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Strategic construction of mRNA vaccine derived from conserved and experimentally validated epitopes of avian influenza type A virus: a reverse vaccinology approach

  • Leana Rich Herrera-Ong
    • Clinical and Experimental Vaccine Research
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    • v.12 no.2
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    • pp.156-171
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    • 2023
  • Purpose: The development of vaccines that confer protection against multiple avian influenza A (AIA) virus strains is necessary to prevent the emergence of highly infectious strains that may result in more severe outbreaks. Thus, this study applied reverse vaccinology approach in strategically constructing messenger RNA (mRNA) vaccine construct against avian influenza A (mVAIA) to induce cross-protection while targeting diverse AIA virulence factors. Materials and Methods: Immunoinformatics tools and databases were utilized to identify conserved experimentally validated AIA epitopes. CD8+ epitopes were docked with dominant chicken major histocompatibility complexes (MHCs) to evaluate complex formation. Conserved epitopes were adjoined in the optimized mVAIA sequence for efficient expression in Gallus gallus. Signal sequence for targeted secretory expression was included. Physicochemical properties, antigenicity, toxicity, and potential cross-reactivity were assessed. The tertiary structure of its protein sequence was modeled and validated in silico to investigate the accessibility of adjoined B-cell epitope. Potential immune responses were also simulated in C-ImmSim. Results: Eighteen experimentally validated epitopes were found conserved (Shannon index <2.0) in the study. These include one B-cell (SLLTEVETPIRNEWGCR) and 17 CD8+ epitopes, adjoined in a single mRNA construct. The CD8+ epitopes docked favorably with MHC peptidebinding groove, which were further supported by the acceptable ∆Gbind (-28.45 to -40.59 kJ/mol) and Kd (<1.00) values. The incorporated Sec/SPI (secretory/signal peptidase I) cleavage site was also recognized with a high probability (0.964814). Adjoined B-cell epitope was found within the disordered and accessible regions of the vaccine. Immune simulation results projected cytokine production, lymphocyte activation, and memory cell generation after the 1st dose of mVAIA. Conclusion: Results suggest that mVAIA possesses stability, safety, and immunogenicity. In vitro and in vivo confirmation in subsequent studies are anticipated.

Effect of Supplemental Corn Dried Distillers Grains with Solubles Fed to Beef Steers Grazing Native Rangeland during the Forage Dormant Season

  • Murillo, M.;Herrera, E.;Ruiz, O.;Reyes, O.;Carrete, F.O.;Gutierrez, H.
    • Asian-Australasian Journal of Animal Sciences
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    • v.29 no.5
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    • pp.666-673
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    • 2016
  • Two experiments were conducted to evaluate the effects of the level of corn dry distillers grains with solubles (CDDGS) supplementation on growing performance, blood metabolites, digestion characteristics and ruminal fermentation patterns in steers grazing dormant forage. In Exp. 1, of growth performance, 120 steers ($204{\pm}5kg$ initial body weight [BW]) were distributed randomly into 3 groups (each of 40 steers), which were provided with the following levels of CDDGS supplement: 0%, 0.25%, or 0.50% BW. All groups of steers were grazed for 30 days in each of 3 grazing periods (March, April, and May). Approximately 1,000 ha of the land was divided with electric fencing into 3 equally sized pastures (333 ha in size). Blood samples were collected monthly from 20 steers in each grazing group for analysis of glucose (G), urea-nitrogen (UN) and non-esterified fatty acids. Final BW, average daily gain (ADG) and supplement conversion (CDDGS-C) increased with increasing levels of CDDGS supplementation (p<0.05).The CDDGS supplementation also increased the plasma G and UN concentrations (p<0.05). In Exp. 2, of digestive metabolism, 9 ruminally cannulated steers ($BW=350{\pm}3kg$) were distributed, following a completely randomized design, into groups of three in each pasture. The ruminally cannulated steers were provided the same levels of CDDGS supplementation as in the growing performance study (0%, 0.25%, and 0.50% BW), and they grazed along with the other 40 steers throughout the grazing periods. The dry matter intake, crude protein intake, neutral detergent fiber intake (NDFI), apparent digestibility of dry matter (ADDM), crude protein (ADCP) and neutral detergent fiber (ADNDF) increased with increasing levels of CDDGS supplementation (p<0.05). The ruminal degradation rates of CP (kdCP), NDF (kdNDF) and passage rate (kp) also increased with increasing levels of CDDGS supplementation (p<0.05). Ruminal ammonia nitrogen ($NH_3$-N) and propionate concentrations also increased with increasing levels of CDDGS supplementation (p<0.05). However, acetate concentrations decreased with increasing levels of CDDGS supplementation (p<0.05). Liquid dilution rate increased with increasing levels of CDDGS supplementation but ruminal liquid volume decreased (p<0.05). On the basis of these findings, we can conclude that CDDGS supplementation enhanced the productive performance of cattle grazing native rangeland without negatively affecting forage intake, glucose and urea-nitrogen blood concentrations, ruminal degradation and ruminal fermentation patterns.

Detection of Clavibacter michiganensis subsp. michiganensis Assisted by Micro-Raman Spectroscopy under Laboratory Conditions

  • Perez, Moises Roberto Vallejo;Contreras, Hugo Ricardo Navarro;Herrera, Jesus A. Sosa;Avila, Jose Pablo Lara;Tobias, Hugo Magdaleno Ramirez;Martinez, Fernando Diaz-Barriga;Ramirez, Rogelio Flores;Vazquez, Angel Gabriel Rodriguez
    • The Plant Pathology Journal
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    • v.34 no.5
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    • pp.381-392
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    • 2018
  • Clavibacter michiganensis subsp. michiganesis (Cmm) is a quarantine-worthy pest in $M{\acute{e}}xico$. The implementation and validation of new technologies is necessary to reduce the time for bacterial detection in laboratory conditions and Raman spectroscopy is an ambitious technology that has all of the features needed to characterize and identify bacteria. Under controlled conditions a contagion process was induced with Cmm, the disease epidemiology was monitored. Micro-Raman spectroscopy ($532nm\;{\lambda}$ laser) technique was evaluated its performance at assisting on Cmm detection through its characteristic Raman spectrum fingerprint. Our experiment was conducted with tomato plants in a completely randomized block experimental design (13 plants ${\times}$ 4 rows). The Cmm infection was confirmed by 16S rDNA and plants showed symptoms from 48 to 72 h after inoculation, the evolution of the incidence and severity on plant population varied over time and it kept an aggregated spatial pattern. The contagion process reached 79% just 24 days after the epidemic was induced. Micro-Raman spectroscopy proved its speed, efficiency and usefulness as a non-destructive method for the preliminary detection of Cmm. Carotenoid specific bands with wavelengths at 1146 and $1510cm^{-1}$ were the distinguishable markers. Chemometric analyses showed the best performance by the implementation of PCA-LDA supervised classification algorithms applied over Raman spectrum data with 100% of performance in metrics of classifiers (sensitivity, specificity, accuracy, negative and positive predictive value) that allowed us to differentiate Cmm from other endophytic bacteria (Bacillus and Pantoea). The unsupervised KMeans algorithm showed good performance (100, 96, 98, 91 y 100%, respectively).

Factors Affecting Sexual Behavior of School Teenagers in Havana, Cuba (쿠바, 아바나 청소년의 성행동 관련 요인)

  • Sanchez, Maybin Herrera;Lee, Ga Ram;Diego, Edith Contreras;Molina, Ruben de Armas;Nam, Eun Woo
    • The Journal of Korean Society for School & Community Health Education
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    • v.21 no.1
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    • pp.47-60
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    • 2020
  • 서론: 청소년기는 여러 생물학적 변화 및 사고방식의 발전과 함께 성행위가 나타나기 시작하는 인간성장의 단계이다. 쿠바를 포함한 많은 라틴아메리카 국가들은 청소년들의 성관계 조기 시작과 같은 위험한 성행동을 보고해 왔다. 이 연구의 목적은 사회 인구 통계학적 요소, 정보 수단 및 쿠바의 십대 학생의 성적 행동에 영향을 미치는 행동 요소 사이의 관계를 확립하는 것이다. 방법: 본 연구를 위해 쿠바 십대들의 생식 및 성적 행동 경향 파악을 위해 설문조사를 실시했다. 전체 260명 중 성경험 여부에 응답한 232명에 대해 인구사회학적 특성의 파악과 성 관련 정보수집수단, 행동적 요인과 성행동 사이의 연관성분석을 위해 회귀분석을 실시했다. 결과: 대상자들의 평균 연령은 16.9세이고 대다수는 여성(64.2%)이었다. 232명 중에 45.3%는 무신론자, 31.2%는 유신론자이었다. 종교의 중요도에 대한 생각은 39.2%가 종교가 중요하지 않다고 응답했고 34.0%는 중요하다고, 14.0%는 종교가 매우 중요하다고 응답했다. 응답자 232명 중 80.5%의 십대들이 성관계 경험이 있다고 하였으며 첫 성관계 평균 연령은 14.65세였다. 피임약을 사용한 비율은 63.8%이었다. 인구사회학적 특성 중에 성별은 성관계와 통계적으로 유의미한 연관성을 나타냈으며, 남성은 여성보다 성관계를 가질 가능성이 3.8 배 더 높았다. 또한, 종교는 성관계와 관련이 없었다. 그러나 나이는 통계적으로 유의미한 것으로 나타났으며, 나이가 한 살 많아질수록 성관계 가능성은 2.29배 증가함을 알 수 있었다. 성 관련 정보수집수단과 관련하여 친구와 학교의 성교육 커리큘럼이 청소년들의 성행동을 결정하는 중요한 요인이었다. 결론: 쿠바 십대들의 연령과 성별은 성행동의 중요한 결정요인으로 나타났다. 또한, 쿠바의 십대들은 친구와 학교로부터 성에 대한 정보를 수집하고 있으므로 학교에서 보다 종합적인 성교육 프로그램을 만드는 등의 십대들의 성행동 문제를 다룰 때 영향요인들을 고려해야 할 것이다.

Acute and Repeated 28-Day Oral Dose Toxicity Studies of Thymus vulgaris L. Essential Oil in Rats

  • Rojas-Armas, Juan;Arroyo-Acevedo, Jorge;Ortiz-Sanchez, Manuel;Palomino-Pacheco, Miriam;Castro-Luna, Americo;Ramos-Cevallos, Norma;Justil-Guerrero, Hugo;Hilario-Vargas, Julio;Herrera-Calderon, Oscar
    • Toxicological Research
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    • v.35 no.3
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    • pp.225-232
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    • 2019
  • Thymus vulgaris L. is widely used as an ingredient in cooking and in herbal medicine. However, there is little information about its toxicity. The present study was performed to evaluate the acute and repeated 28-day oral dose toxicity of thyme essential oil in rats. For the acute toxicity test, two groups of three rats were used. The rats received a single dose of essential oil: 300 or 2,000 mg/kg of body weight (bw). The rats were observed individually during the first four hours, and then daily until day 14. For the toxicity test with repeated doses, four groups of 10 rats were used. Doses of 100, 250, and 500 mg/kg/day were tested for 28 days. At the end of the experiment, blood was collected and the animals were sacrificed. Histopathological examination showed that in the lungs of rats given the 2,000 mg/kg bw dose, polymorph nuclear infiltrates, hemosiderin macrophages, and interstitial space thickening were present. In the repeated dose study, all rats survived the 28-day treatment period and apparently showed no signs of toxicity. The hematological and biochemical parameters were not altered. The histopathological study of the organs showed severe changes in the lung, with the dose of 500 mg/kg/day; in the other organs, no alterations were observed or the changes were slight. The body weight was only altered in male rats given the 500 mg/kg dose. The relative weight of the organs did not show any significant changes. Our studies revealed that the essential oil of Thymus vulgaris has moderate oral toxicity according to the results of the acute test, whereas the results of the 28-day oral toxicity test suggest that the no-observed-adverse effect level (NOAEL) is greater than 250 mg/kg/day.

Growth performance and health of nursing lambs supplemented with inulin and Lactobacillus casei

  • Ayala-Monter, Marco A;Hernandez-Sanchez, David;Gonzalez-Munoz, Sergio;Pinto-Ruiz, Rene;Martinez-Aispuro, Jose A;Torres-Salado, Nicolas;Herrera-Perez, Jeronimo;Gloria-Trujillo, Adrian
    • Asian-Australasian Journal of Animal Sciences
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    • v.32 no.8
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    • pp.1137-1144
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    • 2019
  • Objective: This experiment was designed to evaluate the effects of Agave tequilana inulin and Lactobacillus casei (L. casei) on growth performace, hematological variables, serum metabolites, and total coliforms in nursing lambs. Methods: The experimental design was completely randomized; treatments were T1, control (pre-starter concentrate, PC), T2: T1+2% inulin, and T3: T1+2% inulin+L. casei; treatments were compared with Tukey test ($p{\leq}0.05$); and 45 new born $Kathadin{\times}Dorset lambs$ ($4.8{\pm}0.8kg$ birth weight) were the experimental units (15 per treatment). The variables were daily weight gain (DWG), dry matter intake and diarrheas incidence (%) during 56 d. Twenty-four hours after birth and at the end of the experiment, blood samples were collected to evaluate hematological variables and serum metabolites. Besides, the populations of total coliforms and lactobacilli were estimated in fecal samples. Results: Addition of agave inulin and L. casei increased ($p{\leq}0.05$) DWG 356, 384, and 415 g/d, weaning weight 24.92, 26.18, and 28.07 kg, as well as lactobacilli population 5.79, 6.32, and $6.48Log_{10}cfu/g$, for T1, T2, and T3, respectively. Lambs fed L. casei had decreased ($p{\leq}0.05$) populations of total coliforms (T1 = 6.18, T2 = 5.77, and $T3=5.07Log_{10}cfu/g$), diarrheas incidence (T1 = 11.67%, T2 = 8.33%, and T3 = 5.0%), and serum cholesterol concentration (11% in T2 and 13% in T3, compared to control). Conclusion: The combination of Agave tequilana inulin and L. casei increases weight gain and improves intestinal health by reducing coliforms and diarrheas incidence in $Katahdin{\times}Dorset$ lambs during the pre-weaning period.

Risk Factors for COVID-19 Infection Among Healthcare Workers. A First Report From a Living Systematic Review and meta-Analysis

  • Dzinamarira, Tafadzwa;Nkambule, Sphamandla Josias;Hlongwa, Mbuzeleni;Mhango, Malizgani;Iradukunda, Patrick Gad;Chitungo, Itai;Dzobo, Mathias;Mapingure, Munyaradzi Paul;Chingombe, Innocent;Mashora, Moreblessing;Madziva, Roda;Herrera, Helena;Makanda, Pelagia;Atwine, James;Mbunge, Elliot;Musuka, Godfrey;Murewanhema, Grant;Ngara, Bernard
    • Safety and Health at Work
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    • v.13 no.3
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    • pp.263-268
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    • 2022
  • Health care workers (HCWs) are more than ten times more likely to be infected with coronavirus infectious disease 2019 (COVID-19) than the general population, thus demonstrating the burden of COVID-19 among HCWs. Factors that expose HCWs to a differentially high-risk of COVID-19 acquisition are important to elucidate, enable appropriate public health interventions to mitigate against high risk and reduce adverse outcomes from the infection. We conducted a systematic review and meta-analysis to summarize and critically analyze the existing evidence on SARS-CoV-2 risk factors among HCWs. With no geographical limitation, we included studies, in any country, that reported (i) the PCR laboratory diagnosis of COVID-19 as an independent variable (ii) one or more COVID-19 risk factors among HCWs with risk estimates (relative risk, odds ratio, or hazard ratio) (iii) original, quantitative study design, and published in English or Mandarian. Our initial search resulted in 470 articles overall, however, only 10 studies met the inclusion criteria for this review. Out of the 10 studies included in the review, inadequate/lack of protective personal equipment, performing tracheal intubation, and gender were the most common risk factors of COVID-19. Based on the random effects adjusted pooled relative risk, HCWs who reported the use of protective personal equipment were 29% (95% CI: 16% to 41%) less likely to test positive for COVID-19. The study also revealed that HCWs who performed tracheal intubations were 34% (95% CI: 14% to 57%) more likely to test positive for COVID-19. Interestingly, this study showed that female HCWs are at 11% higher risk (RR 1.11 95% CI 1.01-1.21) of COVID-19 than their male counterparts. This article presents initial findings from a living systematic review and meta-analysis, therefore, did not yield many studies; however, it revealed a significant insight into better understanding COVID-19 risk factors among HCWs; insights important for devising preventive strategies that protect them from this infection.