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1.
Fungal Genet Biol ; 162: 103728, 2022 09.
Artigo em Inglês | MEDLINE | ID: mdl-35932991

RESUMO

Ceratocystis fimbriata is an important pathogen that causes wilt in several plant species. Despite the importance of this pathogen, knowledge about its karyotypic polymorphism and genomic architecture is limited. The main objective of this study was to investigate the karyotype of isolates of the C. fimbriata species complex from different host plants and geographical origins in Brazil. First, the identity of the isolates was confirmed conducting multilocus sequence analysis (MLSA) phylogeny using ß-tubulin (TUBB), translation elongation factor 1α (TEF-1α) and mating-type (MAT1 and MAT2) gene sequences. To investigate the chromosomal polymorphism, two conditions of pulsed-field gel electrophoresis (PFGE) were used and the karyotypes of the isolates obtained. The retrotransposon-microsatellite amplified polymorphism (REMAP) molecular marker was utilized to assess the genetic variability among isolates. In the MLSA utilizing the concatenated gene sequences, Ceratocystis cacaofunesta and C. fimbriata formed separate clades, but considerable variation among C. fimbriata isolates was observed. Polymorphism in chromosome number and size was found, indicating the existence of genomic differences among isolates and occurrence of chromosomal rearrangements in the species complex. The number of chromosomes varied from seven to nine and the estimated minimum chromosome sizes were estimated to be between 2.7 and 6.0 Mbp. Small polymorphic chromosomes ranging from 1.2 to 1.8 Mbp were observed in all isolates, raising the hypothesis that they could be supernumerary chromosomes. REMAP analysis revealed a high genetic variability and that isolates from the same host tend to group together in a same cluster. Our results bring new insights into the chromosomal diversity and genome organization of the C. fimbriata complex.


Assuntos
Ascomicetos , Brasil , Ceratocystis , Cromossomos , DNA Fúngico/genética , Variação Genética/genética , Doenças das Plantas/genética
2.
BMC Med Res Methodol ; 22(1): 216, 2022 08 06.
Artigo em Inglês | MEDLINE | ID: mdl-35933340

RESUMO

BACKGROUND: The Randomised Evaluation of COVID-19 Therapy (RECOVERY) trial is aimed at addressing the urgent need to find effective treatments for patients hospitalised with suspected or confirmed COVID-19. The trial has had many successes, including discovering that dexamethasone is effective at reducing COVID-19 mortality, the first treatment to reach this milestone in a randomised controlled trial. Despite this, it continues to use standard or 'fixed' randomisation to allocate patients to treatments. We assessed the impact of implementing response adaptive randomisation within RECOVERY using an array of performance measures, to learn if it could be beneficial going forward. This design feature has recently been implemented within the REMAP-CAP platform trial. METHODS: Trial data was simulated to closely match the data for patients allocated to standard care, dexamethasone, hydroxychloroquine, or lopinavir-ritonavir in the RECOVERY trial from March-June 2020, representing four out of five arms tested throughout this period. Trials were simulated in both a two-arm trial setting using standard care and dexamethasone, and a four-arm trial setting utilising all above treatments. Two forms of fixed randomisation and two forms of response-adaptive randomisation were tested. In the two-arm setting, response-adaptive randomisation was implemented across both trial arms, whereas in the four-arm setting it was implemented in the three non-standard care arms only. In the two-arm trial, randomisation strategies were performed at the whole trial level as well as within three pre-specified patient subgroups defined by patients' respiratory support level. RESULTS: All response-adaptive randomisation strategies led to more patients being given dexamethasone and a lower mortality rate in the trial. Subgroup specific response-adaptive randomisation reduced mortality rates even further. In the two-arm trial, response-adaptive randomisation reduced statistical power compared to FR, with subgroup level adaptive randomisation exhibiting the largest power reduction. In the four-arm trial, response-adaptive randomisation increased statistical power in the dexamethasone arm but reduced statistical power in the lopinavir arm. Response-adaptive randomisation did not induce any meaningful bias in treatment effect estimates nor did it cause any inflation in the type 1 error rate. CONCLUSIONS: Using response-adaptive randomisation within RECOVERY could have increased the number of patients receiving the optimal COVID-19 treatment during the trial, while reducing the number of patients needed to attain the same study power as the original study. This would likely have reduced patient deaths during the trial and lead to dexamethasone being declared effective sooner. Deciding how to balance the needs of patients within a trial and future patients who have yet to fall ill is an important ethical question for the trials community to address. Response-adaptive randomisation deserves to be considered as a design feature in future trials of COVID-19 and other diseases.


Assuntos
Tratamento Farmacológico da COVID-19 , Dexametasona/uso terapêutico , Humanos , Lopinavir/uso terapêutico , SARS-CoV-2 , Resultado do Tratamento
3.
Mol Biol Rep ; 49(1): 31-38, 2022 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-34773551

RESUMO

BACKGROUND: Catharanthus roseus (L.) G. Donis a medicinal plant species belonging to the Apocynaceae family, which produces vinblastine and vincristine along with 100 other monoterpenoid indole alkaloids. The process of biosynthesis of C. roseus alkaloids is complex, in which many genes, enzymes, and regulators are involved. Induced mutations may be considered as a potential source for producing a higher amount of vinblastine and vincristine in this plant species. Therefore, the objective of the present study was to examine the effects of different treatments utilized on the induced genetic changes in C. roseus plants and enzyme activities. METHODS AND RESULTS: Spermine, jasmonic acid, methyjasmonate, putrescine, and cold plasma treatments were used for seed treatments. Different molecular markers, namely inter simple sequence repeat, inter retrotransposon amplified polymorphism, and retrotransposon microsatellite amplified polymorphism were employed to reveal the induced genetic changes. Antioxidant enzyme activities were also studied. The treated plants showed genetic variability and a significant increase in antioxidant enzyme activity compared to the control plants. The putrescine treatment resulted in the highest level of activity in superoxidase. A significant positive correlation occurred between the molecular markers data and antioxidant enzyme activities in treated plants. CONCLUSION: Our data revealed that the different phytohormones and cold plasma treatments could induce both genetic and chemical content changes in C. roseus plants.


Assuntos
Catharanthus/crescimento & desenvolvimento , Repetições de Microssatélites , Reguladores de Crescimento de Plantas/farmacologia , Gases em Plasma/farmacologia , Retroelementos , Acetatos/farmacologia , Catharanthus/efeitos dos fármacos , Catharanthus/genética , Catharanthus/metabolismo , Ciclopentanos/farmacologia , Regulação da Expressão Gênica de Plantas/efeitos dos fármacos , Oxilipinas/farmacologia , Proteínas de Plantas/metabolismo , Plantas Medicinais/efeitos dos fármacos , Plantas Medicinais/genética , Plantas Medicinais/crescimento & desenvolvimento , Plantas Medicinais/metabolismo , Putrescina/farmacologia , Sementes/efeitos dos fármacos , Sementes/genética , Sementes/crescimento & desenvolvimento , Sementes/metabolismo , Espermina/farmacologia , Superóxido Dismutase/metabolismo
4.
Mol Biol Rep ; 47(10): 7593-7606, 2020 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-32949305

RESUMO

Understanding the genetic diversity and relationships between genotypes is an effective step in designing effective breeding programs. Insertional polymorphisms of retrotransposons were studied in 75 cultivated and wild grape genotypes using retrotransposon-microsatellite amplified polymorphism (REMAP) technique. In the morphological part of work, seven pomological traits with a high breeding interest were also analyzed in the cultivated genotypes. A total of 328 markers were produced by 42 primer pairs, out of which 313 markers (95.43%) were polymorphic. Number of markers ranged from 4 in loci Tvv1Fa-873, Vine1-811, Gret1Ra-855 and Tvv1Fa-890 to 12 in locus Vine1Ra-841 with an average value of 7.45. Similarity values based on Dice's coefficient among all 75 grapevine genotypes varied from 0.41 to 0.77. Classification of genotypes using unweighted pair-group method using complete-linkage clustering led to six distinct groups. Some wild and cultivated varieties placed in the same groups. It seems there are close relationship between wild and cultivated genotypes and maybe wild genotypes are ancestor of native grapevines. Grouping of grapevine genotypes based on molecular marker data was not in agreement with clustering by agro-morphological data indicating that the most of multiplied sequences are confined to the non-coding regions of transposon elements. Results showed a substantial level of genetic diversity at molecular and pomological level and the potential of this diversity for future grape breeding programs.


Assuntos
DNA de Plantas/genética , Repetições de Microssatélites , Polimorfismo Genético , Característica Quantitativa Herdável , Vitis/genética , Irã (Geográfico)
5.
Mol Genet Genomics ; 291(1): 155-68, 2016 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-26231981

RESUMO

The completion of the genome assembly for the economically important coffee plant Coffea canephora (Rubiaceae) has allowed the use of bioinformatic tools to identify and characterize a diverse array of transposable elements (TEs), which can be used in evolutionary studies of the genus. An overview of the copy number and location within the C. canephora genome of four TEs is presented. These are tested for their use as molecular markers to unravel the evolutionary history of the Millotii Complex, a group of six wild coffee (Coffea) species native to Madagascar. Two TEs from the Gypsy superfamily successfully recovered some species boundaries and geographic structure among samples, whereas a TE from the Copia superfamily did not. Notably, species occurring in evergreen moist forests of eastern and southeastern Madagascar were divergent with respect to species in other habitats and regions. Our results suggest that the peak of transpositional activity of the Gypsy and Copia TEs occurred, respectively, before and after the speciation events of the tested Madagascan species. We conclude that the utilization of active TEs has considerable potential to unravel the evolutionary history and delimitation of closely related Coffea species. However, the selection of TE needs to be experimentally tested, since each element has its own evolutionary history. Different TEs with similar copy number in a given species can render different dendrograms; thus copy number is not a good selection criterion to attain phylogenetic resolution.


Assuntos
Coffea/genética , Café/genética , Elementos de DNA Transponíveis/genética , Genoma de Planta/genética , DNA de Plantas/genética , Evolução Molecular , Dosagem de Genes/genética , Filogenia
6.
Genome ; 59(8): 551-64, 2016 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-27458741

RESUMO

Lilium lancifolium Thunb. (2n = 2x = 24) is a cytologically conspicuous species with both diploids and triploids in nature. Cytological and molecular genetic analyses were carried out in both diploids and triploids that were collected from 55 geographical locations in Korea, Japan, and China. While the 5S rRNA gene loci were located at duplicated loci on the long arm of chromosome 2, the 45S rRNA gene loci were present in chromosomes 1, 2, 4, 6, 7, and 11. While the loci on chromosomes 1 and 7 were constant, the loci on chromosomes 2, 4, 6, 7, and 11 were variable in some plants so that the L. lancifolium accessions were grouped into 7 cytotypes in diploids and 12 cytotypes in triploids. REMAP marker analysis revealed that the diploids were classified into seven clusters, and the triploids were classified into a large cluster. Geographic, cytological, and genetic differentiations were not related in both the diploid and triploid accessions of L. lancifolium. Thus, current genetic variations occurred prior to the geographic differentiation in both diploids and triploids, and the 45S rDNA cytotype variations occurred after geographic differentiation in the current habitats of L. lancifolium.


Assuntos
DNA Ribossômico/genética , Lilium/genética , Sequência de Bases , Cromossomos de Plantas , Primers do DNA , Diploide , Loci Gênicos , Variação Genética , Lilium/citologia , Filogenia , RNA Ribossômico/genética , RNA Ribossômico 5S/genética , Retroelementos , Especificidade da Espécie , Triploidia
7.
J Appl Microbiol ; 120(2): 388-401, 2016 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-26541097

RESUMO

AIMS: The goals of the present study were to identify, to analyse the phylogenetic relations and to evaluate the genetic variability in Diaporthe endophytic isolates from common bean. METHODS AND RESULTS: Diaporthe sp., D. infecunda and D. phaseolorum strains were identified using multilocus phylogeny (rDNA ITS region; EF1-α, ß-tubulin, and calmodulin genes). IRAP (Inter-Retrotransposon Amplified Polymorphism) and REMAP (Retrotransposon-Microsatellite Amplified Polymorphism) molecular markers reveal the existence of high genetic variability, especially among D. infecunda isolates. CONCLUSIONS: It was concluded that the multilocus phylogenetic approach was more effective than individual analysis of ITS sequences, in identifying the isolates to species level, and that IRAP and REMAP markers can be used for studying the genetic variability in the genus Diaporthe particularly at the intraspecific level. SIGNIFICANCE AND IMPACT OF THE STUDY: The combined use of molecular tools such as multilocus phylogenetic approach and molecular markers, as performed in this study, is the best way to distinguish endophytic strains of Diaporthe isolated from common bean (Phaseolus vulgaris L.).


Assuntos
Ascomicetos/genética , Endófitos/genética , Variação Genética , Phaseolus/microbiologia , Ascomicetos/classificação , Ascomicetos/isolamento & purificação , Brasil , Endófitos/classificação , Endófitos/isolamento & purificação , Repetições de Microssatélites , Dados de Sequência Molecular , Filogenia , Polimorfismo Genético , Tubulina (Proteína)/genética
8.
Hereditas ; 153: 5, 2016.
Artigo em Inglês | MEDLINE | ID: mdl-28096767

RESUMO

BACKGROUND: Timothy (Phleum pratense L.), a cool-season hexaploid perennial, is the most important forage grass species in Nordic countries. Earlier analyses of genetic diversity in a collection of 96 genebank accessions of timothy with SSR markers demonstrated high levels of diversity but could not resolve population structure. Therefore, we examined a subset of 51 accessions with REMAP markers, which are based on retrotransposons, and compared the diversity results with those obtained with SSR markers. RESULTS: Using four primer combinations, 533 REMAP markers were analyzed, compared with 464 polymorphic alleles in the 13 SSR loci previously. The average marker index, which describes information obtained per experiment (per primer combination or locus) was over six times higher with REMAPs. Most of the variation found was within accessions, with somewhat less, 89 %, for REMAPs, than for SSR, with 93 %. CONCLUSIONS: SSRs revealed differences in the level of diversity slightly better than REMAPs but neither marker type could reveal any clear clustering of accessions based on countries, vegetation zones, or different cultivar types. In our study, reliable evaluation of SSR allele dosages was not possible, so each allele had to be handled as a dominant marker. SSR and REMAP, which report from different mechanisms of generating genetic diversity and from different genomic regions, together indicate a lack of population structure. Taken together, this likely reflects the outcrossing and hexaploid nature of timothy rather than failures of either marker system.


Assuntos
Variação Genética , Phleum/genética , Retroelementos , Alelos , DNA de Plantas/genética , Marcadores Genéticos , Genoma de Planta , Repetições de Microssatélites , Países Escandinavos e Nórdicos
9.
Electrophoresis ; 35(12-13): 1921-7, 2014 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-24634146

RESUMO

Maize is one of the most important crops and also a model for grass genome research. Transposable elements comprise over 78% of the maize genome and their ability to generate new copies makes them good potential markers. Interretrotransposon-amplified polymorphism (IRAP) and retrotransposon microsatellite amplified polymorphism (REMAP) protocols were used for the first time in maize to study the genetic variability between maize cultivars. Ten PCR primers were selected based on a systematic analysis of the sequence conservation in the extremities of different high copy number transposable elements, whereas one primer was chosen based on a microsatellite sequence. Of the 16 primer combinations tested, 14 produced polymorphic bands. These markers were used to identify genetic similarity among 20 maize cultivars selected by their different kernel oil content. Genetic similarity analysis was performed based on the polymorphic band profiles and dendrograms were developed by the unweighted pair-group method with arithmetic averages. Clustering technique revealed that samples were grouped into three clusters that differed in their kernel oil content and size, and in their relative embryo size. In the current investigation, there is evidence that IRAP/REMAP may be useful as markers in maize.


Assuntos
DNA de Plantas/genética , Variação Genética , Zea mays/genética , Análise por Conglomerados , Retroelementos , Sementes/genética
10.
Protoplasma ; 261(3): 581-592, 2024 May.
Artigo em Inglês | MEDLINE | ID: mdl-38191719

RESUMO

Overdoses of pesticides lead to a decrease in the yield and quality of plants, such as beans. The unconscious use of deltamethrin, one of the synthetic insecticides, increases the amount of reactive oxygen species (ROS) by causing oxidative stress in plants. In this case, plants tolerate stress by activating the antioxidant defense mechanism and many genes. 5-Aminolevulinic acid (ALA) improves tolerance to stress by acting exogenously in low doses. There are many gene families that are effective in the regulation of this mechanism. In addition, one of the response mechanisms at the molecular level against environmental stressors in plants is retrotransposon movement. In this study, the expression levels of superoxide dismutase (SOD), ascorbate peroxidase (APX), catalase (CAT), glutathione reductase (GR), and stress-associated protein (SAP) genes were determined by Q-PCR in deltamethrin (0.5 ppm) and various doses (20, 40, and 80 mg/l) of ALA-treated bean seedlings. In addition, one of the response mechanisms at the molecular level against environmental stressors in plants is retrotransposon movement. It was determined that deltamethrin increased the expression of SOD (1.8-fold), GPX (1.4-fold), CAT (2.7-fold), and SAP (2.5-fold) genes, while 20 and 40 mg/l ALA gradually increased the expression of these genes at levels close to control, but 80 mg/l ALA increased the expression of these genes almost to the same level as deltamethrin (2.1-fold, 1.4-fold, 2.6-fold, and 2.6-fold in SOD, GPX, CAT, and SAP genes, respectively). In addition, retrotransposon-microsatellite amplified polymorphism (REMAP) was performed to determine the polymorphism caused by retrotransposon movements. While deltamethrin treatment has caused a decrease in genomic template stability (GTS) (27%), ALA treatments have prevented this decline. At doses of 20, 40, and 80 mg/L of ALA treatments, the GTS ratios were determined to be 96.8%, 74.6%, and 58.7%, respectively. Collectively, these findings demonstrated that ALA has the utility of alleviating pesticide stress effects on beans.


Assuntos
Ácido Aminolevulínico , Nitrilas , Praguicidas , Piretrinas , Ácido Aminolevulínico/farmacologia , Ácido Aminolevulínico/metabolismo , Plântula/metabolismo , Retroelementos/genética , Praguicidas/metabolismo , Praguicidas/farmacologia , Antioxidantes/metabolismo , Catalase/metabolismo , Estresse Oxidativo , Espécies Reativas de Oxigênio/metabolismo , Superóxido Dismutase/metabolismo , Expressão Gênica , Glutationa/metabolismo , Ascorbato Peroxidases/genética , Ascorbato Peroxidases/metabolismo
11.
Cureus ; 16(4): e58530, 2024 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-38770468

RESUMO

Portal vein stenting is a treatment option for portal hypertension caused by extrahepatic portal vein obstruction or stenosis. However, limited pathways to approach the portal vein are available, hindering re-intervention in the portal vein. Portal vein puncture through the transjugular intrahepatic portosystemic shunt route is less invasive and considered suitable for portal vein stenting. Furthermore, transjugular intrahepatic portosystemic shunting facilitates repeat approaches to the portal vein. However, a transjugular intrahepatic portosystemic shunt stent is not recommended unless necessary because of adverse events, and cannot be retrieved, once placed. Herein, we report on a novel approach using the repeatable microcatheter access port: ReMAP™ (Toray, Tokyo, Japan), a central vein port into which a 2.9 Fr catheter can be inserted. We used it for a repeat approach to the portal vein with only one puncture and without placing a transjugular intrahepatic portosystemic shunt stent.

12.
J Genet Eng Biotechnol ; 20(1): 86, 2022 Jun 13.
Artigo em Inglês | MEDLINE | ID: mdl-35696009

RESUMO

BACKGROUND: Aegilops tauschii Coss. as a donor of wheat D genome has an important role in wheat breeding programs. Genetic and phylogeographic diversity of 79 Ae. tauschii accessions collected from north and northwest of Iran were analyzed based on retroelement insertional polymorphisms using inter-retrotransposon amplified polymorphism (IRAP) and retrotransposon-microsatellite amplified polymorphism (REMAP) markers. RESULTS: In total, 306 and 151 polymorphic bands were amplified in IRAP and REMAP analyses, respectively. As a result, a high level of polymorphism was observed among the studied accessions as revealed by an average of 25.5 bands per primer/primer combination and mean PIC value of 0.47 in IRAP and an average of 25.16 bands per primer combination and mean PIC value of 0.47 in REMAP. Genetic relationships of the accessions were analyzed using distance- and model-based cluster analyses. CONCLUSION: The result showed that genetic distance did not seem to be related to geographic distribution, and the accessions could be divided into three groups, which was further supported by principal coordinate analysis. These results on genetic diversity and population structure of Ae. tauschii in Iran should provide important knowledge on genetic resources and their applications in wheat breeding programs.

13.
Eur J Intern Med ; 96: 1-4, 2022 02.
Artigo em Inglês | MEDLINE | ID: mdl-34801401

RESUMO

Systems tend toward inertia until an external pressure pushes them toward change; thus, a situation of crisis such as the COVID-19 pandemic represents an opportunity for technological innovation. The prevailing need for treatments and vaccines has impelled innovation in the world of randomized clinical trials (RCT), resorting to ideas that had been floating around for a while. Is this merely a circumstantial phenomenon or are new methods here to stay?


Assuntos
COVID-19 , Humanos , SARS-CoV-2
14.
Braz J Microbiol ; 52(4): 2179-2192, 2021 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-34491570

RESUMO

The composition of endophytic communities is dynamic and demonstrates host specificity; besides, they have great intra- and interspecific genetic variability. In this work, we isolated leaf endophytic fungi from Serjania laruotteana, identify them using multilocus analysis, and evaluate the genetic variability using IRAP (inter-retrotransposon amplified polymorphism) and REMAP (retrotransposon-microssatellite amplified polymorphism). A total of 261 fungi were isolated and 58 were identified. Multilocus phylogenetic analysis using the partial sequences from the ITS1-5.8S-ITS2 regions, elongation factor 1-alpha, ß-tubulin, actin, glyceraldehyde-3-phosphate dehydrogenase, and calmodulin genes identify that most strains belonged to the Colletotrichum and Diaporthe genera, other isolated genera were Xylaria, Phyllosticta, Muyocopron, Fusarium, Nemania, Plectosphaerella, Corynespora, Bipolaris, and Curvularia. The IRAP and REMAP analyzes were performed with Colletotrichum and Diaporthe genera and showed 100% of polymorphism and high intra- and interspecific variability. This is the first report of the diversity of endophytic fungi from S. laruotteana. In addition, it demonstrated that the IRAP and REMAP can be used to distinguish morphologically similar lineages, revealing differences even strains of the same species.


Assuntos
Biodiversidade , Fungos , Sapindaceae , Fungos/classificação , Fungos/genética , Tipagem de Sequências Multilocus , Filogenia , Retroelementos/genética , Sapindaceae/microbiologia
15.
Monoclon Antib Immunodiagn Immunother ; 40(4): 156-161, 2021 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-34283655

RESUMO

CD44 functions as a major hyaluronan receptor on most cell types, with roles in cell adhesion, migration, proliferation, differentiation, and survival. The CD44 gene comprises 20 exons, with alternative splicing producing many different isoforms. CD44 variant isoforms exhibit tissue-specific expression patterns and have been studied as therapeutic targets for several cancers; therefore, anti-CD44 monoclonal antibodies (mAbs) are useful for investigating CD44 expression in various cancers. Previously, we established an anti-CD44 mAb, C44Mab-46 (IgG1, κ), by immunizing mice with the CD44v3-10 ectodomain. Although C44Mab-46 recognized all CD44 isoforms, the binding epitope of C44Mab-46 has not been determined. In this study, we first checked the reactivity of C44Mab-46 to several CD44v3-10 deletion mutants such as dN79, dN124, dN147, and dN224. We found the N-terminus of the C44Mab-46-binding epitope between residues 147 and 224 of CD44v3-10. We next investigated this epitope using a novel mapping system: RIEDL insertion for epitope mapping (REMAP) method. We constructed 31 CD44 standard (CD44s) mutants where the RIEDL tag was inserted into the expected epitope region in CD44s. We observed that the C44Mab-46 epitope constituted five amino acids: 174-TDDDV-178 of CD44s. Thus, the REMAP method could be used to determine mAb binding epitopes for membrane proteins.


Assuntos
Receptores de Hialuronatos , Neoplasias , Animais , Anticorpos Monoclonais , Mapeamento de Epitopos , Receptores de Hialuronatos/genética , Camundongos , Isoformas de Proteínas/genética
16.
Methods Mol Biol ; 2222: 263-286, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-33301099

RESUMO

Retrotransposable elements (RTEs) are highly common mobile genetic elements that are composed of several classes and make up the majority of eukaryotic genomes. The "copy-out and paste-in" life cycle of replicative transposition in these dispersive and ubiquitous RTEs leads to new genome insertions without excision of the original element. RTEs are important drivers of species diversity; they exhibit great variety in structure, size, and mechanisms of transposition, making them important putative components in genome evolution. Accordingly, various applications have been developed to explore the polymorphisms in RTE insertion patterns. These applications include conventional or anchored polymerase chain reaction (PCR) and quantitative or digital PCR with primers designed for the 5' or 3' junction. Marker systems exploiting these PCR methods can be easily developed and are inexpensively used in the absence of extensive genome sequence data. The main inter-repeat amplification polymorphism techniques include inter-retrotransposon amplified polymorphism (IRAP), retrotransposon microsatellite amplified polymorphism (REMAP), and Inter-Primer Binding Site (iPBS) for PCR amplification with a single or two primers.


Assuntos
Impressões Digitais de DNA/métodos , Variação Genética , Retroelementos , Primers do DNA , Marcadores Genéticos , Repetições de Microssatélites , Reação em Cadeia da Polimerase , Polimorfismo Genético
17.
Methods Mol Biol ; 2356: 211-224, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34236689

RESUMO

Retrotransposons are major components of the Magnaporthe oryzae genome; their high copy number and property of stable insertion in genome make them ideal tools to develop molecular markers. Retrotransposon-based marker techniques mainly rely on the amplification of DNA sequences present between the retrotransposon termini and some component of flanking genomic DNA. In this chapter, two marker systems known as inter-retrotransposon amplified polymorphism (IRAP) and retrotransposon-microsatellite amplified polymorphism (REMAP) are described for genetic diversity studies in M. oryzae. In the IRAP method, DNA profiles are generated using outward-facing primers from two nearby retrotransposons, while REMAP produces DNA profiles from genomic segments present in retrotransposons and microsatellite repeats. These marker techniques are simple, cost-effective, and easy to develop for polymorphism studies among M. oryzae isolates, races, or populations. In addition, the chapter also describes the utility of these retrotransposon-based DNA markers to study stress-induced genomic instabilities in M. oryzae.


Assuntos
Instabilidade Genômica , Ascomicetos , Variação Genética , Humanos , Repetições de Microssatélites/genética , Polimorfismo Genético , Retroelementos/genética
18.
Biology (Basel) ; 10(11)2021 Oct 25.
Artigo em Inglês | MEDLINE | ID: mdl-34827091

RESUMO

In our study, two aggressive-invasive species, Lupinus polyphyllus Lindl. and Heracleum sosnowskyi Manden. from Russia and Ukraine, were investigated. The success in naturalization of both species is associated with human activities, since they have been used in agriculture and floriculture and both have qualities such as environmental tolerance, high fertility and phenotypic plasticity. The purpose of this study was to determine the level of genetic diversity of both species. For Heracleum sosnowskyi Manden., genetic diversity was compared in invasive and native populations. For Lupinus polyphyllus Lindl., the genetic diversity was compared in variety, feral and invasive populations. A genetic diversity was formulated using RAPD, ISSR and REMAP. For Heracleum sosnowskyi Manden., the average genetic diversity within the invasive population was similar (0.432), but slightly less (0.502) than within the native Caucasian population. This may suggest the successful naturalization of invaders and almost complete reconstruction of their genetic diversity. For Lupinus polyphyllus Lindl., the genetic diversity for the invasive population was the highest, with an average of 0.294, while for variety, it was the lowest, with an average of 0.194. The feral population had an intermediate place with an average of 0.248, which could suggest an increase of diversity in the process of naturalization.

19.
J Intensive Care ; 9(1): 34, 2021 Apr 14.
Artigo em Inglês | MEDLINE | ID: mdl-33853684

RESUMO

REMAP-CAP, a randomized, embedded, multifactorial adaptive platform trial for community-acquired pneumonia, is an international clinical trial that is rapidly expanding its scope and scale in response to the COVID-19 pandemic. Japan is now joining REMAP-CAP with endorsement from Japanese academic societies. Commitment to REMAP-CAP can significantly contribute to population health through timely identification of optimal COVID-19 therapeutics. Additionally, it will promote the establishment of a national and global network of clinical trials to tackle future pandemics of emerging and re-emerging infectious diseases, in collaboration with multiple stakeholders, including front-line healthcare workers, governmental agencies, regulatory authorities, and academic societies.

20.
Biol Futur ; 71(1-2): 123-130, 2020 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-34554519

RESUMO

Retrotransposon activity and genomic template stability (GTS) are one of the most significant rearranging mechanisms in environmental stress. Therefore, in this study, it is aimed to elucidate effecting of Cobalt (Co) on the instability of genomes and Long Terminal Repeat retrotransposon polymorphism in Zea mays and whether humic acid (HA) has any role on these parameters. For this purpose, Retrotransposon-microsatellite amplified polymorphism (REMAP) and Inter-Retrotransposon Amplified Polymorphism (IRAP) markers were applied to evaluate retrotransposon polymorphism and the GTS levels. It was found that IRAP and REMAP primers generate unique polymorphic band structures on maize plants treated with various doses of Co. Retrotransposon polymorphism increased and GTS decreased while increasing Co concentration. On the other hand, there was a reduction in negative effects of Co on retrotransposon GTS and polymorphism after treatment with HA. The results indicate that HA may be used effectively for the protection of maize seedlings from the destructive effects of Co toxicity.


Assuntos
Cobalto/toxicidade , Genoma de Planta , Substâncias Húmicas , Polimorfismo Genético/efeitos dos fármacos , Zea mays/efeitos dos fármacos , Zea mays/genética , Cobalto/administração & dosagem , Relação Dose-Resposta a Droga , Retroelementos
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