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1.
Methods Mol Biol ; 2061: 347-358, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-31583671

RESUMO

In this chapter we describe protocols for immunolabeling and FISH of pollen grains undergoing postmeiotic mitosis using Aegilops speltoides, Secale cereale, and Hordeum vulgare as models. Tissue sectioning of pollen overcomes the problem of the pollen grain wall impermeability that interferes with immunolocalization and in situ hybridization. The crucial element of the protocol is the generation and immobilization of tissue sections. Our method facilitates the investigation of the microspore cell divisions and pollen grain maturation.

2.
Org Lett ; 2019 Nov 05.
Artigo em Inglês | MEDLINE | ID: mdl-31689112

RESUMO

Rhodium(III)-catalyzed annulative coupling of sulfoxonium ylides with allenoates was achieved, forming highly functionalized cyclopropanes with a quaternary carbon center by means of the sulfoxonium ylide functionality as a traceless bifunctional directing group and C4 synthon via an arene C-H activation and cyclopropanation cascade. The protocol features simultaneous formation of three new C-C bonds in one pot with excellent diastereoselectivity. The resultant cyclopropanation products could be further transformed to diverse synthetically useful compounds.

3.
Odontology ; 2019 Nov 07.
Artigo em Inglês | MEDLINE | ID: mdl-31701299

RESUMO

Periodontitis is a disease caused by periodontopathogens and is characterized by periodontal inflammation and alveolar bone resorption. As has been proven, host immune responses incite the development and progression of periodontitis. The present study sought to establish B10 cell functions and mechanisms in regulating host immunity during periodontitis. Periodontopathogen-specific B10 cells were purified and then injected into recipients to create the adoptive transfer models. We compared inflammatory cytokines and regulatory T (Treg)/Th17 cell expression in a healthy, normal model, an experimental periodontitis model, and experimental periodontitis model adoptively transferred with B10 cells. Compared with experimental periodontitis animals, our results showed that transfer of B10 cells alleviated alveolar bone resorption (P < 0.05) by reducing periodontal osteoclastogenesis (P < 0.05). Additionally, we found that B10 cell transfer into the experimental periodontitis ones resulted in increased IL-10 (P < 0.05), but decreased IL-17 (P < 0.05) and receptor activator for nuclear factor κB ligand (RANKL) (P < 0.05) gene and protein expression in local lesions. Moreover, adoptive transfer of B10 cells reduced the proportion of Th17 cells (P < 0.05) in the gingiva. The results of our study confirmed that B10 cells can modulate local host immune responses and prevent inflammatory damage of alveolar bone by reducing pro-inflammatory cytokine expression and decreasing local proliferation of Th17 cells.

4.
J Org Chem ; 2019 Nov 08.
Artigo em Inglês | MEDLINE | ID: mdl-31698899

RESUMO

An efficient copper-catalyzed cascade alkynylation/cyclization/isomerization reactions of aurone-derived azadienes with terminal alkynes has been developed, giving a series of 1,2-dihydrobenzofuro[3,2-b]pyridines with excellent yields. The obtained 1,2-dihydrobenzofuro[3,2-b]pyridines can be conveniently transformed to the corresponding benzofuro[3,2-b]pyridines under basic condition. Additionally, benzofuro[3,2-b]pyridines can also be prepared from azadienes and terminal alkynes in one-pot reaction. The synthetic utility was demonstrated by the synthesis of three bioactive molecules with potent topoisomerase inhibition in high yields. This strategy provides a facile approach to 1,2-dihydrobenzofuro[3,2-b]pyridines and benzofuro[3,2-b]pyridines.

5.
Mol Nutr Food Res ; : e1900751, 2019 Oct 04.
Artigo em Inglês | MEDLINE | ID: mdl-31584249

RESUMO

SCOPE: To assess the existing evidence of associations between consumption of soy and isoflavone and multiple health outcomes. METHODS AND RESULTS: This is an umbrella review of meta-analyses and systematic reviews of randomized trials and observational studies in humans. 114 Meta-analyses and systematic reviews are identified with 43 unique outcomes. Soy and isoflavone consumption seems more beneficial than harmful for a series of health outcomes. Beneficial associations are identified for cancers, cardiovascular disease, gynecological, metabolic, musculoskeletal, endocrine, neurological, and renal outcomes, particularly in perimenopausal women. Harmful association is only found for gastric cancer (RR: 1.17, 95% CI: 1.02-1.36) for high intake of miso soup (1-5 cups per day) in male. CONCLUSION: Generally, soy and isoflavone consumption is more beneficial than harmful. The results herein support promoting soy intake as part of a healthy diet. Randomized controlled trials are necessary to confirm this finding.

6.
Mol Cell ; 2019 Oct 03.
Artigo em Inglês | MEDLINE | ID: mdl-31606273

RESUMO

The Ras GTPases are frequently mutated in human cancer, and, although the Raf kinases are essential effectors of Ras signaling, the tumorigenic properties of specific Ras-Raf complexes are not well characterized. Here, we examine the ability of individual Ras and Raf proteins to interact in live cells using bioluminescence resonance energy transfer (BRET) technology. We find that C-Raf binds all mutant Ras proteins with high affinity, whereas B-Raf exhibits a striking preference for mutant K-Ras. This selectivity is mediated by the acidic, N-terminal segment of B-Raf and requires the K-Ras polybasic region for high-affinity binding. In addition, we find that C-Raf is critical for mutant H-Ras-driven signaling and that events stabilizing B-Raf/C-Raf dimerization, such as Raf inhibitor treatment or certain B-Raf mutations, can allow mutant H-Ras to engage B-Raf with increased affinity to promote tumorigenesis, thus revealing a previously unappreciated role for C-Raf in potentiating B-Raf function.

7.
Hum Genet ; 2019 Oct 12.
Artigo em Inglês | MEDLINE | ID: mdl-31606751

RESUMO

Pluripotent stem cell (PSC) cultures form an integral part of biomedical and medical research due to their capacity to rapidly proliferate and differentiate into hundreds of highly specialized cell types. This makes them a highly useful tool in exploring human physiology and disease. Genomic editing of PSC cultures is an essential method of attaining answers to basic physiological functions, developing in vitro models of human disease, and exploring potential therapeutic strategies and the identification of drug targets. Achieving reliable and efficient genomic editing is an important aspect of using large-scale PSC cultures. The CRISPR/Cas9 genomic editing tool has facilitated highly efficient gene knockout, gene correction, or gene modifications through the design and use of single-guide RNAs which are delivered to the target DNA via Cas9. CRISPR/Cas9 modification of PSCs has furthered the understanding of basic physiology and has been utilized to develop in vitro disease models, to test therapeutic strategies, and to facilitate regenerative or tissue repair approaches. In this review, we discuss the benefits of the CRISPR/Cas9 system in large-scale PSC cultures.

8.
Mucosal Immunol ; 2019 Oct 18.
Artigo em Inglês | MEDLINE | ID: mdl-31628427

RESUMO

MicroRNA (miR)-219a-5p has been implicated in the development of numerous progression of carcinoma and autoimmune diseases. However, whether miR-219a-5p is involved in the pathogenesis of inflammatory bowel disease (IBD) remains elusive. In this study, we demonstrated that miR-219a-5p expression was significantly decreased in the inflamed intestinal mucosa and peripheral blood (PB)-CD4+ T cells from patients with IBD. Proinflammatory cytokines (e.g., IL-6, IL-12, IL-23 and TNF-α) inhibited miR-219a-5p expression in CD4+ T cells in vitro. Lentivirus-mediated miR-219a-5p downregulation facilitated Th1/Th17 cell differentiation, whereas miR-219a-5p overexpression exerted an opposite effect. Luciferase assays confirmed that ETS variant 5 (ETV5) was a functional target of miR-219a-5p and ETV5 expression was significantly increased in the inflamed intestinal mucosa and PB-CD4+ T cells from IBD patients. ETV5 overexpression enhanced Th1/Th17 immune response through upregulating the phosphorylation of STAT3 and STAT4. Importantly, supplementation of miR-219a-5p ameliorated TNBS-induced intestinal mucosal inflammation, characterized by decreased IFN-γ+ CD4+ T cells and IL-17A+ CD4+ T cells infiltration in the colonic lamina propria. Our data thus reveal a novel mechanism whereby miR-219a-5p suppresses intestinal inflammation through inhibiting Th1/Th17-mediated immune responses. miR-219a-5p might be a target for the treatment of IBD.

9.
Int J Mol Med ; 44(6): 2213-2222, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31661121

RESUMO

Acute lung injury (ALI) is a severe health issue with significant morbidity and mortality. Artemisinin is used for the treatment of fever and malaria in clinical practice. Dihydroartemisinin (DHA), the major active metabolite of artemisinin, plays a role in anti­organizational fibrosis and anti­neuronal cell death. However, whether DHA can attenuate ALI remains unclear. The current study thus examined the effects of DHA on ALI and primary macrophages. The results revealed that DHA attenuated lipopolysaccharide (LPS)­induced pulmonary pathological damage. DHA suppressed the LPS­induced infiltration of inflammatory cells, the elevation of myeloperoxidase activity, oxidative stress and the production of pro­inflammatory cytokines, including interleukin (IL)­1ß, tumor necrosis factor­α, and IL­6. Furthermore, DHA reduced the LPS­induced inflammatory response by suppressing the degradation of I­κB and the nuclear translocation of nuclear factor κ­light­chain­enhancer of activated B cells (NF­κB)/p65 in vivo and in vitro. DHA activated the nuclear factor­erythroid 2 related factor 2 (Nrf2) pathway, which was suppressed by LPS treatment. The Nrf2 inhibitor, ML385, diminished the protective effects of DHA against LPS­induced inflammation in macrophages. On the whole, the findings of this study demonstrate that DHA exerts therapeutic effects against LPS­induced ALI by inhibiting the Nrf2­mediated NF­κB activation in macrophages. The present study also confirmed the therapeutic effects of DHA in mice with LPS­induced ALI. Thus, these findings demonstrate that DHA exhibits anti­inflammatory activities and may be a therapeutic candidate for the treatment of ALI.

10.
Mol Med Rep ; 20(6): 5172-5182, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31661137

RESUMO

Increasing evidence suggests that microRNAs (miRNAs or miRs) serve a critical role in tumor development. However, the role of miRNAs in non­small cell lung cancer (NSCLC) progression remains largely unknown. The present study observed that miR­593 was significantly impaired in patients with NSCLC and was a novel regulator of NSCLC progression. Patients whose tumors had high expression levels of miR­593 had longer overall survival than patients whose tumors had low levels of miR­593 expression (P=0.0219). miR­593 expression levels were inversely correlated with zinc finger protein SNAI2 (SLUG) messenger RNA (mRNA) levels in 87 clinical tissue specimens of NSCLC (P<0.001). A luciferase assay demonstrated that miR­593 interacted with the binding sites present in the SLUG 3'­untranslated region and reduced the expression of SLUG. Introduction of a miR­593 mimic suppressed cell proliferation by inactivating the SLUG/protein kinase B (Akt)/cyclin D1/CDK4 or CDK6 signaling pathway, while it induced apoptosis by activating the SLUG/Akt/Bcl­2/BAX signaling pathway. Furthermore, introduction of a miR­593 mimic recovered the expression of E­cadherin at the protein and mRNA level, and inhibited cell migration and invasion. In conclusion, these results indicated that miR­593 may act as a tumor suppressor in NSCLC to decelerate cancer aggressiveness by inhibiting SLUG expression.

11.
Ann Vasc Surg ; 2019 Oct 18.
Artigo em Inglês | MEDLINE | ID: mdl-31634599

RESUMO

OBJECTIVE: Inferior vena cava (IVC) filters are effective in preventing pulmonary embolism in patients at risk. This study aimed to investigate whether the dwell time of retrievable IVC filters have impact on IVC lumen diameter. METHODS: The clinical data of 36 patients treated with retrievable IVC filters from January 2016 to November 2018 were retrospectively collected. A total of 33 filters were successfully removed. At times of filter placement and removal, the IVC lumen diameter (at upper, middle, and lower levels of the filter), distance between the filter upper end and the right renal vein opening, and degree of filter tilt were measured. RESULTS: IVC filters were placed because of deep vein thrombosis in the lower limbs after fractures in 26 patients. The median dwell time of the IVC filters was 18 days. From the time of filter placement to that of removal, the IVC diameter decreased significantly at the middle (28.07 ± 5.92 vs 25.73 ± 7.33 mm, p = 0.002) and lower levels (27.48 ± 4.73 vs 26.36 ± 4.72 mm, p = 0.003) of the filters. No significant difference was noticed in the IVC diameter at the upper levels of the filters (27.78 ± 6.43 vs 27.11 ± 6.63 mm, p = 0.082). Positive correlation was noticed between filter dwell time and IVC diameter changes at the upper (r = 0.381, p = 0.029) and middle (r = 0.555, p = 0.001) levels of the filters. No significant change was noticed in the distance from the filter upper end to the right renal vein opening and the degree of filter tilt. CONCLUSION: Retrievable IVC filters are associated with IVC stenosis. The severity of IVC stenosis is positively correlated with the dwell time of filters.

12.
Medicine (Baltimore) ; 98(39): e16965, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31574795

RESUMO

This study presents the postoperative pregnancy rate of women with recurrent endometriosis and evaluates the predictive value of the endometriosis fertility index (EFI) for the pregnancy.A total of 107 women who wished to conceive after surgery for recurrent endometriosis from January 2007 to December 2016 were included. The EFI score was calculated postoperatively. The receiver operator characteristic (ROC) curve was plotted to determine the most promising contributor to predicting pregnancy, and Kaplan-Meier (K-M) analysis was used to estimate the cumulative pregnancy rate (CPR).A total of 61 pregnancies were registered in 58 women and the remaining 49 patients failed to become pregnant. The EFI score was strongly associated with the postoperative fertility prognosis. The CPRs during the first 2 and 3 years postoperatively were 51.86% and 66.38%, respectively, and increased to 71.98% within the first 5 years postoperatively in patients with EFI scores ≥5. However, the CPR was 26.00% during the first 2 years after surgery in individuals with EFI scores <5, and there was no increase in the CRP thereafter.Women suffering from recurrent endometriosis still experienced a probability of natural pregnancy, especially patients with EFI scores ≥5. The EFI score had good predictive power for postoperative pregnancy in these patients.


Assuntos
Endometriose/cirurgia , Fertilização , Índice de Massa Corporal , Endometriose/complicações , Feminino , Seguimentos , Humanos , Infertilidade Feminina/diagnóstico , Infertilidade Feminina/etiologia , Infertilidade Feminina/terapia , Estimativa de Kaplan-Meier , Valor Preditivo dos Testes , Gravidez , Taxa de Gravidez , Curva ROC , Recidiva , Técnicas de Reprodução Assistida , Estudos Retrospectivos
13.
Int J Nanomedicine ; 14: 7987-8000, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31632013

RESUMO

Background: The hierarchical porous structure and surface topography of calcium phosphate (CaP) bioceramics have a crucial impact on their osteoinductivity. Purpose: To fabricate a biomimetic bone graft with an interconnected porous structure analogous to that of trabecular bone and a bioactive nanostructured surface with excellent osteoinductive potential. Materials and methods: A biphasic CaP (BCP) substrate with highly porous structure was fabricated by an improved sponge replication method. Surface modification was performed by uniformly depositing a hydroxyapatite (HA) nanoparticle layer to create nHA-coated BCP scaffolds. The effects of these scaffolds on osteogenic differentiation of murine bone marrow-derived stem cells (BMSCs) were investigated in vitro, and their osteoinductivity was further assessed in vivo. Results: The BCP and nHA-coated BCP scaffolds had similar trabecular bone-like architectures but different surface structures, with mean grain sizes of ~55 nm and ~1 µm, respectively. Compared with the BCP substrate, the nHA-coated BCP scaffolds favored cell adhesion and promoted osteogenic differentiation of BMSCs, as evidenced by upregulated expression of osteogenic genes, enhanced alkaline phosphatase activity, and increased osteocalcin production. This could be attributed to activation of the BMP/Smad signaling pathway, as significantly higher expression levels of BMPRI, Smad1, Smad4, and Smad5 were observed in the nHA-coated BCP group. The nHA-coated BCP scaffold not only maintained scaffold integrity but also induced ectopic bone formation when implanted into rabbit dorsal muscle in vivo for 90 days, whereas the BCP substrate underwent marked biodegradation that led to severe inflammation with no sign of osteogenesis. Conclusion: The present study demonstrates the potential of this biomimetic bone graft with a trabecular framework and nanotopography for use in orthopedic applications.

14.
BMC Genomics ; 20(1): 748, 2019 Oct 16.
Artigo em Inglês | MEDLINE | ID: mdl-31619177

RESUMO

BACKGROUND: The homeodomain-leucine zipper (HD-Zip) gene family is one of the plant-specific transcription factor families, involved in plant development, growth, and in the response to diverse stresses. However, comprehensive analysis of the HD-Zip genes, especially those involved in response to drought and salinity stresses is lacking in sesame (Sesamum indicum L.), an important oil crop in tropical and subtropical areas. RESULTS: In this study, 45 HD-Zip genes were identified in sesame, and denominated as SiHDZ01-SiHDZ45. Members of SiHDZ family were classified into four groups (HD-Zip I-IV) based on the phylogenetic relationship of Arabidopsis HD-Zip proteins, which was further supported by the analysis of their conserved motifs and gene structures. Expression analyses of SiHDZ genes based on transcriptome data showed that the expression patterns of these genes were varied in different tissues. Additionally, we showed that at least 75% of the SiHDZ genes were differentially expressed in responses to drought and salinity treatments, and highlighted the important role of HD-Zip I and II genes in stress responses in sesame. CONCLUSIONS: This study provides important information for functional characterization of stress-responsive HD-Zip genes and may contribute to the better understanding of the molecular basis of stress tolerance in sesame.

15.
J Chem Phys ; 151(15): 154201, 2019 Oct 21.
Artigo em Inglês | MEDLINE | ID: mdl-31640389

RESUMO

Lead-halide perovskite nanocrystals (NCs) have emerged as a novel type of semiconductor nanostructure, attracting great research interests in both fundamental science and practical applications. Here, we compare the optical properties of single CsPbI3 NCs under both one-photon and two-photon excitations, mainly including the photoluminescence (PL) blinking and PL decay dynamics. By means of the PL saturation effect caused by multi-exciton Auger recombination, we have also estimated a two-photon absorption cross section of ∼6.8 × 106 GM for single CsPbI3 NCs. The ability to realize efficient two-photon excitation of single perovskite NCs with strongly suppressed background fluorescence will help not only to promote their bio-imaging and biolabeling applications but also to reveal and manipulate their delicate electronic structures for potential usage in quantum information processing.

16.
Cancer Med ; 2019 Oct 23.
Artigo em Inglês | MEDLINE | ID: mdl-31643147

RESUMO

BACKGROUND: Many studies have documented the abnormal concentrations of major/trace elements in serum or malignant tissues of patients, but very few works systematically tested the concentrations of elements in tumor tissues in comparison with paired adjacent normal tissues from the same patients. METHODS: Tumor and adjacent normal lung tissues were obtained from 93 patients with previously untreated NSCLC, and 43 patients whose tumor and paired normal lung tissues reached 200 mg or more were selected for measurement of the elements' concentrations using an inductively coupled plasma-atomic emission spectrometer. RESULTS: We found that the concentrations of the 52 elements varied from 0.4 ng/g tissue (Lu, Pd, and Tm) to 1 658 000 ng/g (Na), 1 951 000 ng/g (P), and 2 495 000 ng/g (K). Thirty eight of the 52 (73.1%) elements showed approximately equal concentrations in tumor and adjacent normal lung tissues of the patients. The concentrations of nine elements (K, P, Mg, Zn, Rb, Cu, Se, Cs, and Tl) in tumor samples were significantly higher than their paired normal lung tissues, and five elements (Na, Fe, Cr, Cd, and Ge) exhibited decreased concentrations in cancer samples compared to counterpart normal lung tissues. Low Fe in tumor samples was associated with smoking history, whereas low Cr was associated with histology (squamous cell carcinoma) of the patients. CONCLUSIONS: Our results demonstrate that measurement of elements' concentrations in both cancer and paired normal tissues is important to get insights into the roles of these elements in carcinogenesis, and therapeutic approaches to normalize the elements are warranted to treat NSCLCs.

17.
J Proteome Res ; 2019 Nov 01.
Artigo em Inglês | MEDLINE | ID: mdl-31618575

RESUMO

Lyme disease results from infection of humans with the spirochete Borrelia burgdorferi. The first and most common clinical manifestation is the circular, inflamed skin lesion referred to as erythema migrans; later manifestations result from infections of other body sites. Laboratory diagnosis of Lyme disease can be challenging in patients with erythema migrans because of the time delay in the development of specific diagnostic antibodies against Borrelia. Reliable blood biomarkers for the early diagnosis of Lyme disease in patients with erythema migrans are needed. Here, we performed selected reaction monitoring, a targeted mass spectrometry-based approach, to measure selected proteins that (1) are known to be predominantly expressed in one organ (i.e., organ-specific blood proteins) and whose blood concentrations may change as a result of Lyme disease, or (2) are involved in acute immune responses. In a longitudinal cohort of 40 Lyme disease patients and 20 healthy controls, we identified 10 proteins with significantly altered serum levels in patients at the time of diagnosis, and we also developed a 10-protein panel identified through multivariate analysis. In an independent cohort of patients with erythema migrans, six of these proteins, APOA4, C9, CRP, CST6, PGLYRP2, and S100A9, were confirmed to show significantly altered serum levels in patients at time of presentation. Nine of the 10 proteins from the multivariate panel were also verified in the second cohort. These proteins, primarily innate immune response proteins or proteins specific to liver, skin, or white blood cells, may serve as candidate blood biomarkers requiring further validation to aid in the laboratory diagnosis of early Lyme disease.

18.
ACS Appl Mater Interfaces ; 11(43): 40916-40922, 2019 Oct 30.
Artigo em Inglês | MEDLINE | ID: mdl-31597426

RESUMO

The paper presents a methodology to control the motion and orientation of suspended reflective cholesteric flakes in a nematic liquid crystal (LC) matrix. The flakes exhibit a dielectric anisotropy which controls their alignment with their in-plane axes parallel to an external electrical dc field. The elastic forces imposed by the LC host affect the switching behavior of the flakes and take care of the realignment to the planar state as soon as the dc field is switched off. When the LC host has a positive dielectric anisotropy, the switching voltage of the flakes is reduced by a factor of 2 in comparison with a LC host with negative dielectric anisotropy or in comparison with an isotropic host. We discovered that the LC host further regulates the back relaxation of cholesteric to return to the planar state upon retrieving the electric field. Whereas, in the isotropic fluid, flakes do not exhibit a preferred orientation when relaxed. Based on this newly proposed principle, we demonstrated its application as an optical switch for smart windows. Depending on the pitch of the cholesteric helix of the flakes, the light of a preset wavelength is reflected. Upon application of an electric field, the embedded flakes rotate their planes perpendicular to the substrate and consequently the incident light becomes fully transmitted without reflection or scattering of light.

19.
FEBS J ; 2019 Oct 24.
Artigo em Inglês | MEDLINE | ID: mdl-31646730

RESUMO

Pulmonary fibrosis (PF) is a senescence-associated disease with poor prognosis. Currently, there is no effective therapeutic strategy for preventing and treating the disease process. Mounting evidence suggests that arachidonic acid (ARA) metabolites are involved in the pathogenesis of various fibrosis. However, the relationship between the metabolism of ARA and PF is still elusive. In this study, we observed a disorder in the cyclooxygenase-2/cytochrome P450 (COX-2/CYP) metabolism of ARA in the lungs of PF mice induced by bleomycin (BLM). Therefore, we aimed to explore the role of COX-2/CYP-derived ARA metabolic disorders in PF. PTUPB, a dual COX-2 and soluble epoxide hydrolase (sEH) inhibitor, was used to restore the balance of COX-2/CYP metabolism. sEH is an enzyme hydrolyzing epoxyeicosatrienoic acids derived from ARA by CYP. We found that PTUPB alleviated the pathological changes in lung tissue and collagen deposition, as well as reduced senescence marker molecules (p16Ink4a and p53-p21Waf1/Cip1 ) in the lungs of mice treated by BLM. In vitro, we found that PTUPB pretreatment remarkably reduced the expression of senescence-related molecules in the alveolar epithelial cells (AECs) induced by BLM. In conclusion, our study supports the notion that the COX-2/CYP-derived ARA metabolic disorders may be a potential therapeutic target for PF via inhibiting the cellular senescence in AECs.

20.
Plant J ; 2019 Sep 08.
Artigo em Inglês | MEDLINE | ID: mdl-31494982

RESUMO

Fluorescence in situ hybridization using probes based on oligonucleotides (oligo-FISH) is a useful tool for chromosome identification and karyotype analysis. Here we developed two oligo-FISH probes that allow the identification of each of the 12 pairs of chromosomes in rice (Oryza sativa). These two probes comprised 25 717 (green) and 25 215 (red) oligos (45 nucleotides), respectively, and generated 26 distinct FISH signals that can be used as a barcode to uniquely label each of the 12 pairs of rice chromosomes. Standard karyotypes of rice were established using this system on both mitotic and meiotic chromosomes. Moreover, dual-color oligo-FISH was used to characterize diverse chromosomal abnormalities. Oligo-FISH analyses using these probes in various wild Oryza species revealed that chromosomes from the AA, BB or CC genomes generated specific and intense signals similar to those in rice, while chromosomes with the EE genome generated less specific signals and the FF genome gave no signal. Together, the oligo-FISH probes we established will be a powerful tool for studying chromosome variations and evolution in the genus Oryza.

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