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1.
Phytomedicine ; 106: 154416, 2022 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-36037770

RESUMEN

BACKGROUND: Anemoside B4 (AB4) is reported to prevent acute colitis when given via intraperitoneal injection by two recent studies. However, whether oral AB4 protects against chronic colitis which resembles the clinical phenotype of ulcerative colitis (UC) and its mechanism of action are largely unknown. PURPOSE: To systemically investigate the effects of oral AB4 against chronic colitis and illustrate the underlying mechanism of action. METHODS: The preventive, therapeutic, and dose-dependent effects of AB4 against UC were examined in mice with acute or chronic relapsing colitis induced by dextran sulfate sodium (DSS). The inflammatory responses, colonic transcriptome, and 16S rDNA sequencing of the intestinal content of mice were analyzed. RESULTS: Oral administration of AB4 alleviated disease severity and colon shortening in mice with chronic relapsing colitis in a dose-dependent manner. The effects of AB4 were comparable to those of two positive-control compounds: tofacitinib and berberine. Unlike tofacitinib, AB4 did not have a deleterious effect on DSS-induced splenic swelling and anemia. Furthermore, AB4 inhibited the inflammatory responses of colitis, as evidenced by in-vivo, ex-vivo, and in-vitro studies. Transcriptomics revealed that AB4 treatment reversed the DSS-mediated decrease in the expression of colonic Pelo, B3gat2 and Mir8010. In addition, AB4 reversed DSS-induced alterations in the intestinal microbiome in mice. Through fecal microbiota transplantation, we proved that AB4 partially exerted its anti-colitis effects by modulating the gut microbiota. CONCLUSIONS: We demonstrated for the first time that AB4 has dose-dependent therapeutic effects against chronic relapsing colitis by modulating the inflammatory response, colonic gene expression, and intestinal microbiota.


Asunto(s)
Berberina , Colitis Ulcerosa , Colitis , Microbioma Gastrointestinal , Animales , Berberina/farmacología , Colitis/inducido químicamente , Colitis/tratamiento farmacológico , Colitis/metabolismo , Colitis Ulcerosa/inducido químicamente , Colitis Ulcerosa/tratamiento farmacológico , Colitis Ulcerosa/metabolismo , Colon , Citocinas/metabolismo , ADN Ribosómico/metabolismo , ADN Ribosómico/farmacología , Sulfato de Dextran/efectos adversos , Modelos Animales de Enfermedad , Ratones , Ratones Endogámicos C57BL , Saponinas , Transcriptoma
2.
Phytomedicine ; 104: 154284, 2022 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-35777121

RESUMEN

BACKGROUND: Lonicera rupicola Hook.f.et Thoms (LRH) is used as a customary medicinal herb in Tibetans. And LRH flavonoids have excellent anti-inflammatory and antioxidant pharmacological activities. However, the specific effects of LRH and its mechanism remain unknown, and there is a deficiency of systematic research, leading to the waste of LRH as a medicinal resource. PURPOSE: In this study, in an attempt to rationalize the development and utilization of Tibetan herbal resources, the therapeutic efficacy and the underlying molecular mechanisms of LRH flavonoids on dextran sulfate sodium (DSS)-induced ulcerative colitis (UC) were investigated, establishing the favorable basis for the pharmacodynamic material basis of LRH and providing a scientific basis for the discovery of new drugs for the treatment of UC. METHODS: Firstly, ultra-performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS) was used for identification and detection of the flavonoid components of LRH. Meanwhile, their potential targets, biological functions and signaling pathways were predicted with the assistance of network pharmacology analysis. Subsequently, pharmacological efficacy of LRH were evaluated by body weight loss, colon length, disease activity index (DAI), histology observation and the expression levels of inflammatory mediators, messenger RNA (mRNA) and tight junction proteins. Moreover, in the present investigation, we also profiled the gut microbiome via high-throughput sequencing of the V3-V4 region of 16S ribosomal DNA (rDNA) for bacterial community composition and diversity by Illumina MiSeq platforms. Finally, the key regulatory proteins in the PI3K/AKT pathways were measured to investigate their underlying molecular mechanisms. RESULTS: A total of 37 LRH flavonoid components were identified and detected by UPLC-MS/MS, and 12 potential active components were obtained after screening. 137 of their common targets with UC were further predicted. GO and KEGG pathway enrichment analysis and molecular docking experiments demonstrated that LRH flavonoids could interfere with UC through "multi-component-multi-target-multi-pathway". In the animal experiments, LRH flavonoids could significantly attenuate UC as demonstrated by reducing the body weight loss and DAI, restoring colon length, decreasing oxidative stress, and improving the intestinal epithelial cell barrier. The mRNA and proteins expression levels of inflammatory mediators were returned to dynamic balance following LRH flavonoids treatment. 16S rDNA sequence analysis indicated that LRH flavonoids promoted the recovery of gut microbiome. And the PI3K/AKT pathway was significantly suppressed by LRH flavonoids. CONCLUSIONS: LRH flavonoids exhibited multifaceted protective effects against DSS-induced UC in mice through mitigating colon inflammation and oxidative stress, restoring epithelial barrier function, and improving the gut microenvironment potentially through modulation of the PI3K/AKT pathway. This finding demonstrated that LRH flavonoids possessed great potential for becoming an excellent drug for the treatment of UC.


Asunto(s)
Colitis Ulcerosa , Colitis , Microbioma Gastrointestinal , Lonicera , Animales , Cromatografía Liquida , Colitis/tratamiento farmacológico , Colitis Ulcerosa/inducido químicamente , Colitis Ulcerosa/tratamiento farmacológico , Colitis Ulcerosa/patología , Colon/patología , ADN Ribosómico/metabolismo , ADN Ribosómico/farmacología , Sulfato de Dextran/efectos adversos , Modelos Animales de Enfermedad , Flavonoides/farmacología , Flavonoides/uso terapéutico , Mediadores de Inflamación/metabolismo , Lonicera/metabolismo , Ratones , Ratones Endogámicos C57BL , Simulación del Acoplamiento Molecular , Fosfatidilinositol 3-Quinasas/metabolismo , Proteínas Proto-Oncogénicas c-akt/metabolismo , ARN Mensajero/metabolismo , Espectrometría de Masas en Tándem , Pérdida de Peso
3.
Biomed Res Int ; 2021: 6649085, 2021.
Artículo en Inglés | MEDLINE | ID: mdl-34136571

RESUMEN

Aging affects the brain function in elderly individuals, and Dushen Tang (DST) is widely used for the treatment of senile diseases. In this study, the protective effect of DST against memory impairment was evaluated through the Morris water maze (MWM) test and transmission electron microscopy (TEM). A joint analysis was also performed using LC-MS metabolomics and the microbiome. The MWM test showed that DST could significantly improve the spatial memory and learning abilities of rats with memory impairment, and the TEM analysis showed that DST could reduce neuronal damage in the hippocampus of rats with memory impairment. Ten potential biomarkers involving pyruvate metabolism, the synthesis and degradation of ketone bodies, and other metabolic pathways were identified by the metabolomic analysis, and it was found that 3-hydroxybutyric acid and lactic acid were involved in the activation of cAMP signaling pathways. The 16S rDNA sequencing results showed that DST could regulate the structure of the gut microbiota in rats with memory impairment, and these effects were manifested as changes in energy metabolism. These findings suggest that DST exerts a good therapeutic effect on rats with memory impairment and that this effect might be mainly achieved by improving energy metabolism. These findings might lead to the potential development of DST as a drug for the treatment of rats with memory impairment.


Asunto(s)
Encéfalo/efectos de los fármacos , Medicamentos Herbarios Chinos/farmacología , Galactosa/química , Metabolómica , Microbiota/efectos de los fármacos , Panax/química , Ácido 3-Hidroxibutírico/química , Animales , Biomarcadores/metabolismo , Cromatografía en Capa Delgada , AMP Cíclico/metabolismo , ADN Ribosómico/metabolismo , Metabolismo Energético , Hipocampo/efectos de los fármacos , Ácido Láctico/química , Masculino , Aprendizaje por Laberinto , Memoria , Microscopía Electrónica de Transmisión , Modelos Animales , Neuronas/metabolismo , Ratas , Ratas Sprague-Dawley , Transducción de Señal
4.
Molecules ; 26(8)2021 Apr 16.
Artículo en Inglés | MEDLINE | ID: mdl-33923441

RESUMEN

Salmonella typhimurium infection is associated with gastrointestinal disorder and cellular injury in the liver of both humans and animals. Cinnamaldehyde, the main component of essential oil from cinnamon, has been reported to have anti-inflammatory, anti-oxidative, and anti-apoptotic effects. However, it remains unknown whether cinnamaldehyde can alleviate Salmonella typhimurium infection-induced liver injury in mice. In the present study, we found that cinnamaldehyde attenuated Salmonella typhimurium-induced body weight loss, the increase of organ (liver and spleen) indexes, hepatocyte apoptosis, and the mortality rate in mice. Further study showed that cinnamaldehyde significantly alleviated Salmonella typhimurium-induced liver injury as shown by activities of alanine transaminase, aspartate transaminase, and myeloperoxidase, as well as malondialdehyde. The increased mRNA level of pro-inflammatory cytokines (IL-1ß, IL-6, TNF-α, and IFN-γ) and chemokines (CCL2 and CCL3) induced by Salmonella typhimurium were significantly abolished by cinnamaldehyde supplementation. These alterations were associated with a regulatory effect of cinnamaldehyde on TLR2, TLR4, and MyD88. 16S rDNA sequence analysis showed that Salmonella typhimurium infection led to upregulation of the abundances of genera Akkermansia, Bacteroides, Alistipes, Muribaculum, and Prevotellaceae UCG-001, and downregulation of the abundances of genera Lactobacillus, Enterorhabdus, and Eggerthellaceae (unclassified). These alterations were reversed by cinnamaldehyde supplementation. In conclusion, cinnamaldehyde attenuated the inflammatory response, oxidative stress, and apoptosis in the liver of Salmonella typhimurium-infected mice. Supplementation of cinnamaldehyde might be a preventive strategy to alleviate liver injury caused by Salmonella typhimurium infection in humans and animals.


Asunto(s)
Acroleína/análogos & derivados , Acroleína/química , Animales , Apoptosis/genética , Apoptosis/fisiología , Western Blotting , ADN Ribosómico/metabolismo , Microbioma Gastrointestinal/genética , Microbioma Gastrointestinal/fisiología , Etiquetado Corte-Fin in Situ , Inflamación/metabolismo , Hígado/metabolismo , Masculino , Ratones , Ratones Endogámicos C57BL , Estrés Oxidativo/fisiología , Salmonella typhimurium/patogenicidad , Transducción de Señal/genética , Transducción de Señal/fisiología
5.
Biomolecules ; 10(10)2020 10 06.
Artículo en Inglés | MEDLINE | ID: mdl-33036123

RESUMEN

Long-term cultivation of Panax ginseng cell lines leads to a decreasing synthesis of the biologically active substances used in traditional medicine. To gain insight into the cellular mechanisms which may influence this process, we analyzed variations within the rDNA cluster of the Oriental ginseng cell lines. The cell lines were cultivated for 6 and 24 years; the number of nucleoli and chromosomes was analyzed. The complete 18S rDNA sequences were cloned and sequenced. The nucleotide polymorphism and phylogenetic relations of the sequences were analyzed, and the secondary structures for separate 18S rRNA regions were modeled. The 18S rDNA accumulated mutations during cell cultivation that correlate well with an increase in the number of chromosomes and nucleoli. The patterns of nucleotide diversity are culture-specific and the increasing polymorphism associates with cytosine methylation sites. The secondary structures of some 18S rRNA regions and their interaction can alter during cultivation. The phylogenetic tree topologies are particular for each cell line.The observed alterations in rDNA clusters are associated with a somaclonal variation, leading to changes in the pattern of intracellular synthesis during cell cultivation. The identified divergent rRNAs could provide additional gene expression regulation in P. ginseng cells by forming heterogeneous ribosomes.


Asunto(s)
Senescencia Celular , ADN de Plantas/metabolismo , ADN Ribosómico/metabolismo , Regulación de la Expresión Génica de las Plantas , Familia de Multigenes , Panax/metabolismo , Células Vegetales/metabolismo , Panax/genética
6.
J Immunol Res ; 2020: 2383970, 2020.
Artículo en Inglés | MEDLINE | ID: mdl-32733972

RESUMEN

Constipation, a gastrointestinal function disorder, is one of the side effects of paclitaxel (PTX) chemotherapy. Shenhuang plaster (SHP) application on the Shenque acupoint promotes gut motility in clinical settings. In this study, we elucidated the ingredients in SHP and evaluated its effects on PTX-induced constipation using a tumour-bearing mouse model. SHP was prepared using the traditional Chinese plaster preparation method. The ingredients were analysed using UPLC-MS/MS and identified via screening in a standard drug database. The gastrointestinal transit was evaluated by the movement of a fluorescein-labelled dextran in the gastrointestinal tract. A histological study of the mucosa was carried out after haematoxylin and eosin staining. mRNA expression was assessed using real-time RT-PCR, and the foetal microbiota composition was elucidated through 16 s rDNA sequencing and BLAST analysis. Our results indicate that the application of SHP attenuated weight gain inhibition by PTX; however, no inhibitory effect was observed on tumour growth. PTX-induced sluggish intestine, villus, and mucosal base layer damage were significantly improved following the application of SHP. Further, SHP enhanced the stimulation efficiency of PTX on TLR4 and its downstream cytokines, as well as on IL-1ß in intestinal cells. SHP combined with PTX reshaped the microbiota, which showed beneficial effects on health. Hence, these results provide evidence that SHP alleviates PTX-induced constipation and intestinal morphological damage but augments the effects of PTX on the expression of cytokines in the TLR4 pathway and IL-1ß. Therefore, we propose that SHP stimulates the host immune response to eradicate cancer cells.


Asunto(s)
Medicamentos Herbarios Chinos/farmacología , Motilidad Gastrointestinal/efectos de los fármacos , Animales , Línea Celular Tumoral , Proliferación Celular/efectos de los fármacos , Cromatografía Liquida/métodos , Citocinas/metabolismo , ADN Ribosómico/metabolismo , Femenino , Interleucina-1beta/metabolismo , Medicina Tradicional China/métodos , Ratones , Ratones Endogámicos BALB C , Paclitaxel/farmacología , ARN Mensajero/metabolismo , Espectrometría de Masas en Tándem/métodos , Receptor Toll-Like 4/metabolismo
7.
Prep Biochem Biotechnol ; 50(7): 697-707, 2020.
Artículo en Inglés | MEDLINE | ID: mdl-32108551

RESUMEN

Although tea seed cake (TSC) possesses high nutritional value, its high content of tea saponin (TS) limits its potential as feed. This study aimed to degrade TS in TSC by saponin-degrading strain and used a multistrains fermentation method to improve its nutritional value and palatability. Three saponin-degrading strains were isolated from Oleum Camelliae mill soil and identified as Citrobacter sp. FCTS301, Pantoea sp. FCTS302, and Enterobacter sp. FCTS303. Single-factor experiment showed that Citrobacter sp. FCTS301 had the highest degradation rate of TS. Response surface analysis for Citrobacter sp. FCTS301 indicated that the optimum culture conditions were as follows: initial pH of 7.2, culture temperature of 34.2 °C, inoculation amount of 7.3%, the agitation rate of 150 rpm, and the TS concentration of 10.0 g/L. Under these conditions, the maximum degradation rate was 82.6%. The fermentation process of TSC was obtained by a multistrains fermentation experiment. Considering the protein content, crude fiber degradation rate, and TS degradation rate of each group, the optimum inoculum amount of strains included Citrobacter sp. FCTS301, Aspergillus oryzae NCUF414, Saccharomyces cersvisiae NCUF306.5, and Lactobacillus plantarum NCUF201.1(5%, 0.5%, 1.0%, and 1.5%). After TS was degraded efficiently, fermented TSC can be presumed a potential feed raw material.


Asunto(s)
Citrobacter/metabolismo , Enterobacter/metabolismo , Microbiología Industrial/métodos , Pantoea/metabolismo , Saponinas/química , Té/química , Aspergillus oryzae , ADN Ribosómico/metabolismo , Fermentación , Concentración de Iones de Hidrógeno , Lactobacillus plantarum , Filogenia , Saccharomyces cerevisiae , Temperatura
8.
Sci Rep ; 9(1): 19575, 2019 12 20.
Artículo en Inglés | MEDLINE | ID: mdl-31862965

RESUMEN

Alzheimer's disease (AD) is a complex disorder influenced by both genetic and environmental components and has become a major public health issue throughout the world. Oxidative stress and inflammation play important roles in the evolution of those major pathological symptoms. Jatrorrhizine (JAT), a main component of a traditional Chinese herbal, coptidis rhizome, has been shown to have neuroprotective effects and we previously showed that it is also able to clear oxygen free radicals and reduce inflammatory responses. In this study, we demonstrated that JAT administration could alleviate the learning and memory deficits in AD. Furthermore, we also found that JAT treatment reduced the levels of Aß plaques in the cortex and hippocampus of APP/PS1 double-transgenic mice. Other studies suggest that there are gut microbiome alterations in AD. In order to explore the underlying mechanisms between gut microbiota and AD, DNA sequencing for 16s rDNA V3-V4 was performed in fecal samples from APP/PS1 transgenic mice and C57BL/6 wild-type (WT) mice. Our results indicated that APP/PS1 mice showed less Operational Taxonomic Units (OTUs) abundance in gut microbiota than WT mice and with different composition. Furthermore, JAT treatment enriched OTUs abundance and alpha diversity in APP/PS1 mice compared to WT mice. High dose of JAT treatment altered the abundance of some specific gut microbiota such as the most predominant phylum Firmicutes and Bacteroidetes in APP/PS1 mice. In conclusion, APP/PS1 mice display gut dysbiosis, and JAT treatment not only improved the memory deficits, but also regulated the abundance of the microbiota. This may provide a therapeutic way to balance the gut dysbiosis in AD patients.


Asunto(s)
Péptidos beta-Amiloides/metabolismo , Berberina/análogos & derivados , Microbioma Gastrointestinal/efectos de los fármacos , Aprendizaje/efectos de los fármacos , Trastornos de la Memoria/tratamiento farmacológico , Péptidos beta-Amiloides/genética , Análisis de Varianza , Animales , Astrocitos/efectos de los fármacos , Astrocitos/metabolismo , Berberina/uso terapéutico , ADN Ribosómico/metabolismo , Masculino , Aprendizaje por Laberinto/efectos de los fármacos , Ratones , Ratones Endogámicos C57BL , Ratones Transgénicos , Placa Amiloide/metabolismo
9.
Mediators Inflamm ; 2016: 2839232, 2016.
Artículo en Inglés | MEDLINE | ID: mdl-27610003

RESUMEN

This study was conducted to explore the effects of interferon tau (IFNT) on the intestinal microbiota and expression of interleukin 17 (IL-17) in the intestine of mice. IFNT supplementation increased microbial diversity in the jejunum and ileum but decreased microbial diversity in the feces. IFNT supplementation influenced the composition of the intestinal microbiota as follows: (1) decreasing the percentage of Firmicutes and increasing Bacteroidetes in the jejunum and ileum; (2) enhancing the percentage of Firmicutes but decreasing Bacteroidetes in the colon and feces; (3) decreasing Lactobacillus in the jejunum and ileum; (4) increasing the percentage of Blautia, Bacteroides, Alloprevotella, and Lactobacillus in the colon; and (5) increasing the percentage of Lactobacillus, Bacteroides, and Allobaculum, while decreasing Blautia in the feces. Also, IFNT supplementation decreased the expression of IL-17 in the intestines of normal mice and of an intestinal pathogen infected mice. In conclusion, IFNT supplementation modulates the intestinal microbiota and intestinal IL-17 expression, indicating the applicability of IFNT to treat the intestinal diseases involving IL-17 expression and microbiota.


Asunto(s)
Microbioma Gastrointestinal/efectos de los fármacos , Interferón Tipo I/farmacología , Interleucina-17/metabolismo , Proteínas Gestacionales/farmacología , Animales , ADN Ribosómico/metabolismo , Ingestión de Alimentos/efectos de los fármacos , Ratones , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa
10.
Int J Mol Sci ; 16(1): 1562-75, 2015 Jan 09.
Artículo en Inglés | MEDLINE | ID: mdl-25584616

RESUMEN

Taraxacum formosanum (TF) is a medicinal plant used as an important component of health drinks in Taiwan. In this study, a rapid, sensitive and specific loop-mediated isothermal amplification (LAMP) assay for authenticating TF was established. A set of four specific LAMP primers was designed based on the nucleotide sequence of the internal transcribed spacer 2 (ITS2) nuclear ribosomal DNA (nrDNA) of TF. LAMP amplicons were successfully amplified and detected when purified genomic DNA of TF was added in the LAMP reaction under isothermal condition (65 °C) within 45 min. These specific LAMP primers have high specificity and can accurately discriminate Taraxacum formosanum from other adulterant plants; 1 pg of genomic DNA was determined to be the detection limit of the LAMP assay. In conclusion, using this novel approach, TF and its misused plant samples obtained from herbal tea markets were easily identified and discriminated by LAMP assay for quality control.


Asunto(s)
Genes de Plantas , Taraxacum/metabolismo , Secuencia de Bases , Cartilla de ADN/metabolismo , ADN Ribosómico/genética , ADN Ribosómico/metabolismo , Análisis Discriminante , Datos de Secuencia Molecular , Técnicas de Amplificación de Ácido Nucleico , Alineación de Secuencia , Temperatura
11.
Proc Natl Acad Sci U S A ; 111(45): 16166-71, 2014 Nov 11.
Artículo en Inglés | MEDLINE | ID: mdl-25344531

RESUMEN

Centromeres mediate chromosome segregation and are defined by the centromere-specific histone H3 variant (CenH3)/centromere protein A (CENP-A). Removal of CenH3 from centromeres is a general property of terminally differentiated cells, and the persistence of CenH3 increases the risk of diseases such as cancer. However, active mechanisms of centromere disassembly are unknown. Nondividing Arabidopsis pollen vegetative cells, which transport engulfed sperm by extended tip growth, undergo loss of CenH3; centromeric heterochromatin decondensation; and bulk activation of silent rRNA genes, accompanied by their translocation into the nucleolus. Here, we show that these processes are blocked by mutations in the evolutionarily conserved AAA-ATPase molecular chaperone, CDC48A, homologous to yeast Cdc48 and human p97 proteins, both of which are implicated in ubiquitin/small ubiquitin-like modifier (SUMO)-targeted protein degradation. We demonstrate that CDC48A physically associates with its heterodimeric cofactor UFD1-NPL4, known to bind ubiquitin and SUMO, as well as with SUMO1-modified CenH3 and mutations in NPL4 phenocopy cdc48a mutations. In WT vegetative cell nuclei, genetically unlinked ribosomal DNA (rDNA) loci are uniquely clustered together within the nucleolus and all major rRNA gene variants, including those rDNA variants silenced in leaves, are transcribed. In cdc48a mutant vegetative cell nuclei, however, these rDNA loci frequently colocalized with condensed centromeric heterochromatin at the external periphery of the nucleolus. Our results indicate that the CDC48A(NPL4) complex actively removes sumoylated CenH3 from centromeres and disrupts centromeric heterochromatin to release bulk rRNA genes into the nucleolus for ribosome production, which fuels single nucleus-driven pollen tube growth and is essential for plant reproduction.


Asunto(s)
Proteínas de Arabidopsis/metabolismo , Arabidopsis/metabolismo , Proteínas de Ciclo Celular/metabolismo , Centrómero/metabolismo , Cromosomas de las Plantas/metabolismo , Heterocromatina/metabolismo , Chaperonas Moleculares/metabolismo , ARN de Planta/biosíntesis , ARN Ribosómico/biosíntesis , Sumoilación/fisiología , ATPasas Asociadas con Actividades Celulares Diversas , Arabidopsis/genética , Proteínas de Arabidopsis/genética , Proteínas de Ciclo Celular/genética , Nucléolo Celular/genética , Nucléolo Celular/metabolismo , Centrómero/genética , Cromosomas de las Plantas/genética , ADN de Plantas/genética , ADN de Plantas/metabolismo , ADN Ribosómico/genética , ADN Ribosómico/metabolismo , Sitios Genéticos/fisiología , Heterocromatina/genética , Humanos , Chaperonas Moleculares/genética , Polen/genética , Polen/metabolismo , ARN de Planta/genética , ARN Ribosómico/genética , Ribosomas/genética , Ribosomas/metabolismo
12.
Cell Death Dis ; 3: e281, 2012 Mar 15.
Artículo en Inglés | MEDLINE | ID: mdl-22419110

RESUMEN

Bidirectional non-protein-coding RNAs are ubiquitously transcribed from the genome. Convergent sense and antisense transcripts may regulate each other. Here, we examined the convergent cis-noncoding rRNAs (nc-rRNAs) in A5 and E9 lung cancer models. Sense nc-rRNAs extending from rDNA intergenic region to internal transcribed spacer of around 10 kb in length were identified. nc-rRNAs in sense direction exhibited in vitro characteristics of ribozymes, namely, degradation upon incubation with MgCl(2) and stabilization by complementary oligonucleotides. Detection of endogenous cleavage-ligation products carrying internal deletion of hundreds to thousands nucleotides by massively parallel sequencing confirmed the catalytic properties. Transfection of oligonucleotides pairing with antisense nc-rRNAs stabilized both target and complementary transcripts, perturbed rRNA biogenesis, and induced massive cell death via apoptotic and/or nonapoptotic mechanisms depending on cell type and treatment. Oligonucleotides targeting cellular sense transcripts are less responsive. Spontaneously detached cells, though rare, also showed accumulation of nc-rRNAs and perturbation of rRNA biogenesis. Direct participation of nc-rRNAs in apoptotic and nonapoptotic death was demonstrated by transfection of synthetic nc-rRNAs encompassing the rDNA promoter. In sum, convergent cis-nc-rRNAs follow a feed-forward mechanism to regulate each other and rRNA biogenesis. This opens an opportunity to disrupt rRNA biogenesis, commonly upregulated in cancers, via inhibition of ribozyme-like activities in nc-rRNAs.


Asunto(s)
Apoptosis , Células Epiteliales/metabolismo , Pulmón/metabolismo , ARN Catalítico/metabolismo , ARN Ribosómico/metabolismo , ARN no Traducido/metabolismo , Adenocarcinoma , Adenocarcinoma del Pulmón , Animales , Secuencia de Bases , Ciclo Celular/genética , Muerte Celular/genética , Línea Celular Tumoral , ADN Ribosómico/genética , ADN Ribosómico/metabolismo , Células Epiteliales/patología , Secuenciación de Nucleótidos de Alto Rendimiento , Pulmón/patología , Neoplasias Pulmonares , Cloruro de Magnesio/metabolismo , Ratones , Datos de Secuencia Molecular , Oligonucleótidos Antisentido/genética , Oligonucleótidos Antisentido/metabolismo , Regiones Promotoras Genéticas , ARN Catalítico/genética , ARN Ribosómico/genética , ARN no Traducido/genética , Transducción de Señal , Transcripción Genética , Transfección
13.
Bioresour Technol ; 105: 67-73, 2012 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-22189078

RESUMEN

An algal-bacterial culture, composed of wastewater-born algae and activated sludge, was cultivated to treat domestic wastewater and accumulate biomass simultaneously. The influence of algae and sludge inoculation ratios on the treatment efficiency and the settleability of the accumulated biomass were investigated. There was no significant effect of the inoculation ratios on the chemical oxygen demand removal. Comparatively, the nutrients removal and related mechanism were varied with different inoculation ratios. The highest nitrogen and phosphorus removal efficiencies were observed with 5:1 (algae/sludge) culture (91.0±7.0% and 93.5±2.5%, respectively) within 10 days, which was 5-40% higher and 2-4 days faster than those with other inoculation ratios. The biomass settleability was improved with the assistance of sludge, and the 1:5 (algae/sludge) culture showed the best settleability. Furthermore, 16S rDNA gene analysis showed that the bacterial communities were varying with different algae and sludge inoculation ratios and some specific bacteria were enriched during operation.


Asunto(s)
Chlorophyta/metabolismo , Aguas del Alcantarillado , Contaminantes Químicos del Agua/aislamiento & purificación , Purificación del Agua/métodos , Biomasa , Biotecnología/métodos , ADN Ribosómico/metabolismo , Restauración y Remediación Ambiental , Concentración de Iones de Hidrógeno , Nitrógeno/análisis , Oxígeno/química , Fósforo/análisis , ARN Ribosómico 16S/metabolismo , Temperatura , Factores de Tiempo
14.
Appl Microbiol Biotechnol ; 91(4): 1193-202, 2011 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-21667276

RESUMEN

Terminal restriction fragment length polymorphism (T-RFLP) can be used to assess how land use management changes the dominant members of bacterial communities. We compared T-RFLP profiles obtained via amplification with forward primers (27, 63F) each coupled with the fluorescently labeled reverse primer (1392R) and multiple restriction enzymes to determine the best combination for interrogating soil bacterial populations in an agricultural soil used for potato production. Both primer pairs provide nearly universal recognition of a 1,400-bp sequence of the bacterial domain in the V(1)-V(3) region of the 16S ribosomal RNA (rRNA) gene relative to known sequences. Labeling the reverse primer allowed for direct comparison of each forward primer and the terminal restriction fragments' relative migration units obtained with each primer pair and restriction enzyme. Redundancy analysis (RDA) and nested multivariate analysis of variance (MANOVA) were used to assess the effects of primer pair and choice of restriction enzyme on the measured relative migration units. Our research indicates that the 63F-1392R amplimer pair provides a more complete description with respect to the bacterial communities present in this potato (Solanum tuberosum L.)-barley (Hordeum vulgare L.) rotation over seeded to crimson clover (Trifolium praense L.). Domain-specific 16S rRNA gene primers are rigorously tested to determine their ability to amplify across a target region of the gene. Yet, variability within or between T-RFLP profiles can result from factors independent of the primer pair. Therefore, researchers should use RDA and MANOVA analyses to evaluate the effects that additional laboratory and environmental variables have on bacterial diversity.


Asunto(s)
Bacterias/clasificación , Bacterias/genética , Técnicas Bacteriológicas/métodos , Biodiversidad , ADN Bacteriano/genética , Polimorfismo de Longitud del Fragmento de Restricción , Microbiología del Suelo , Cartilla de ADN/genética , Enzimas de Restricción del ADN/metabolismo , ADN Bacteriano/metabolismo , ADN Ribosómico/genética , ADN Ribosómico/metabolismo , Hordeum/crecimiento & desarrollo , ARN Ribosómico 16S/genética , Solanum tuberosum/crecimiento & desarrollo , Trifolium/crecimiento & desarrollo
15.
Biodegradation ; 20(5): 643-50, 2009 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-19214760

RESUMEN

Three pentachlorophenol (PCP) degrading bacterial strains were isolated from sediment core of pulp and paper mill effluent discharge site. The strains were continuously enriched in mineral salts medium supplemented with PCP as sole source of carbon and energy. One of the acclimated strains with relatively high PCP degradation capability was selected and characterized in this study. Based on morphology, biochemical tests, 16S rDNA sequence analysis and phylogenetic characteristics, the strains showed greatest similarity with Acinetobacter spp. The strain was identified as Acinetobacter sp. ISTPCP-3. The physiological characteristics and optimum growth conditions of the bacterial strain were investigated. The results of optimum growth temperature revealed that it was a mesophile. The optimum growth temperature for the strain was 30 degrees C. The preferential initial pH for the strain was ranging at 6.5-7.5, the optimum pH was 7. The bacterium was able to tolerate and degrade PCP up to a concentration of 200 mg/l. Increase in PCP concentration had a negative effect on biodegradation rate and PCP concentration above 250 mg/l was inhibitory to its growth. Acinetobacter sp. ISTPCP-3 was able to utilize PCP through an oxidative route with ortho ring-cleavage with the formation of 2,3,5,6-tetrachlorohydroquinone and 2-chloro-1,4-benzenediol, identified using gas chromatograph-mass spectrometric (GC-MS) analysis. The degradation pathway followed by isolated bacterium is different from previously characterized pathway.


Asunto(s)
Acinetobacter/aislamiento & purificación , Acinetobacter/metabolismo , Biodegradación Ambiental , Pentaclorofenol/metabolismo , Acinetobacter/crecimiento & desarrollo , ADN Ribosómico/metabolismo , Cromatografía de Gases y Espectrometría de Masas , Residuos Industriales , Filogenia , Eliminación de Residuos Líquidos
16.
J Integr Plant Biol ; 50(3): 360-7, 2008 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-18713369

RESUMEN

The complete process of meiosis was investigated in Pinus wallichiana, P. strobus and their artificial hybrid (F1) using microsporocytes. It is revealed that there were slightly lower chiasma frequency, lower ring bivalent frequency, lower meiotic index and distinctly higher frequency of aberrance (chromosomal bridges, fragments or micronuclei) in pollen mother cells (PMCs) of the hybrid (F1) than those of the parental species, which showed a certain degree of differentiation between homologous chromosomes of the two parents. However, relatively higher frequency of ring bivalents and higher meiotic index in all the three entities indicate the great stability of genomes of parental species, and the differentiation of genomes between the two parents must have been slight. Total nineteen signal loci of 18S rDNA were observed in nine bivalents of the hybrid (F1), among which one bivalent bears two loci, while the others have only one. It is suggested that distinct differentiation at genetic level existed in homologous chromosomes of the two parental species, whereas only slight differentiation at karyotypic and genomic levels take place between the parent species.


Asunto(s)
Cromosomas de las Plantas/metabolismo , ADN Ribosómico/metabolismo , Hibridación Genética , Hibridación Fluorescente in Situ , Meiosis , Pinus/citología , Polen/citología , Anafase , Cruzamientos Genéticos , Metafase , Pinus/metabolismo , Polen/metabolismo , Profase , Telofase
17.
Mol Ecol ; 17(5): 1375-86, 2008 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-18302695

RESUMEN

Female Mediterranean fruit flies (Ceratitis capitata) oviposit in fruits, within which the larvae develop. This development is associated with rapid deterioration of the fruit, and frequently with invasion by secondary pests. Most research on the associations between medflies and microorganisms has focused on the bacteria inhabiting the digestive system of the adult fly, while the role of the fruit in mediating, amplifying or regulating the fruit fly microflora has been largely neglected. In this study, we examine the hypothesis that the host fruit plays a role in perpetuating the fly-associated bacterial community. Using direct and cultured-based approaches, we show that this community is composed in its very large majority of diazotrophic and pectinolytic Enterobacteriaceae. Our data suggest that this fly-associated enterobacterial community is vertically transmitted from the female parent to its offspring. During oviposition, bacteria are transferred to the fruit, establish and proliferate within it, causing its decay. These results show that the host fruit is indeed a central partner in the fruit fly-bacterial interaction as these transmitted bacteria are amplified by the fruit, and subsequently maintained throughout the fly's life. This enterobacterial community may contribute to the fly's nitrogen and carbon metabolism, affecting its development and ultimately, fitness.


Asunto(s)
Fenómenos Fisiológicos Bacterianos , Ceratitis capitata/microbiología , Frutas/parasitología , Psidium/parasitología , Animales , Ceratitis capitata/genética , Ceratitis capitata/crecimiento & desarrollo , ADN Ribosómico/metabolismo , Femenino , Tracto Gastrointestinal/microbiología , Regulación Enzimológica de la Expresión Génica , Oxidorreductasas/genética , Oxidorreductasas/metabolismo , Pectinas/metabolismo , Filogenia , Reacción en Cadena de la Polimerasa , ARN Mensajero/genética , ARN Mensajero/metabolismo
18.
J Basic Microbiol ; 47(1): 31-9, 2007 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-17304616

RESUMEN

As the first step in an investigation of the problem with quality deterioration seen in the Neungee mushroom (Sarcodon aspratus) due to bacterial overgrowth during its storage, an attempt to isolate bacterial strains was made using infected gills of Sarcodon aspratus. Five bacterial strains were isolated; one phototrophic cyanobacterial species and four heterotrophic Gram negative rods. The four heterotrophic bacterial isolates (strains P, S, R, and MK1) were subjected to identification based on biochemical characteristics using the Biolog system, cellular fatty acid analysis using the MIDI system, cytology by scanning microscopy, and 16s rDNA sequence analysis. A slow grower, the P strain (ca. 0.7 microm x 1.5 microm), which forms pink colonies on Tryptic Soy agar (TSA) and glucose minimal salt medium containing thiamine (MT medium), belongs to genus Methylobacterium, and is likely M. radiotolerans. The methanol-utilizing capacity of the P strain was confirmed by growth on methanol-supplemented medium as a sole carbon source. Both the S and R strains (ca. 0.5 microm x 0.8 microm) produced smooth and slightly rough white colonies, respectively, on TSA, MT, and potato dextrose (PD) agar are members of the Burkholderia cepacia complex. Although both strains showed some differences from each other in colony morphology, nitrogen fixation capacity, and denitrification, they were considered to be Burkholderia stabilis because their 16s rDNA sequences showed 99.93% similarity with those of B. stabilis LMG 14294T (NCBI AF 148554). The MK1 strain, a rod-shaped bacterium with a tapered end (ca. 0.6 microm x 1.8 microm), produces a copious mucoid substance on MT and PD agar, but not on TSA. Despite extensive identification studies, the M strain is not currently identifiable, which suggests that it is a novel bacterium.


Asunto(s)
Basidiomycota , Burkholderia/aislamiento & purificación , Methylobacterium/aislamiento & purificación , Técnicas de Tipificación Bacteriana , Basidiomycota/metabolismo , Basidiomycota/ultraestructura , Burkholderia/clasificación , ADN Bacteriano/metabolismo , ADN Ribosómico/metabolismo , Ácidos Grasos/análisis , Methylobacterium/clasificación , Datos de Secuencia Molecular , Filogenia , Homología de Secuencia de Ácido Nucleico
19.
Genome ; 49(1): 91-6, 2006 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-16462906

RESUMEN

Variability in the organization of repeats of 5S rDNA is useful for phylogenetic studies in various crops. We found variable repeats of 5S rDNA gene in the genome of tea (Camellia sinensis (L.) O. Kuntze) during Southern hybridization. Variability in the repeats of 5S rDNA with specific restriction endonucleases (Sau3AI, BamHI, and ApoI) was analyzed in 28 different tea clones representing 3 types of tea. Our results clearly show that the 5S rDNA gene in tea could be used as a molecular marker to distinguish C. sinensis Chinary tea from the other important types of tea, namely Assamica and Cambod. Upon analysis with restriction endonucleases, the 5S rDNA gene in the tea genome was found to be heavily methylated.


Asunto(s)
Camellia sinensis/clasificación , Camellia sinensis/genética , ADN Ribosómico/análisis , ARN Ribosómico 5S/genética , Metilación de ADN , ADN Ribosómico/metabolismo , Genes de Plantas , Variación Genética , Filogenia , Polimorfismo de Longitud del Fragmento de Restricción
20.
Genome ; 48(2): 352-4, 2005 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-15838559

RESUMEN

Amplification by polymerase chain reaction of the 5S rDNA repeat units of Beta vulgaris subsp. maritima resulted in a 350-bp product corresponding to the full-length 5S unit, but also revealed 4 abridged unit classes, each with a deletion that removed most of the spacer and 12-76 bp of the coding sequence. Each abridged type lacks at least 1 of the conserved elements involved in transcription of the 5S gene, and so appear to be nonfunctional. Network analysis revealed that the abridged units are evolving in the same manner as the full-length versions.


Asunto(s)
Beta vulgaris/genética , ADN de Plantas/química , ADN Ribosómico/química , ADN Ribosómico/metabolismo , ARN Ribosómico 5S/genética , ADN de Plantas/genética , ADN de Plantas/metabolismo , ARN Ribosómico 5S/metabolismo , Eliminación de Secuencia
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