Determination of the relationship between olfactory function threshold and quality of life among adult Filipinos with perceived olfactory dysfunction

Objective@#To determine the relationship between olfactory function threshold and quality of life (QOL) among adult Filipinos with perceived olfactory dysfunction (OD).@*Methods@#Design:Cross - Sectional Study. Setting:Tertiary Government Training Hospital. Participants: 98 adults who had self-perceived olfactory dysfunction described as “poor” or “no sense” of smell@*Results@#We analyzed data from 98 participants, with a mean age of 35.91 + 12.58 years old, composed of 46 men (47%) and 52 women (53%), with 82 normosmic, six hyposmic and ten anosmic as categorized by their BTT scores. Twenty-seven percent (27%) identified themselves as having poor QOL based on Fil 17 QODNS. Differences were exhibited between sexes’ BTT scores - [t(96) = -2.32; p = .022; females, M: 9.25; SD: 2.33 vs. males, M: 7.76; SD : 3.91], civil status - Fil17QODNS scores [t(96)= 3.05, p < .003; married M: 11.72, SD: 13.74 vs. single, M: 4.71; SD: 8.66), and the presence of ENT symptoms BTT [t(96) = -7.15; p < .0001; symptomatic, M: 5.62; SD: 4.54, vs. asymptomatic, M: 9.78; SD: 1.14] and Fil 17 QODNS scores [t(96)= 3.94; P < .00001; symptomatic, M: 14.86; SD: 13.97] vs. asymptomatic, M: 5.217; SD: 9.60]. Significant risk factors were the presence of ENT symptoms [OR= 0.15; 95% CI: 0.02-0.97; P = .046] for poor smell threshold, and comorbidities [OR= 3.36; 95% CI: 1.04-10.85; P = .043] for poor QOL. A negative correlation was observed between Fil-QOD-NS scores and BTT scores [r = -0.477, p < .001; rs = -0.292, p = .004], signifying that the presence of olfactory dysfunction has an inverse relationship with the quality of life.@*Conclusion@#Individuals with olfactory dysfunction in this study population had poor quality of life as determined by the translated Filipino 17-item QOD-NS which has an inverse relationship with their smell threshold as represented by the BTT results. Among the factors studied, significant differences were found between sexes, civil status and presence of ENT symptoms in the BTT and Fil 17 QODNS scores. However, only presence of comorbidities and ENT symptoms are significant risk factors for quality of life and smell threshold, respectively, in this population.

Predominant amino acid substitutions in NS5B gene of hepatitis C virus in blood donors and treatment-naïve hepatitis and HIV patients in Nigeria

Background: Hepatitis C virus (HCV) genome undergoes high rate of mutation, which results in generation of genetically diverse HCV isolates. There is paucity of data on mutations in the nonstructural 5b (NS5b) gene of circulating HCV and their implications in the Nigerian population. Here, we identified clinically-important mutations in HCV isolates, which may influence response to therapy and disease prognosis. Methodology: HCV RNA was extracted from a total of 301 blood samples collected from 99 symptomatic treatment-naïve hepatitis patients, 125 HIV-infected individuals and 77 asymptomatic blood donors in Ibadan, Nigeria. The RNA was reverse­transcribed to complimentary DNA and HCV NS5B gene amplified by nested PCR. The amplified products of 42 HCV were sequenced and sequences were aligned with those from GenBank and HCV databases in MEGA 7.0. Nucleotide sequences were translated to amino acids while substitutions in the amino acids were analyzed with reference to H77 prototype strain of HCV. Results: A total of 10 amino acid polymorphisms were observed from the 42 sequenced NS5B gene, with the major clinically-important amino acid mutations being S15G in 28 (66.7%) participants, T7N (24, 57.1%), G61R (23, 54.8%), S54L (22, 52.4%), G89E (14, 33.3%), T79M (12, 28.6%), and T711 (11, 26.2%). Others were Q67R (7, 16.7%), Q47H (7, 16.7%) and S84F (2, 4.8%). S15G/A/V mutations were more predominant in patients with HIV (76.9%, 10/13) followed by patients with clinical hepatitis (75.0%, 12/16) and blood donors (46.1%, 6/13). Q67R and T71I mutations were not predominant in patients with clinical hepatitis as they were detected in only 31.3% (5/16) and 43.8% (7/16) participants respectively, compared to S15G (75.0%, 12/16), S54L (68.8%, 11/16), G61R/E (68.8%, 11/16) and T7N/S (56.3%, 9/16). There was no statistically significant difference in the distribution of each of the 10 amino acid polymorphisms detected within patients with symptomatic clinical hepatitis (x 2=9.311, p=0.409), HIV-infected patients (x 2=13.431, p=0.1440) and asymptomatic blood donors (x 2=3.775, p=0.9256). Similarly, there was no significant difference in the distribution between the 3 categories of the study participants except for T79M mutation, which was significantly higher in HIV-infected patients (61.5%, 8/13) compared to patients with clinical hepatitis (18.8%, 3/16) and asymptomatic blood donors (7.7%, 1/13) (x 2=10.456, p=0.0054). Conclusion: Mutations in the NS5B gene could be associated with worse prognosis of the disease or antiviral failure due to viral resistance in patients undergoing therapy. The absence of Q47H mutations in majority of the study participants in our study implies that they will not respond well to daprevir and mericitabine. Screening of patients for pre-existing resistant mutations before commencement of therapy and monitoring during and after therapy are recommended.

Elevated NS1 serum levels reduce CD119 expression and CXCL-10 synthesis in patients with dengue hemorrhagic fever

ABSTRACT Background: The intensity of dengue virus (DV) replication and circulating non-structural protein 1 (NS1) levels may promote changes in the human immune response and favor severe forms of infection. We investigated the correlations between NS1 with CXCL-8, CXCL-10, IFN-γ, and IL-12p40 serum levels, and IFN-γ receptor α chain (CD119) expression, and CXCL10 production by peripheral blood mononuclear cells (PBMCs) stimulated with recombinant IFN-γ in DV-infected patients with different clinical forms. Methods: Dengue virus NS1, CXCL-8, CXCL-10, IFN-γ, and IL-12p40 serum levels were measured in 152 DV-infected patients with different clinical forms and 20 non-infected individuals (NI) using enzyme-linked immunosorbent assay (ELISA). In addition, we investigated the CXCL-10 production after in vitro IFN-γ stimulation of PBMCs from 48 DV-infected individuals (with different clinical forms of dengue fever) and 20 NI individuals using ELISA, and CD119 expression on CD14+ cells with flow cytometry. Results: Patients with dengue hemorrhagic fever (DHF) had significantly higher NS1, CXCL-8, and CXCL-10 serum levels than those with classic dengue fever (DF). The response of PBMCs to IFN-γ stimulation was lower in patients with DHF than in those with DF or dengue with complications (DWC), with lower CD119 expression and reduced CXCL-10 synthesis. In addition, these alterations are associated with high NS1 serum levels. Conclusions: Patients with DHF reported high NS1 levels, low CD119 expression, and low CXCL-10 synthesis in PBMCs, which may be associated with infection progression and severity.

Comparative screening of anti-dengue activity in aqueous and ethanol extracts of mangrove plants from Sabah

Aims@#Dengue virus is a global pathogen that lacks an effective vaccine or therapy. Screening medicinal plants for anti-dengue properties provides a promising avenue to identify potent compounds. Mangroves, known for their resilience in harsh conditions, produce a diverse range of natural products with unique biochemical profiles, which hold potential for anti-dengue treatments. This study aims to evaluate the anti-dengue activity of selected mangrove plant species from Sabah against DENV2 NS2B-NS3pro, utilizing an enzymatic protease assay. @*Methodology and results @#Six mangrove species (Avicennia marina, Bruguiera gymnorrhiza, Ceriops tagal, Rhizophora apiculata, Rhizophora mucronata and Xylocarpus granatum) were investigated, with various plant parts subjected to aqueous and ethanol extraction. The results demonstrated significant anti-dengue activity in both aqueous and ethanolic extracts of the mangroves against DENV2 NS2B-NS3pro, with IC50 values ranging from 0.95 µg/mL to 6.24 µg/mL. Notably, the ethanolic extract of R. apiculata leaves exhibited the highest inhibition, with an IC50 value of 0.95 µg/mL. @*Conclusion, significance and impact of study@#These findings suggest that the ethanolic extracts from R. apiculata leaves hold promise as potential candidates for dengue treatment. This study underscores the importance of natural products as valuable sources for the development of novel anti-dengue treatments, highlighting the need to explore mangroves in the quest for effective therapeutic options.

Optimization of expression conditions of dengue virus NS2B-NS3 protease in E.coli and determination of enzyme activity / 中国生物制品学杂志

@#Objective To express dengue virus(DENV)NS2B-NS3 protease in E.coli,optimize the expression conditions and determine the enzyme activity,so as to lay a foundation of screening and discovering of lead compounds targeting DENV.Methods Codon-optimized NS2B-NS3 gene was inserted into pET-28a vector to construct recombinant prokaryotic expression plasmid pET-28a-NS2B-NS3,which was transformed E.coli Rosetta(DE3)competent cells and induced by IPTG to express NS2B-NS3 protease. The optimal expression conditions of NS2B-NS3 protease in E.coli were determined by optimizing induction length,induction temperature and IPTG concentration. NS2B-NS3 protease was isolated and purified by HisTrap~(TM) affinity chromatography column and measured for the protease activity by fluorescence resonance energy transfer(FRET)assay.Results The recombinant prokaryotic expression plasmid pET-28a-NS2B-NS3 was constructed correctly as identified by restriction analysis(NheⅠ/XhoⅠ)and sequencing. The optimal expression conditions of NS2BNS3 protease in E.coli were as follows:induction temperature of 20 ℃,induction length of 10 h and IPTG concentration of0. 2 mmol/L. The purified NS2B-NS3 protease showed a purity of more than 90% with a exhibited a of 20 mg/L,which bound to mouse monoclonal antibody against His-tag specifically and had good hydrolytic activity with a specific activity of 16. 111 U/mg,a K_m of 16. 46 μmol/L and a k_(cat) of 0. 028/s.Conclusion DENV NS2B-NS3 protease with high purity and activity was successfully prepared,which laid an experimental foundation of the establishment of high-throughput screening model for inhibitors targeting NS2B-NS3 protease.

Design, synthesis, and activity of 5-cyano-2-thiacetyl aromatic pyrimidinone as DENV NS5 RdRp inhibitors / 药学学报

To screen novel anti-dengue virus (DENV) NS5 RdRp enzyme inhibitors, a series of 5-cyano-2-thiacetoaryl pyrimidinone compounds were designed and synthesized by molecular hybridization method with HCV NS5B RdRp inhibitor 3jc and ZIKV NS5 RdRp inhibitor 4w as lead compounds. The anti-DENV activity of these compounds was evaluated by MTT assay and plaque assay and five compounds showed anti-DENV activity. The most active compound 7a'k showed better anti-DENV activity than that of the positive control ribavirin (EC50 = 7.86 μmol·L-1 vs EC50 = 18.07 μmol·L-1), and the other four compounds showed almost the same anti-DENV activity as ribavirin. Finally, the prediction and simulation of the binding mode through molecular provided new ideas for the further development of this new DENV NS5 RdRp inhibitor.

Toxicity and larvicidal activity on Aedes aegypti of citronella essential oil submitted to enzymatic esterification / Toxicidade e atividade larvicida sobre Aedes aegypti de óleo essencial de citronella submetido à esterificação enzimática

Abstract The essential oil of citronella (Cymbopogon winterianus) has several biological activities, among them the insect repellent action. Some studies showed that cinnamic acid esters can be applied as natural pesticides, insecticides and fungicides. In this context, the objective of the present work was to evaluate the production of esters from citronella essential oil with cinnamic acid via enzymatic esterification. Besides, the essential oil toxicity before and after esterification against Artemia salina and larvicidal action on Aedes aegypti was investigated. Esters were produced using cinnamic acid as the acylating agent and citronella essential oil (3:1) in heptane and 15 wt% NS 88011 enzyme as biocatalysts, at 70 °C and 150 rpm. Conversion rates of citronellyl and geranyl cinnamates were 58.7 and 69.0% for NS 88011, respectively. For the toxicity to Artemia salina LC50 results of 5.29 μg mL-1 were obtained for the essential oil and 4.36 μg mL-1 for the esterified oils obtained with NS 88011. In the insecticidal activity against Aedes aegypti larvae, was obtained LC50 of 111.84 μg mL-1 for the essential oil of citronella and 86.30 μg mL-1 for the esterified oils obtained with the enzyme NS 88011, indicating high toxicity of the esters. The results demonstrated that the evaluated samples present potential of application as bioinsecticide.

Resumo O óleo essencial de citronela (Cymbopogon winterianus) possui diversas atividades biológicas, entre elas a ação repelente a insetos. Alguns estudos mostraram que os ésteres do ácido cinâmico podem ser aplicados como pesticidas naturais, inseticidas e fungicidas. Nesse contexto, o objetivo do presente trabalho foi avaliar a produção de ésteres a partir do óleo essencial de citronela com ácido cinâmico via esterificação enzimática. Além disso, foi investigada a toxicidade do óleo essencial antes e após a esterificação contra Artemia salina e a ação larvicida sobre Aedes aegypti. Os ésteres foram produzidos utilizando ácido cinâmico como agente acilante e óleo essencial de citronela (3: 1) em heptano e 15% em peso da enzima NS 88011 como biocatalisadores, a 70 ° C e 150 rpm. As taxas de conversão de cinamatos de citronelil e geranil foram 58,7 e 69,0% para NS 88011, respectivamente. Para a toxicidade sobre Artemia salina foram obtidos CL50 de 5,29 μg mL-1 para o óleo essencial e 4,36 μg mL-1 para os óleos esterificados com NS 88011. Na atividade inseticida contra larvas de Aedes aegypti, obteve-se CL50 de 111,84 μg mL-1 para o óleo essencial de citronela e 86,30 μg mL-1 para os óleos esterificados com a enzima NS 88011, indicando alta toxicidade dos ésteres. Os resultados demonstraram que as amostras avaliadas apresentam potencial de aplicação como bioinseticida.

Toxicity and larvicidal activity on Aedes aegypti of citronella essential oil submitted to enzymatic esterification / Toxicidade e atividade larvicida sobre Aedes aegypti de óleo essencial de citronella submetido à esterificação enzimática

The essential oil of citronella (Cymbopogon winterianus) has several biological activities, among them the insect repellent action. Some studies showed that cinnamic acid esters can be applied as natural pesticides, insecticides and fungicides. In this context, the objective of the present work was to evaluate the production of esters from citronella essential oil with cinnamic acid via enzymatic esterification. Besides, the essential oil toxicity before and after esterification against Artemia salina and larvicidal action on Aedes aegypti was investigated. Esters were produced using cinnamic acid as the acylating agent and citronella essential oil (3:1) in heptane and 15 wt% NS 88011 enzyme as biocatalysts, at 70 °C and 150 rpm. Conversion rates of citronellyl and geranyl cinnamates were 58.7 and 69.0% for NS 88011, respectively. For the toxicity to Artemia salina LC50 results of 5.29 μg mL-¹ were obtained for the essential oil and 4.36 μg mL-¹ for the esterified oils obtained with NS 88011. In the insecticidal activity against Aedes aegypti larvae, was obtained LC50 of 111.84 μg mL-¹ for the essential oil of citronella and 86.30 μg mL-¹ for the esterified oils obtained with the enzyme NS 88011, indicating high toxicity of the esters. The results demonstrated that the evaluated samples present potential of application as bioinsecticide.

O óleo essencial de citronela (Cymbopogon winterianus) possui diversas atividades biológicas, entre elas a ação repelente a insetos. Alguns estudos mostraram que os ésteres do ácido cinâmico podem ser aplicados como pesticidas naturais, inseticidas e fungicidas. Nesse contexto, o objetivo do presente trabalho foi avaliar a produção de ésteres a partir do óleo essencial de citronela com ácido cinâmico via esterificação enzimática. Além disso, foi investigada a toxicidade do óleo essencial antes e após a esterificação contra Artemia salina e a ação larvicida sobre Aedes aegypti. Os ésteres foram produzidos utilizando ácido cinâmico como agente acilante e óleo essencial de citronela (3: 1) em heptano e 15% em peso da enzima NS 88011 como biocatalisadores, a 70 ° C e 150 rpm. As taxas de conversão de cinamatos de citronelil e geranil foram 58,7 e 69,0% para NS 88011, respectivamente. Para a toxicidade sobre Artemia salina foram obtidos CL50 de 5,29 μg mL-¹ para o óleo essencial e 4,36 μg mL-¹ para os óleos esterificados com NS 88011. Na atividade inseticida contra larvas de Aedes aegypti, obteve-se CL50 de 111,84 μg mL-¹ para o óleo essencial de citronela e 86,30 μg mL-¹ para os óleos esterificados com a enzima NS 88011, indicando alta toxicidade dos ésteres. Os resultados demonstraram que as amostras avaliadas apresentam potencial de aplicação como bioinseticida.

Toxicity and larvicidal activity on Aedes aegypti of citronella essential oil submitted to enzymatic esterification

Abstract The essential oil of citronella (Cymbopogon winterianus) has several biological activities, among them the insect repellent action. Some studies showed that cinnamic acid esters can be applied as natural pesticides, insecticides and fungicides. In this context, the objective of the present work was to evaluate the production of esters from citronella essential oil with cinnamic acid via enzymatic esterification. Besides, the essential oil toxicity before and after esterification against Artemia salina and larvicidal action on Aedes aegypti was investigated. Esters were produced using cinnamic acid as the acylating agent and citronella essential oil (3:1) in heptane and 15 wt% NS 88011 enzyme as biocatalysts, at 70 °C and 150 rpm. Conversion rates of citronellyl and geranyl cinnamates were 58.7 and 69.0% for NS 88011, respectively. For the toxicity to Artemia salina LC50 results of 5.29 g mL-1 were obtained for the essential oil and 4.36 g mL-1 for the esterified oils obtained with NS 88011. In the insecticidal activity against Aedes aegypti larvae, was obtained LC50 of 111.84 g mL-1 for the essential oil of citronella and 86.30 g mL-1 for the esterified oils obtained with the enzyme NS 88011, indicating high toxicity of the esters. The results demonstrated that the evaluated samples present potential of application as bioinsecticide.

Resumo O óleo essencial de citronela (Cymbopogon winterianus) possui diversas atividades biológicas, entre elas a ação repelente a insetos. Alguns estudos mostraram que os ésteres do ácido cinâmico podem ser aplicados como pesticidas naturais, inseticidas e fungicidas. Nesse contexto, o objetivo do presente trabalho foi avaliar a produção de ésteres a partir do óleo essencial de citronela com ácido cinâmico via esterificação enzimática. Além disso, foi investigada a toxicidade do óleo essencial antes e após a esterificação contra Artemia salina e a ação larvicida sobre Aedes aegypti. Os ésteres foram produzidos utilizando ácido cinâmico como agente acilante e óleo essencial de citronela (3: 1) em heptano e 15% em peso da enzima NS 88011 como biocatalisadores, a 70 ° C e 150 rpm. As taxas de conversão de cinamatos de citronelil e geranil foram 58,7 e 69,0% para NS 88011, respectivamente. Para a toxicidade sobre Artemia salina foram obtidos CL50 de 5,29 g mL-1 para o óleo essencial e 4,36 g mL-1 para os óleos esterificados com NS 88011. Na atividade inseticida contra larvas de Aedes aegypti, obteve-se CL50 de 111,84 g mL-1 para o óleo essencial de citronela e 86,30 g mL-1 para os óleos esterificados com a enzima NS 88011, indicando alta toxicidade dos ésteres. Os resultados demonstraram que as amostras avaliadas apresentam potencial de aplicação como bioinseticida.

Rendimiento diagnóstico de la prueba rápida para la detección del antígeno NS1 y anticuerpos IgM e IgG contra el virus del dengue / Diagnostic performance of the rapid test for the detection of NS1 antigen and IgM and IgG anti-antibodies against dengue virus

Objetivos. Determinar el rendimiento diagnóstico de la prueba rápida SD dengue DUO (Inyecta) para la detección de NS1, IgM e IgG en comparación con la prueba de ELISA. Materiales y métodos. Es una evaluación de prueba diagnóstica que incluyó 286 muestras de suero de pacientes con sintomatología atribuible a dengue de zonas endémicas del Perú. Las muestras se analizaron por ELISA y la prueba rápida SD dengue DUO (Inyecta) para IgM, NS1 e IgG en el Instituto de Investigación Nutricional en Lima. Resultados. La sensibilidad de la prueba rápida fue de 68% para NS1 e IgM, y 86% para IgG, mejorando este parámetro a 75% y 81% para NS1 e IgM, respectivamente, en los tres primeros días. La especificidad para los tres analitos fue mayor a 87%. La concordancia de los resultados obtenidos medidos por el coeficiente Kappa para los tres analitos fue buena y no se encontró reacción cruzada con otros arbovirus. Conclusiones. La prueba rápida SD Dengue DUO permite detectar con una adecuada sensibilidad y especificidad NS1, IgM e IgG. La sensibilidad para IgM y NS1 aumenta cuando se detecta en los tres primeros días de síntomas, por lo que se recomienda su implementación en los centros de primer nivel de atención para un diagnóstico temprano y oportuno.

Objectives . To assess the diagnostic performance of the SD dengue DUO rapid test (Inyecta) for the detection of NS1, IgM and IgG in comparison to the ELISA test. Materials and methods . This is a diagnostic test evaluation that included 286 serum samples from patients with symptomatology attributable to dengue from endemic areas of Peru. The samples were analyzed by ELISA and the SD dengue DUO rapid test (Inyecta) for IgM, NS1 and IgG at the Instituto de Investigación Nutricional in Lima. Results . The sensitivity of the rapid test was 68.0% for NS1 and IgM, and 86.0% for IgG, improving to 75.0% and 81.0% for NS1 and IgM, respectively, during the first three days. The specificity for all three analytes was greater than 87.0%. The concordance of the results, measured by the Kappa coefficient for the three analytes, was good and no cross-reaction with other arboviruses was found. Conclusions . The SD dengue DUO rapid test allows detection of NS1, IgM and IgG with adequate sensitivity and specificity. Sensitivity for IgM and NS1 increases when detected during the first three days of symptoms. Therefore, we recommend its implementation in primary care centers for early and timely diagnosis.

Prevalencia del virus de dengue y factores de riesgo en pacientes que asistieron a las unidades de salud del cantón esmeraldas en el 2019 / Prevalence of the dengue virus and risk factors in patients who attended the health units of the Esmeraldas canton in 2019

El dengue es una enfermedad infecciosa ocasionada por alguno de los cuatro serotipos del virus análogo, que se transmite a los humanos a través de la picadura de mosquitos infectados del género Aedes Objetivo: Identificar la prevalencia del virus de dengue y los factores de riesgo en los pacientes que asistieron a las unidades de salud del cantón Esmeraldas en el 2019. Materiales y Métodos: La investigación se enmarca dentro del diseño de investigación no experimental, de tipo retrospectivo, descriptivo, basado en la información de pacientes con diagnóstico de dengue positivo atendidos en las unidades de Salud del cantón Esmeraldas en el 2019.Resultados: En este estudio se encontró a 247 pacientes registrados con diagnóstico presuntivo de dengue, los cuales tomamos como población, se excluyeron a 4 muestras de pacientes, no procesadas, y a 121 muestras de pacientes con resultado negativo, quedándonos 121 muestras de pacientes con diagnóstico de dengue confirmadas, tomando a esta cantidad de pacientes como muestra para nuestro estudio, lo que representa el 49% como prevalencia de virus del dengue en pacientes que asistieron a las unidades de salud del cantón Esmeraldas durante el 2019.Conclusión: La prevalencia de dengue en pacientes que asistieron a las unidades de salud del cantón Esmeraldas es del 49%, la edad que más predomino fue la comprendida entre los 5 y 9 años, un número significativo de pacientes, provienen de la parroquia Simón Plata Torres, sectores con falta de infraestructura de servicios básicos, jardines y patios con malezas y vegetación, arbustos y árboles que rodean a las viviendas, aumentando las posibilidades de cría de mosquitos vectores de esta afección(AU)

Dengue is an infectious disease caused by one of the four serotypes of the analogous virus, which is transmitted to humans through the bite of infected mosquitoes of the genus Aedes Objective: To identify the prevalence of dengue virus and risk factors in the patients who attended the health units of the Esmeraldas canton in 2019. Materials and Methods: The research is part of the non-experimental, retrospective, descriptive research design, based on the information of patients with a positive dengue diagnosis. treated in the health units of the Esmeraldas canton in 2019. Results: In this study, 247 registered patients with a presumptive diagnosis of dengue were found, which we took as a population, 4 samples of patients, not processed, and 121 samples were excluded. of patients with a negative result, leaving us with 121 samples of patients with a confirmed diagnosis of dengue, taking to this number of patients as a sample for our study, which represents 49% as the prevalence of dengue virus in patients who attended the health units of the Esmeraldas canton during 2019. Conclusion: The prevalence of dengue in patients who attended the health units of the Esmeraldas canton is 49%, the most predominant age was between 5 and 9 years, a significant number of patients come from the Simón Plata Torres parish, sectors with a lack of basic services infrastructure, gardens and patios with weeds and vegetation, bushes and trees that surround the houses, increasing the chances of breeding mosquito vectors of this condition(AU)

Evidence of dengue virus in eviscerated specimens of panophthalmitis secondary to dengue fever: A possible cause-effect phenomenon

Purpose: To report a retrospective series of three cases of infectious panophthalmitis post?dengue fever with ex vivo confirmation of dengue virus ribonucleic acid (RNA) in the tissues of the eye. Methods: Four eyes of three patients, who were diagnosed with panophthalmitis following dengue fever and who underwent evisceration, were included. All demographic and clinical data were recorded. The eviscerated samples were subjected to direct microscopy, culture for bacteria, fungi, and parasites, and molecular virology (dengue virus [DENV] NS1?specific reverse transcription loop?mediated isothermal amplification (RT?LAMP) assay). Results: The time from the development of dengue fever to the occurrence of ocular symptoms was 4.33 ± 1.15 (median 5) days. DENV NS1 RNA, suggestive of the presence of the dengue virus, was confirmed in all evisceration specimens (uveal tissue, cornea). All the patients recovered completely from dengue fever and on follow?up had healthy eviscerated sockets. Conclusion: Demonstration of the DENV RNA in the eviscerated specimens of panophthalmitis following dengue fever implicates the DENV in the pathophysiology of the ocular infection.

Detection of recombinant dengue virus 2 NS1 protein in Aedes aegypti mosquitoes using commercial Dengue NS1 ELISA kit

Dengue, a vector-borne disease remains as one of the most serious public health problems globally. Incidence of this disease is on an increasing trend and currently over a billion people in tropical and subtropical regions are at risk. In the absence of an operational vaccine, prevention of dengue virus (DENV) is primarily focused upon controlling mosquito vectors. Mosquito vector surveillance programmes require simple and rapid tools to detect mosquitoes infected with DENV. Here, we tested the commercially available DENV Detect™ NS1 ELISA kit (InBios International, Inc.) for detection of recombinant DENV-NS1 protein in Aedes mosquito samples. The kit was evaluated to find out the minimum detection limit of recombinant DENV-2 NS1 protein following the manufacturer’s instructions. Initially, the NS1 protein detection threshold of the kit was determined and later the assay was standardized for detection of NS1 protein in Aedes aegypti mosquito pools containing 5, 10 and 25 mosquitoes. The ELISA kit displayed high sensitivity towards detection of recombinant dengue virus-2 NS1 protein in mosquito pools (up to 25 mosquitoes per pool) at 25 pico gram concentration. Since the commercial NS1 ELISA is highly sensitive and follows a very simple procedure, it could be employed for DENV surveillance in Aedes aegypti mosquitoes, after carrying out laboratory and field bioassays with DENV infected specimens.

Quality of life among SARS-CoV-2 (COVID-19) positive patients with anosmia using the short version questionnaire of olfactory disorders - Negative statements translated in Filipino (sQOD-NS Ph)

@#<p><strong>Objective:</strong> To determine the quality of life among SARS-CoV-2 (COVID-19) positive patients with anosmia using the Short Version Questionnaire of Olfactory Disorders - Negative Statements translated in Filipino (sQOD-NS Ph).</p><p><strong>Methods: </strong><strong>Design:</strong> Cross-Sectional Study</p><p><strong>                 Setting:</strong> Tertiary Government Training Hospital</p><p><strong>                 Participants:</strong> SARS-CoV-2 (COVID-19) RT-PCR positive patients aged 18 years old and above with COVID-19 symptoms and anosmia in a tertiary government hospital who consulted from March 2020 to August 2021 answered the short version of sQOD-NS Ph.</p><p><strong>Results:</strong> Out of 108 participants with positive SARS-CoV-2 (COVID-19) RT-PCR test, 72 (66%) presented with anosmia, and sQOD-NS Ph scores ranged from 1 to 21 with a mean of 14.78. Thirty two (44%) encountered problems in eating while 21 (29%) had feelings of isolation due to loss of smell. There was an inverse correlation of -0.478 between recovery time of olfaction and QoL score, hence the longer the recovery time, the lower the QoL score, while the shorter the recovery time, the higher the QoL score (p < .0001).</p><p><strong>Conclusion:</strong> Majority of COVID-19 patients with anosmia had mild or negligible impairment, while a small percentage had impaired quality of life. The low percentage may be due to high number of patients who may have recovered their sense of smell along the course of the disease.</p>

Epidemiology of Dengue in patients with febrile syndrome at Saint Camille Hospital, Ouagadougou, Burkina Faso from 2020 to 2

: Dengue is still a public health problem in tropical countries. This disease, which had almost disappeared in some areas of the world, has become re-emergent in certain parts of the world including Africa.The aim of this study is to determine the seroprevalence and evolution of Dengue virus (DENV) infection from 2020 to 2021 at the Hospital Saint Camille de Ouagadougou (HOSCO), Burkina Faso. Methodology: This was a descriptive analytical study of patients seen in general practice with febrile syndrome referred for serological diagnosis of Dengue at the HOSCO laboratory over a period of 2 years (January 1, 2020 ­ December 31, 2021). The "Dengue Duo (AgNS1/IgM/IgG)" kit from SD Bioline was used for the rapid diagnosis through the detection of NS1 antigen and IgM/IgG antibodies in plasma. Data were analysed with SPSS version 20.0 software. Association between demographic data and prevalence of DENV infection was determined by Chisquare test and odds ratio (with 95% confidence interval). P value less than 0.05 was considered statistical significance. Results: A total of 2957 patients aged 0-94 years were referred for serological diagnosis of DENV infection at the HOSCO laboratory over the period 2020-2021, comprising 56.3% females and 43.7% males. The overall prevalence of acute DENV infection (NS1Ag positive) was 5.4% (159/2957), with 2.4% (41/1700) in 2020 and 9.4% (118/1257) in 2021 (OR=4.192, 95% CI=2.915-6.028, p<0.0001). The prevalence of acute DENV infection of 7.0% (91/1292) in the males was significantly higher than 4.1% (68/1665) in the females (OR=1.779, 95% CI=1.288-2.458, p=0.0005), and also significantly higher in age groups 20-29 years (7.6%), 10-19 years (6.9%) and 40-49 years (5.8%) than other age groups (X 2=14.928, p=0.0107). The overall prevalence of DENV IgM and IgG antibodies was 3.2% and 37.3% respectively. The prevalence of DENV IgG antibodies was significantly higher in males (44.0%) than females (32.1%) (OR=1.667, 95%CI=1.434-1.938, p<0.0001) and in age groups 30-39 (43.4%), 40-49 (44.0%) and >50 years (49.3%) than other age groups (X2=121.0, p<0.0001), indicating that past exposure to DENV infection is higher among males and older age groups. The peak of DENV infection was between October and November with 84.3% (134/159) of NS1Ag positivity occurring during this period. Conclusion: The present study reports a high prevalence of acute Dengue virus infection in patients from October to November. To eradicate Dengue which has become a tropical silent epidemic, interventions such as vector control, availability of and accessibility to diagnostic tests, and good therapeutic management are of great importance

Bixinoids Derived from Bixa orellana as a Potential Zika Virus Inhibitor Using Molecular Simulations. Antiviral Effect on the Zika Virus of Bixinoids

Abstract Zika fever is a viral infection of great relevance in public health, especially in tropic regions, in which there is a predominance of mosquitoes of the genus Aedes, vectors of the disease. Microcephaly in neonatal children and Guillain-Barré syndrome in adults can be caused by the action of the Zika virus (ZIKV). Non-structural proteins, such as NS2B, NS3 and NS5, are important pharmacological targets, due to their action in the life cycle. The absence of anti-Zika drugs raises new research, including prospecting for natural products. This work investigated the in silico antiviral activity of bixin and six other derived molecules against the Zika viral proteins NS2B-NS3 and NS5. The optimized structure was subjected to molecular docking to characterize the interaction between bixinoids and ZIKV non-structural proteins, where significant interactions were observed with amino acid residues in the catalytic site in each enzyme. These results suggest that bixin and ethyl bixin has the potential to interfere with the enzymatic activity of NS2B, NS3 and NS5, thus being an indication of being a promising anti-Zika agent.

Characterization of primary direct-acting antiviral (DAA) drugs resistance mutations in NS5A/NS5B regions of hepatitis C virus with genotype 1a and 1b from patients with chronic hepatitis

ABSTRACT The Hepatitis C virus (HCV) infection is a public health problem. The high level of HCV replication and its lack of post-transcriptional correction mechanisms results in the emergence of viral variants and the difficulty in determining polymorphisms and variants that contain the substitutions associated with resistance towards new antivirals. The main focus of this study was to map the NS5A and NS5B polymorphisms and resistance mutations to new antiviral drugs in HCV strains genotype 1 from patients with chronic hepatitis C infection. Serum samples were collected from patients who underwent routine viral load tests at the Instituto Adolfo Lutz, Sao Paulo city, Brazil. A total of 698 and 853 samples were used for the characterization of NS5A and NS5B regions, respectively, which comprise the HCV genotypes 1a and 1b. The prevalence of resistance mutations found in the NS5A region was 6.4%, with Y93H, L31M, Q30R, and Y93N as the main resistance-associated substitutions (RAS). No NS5B-associated RAS was observed for any of the analyzed drugs. These findings support that the RAS test should be offered to individuals with poor response to double combination regimens prior to treatment initiation, thereby assisting strain vigilance and selection of effective treatment or retreatment options using DAA regimens.

Natural DENV-2 NS2B/NS3 protease inhibitors from Myristica cinnamomea King

@#The NS2B/NS3 protease is crucial for the pathogenesis of the DENV. Therefore, the inhibition of this protease is considered to be the key strategy for the development of new antiviral drugs. In the present study, malabaricones C (3) and E (4), acylphenols from the fruits of Myristica cinnamomea King, have been respectively identified as moderate (27.33 ± 5.45 μM) and potent (7.55 ± 1.64 μM) DENV-2 NS2B/NS3 protease inhibitors, thus making this the first report on the DENV-2 NS2B/NS3 protease inhibitory activity of acylphenols. Based on the molecular docking studies, compounds 3 and 4 both have π-π interactions with Tyr161. While compound 3 has hydrogen bonding interactions with Gly151, Gly153 and Tyr161, compound 4 however, forms hydrogen bonds with Ser135, Asp129, Phe130 and Ile86 instead. The results from the present study suggests that malabaricones C (3) and E (4) could be employed as lead compounds for the development of new dengue antivirals from natural origin.

Phytochemical analysis of Berberis lyceum methanolic extract and its antiviral activity through the restoration of MAPK signaling pathway modulated by HCV NS5A

Objective: To evaluate the antiviral activity and phytochemicals of selected plant extracts and their effect on the mitogen-activated protein kinase (MAPK) signaling pathway modulated by hepatitis C virus (HCV) nonstructural protein 5A (NS5A). Methods: A total of ten plant extracts were initially screened for their toxicities against HepG2 cells. The non-toxic plants were tested for their inhibitory effect on the expression of HCV NS5A at both mRNA and protein levels using real-time PCR and Western blotting assays, respectively. The differential expression of the genes associated with MAPK pathway in the presence of NS5A gene and plant extract was measured through real-time PCR. Subsequently, the identification of secondary metabolites was carried out by phytochemical and HPLC analysis. Results: The phytochemical profiling of Berberis lyceum revealed the presence of alkaloids, phenols, saponins, tannins, flavonoids, carbohydrates, terpenoids, steroids, and glycosides. Similarly, quercetin, myricetin, gallic acid, caffeic acid, and ferulic acid were identified through HPLC analysis. The methanolic extract of Berberis lyceum strongly inhibited HCV RNA replication with an IC50 of 11.44 μg/mL. RT-PCR and Western blotting assays showed that the extract reduced the expression of HCV NS5A in a dose-dependent manner. Berberis lyceum extract also attenuated NS5A-induced dysregulation of the MAPK signaling pathway. Conclusions: Our findings suggest that Berberis lyceum extract strongly inhibits HCV propagation by reducing HCV NS5A-induced perturbation of MAPK signaling.