Diseño de casetes de expresión que confieran tolerancia a sequía y a glufosinato en maíz (Zea mays)

Alejandro Chaparro-Giraldo; Andrea Carreño-Venegas

doi:10.15446/abc.v21n3.51170

Best Molecular Tools to Investigate Coronavirus Diversity in Mammals: A Comparison

Viruses ◽

10.3390/v13101975 ◽

2021 ◽

Vol 13 (10) ◽

pp. 1975

Author(s):

Petra Drzewnioková ◽

Francesca Festa ◽

Valentina Panzarin ◽

Davide Lelli ◽

Ana Moreno ◽

...

Keyword(s):

Literature Review ◽

In Silico ◽

Host Shift ◽

Molecular Tools ◽

Rt Pcr ◽

Pcr Assay ◽

Field Samples ◽

Low Sensitivity

Coronaviruses (CoVs) are widespread and highly diversified in wildlife and domestic mammals and can emerge as zoonotic or epizootic pathogens and consequently host shift from these reservoirs, highlighting the importance of veterinary surveillance. All genera can be found in mammals, with α and β showing the highest frequency and diversification. The aims of this study were to review the literature for features of CoV surveillance in animals, to test widely used molecular protocols, and to identify the most effective one in terms of spectrum and sensitivity. We combined a literature review with analyses in silico and in vitro using viral strains and archive field samples. We found that most protocols defined as pan-coronavirus are strongly biased towards α- and β-CoVs and show medium-low sensitivity. The best results were observed using our new protocol, showing LoD 100 PFU/mL for SARS-CoV-2, 50 TCID50/mL for CaCoV, 0.39 TCID50/mL for BoCoV, and 9 ± 1 log2 ×10−5 HA for IBV. The protocol successfully confirmed the positivity for a broad range of CoVs in 30/30 field samples. Our study points out that pan-CoV surveillance in mammals could be strongly improved in sensitivity and spectrum and propose the application of a new RT-PCR assay, which is able to detect CoVs from all four genera, with an optimal sensitivity for α-, β-, and γ-.

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Importance of miRNA in SARS-CoV2 infection

Gaceta Médica de Caracas - Supplement 1 ◽

10.47307/gmc.2020.128.s1.3 ◽

2020 ◽

Vol 128 (S1) ◽

pp. 7-22

Author(s):

Juan Bautista De Sanctis ◽

Alexis García ◽

Jenny Garmendia ◽

Dolores Moreno ◽

Marian Hajduch ◽

...

Keyword(s):

In Silico ◽

Expresión Génica

Al final del 2019, un nuevo patógeno, el coronavirus SARS-CoV2, fue identificado. El virus ha infectado a más de 30 millones de personas en el mundo entero con una letalidad cercana al 5 %. El SARS-CoV2 es un ARN virus. El genoma viral contiene 29 891 nucleótidos que codifican para 9 889 aminoácidos; 5´-replicasa (orf1/ab), cuatro proteínas estructurales principales [espiga (S)-Envoltura (E)-Membrana (M)-Nucleocápside (N)] y las proteínas de la región terminal ORF. Los microARN (miARN) son potentes reguladores pos-transcripcionales de la expresión génica y por ende pueden controlar la infección y replicación viral. Estudios in silico y bioinformática revelaron los miARN del huésped que afectan la replicación viral (15b-5p, 15a-5p, 197-5p, 548c-5p, 548d-5p, 409-3p, 30b-5p, 505-3p). Además hay miRNA virales que son compartidos con el hospedero, (8066, 5197, 3611, 3934-3p, 1307-3p, 3691-3p, 1468-5p) y son importantes en la infección por SARS-CoV2. Aún cuando esas moléculas deben ser validadas en estudios in vitro e in vivo, hay un alto potencial terapéutico involucrado que ya ha sido propuesto y usado en otras infecciones virales. Más estudios de los mecanismos moleculares de esta compleja infección viral son necesarios para entender la patogénesis viral.

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Evaluation of 2-Thioxoimadazolidin-4-One Derivatives as Potent Anti-Cancer Agents through Apoptosis Induction and Antioxidant Activation: In Vitro and In Vivo Approaches

Molecules ◽

10.3390/molecules27010083 ◽

2021 ◽

Vol 27 (1) ◽

pp. 83

Author(s):

Mohamed S. Nafie ◽

Ahmed I. Khodair ◽

Hebat Allah Y. Hassan ◽

Noha M. Abd El-Fadeal ◽

Hanin A. Bogari ◽

...

Keyword(s):

Cell Cycle ◽

In Silico ◽

Hepg2 Cells ◽

Flow Cytometric Analysis ◽

Apoptosis Induction ◽

Rt Pcr ◽

Anti Cancer ◽

Ic50 Values

Background: Hepatocellular carcinoma (HCC) is one of the most widespread malignancies and is reported as the fourth most prevalent cause of cancer deaths worldwide. Therefore, we aimed to investigate the probable mechanistic cytotoxic effect of the promising 2-thioxoimidazolidin-4-one derivative on liver cancer cells using in vitro and in vivo approaches. The compounds were tested for the in vitro cytotoxic activity using MTT assay, and the promising compound was tested in colony forming unit assay, flow cytometric analysis, RT-PCR, Western blotting, in vivo using SEC-carcinoma and in silico to highlight the virtual mechanism of action. Both compounds 4 and 2 performed cytotoxic effects against HepG2 cells with IC50 values of 0.017 and 0.18 μM, respectively, compared to Staurosporine and 5-Fu as reference drugs with IC50 values of 5.07 and 5.18 µM, respectively. Compound 4 treatment revealed apoptosis induction by 19.35-fold (11.42% compared to 0.59% in control), arresting the cell cycle at G2/M phase. Moreover, studying gene expression that plays critical roles in cell cycle and apoptosis by RT-PCR demonstrated that compound 4 enhances the expression of the pro-apoptotic genes p53, PUMA, and Caspase 3, 8, and 9, and impedes the anti-apoptotic Bcl-2 gene in the HepG2 cells. It can also inhibit the PI3K/AKT pathway at both gene and protein levels, which was reinforced by the in silico predictions of the molecular docking simulations towards the PI3K/AKT proteins. Finally, in vivo study verified that compound 4 has a promising anti-cancer activity through activating antioxidant levels (CAT, SOD and GSH) and ameliorating hematological, biochemical, and histopathological findings.

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Thinking Out of the Box: On the Ability of Zea mays L. to Biotrasform Aflatoxin B1 Into Its Modified Forms

Frontiers in Plant Science ◽

10.3389/fpls.2020.599158 ◽

2021 ◽

Vol 11 ◽

Author(s):

Laura Righetti ◽

Enrico Rolli ◽

Luca Dellafiora ◽

Gianni Galaverna ◽

Michele Suman ◽

...

Keyword(s):

Zea Mays ◽

Phase I ◽

Aflatoxin B1 ◽

In Silico ◽

Zea Mays L ◽

Limited Information ◽

Maize Plants ◽

Modified Forms

While aflatoxin metabolism in animals has been clarified, very limited information is so far available on the possible biotransformation occurring in plants. Therefore, this work aimed at investigating whether AFB1 metabolites could occur in field-grown infected maize and the putative role of Zea mays L. metabolism in their production. For such scope, asymptomatic in vitro–grown plantlets and in silico evaluations of plant transforming enzymes were used to pinpoint how plants may handle these compounds. Our data demonstrated the role of maize plants in the production of Phase I hydroxylated aflatoxins, including, among others, AFM1, AFM2, and aflatoxicol, and suggest that plant cytochromes may be involved in this biotransformation of AFB1.

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Actividad antihipertensiva de péptidos de zeína extraídos de maíz (Zea mays L.) criollo (azul y rojo) del Estado de México

RECUBRIMIENTO COMESTIBLE CON BASE DE PIÑON MEXICANO (Jatropha curcas) SOBRE LA CALIDAD DE CHAYOTE MINIMAMENTE PROCESADO ◽

10.32854/agrop.vi.1727 ◽

2020 ◽

Vol 13 (7) ◽

Author(s):

Alan Uriel García-Campos ◽

Mayra Díaz-Ramírez ◽

Georgina Calderón-Domínguez ◽

Rosy Gabriela Cruz-Monterrosa ◽

Adolfo Armando Rayas-Amor ◽

...

Keyword(s):

Zea Mays ◽

In Silico ◽

Zea Mays L

Objetivo: Evaluar la actividad antihipertensiva de péptidos extraídos de zeína de maíz criollo azul y rojo nixtamalizado y sin nixtamalizar del Estado de México. Diseño/metodología/aproximación: Se emplearon herramientas bioinformáticas, tales como la base de datos NCBI para la búsqueda de secuencias primarias de la proteína zeína del maíz. La metodología incluyó una predicción de los péptidos con actividad antihipertensiva a través de diversos servidores bioinformáticos. La actividad inhibitoria se determinó mediante la regresión del porcentaje. Se realizó un análisis estadístico para evaluar las posibles diferencias significativas utilizando la prueba de Tukey (p<0.05). Resultados: Por medio de la hidrólisis in sílico se encontraron los siguientes péptidos: IFSILMLLA, LSACVLDATI, TASVCENPTL, LPLSPLLFQQ, SPALSLVQSL, LSPYSQQQQF, PFSQLATAY, LPFYQQFSAN y PAAFYQQHII que mostraron actividad de inhibición de la ECA. Los resultados mostraron que el GH de la zeína fue mayor en maíz azul (13.96±0.02) que en maíz rojo (10.38±0.79) sin nixtamalizar, mientras que, con el proceso de nixtamalización, este parámetro también fue mayor en el maíz azul (10.52±0.57) que en el maíz rojo (8.96±0.01). Limitaciones del estudio/implicaciones: La enzima utilizada para la hidrólisis enzimática genera oligopéptidos que tal vez no puedan ser tan afines a la enzima convertidora de angiotensina. Hallazgos/conclusiones: La actividad antihipertensiva in vitro del maíz azul sin hidrolizar mostraron un menor porcentaje de inhibición de la ECA con respecto al maíz rojo sin hidrolizar. La hidrólisis con tripsina solo incrementa la actividad inhibitoria de la ECA en maíz azul a concentración de 0.3 mg/mL sin nixtamalizar.

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Ekspresi fenotipe gen APETALA1 kakao (TcAP1) pada eksplan tembakau Phenotypic expression of cacao APETALA1 (TcAP1) in tobacco explant

E-Journal Menara Perkebunan ◽

10.22302/ppbbi.jur.mp.v74i1.116 ◽

2016 ◽

Vol 74 (1) ◽

Author(s):

Tetty CHAIDAMSARI ◽

. SAMANHUDI ◽

Asmini BUDIANI ◽

Roedhy POERWANTO ◽

Djoko SANTOSO

Keyword(s):

Agrobacterium Tumefaciens ◽

Tobacco Plant ◽

Phenotypic Expression ◽

Plant Cells ◽

Leaf Explants ◽

Leaf Disk ◽

Rt Pcr ◽

Set Up ◽

Morphological Phenotype

Summary APETALA1 (AP1) is one of flowering identity genes that determines the formation of sepal and petal tissues. An AP1 homologue was cloned from cacao flowers by bio-techniques coupled with bio-informatics. Examination of phenotypic expression was conducted with transgenesis of the 35S-TcAP1 construct using leaf disk technique of tobacco leaf explants mediated by Agrobacterium tumefaciens. PCR specific to TcAP1 demonstrated that the technique is effective in introducing the 35S-TcAP1 construct into tobacco plant cells. RT-PCR with total RNA from the leaves of transgenic tobacco plantlets showed that expression levels of the TcAP1 events varied. The variation of the transcript levels was comparable to the morphological phenotype of the tobacco plantlets grown in vitro. The cultures expressing TcAP1 at moderate levels, have developed into intact plantlets and set up flowers in vitro.Ringkasan APETALA1 (AP1) diketahui merupakan salah satu gen identitas pembungaan yang mengendalikan terbentuknya jaringan sepal dan petal. Homolog AP1 telah diklon dari organ bunga kakao (TcAP1) dengan kombinasi bio-techniques dan bio-informatics. Pengujian ekspresi fenotipe TcAP1 dilakukan dengan transgenesis konstruk konstitutif 35S-TcAP1 menggunakan teknik leaf disk eksplan daun tembakau dan mediasi Agrobacterium tumefaciens. Pengujian PCR spesifik TcAP1 menunjukkan bahwa teknik tersebut cukup efektif dalam mengintroduksikan konstruk 35S-TcAP1 ke dalam sel tanaman tembakau. RT-PCR dari daun planlet tembakau trangenik membuktikan bahwa tingkat ekspresi TcAP1 tersebut bervariasi. Perbedaan level ekspresi TcAP1 ini memberikan pengaruh yang nampak sebanding terhadap perkembangan morfologis planlet tembakau in vitro. Kultur yang mengekspresikan TcAP1 pada level sedang mampu beregenerasi menjadi planlet sempurna dan membentuk bunga in vitro.

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Anti-dengue activity of super critical extract and isolated oleanolic acid of Leucas cephalotes using in vitro and in silico approach

BMC Complementary Medicine and Therapies ◽

10.1186/s12906-021-03402-2 ◽

2021 ◽

Vol 21 (1) ◽

Cited By ~ 1

Author(s):

Sulochana Kaushik ◽

Lalit Dar ◽

Samander Kaushik ◽

Jaya Parkash Yadav

Keyword(s):

Nuclear Magnetic Resonance ◽

Magnetic Resonance ◽

Oleanolic Acid ◽

Plant Extract ◽

In Silico ◽

Rt Pcr ◽

Ft Ir ◽

Isolated Compound ◽

Layer Chromatography

Abstract Backgrounds Leucas cephalotes is a common ethnomedicinal plant widely used by traditional healers for the treatment of Malaria and other types of fever. Oleanolic acid and its derivatives have been reported for various types of pharmacological activities, such as anti-inflammatory, antioxidant, anticancer, hepatoprotective, anti-HIV and anti-HCV activity. Methods L.cephalotes plant extracts were prepared by supercritical fluid extraction (SFE) method and oleanolic acid was isolated by preparatory thin-layer chromatography. The compound was identified and characterize by using ultraviolet-visible spectroscopy (UV-VIS), Fourier transform infra-Red spectroscopy (FT-IR) and high-performance thin-layer chromatography (HPTLC). The structure of the compound was elucidated by proton nuclear magnetic resonance (1HNMR) and carbon nuclear magnetic resonance (1CNMR) and the purity checked by differential scanning calorimetry (DSC). The MTT assay was used to determine the toxicity of plant extract and oleanolic acid using a microplate reader at 595 nm. The anti-dengue activity of plant extract and oleanolic acid was tested in vitro and in silico using real-time RT-PCR. Results The optimum yield of the extract was obtained at 40 °C temperature and 15Mpa pressure. The maximum non-toxic dose (MNTD) of plant extract and oleanolic acid were found as 46.87 μg/ml and 93.75 μg/ml, respectively in C6/36 cell lines. UV spectrophotometer curve of the isolated compound was overlapped with standard oleanolic acid at 232 nm. Superimposed FT-IR structure of the isolated compound was indicated the same spectra at 3433, 2939, 2871, 1690, 1500,1463, 1387, 1250, 1209, 1137 and 656 position as per marker compound. HPTLC analysis showed the retention factor of L. cephalotes extract was 0.19 + 0.06 as similar to the standard oleanolic acid chromatogram. The NMR structure of the isolated compound was identified as similar to the marker oleanolic acid structure. DSC analysis revealed the purity of isolated oleanolic acid was 98.27% with a melting point of 311.16 °C. Real-time RT PCR results revealed that L. cephalotes supercritical extract and isolated oleanolic acid showed 100 and 99.17% inhibition against the dengue − 2 virus when treated with MNTD value of plant extract (46.87 μg/ml) and the test compound (93.75 μg/ml), respectively. The molecular study demonstrated the binding energy of oleanolic acid with NS1and NS5 (non-structural protein) were − 9.42 & -8.32Kcal/mol, respectively. Conclusions The SFE extract L. cephalotes and its active compound, oleanolic acid inhibiting the activity of dengue-2 serotype in the in vitro and in silico assays. Thus, the L.cephalotes plant could be an excellent source for drug design for the treatment of dengue infection.

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Naringin inhibits the Zea mays coniferyl aldehyde dehydrogenase: an in silico and in vitro approach

Journal of Plant Biochemistry and Biotechnology ◽

10.1007/s13562-020-00561-0 ◽

2020 ◽

Vol 29 (3) ◽

pp. 484-493

Author(s):

Ana Paula Ferro ◽

Angela Valderrama Parizotto ◽

Wanderley Dantas dos Santos ◽

Rogério Marchiosi ◽

Flavio Augusto Vicente Seixas ◽

...

Keyword(s):

Zea Mays ◽

Aldehyde Dehydrogenase ◽

In Silico

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Ekspresi fenotipe gen APETALA1 kakao (TcAP1) pada eksplan tembakau Phenotypic expression of cacao APETALA1 (TcAP1) in tobacco explant

E-Journal Menara Perkebunan ◽

10.22302/iribb.jur.mp.v74i1.116 ◽

2016 ◽

Vol 74 (1) ◽

Author(s):

Tetty CHAIDAMSARI ◽

. SAMANHUDI ◽

Asmini BUDIANI ◽

Roedhy POERWANTO ◽

Djoko SANTOSO

Keyword(s):

Agrobacterium Tumefaciens ◽

Tobacco Plant ◽

Phenotypic Expression ◽

Plant Cells ◽

Leaf Explants ◽

Leaf Disk ◽

Rt Pcr ◽

Set Up ◽

Morphological Phenotype

Summary APETALA1 (AP1) is one of flowering identity genes that determines the formation of sepal and petal tissues. An AP1 homologue was cloned from cacao flowers by bio-techniques coupled with bio-informatics. Examination of phenotypic expression was conducted with transgenesis of the 35S-TcAP1 construct using leaf disk technique of tobacco leaf explants mediated by Agrobacterium tumefaciens. PCR specific to TcAP1 demonstrated that the technique is effective in introducing the 35S-TcAP1 construct into tobacco plant cells. RT-PCR with total RNA from the leaves of transgenic tobacco plantlets showed that expression levels of the TcAP1 events varied. The variation of the transcript levels was comparable to the morphological phenotype of the tobacco plantlets grown in vitro. The cultures expressing TcAP1 at moderate levels, have developed into intact plantlets and set up flowers in vitro.Ringkasan APETALA1 (AP1) diketahui merupakan salah satu gen identitas pembungaan yang mengendalikan terbentuknya jaringan sepal dan petal. Homolog AP1 telah diklon dari organ bunga kakao (TcAP1) dengan kombinasi bio-techniques dan bio-informatics. Pengujian ekspresi fenotipe TcAP1 dilakukan dengan transgenesis konstruk konstitutif 35S-TcAP1 menggunakan teknik leaf disk eksplan daun tembakau dan mediasi Agrobacterium tumefaciens. Pengujian PCR spesifik TcAP1 menunjukkan bahwa teknik tersebut cukup efektif dalam mengintroduksikan konstruk 35S-TcAP1 ke dalam sel tanaman tembakau. RT-PCR dari daun planlet tembakau trangenik membuktikan bahwa tingkat ekspresi TcAP1 tersebut bervariasi. Perbedaan level ekspresi TcAP1 ini memberikan pengaruh yang nampak sebanding terhadap perkembangan morfologis planlet tembakau in vitro. Kultur yang mengekspresikan TcAP1 pada level sedang mampu beregenerasi menjadi planlet sempurna dan membentuk bunga in vitro.

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