for successful vaccine development.ResultsIn this study, the pan-epitope peptide TBT was combined with adjuvant CpG to form the TBT-CpG nanovaccine (TBT-CpG NaVs), which was found to be spherical, uniform in shape, and demonstrated strong serum stability. In vitro studies showed that the TBT-CpG NaVs were efficiently taken up and internalized by bone marrow-derived dendritic cells (BMDCs). Flow cytometry and transcriptomic analysis indicated that the antigens were effectively presented to antigen-presenting cells (APCs) via the MHC II pathway, which facilitated BMDCs maturation and promoted the release of pro-inflammatory cytokines IL-1β, TNF-α, and IL-6. In vivo studies confirmed that TBT-CpG NaVs enhanced antigen-specific IgG levels, significantly increased IFN-γ and IL-4 expression in spleen cells, and offered protective effects against Dengue virus (DENV) and Zika virus (ZIKV) infections. Safety evaluations revealed no hepatotoxicity and no significant organ damage in immunized mice.ConclusionThe self-assembled candidate nanovaccine TBT-CpG NaVs effectively activates BMDCs and triggers a targeted immune response, providing antiviral effects against DENV and ZIKV. This vaccine demonstrates good immunogenicity and safety, establishing a promising foundation and a new strategy for the development of safe and effective vaccines.Graphical

Sustained antiviral insulin signaling during West Nile virus infection results in viral mutations

Arthropod-borne viruses or arboviruses, including West Nile virus (WNV), dengue virus (DENV), and Zika virus (ZIKV) pose significant threats to public health. It is imperative to develop novel methods to control these mosquito-borne viral infections. We previously showed that insulin/insulin-like growth factor-1 signaling (IIS)-dependent activation of ERK and JAK-STAT signaling has significant antiviral activity in insects and human cells. Continuous immune pressure can lead to adaptive mutations of viruses during infection. We aim to elucidate how IIS-signaling in mosquitoes selects for West Nile virus escape variants, to help formulate future transmission blocking strategies. We hypothesize that passage of WNV under activation of IIS will induce adaptive mutations or escape variants in the infecting virus. To test our hypothesis, WNV was serially passaged through Culex quinquefasciatus Hsu cells in the presence or absence of bovine insulin to activate IIS antiviral pressure. We sequenced WNV genes encoding for E, NS2B, NS3, and NS5 and identified variants in E and NS5 arising from IIS antiviral pressure. In parallel to the genetic analyses, we also report differences in the levels of virus replication and Akt activation in human cells and mosquitoes using virus passaged in the presence or absence of insulin. Finally, using adult Culex quinquefasciatus, we demonstrated the enhancement of immune response gene expression in virus-infected mosquitoes fed on insulin, compared to control. Notably, virus collected from insulin-fed mosquitoes contained a non-synonymous mutation in NS3. These results contribute towards achieving our long-term goal of manipulating mosquito IIS-dependent antiviral immunity to reduce WNV or other flavivirus transmission to mammalian hosts.

Effects of copper nanoparticles synthesized from the entomopathogen Metarhizium robertsii against the dengue vector Aedes albopictus (Skuse, 1894).

Aedes albopictus, known as the Asian tiger mosquito, is a significant vector for dengue fever, chikungunya, zika virus, yellow fever. Current control methods rely on chemical insecticides, which face challenges such as resistance, environmental harm, and impact on non-target species Eudrilus eugeniae and Artemia salina. This study evaluates the toxic effects of biogenic copper nanoparticles (CuNPs) synthesized using Metarhizium robertsii intracellular extract obtained from our previous research. The CuNPs were tested against A. albopictus and non-target species at 24 and 48 hours post-treatment. Results demonstrated that entomopathogenic fungi-derived CuNPs exhibited potent mosquitocidal activity, resulting in 97.33% mortality in larvae, 93.33% in pupae, and 74.66% in adults at 48 hours post-treatment. The CuNPs derived from M. robertsii showed lower LC50 values of 74.873 mg/L in larvae, 76.101 mg/L in pupae, and 136.645 mg/L in adults at 48 hours post-treatment. Additionally, 12 hours post-treatment, catalase (an antioxidant enzyme) activity decreased 1.5-fold in a dose-dependent manner, while glutathione S-transferase (a detoxification enzyme) activity increased 7.8-fold. CuNPs demonstrated lower toxicity to non-target species, with 24% mortality in A. salina and 24.44% mortality in E. eugeniae at 24 hours post-treatment. The LC50 values were 634.747 mg/L for A. salina and 602.494 mg/L for E. eugeniae at 24 hours post-treatment. These findings indicate that entomopathogenic fungi-derived CuNPs are a promising, target-specific candidate for controlling A. albopictus at various life stages (larvae, pupae, and adults).

Analysis of the heterologous expression, localization, and cellular response to the Zika virus E protein in vitro.

Zika virus (ZIKV) infection has been associated with damage to neural stem cells in microcephaly in newborns. The virus possesses specific tropism for glioma stem cells mediated by the ZIKV E protein. This infection causes endoplasmic reticulum stress and activation of the unfolded protein response (UPR). However, the cellular response to the expression of the ZIKV E protein alone is unknown. Therefore, in this study, we determined the effect of the expression of the ZIKV E protein on cellular responses and its subcellular localization in HEK-293T cells, due to their use as a biotechnological tool for cellular and lentiviral therapy. We observed that the ZIKV E protein is synthesized in the cytoplasm and inserted into the endoplasmic reticulum (ER), without causing activation of the UPR or cell death, and it is finally transported and located in the cell membrane. Moreover, the expression of the ZIKV E protein does not induce UPR or apoptosis in glioma cells. These results help us to better understand the characteristics of this protein and its possible use as a biotechnological tool for the creation of different gene therapy strategies, vaccines, and synthetic vectors with tropism for neural and glioma stem cells.