Sequential nucleophilic aromatic substitutions on cyanuric chloride: synthesis of BODIPY derivatives and mechanistic insights.

Herein we report a study on the sequential substitution of different nucleophiles on cyanuric chloride to obtain potential candidates for metal sensors (5a-c). The set of nucleophiles on the 1,3,5-triazine ring includes a phenolic BODIPY, an aminoalkyl pyridine and aminoalkyl phosphoramidates, each one designed to play a specific role in the final fluoroionophore. Three new triazine triads were synthesized in similar yields: 5a (45%), 5b (43%) and 5c (52%) after a methodical sequential combination of the nucleophiles via thermodependent nucleophilic aromatic substitution of the three chlorine atoms of cyanuric chloride. To ratify the synthetic results we simulated the reaction mechanisms for the different nucleophiles, aiming to address the distinctive orthogonality and temperature control inherent in this process, identifying and providing a sound rationale for any preferential sequence of nucleophiles inserted into the triazine core. According to our experimental and computational analysis (thermo- and kinetic preferences), we have identified the following preferential order for the sequential substitution: p-hydroxybenzaldehyde > 2-(pyridin-2-yl)ethanamine > aminoalkyl phosphoramidate, indicating that all steps follow a single-step process (concerted) in two stages, where nucleophilic addition precedes leaving group dissociation. The Meisenheimer σ-complex was identified as a transition state structure, with insufficient stability to exist as an intermediate. We observed a consistent and progressive increase in barrier height: 2-8 kcal mol-1 for the first step, 9-15 kcal mol-1 for the second step, and >15 kcal mol-1 for the third substitution. These findings align with the experimental observation of thermodependency in the sequential substitution.

Photocatalytic removal of 17ß-estradiol from water using a novel bimetallic NiCu/Nb2O5 catalyst.

The development of effective photocatalytic materials is essential for removing emerging pollutants from aqueous media, such as the hormone 17ß-estradiol (E2). In this study, a novel photocatalyst based on niobium pentoxide (Nb2O5) functionalized with nickel (Ni) and copper (Cu) was synthesized for E2 removal. The NiCu/Nb2O5 photocatalyst was prepared using a facile wet impregnation method and characterized by various techniques. The incorporation of Ni and Cu into Nb2O5 reduced the band gap energy from 3.3 to 2.8 eV, enabling efficient utilization of visible light. Moreover, NiCu/Nb2O5 exhibited the highest E2 removal efficiency (82%) under UV-A-assisted conditions at a concentration of 1.5 g L-1. The reaction kinetics were found to follow a second-order model with a rate constant of k = 0.0020 L g-1 min-1, and a plausible reaction mechanism was proposed. Through the study of radical elimination, it was proven that the radical oxidation reaction mechanism predominated in the reaction. The results of the toxicity assays, combined with the TOC parameter, demonstrated the efficacy of photocatalytic degradation in reducing E2. These findings demonstrate the great potential of the NiCu/Nb2O5 photocatalyst for removing persistent pollutants.

Evaluation of SARS-CoV-2 ORF7a Deletions from COVID-19-Positive Individuals and Its Impact on Virus Spread in Cell Culture.

The spread of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), causing the COVID-19 outbreak, posed a primary concern of public health worldwide. The most common changes in SARS-CoV-2 are single nucleotide substitutions, also reported insertions and deletions. This work investigates the presence of SARS-CoV-2 ORF7a deletions identified in COVID-19-positive individuals. Sequencing of SARS-CoV-2 complete genomes showed three different ORF7a size deletions (190-nt, 339-nt and 365-nt). Deletions were confirmed through Sanger sequencing. The ORF7a∆190 was detected in a group of five relatives with mild symptoms of COVID-19, and the ORF7a∆339 and ORF7a∆365 in a couple of co-workers. These deletions did not affect subgenomic RNAs (sgRNA) production downstream of ORF7a. Still, fragments associated with sgRNA of genes upstream of ORF7a showed a decrease in size when corresponding to samples with deletions. In silico analysis suggests that the deletions impair protein proper function; however, isolated viruses with partial deletion of ORF7a can replicate in culture cells similarly to wild-type viruses at 24 hpi, but with less infectious particles after 48 hpi. These findings on deleted ORF7a accessory protein gene, contribute to understanding SARS-CoV-2 phenotypes such as replication, immune evasion and evolutionary fitness as well insights into the role of SARS-CoV-2_ORF7a in the mechanism of virus-host interactions.

A three-dimensionally printed otological model for cholesteatoma mastoidectomy training.

PURPOSE: To relate the creation and expert validation (face and content validity) of an affordable three-dimensional (3-D) printed model of temporal bones with chronic otitis media with cholesteatoma (COMC) as a simulator for mastoidectomy. METHODS: We performed computed tomography (CT) of the temporal bones of a patient with COMC followed at the University of São Paulo (USP) Hospital with 3-D Slicer to create a 3-D model of the affected bone using light-curing resin and silicone (cholesteatoma). The final 3-D printed images were scored by 10 otologists using a customized version of the Michigan Standard Simulation Scale Experience (MiSSES). Internal consistency and inter-rater reliability were assessed using Cronbach's α and intraclass correlations. RESULTS: Otologists consistently scored the model positively for fidelity, educational value, reactions, and the overall model quality. Nine otologists agreed that the model was a good educational device for surgical training of COMC. All experts deemed the model ready-or nearly ready-for use. The final cost of the model, including raw materials and manufacturing, was 120 USD. CONCLUSIONS: Using 3-D printing technology, we created the first anatomically accurate, low-cost, disease-reproducing 3-D model of temporal bones for mastoidectomy training for cholesteatoma.

Chloroquinolone Carboxamide Derivatives as New Anti-HSV-1 Promising Drugs.

BACKGROUND: Since the emergence of HSV resistant strains, new antiviral agents have emerged and still are urgently needed, especially those with alternative targets. OBJECTIVE: In this work, we evaluated new quinolone derivatives as anti-HSV. METHODS: For this study, cells were infected and treated with different components to evaluate the profile of HSV replication in vitro. In addition, studies were performed to determine the pharmacokinetic toxicity and profile of the compound. RESULTS: Indeed the EC50 values of these promising molecules ranged between 8 µM and 32 µM. We have also showed that all compounds inhibited the expression of ICP27 viral proteins, which gives new insights in the search for new target for antiherpetic therapy. Chlorine in positions C6 and phosphonate in position C1 have shown to be important for viral inhibition. The chloroquinolone carboxamide derivatives fulfilled "Lipinsky Rule of Five" for good oral bioavailability and showed higher intestinal absorption and blood brain barrier penetration, as well as lower toxicity profile. CONCLUSION: Although the inhibition activities of chloroquinolone carboxamide derivatives were lower than acyclovir, they showed different modes of action in comparison to the drugs currently available. These findings encourage us to continue pre-clinical studies for the development of new anti-HSV-1 agents.

A SARS-CoV-2 Negative Antigen Rapid Diagnostic in RT-qPCR Positive Samples Correlates With a Low Likelihood of Infectious Viruses in the Nasopharynx.

Severe acute respiratory syndrome-related coronavirus (SARS-CoV-2) transmission occurs even among fully vaccinated individuals; thus, prompt identification of infected patients is central to control viral circulation. Antigen rapid diagnostic tests (Ag-RDTs) are highly specific, but sensitivity is variable. Discordant RT-qPCR vs. Ag-RDT results are reported, raising the question of whether negative Ag-RDT in positive RT-qPCR samples could imply the absence of infectious viruses. To study the relationship between negative Ag-RDT results with virological, molecular, and serological parameters, we selected a cross-sectional and a follow-up dataset and analyzed virus culture, subgenomic RNA quantification, and sequencing to determine infectious viruses and mutations. We demonstrated that RT-qPCR positive while SARS-CoV-2 Ag-RDT negative discordant results correlate with the absence of infectious virus in nasopharyngeal samples. A decrease in sgRNA detection together with an expected increase in detectable anti-S and anti-N IgGs was also verified in these samples. The data clearly demonstrate that a negative Ag-RDT sample is less likely to harbor infectious SARS-CoV-2 and, consequently, has a lower transmissible potential.

Analysis of the clinical practice of the pharmacist in a community pharmacy: A Cross-sectional Study from Brazil.

Background: The pharmaceutical profession has experienced renewals over time. In community pharmacies, pharmaceutical services contribute to the public health system in Brazil. The development of these tasks, in collaboration with professionals from the multidisciplinary team, demonstrates the involvement with the well-being, health and improvement of the patient's life. Objectives: This study aimed to identify the professional practice of pharmacists, their understandings and attitudes towards clinical practice in community pharmacies in northern Brazil. Methods: This is a cross-sectional descriptive study, based on a questionnaire carried out with pharmacists for seven months in the municipality of Belém, in the state of Pará. Data were expressed using descriptive statistics and the results were shown as a percentage. Results: 182 pharmacists participated. Females were predominant (80.2%) and the average age of participants was 34.2 years. 77.4% graduated from private institutions and 59.3% already have a specialization. 38.4% hold the position of technical director. 50.5% of respondents say that community pharmacies have a reserved place for service. The most used clinical services were pharmacotherapy review (89.5%), health education (60.9%), dispensing (34%), therapeutic monitoring (25.8%) and pharmacotherapeutic follow-up (4.9%). In the study, it was realized that community pharmacies should not be seen as a commercial place but rather as a health care facility. Conclusions: Therefore, these establishments must adapt at a structural and professional level, to meet an increasingly growing demand of a population in need of services offered with quality health care.

A new species of Progomphus Selys, 1854 (Odonata: Anisoptera: Gomphidae) from Minas Gerais state, Southeastern Brazil.

Progomphus teolitavius sp. nov. (Odonata: Anisoptera: Gomphidae) is described and diagnosed based on a specimen collected in a gallery forest of Cerrado from Barroso municipality, Minas Gerais state, Brazil (-21.2238, -43.9895, 1033 m, 30.iii.2021, G.S. Santos leg.). The new species can be distinguished from congeners by its enlarged basal externo-lateral dilatation of cerci (which bears large teeth), and epiproct morphology.

Natural Areas of Cerrado Foster Wasp (Hymenoptera) Diversity in Human Modified Landscapes.

Land use changes from native vegetation to agriculture, livestock grazing, and urban development are among the main problems related to biodiversity loss worldwide. In this paper we evaluate how land use changes (eucalypt plantation and pasture) affect the richness and assemblage of wasps (Braconidae, Ichneumonidae, Pompilidae, and Vespidae), in comparison with nearby areas with native vegetation in the Cerrado. Specimens were collected at six points, with two Malaise traps at each location. The collections were performed monthly for 10 d, for 12 mo. A total of 773 hymenopterans of the selected groups were collected, representing 253 species or morphospecies. Richness of the families Ichneumonidae and Pompilidae between the areas did not present significant differences. For the families Braconidae and Vespidae, the richness was greater in the eucalypt plantation and pasture areas compared to the native area. Species composition in the native habitat was different from either of the managed habitats in the studied environment. Furthermore, the composition of wasps in native areas varied less throughout the sampling campaigns when compared with the pasture and eucalyptus sites. In native areas, 85 exclusive morphospecies were found. Thus, changes in land use may cause changes in the composition of wasp species, since areas with native vegetation presented more heterogeneous and stable environments than the other land uses. The maintenance of native areas, even if close to planted forest and/or pasture areas, could be the best way to combine forest productivity with biodiversity conservation.

Three-dimensional printing in otolaryngology education: a systematic review.

PURPOSE: The progressive expansion of the technology that facilitates the development of three-dimensional (3D) printing within the field of otorhinolaryngology has opened up a new study front in medicine. The objective of this study is to systematically review scientific publications describing the development of 3D models having applications in otorhinolaryngology, with emphasis on subareas with a large number of publications, as well as the countries in which the publications are concentrated. METHODS: In this literature review, specific criteria were used to search for publications on 3D models. The review considered articles published in English on the development of 3D models to teach otorhinolaryngology. The studies with presurgical purposes or without validation of the task by surgeons were excluded from this review. RESULTS: This review considered 39 articles published in 10 countries between 2012 and 2021. The works published prior to 2012 were not considered as per the inclusion criteria for the research. Among the 39 simulators selected for review, otology models comprised a total of 15 publications (38%); they were followed by rhinology, with 12 (31%); laryngology, with 8 (21%); and head and neck surgery, with 4 publications (10%). CONCLUSION: The use of 3D technology and printing is well established in the context of surgical education and simulation models. The importance of developing new technological tools to enhance 3D printing and the current limitations in obtaining appropriate animal and cadaver models signify the necessity of investing more in 3D models.

A Novel Host of an Emerging Disease: SARS-CoV-2 Infection in a Giant Anteater (Myrmecophaga tridactyla) Kept Under Clinical Care in Brazil.

A young male free-ranging giant anteater (Myrmecophaga tridactyla) was found with paralysis of pelvic limbs on a highway and kept under human care. Radiographs confirmed multiple incomplete fractures in the thoracolumbar vertebrae. Due to the poor prognosis, euthanasia was chosen. The infection was established by viral SARS-CoV-2 RNA detection in the rectal swab, spleen and kidney samples. Immunohistochemistry detected the viral nucleocapsid protein in sections of the lungs, liver, spleen, lymph nodes, and large intestine sections, and spike protein antigen in the lung tissue. Pilosa order species should be included as potential hosts of natural infection of SARS-CoV-2.

Complete experimental and theoretical characterization of nonlinear concentration gradient generator microfluidic device for analytical purposes.

Microfluidic devices that generate stable concentration gradients are efficient instruments for automated calibration for analytical and bioanalytical systems. However, little attention has been paid to the development of reusable microfluidic concentration gradient generators, which can be useful for a range of species through mathematical characterization. In this work, we develop a microfluidic device based on three steps of serial dilution that were able to generate nonlinear concentration gradient for dyes and biomolecules. The microfluidic device was described mathematically, statistically and was suitable for reusable analytical and bioanalytical analysis. The device reproducibility was assessed by experimental tests, which have shown the same gradient concentration profile for different dyes and statistical reproducibility with 95% confidence interval for bovine serum albumin (BSA). Moreover, the experimental data converged well with those  obtained by computational fluid dynamics simulation. Applicability was verified by coupling the microfluidic device to a surface plasmon resonance (SPR) biosensor, based on nanohole arrays with sensitivity of 358.7 nm RIU-1 determined by white-light SPR excitation exposed to different D-(+)-glucose aqueous solutions with 1.3361-1.4035 refractive index interval. The transmission light intensities obtained by the array of images allowed to quantify a pseudo-unknown BSA sample (160 µg mL-1) at 138 µg mL-1. The SPR analysis has been validated in parallel by fluorescence emissions, which showed a concentration of 154.8 ± 16.6 µg mL-1.

Prolonged SARS-CoV-2 Positivity in Immunocompetent Patients: Virus Isolation, Genomic Integrity, and Transmission Risk.

Current guidelines for patient isolation in COVID-19 cases recommend a symptom-based approach, averting the use of control real-time reverse transcription PCR (rRT-PCR) testing. However, we hypothesized that patients with persistently positive results by RT-PCR for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) could be potentially infectious for a prolonged time, even if immunocompetent and asymptomatic, which would demand a longer social isolation period than presently recommended. To test this hypothesis, 72 samples from 51 mildly symptomatic immunocompetent patients with long-lasting positive rRT-PCR results for SARS-CoV-2 were tested for their infectiousness in cell culture. The serological response of samples from those patients and virus genomic integrity were also analyzed. Infectious viruses were successfully isolated from 34.38% (22/64) of nasopharynx samples obtained 14 days or longer after symptom onset. Indeed, we observed successful virus isolation up to 128 days. Complete SARS-COV-2 genome integrity was demonstrated, suggesting the presence of replication-competent viruses. No correlation was found between the isolation of infectious viruses and rRT-PCR cycle threshold values or the humoral immune response. These findings call attention to the need to review current isolation guidelines, particularly in scenarios involving high-risk individuals. IMPORTANCE In this study, we evaluated mildly symptomatic immunocompetent patients with long-lasting positive rRT-PCR results for SARS-CoV-2. Infectious viruses were successfully isolated in cell cultures from nasopharynx samples obtained 14 days or longer after symptom onset. Indeed, we observed successful virus isolation for up to 128 days. Moreover, SARS-CoV-2 genome integrity was demonstrated by sequencing, suggesting the presence of replication-competent viruses. These data point out the risk of continuous SARS-CoV-2 transmission from patients with prolonged detection of SARS-CoV-2 in the upper respiratory tract, which has important implications for current precaution guidelines, particularly in settings where vulnerable individuals may be exposed (e.g., nursing homes and hospitals).

Camouflaged Nests of Mischocyttarus mirificus (Hymenoptera, Vespidae).

Social wasps present various architectural patterns for their nests, which may differ in shape, size, color, and material used in construction. The distribution pattern of comb cells presented by Mischocyttarus mirificus (Zikán, 1935) is vertical with a single cell of width, resulting in a filiform shape that camouflages in the middle of the substrate. There are few studies regarding this architectural pattern for social wasps and their role in camouflage, and this study aims to detail the nesting habits of this species. In total, 40 colonies were analyzed in their natural habitat and six were dissected in the laboratory. Aspects about the construction of the nests and nesting environment have been described. A test was carried out with nest photographs, to quantify their camouflage percentage within the nesting substrate. The colonies were found mainly in riparian forests where there were elements in the environment that favored their camouflage. According to the nest photograph analysis, their camouflage levels can vary according to the position from which the photograph is taken. It is evident that M. mirificus founders select sites where they can camouflage their colonies, and this gives them advantages in exploring a new environment.

How chemical engineers can contribute to fight the COVID-19.

The SARS-CoV-2 virus, promoter of COVID-19, already infected millions of people around the world, resulting in thousands of fatal victims. Facing this unprecedented crisis in human history, several research groups, industrial companies and governments have been spending efforts to develop vaccines and medications. People from distinct knowledge fields are doing their part in order to overcome this crisis. Chemical Engineers are also contributing in the development of actions to control the SARS-CoV-2 virus. However, many chemical engineers still do not know how to use the knowledge acquired from Chemical Engineering school to collaborate in the fight against the COVID-19. In this context, the present paper aims to discuss several knowledge fields within the Chemical Engineering and correlated areas successfully applied to create innovative and effective solutions in the fight against the COVID-19.

Multi-volume modeling of Eucalyptus trees using regression and artificial neural networks.

The stem volume of commercial trees is an important variable that assists in decision making and economic analysis in forest management. Wood from forest plantations can be used for several purposes, which makes estimating multi-volumes for the same tree a necessary task. Defining its exploitation and use potential, such as the total and merchantable volumes (up to a minimum diameter of interest), with or without bark, is a possible work. The goal of this study was to use different strategies to model multi-volumes of the stem of eucalyptus trees. The data came from rigorous scaling of 460 felled trees stems from four eucalyptus clones in high forest and coppice regimes. The diameters were measured at different heights, with the volume of the sections obtained by the Smalian method. Data were randomly separated into fit and validation data. The single multi-volume model, volume-specific models, and the training of artificial neural networks (ANNs) were fitted. The evaluation criteria of the models were: coefficient of determination, root mean square error, mean absolute error, mean bias error, as well as graphical analysis of observed and estimated values and distribution of residuals. Additionally, the t-test (α = 0.05) was performed between the volume obtained in the rigorous scaling and estimated by each strategy with the validation data. Results showed that the strategies used to model different tree stem volumes are efficient. The actual and estimated volumes showed no differences. The multi-volume model had the most considerable advantage in volume estimation practicality, while the volume-specific models were more efficient in the accuracy of estimates. Given the conditions of this study, the ANNs are more suitable than the regression models in the estimation of multi-volumes of eucalyptus trees, revealing greater accuracy and practicality.

Host Retromer Protein Sorting Nexin 2 Interacts with Human Respiratory Syncytial Virus Structural Proteins and is Required for Efficient Viral Production.

Human respiratory syncytial virus (HRSV) envelope glycoproteins traffic to assembly sites through the secretory pathway, while nonglycosylated proteins M and N are present in HRSV inclusion bodies but must reach the plasma membrane, where HRSV assembly happens. Little is known about how nonglycosylated HRSV proteins reach assembly sites. Here, we show that HRSV M and N proteins partially colocalize with the Golgi marker giantin, and the glycosylated F and nonglycosylated N proteins are closely located in the trans-Golgi, suggesting their interaction in that compartment. Brefeldin A compromised the trafficking of HRSV F and N proteins and inclusion body sizes, indicating that the Golgi is important for both glycosylated and nonglycosylated HRSV protein traffic. HRSV N and M proteins colocalized and interacted with sorting nexin 2 (SNX2), a retromer component that shapes endosomes in tubular structures. Glycosylated F and nonglycosylated N HRSV proteins are detected in SNX2-laden aggregates with intracellular filaments projecting from their outer surfaces, and VPS26, another retromer component, was also found in inclusion bodies and filament-shaped structures. Similar to SNX2, TGN46 also colocalized with HRSV M and N proteins in filamentous structures at the plasma membrane. Cell fractionation showed enrichment of SNX2 in fractions containing HRSV M and N proteins. Silencing of SNX1 and 2 was associated with reduction in viral proteins, HRSV inclusion body size, syncytium formation, and progeny production. The results indicate that HRSV structural proteins M and N are in the secretory pathway, and SNX2 plays an important role in the traffic of HRSV structural proteins toward assembly sites.IMPORTANCE The present study contributes new knowledge to understand HRSV assembly by providing evidence that nonglycosylated structural proteins M and N interact with elements of the secretory pathway, shedding light on their intracellular traffic. To the best of our knowledge, the present contribution is important given the scarcity of studies about the traffic of HRSV nonglycosylated proteins, especially by pointing to the involvement of SNX2, a retromer component, in the HRSV assembly process.

Anti-EGFR Fibronectin Bispecific Chemically Self-Assembling Nanorings (CSANs) Induce Potent T Cell-Mediated Antitumor Responses and Downregulation of EGFR Signaling and PD-1/PD-L1 Expression.

Overexpression of the epidermal growth factor receptor (EGFR) on various cancers makes it an important target for cancer immunotherapy. We recently demonstrated that single-chain variable fragment-based bispecific chemically self-assembled nanorings (CSANs) can successfully modify T cell surfaces and function as prosthetic antigen receptors (PARs) allowing selective targeting of tumor antigens while incorporating a dissociation mechanism of the rings. Here, we report the generation of anti-EGFR fibronectin (FN3)-based PARs with high yield, rapid protein production, predicted low immunogenicity, and increased protein stability. We demonstrated the cytotoxicity of FN3-PARs successfully while evaluating FN3 affinities, CSAN valencies, and antigen expression levels. Using an orthotopic breast cancer model, we showed that FN3-PARs can suppress tumor growth with no adverse effects and FN3-PARs reduced immunosuppressive programmed cell death ligand-1 (PD-L1) expression by downregulating EGFR signaling. These results demonstrate the potential of FN3-PARs to direct selective T cell-targeted tumor killing and to enhance antitumor T cell efficacy by modulating the tumor microenvironment.

Chikungunya Virus: An Emergent Arbovirus to the South American Continent and a Continuous Threat to the World.

Chikungunya virus (CHIKV) is an arthropod-borne virus (arbovirus) of epidemic concern, transmitted by Aedes ssp. mosquitoes, and is the etiologic agent of a febrile and incapacitating arthritogenic illness responsible for millions of human cases worldwide. After major outbreaks starting in 2004, CHIKV spread to subtropical areas and western hemisphere coming from sub-Saharan Africa, South East Asia, and the Indian subcontinent. Even though CHIKV disease is self-limiting and non-lethal, more than 30% of the infected individuals will develop chronic disease with persistent severe joint pain, tenosynovitis, and incapacitating polyarthralgia that can last for months to years, negatively impacting an individual's quality of life and socioeconomic productivity. The lack of specific drugs or licensed vaccines to treat or prevent CHIKV disease associated with the global presence of the mosquito vector in tropical and temperate areas, representing a possibility for CHIKV to continually spread to different territories, make this virus an agent of public health burden. In South America, where Dengue virus is endemic and Zika virus was recently introduced, the impact of the expansion of CHIKV infections, and co-infection with other arboviruses, still needs to be estimated. In Brazil, the recent spread of the East/Central/South Africa (ECSA) and Asian genotypes of CHIKV was accompanied by a high morbidity rate and acute cases of abnormal disease presentation and severe neuropathies, which is an atypical outcome for this infection. In this review, we will discuss what is currently known about CHIKV epidemics, clinical manifestations of the human disease, the basic concepts and recent findings in the mechanisms underlying virus-host interaction, and CHIKV-induced chronic disease for both in vitro and in vivo models of infection. We aim to stimulate scientific debate on how the characterization of replication, host-cell interactions, and the pathogenic potential of the new epidemic viral strains can contribute as potential developments in the virology field and shed light on strategies for disease control.

In Vitro and In Vivo Characterization of the Anti-Zika Virus Activity of ProTides of 2'-C-ß-Methylguanosine.

The ProTide approach has emerged as a powerful tool to improve the intracellular delivery of nucleotide analogs with antiviral and anticancer activity. Here, we characterized the anti-ZIKV (ZIKV, Zika virus) activity of two ProTides of 2'-C-ß-methylguanosine. ProTide UMN-1001 is a 2'-C-ß-methylguanosine tryptamine phosphoramidate monoester, and ProTide UMN-1002 is a 2-(methylthio)-ethyl-2'-C-ß-methylguanosine tryptamine phosphoramidate diester. UMN-1002 undergoes stepwise intracellular activation to the corresponding nucleotide monophosphate followed by P-N bond cleavage by intracellular histidine triad nucleotide binding protein 1 (Hint1). UMN-1001 is activated by Hint1 but is less cell-permeable than UMN-1002. UMN-1001 and UMN-1002 were found to be more potent than 2'-C-ß-methylguanosine against ZIKV in human-derived microvascular endothelial and neuroblastoma cells and in reducing ZIKV RNA replication. Studies with a newborn mouse model of ZIKV infection demonstrated that, while treatment with 2'-C-ß-methylguanosine and UMN-1001 was lethal, treatment with UMN-1002 was nontoxic and significantly reduced ZIKV infection. Our data suggests that anchimeric activated ProTides of 2'-C-ß-methyl nucleosides should be further investigated for their potential as anti-ZIKV therapeutics.