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1.
Medicine (Baltimore) ; 100(31): e26844, 2021 Aug 06.
Artigo em Inglês | MEDLINE | ID: mdl-34397857

RESUMO

RATIONALE: Primary signet ring cell carcinoma of the uterine cervix is extremely rare and the clinical characteristics and prognosis are not well known and there are no specific guidelines for treatment. PATIENT CONCERNS: A 43-year-old woman was referred to our hospital for abnormal uterine bleeding lasting 1 month. DIAGNOSES: Histological examination revealed a signet ring cell carcinoma of the uterine cervix. After evaluation of extragenital origin, the patient was diagnosed International Federation of Gynecology and Obstetrics stage IIIC1 primary signet ring cell carcinoma or the uterine cervix. INTERVENTION: The patient was prescribed concomitant chemo-radiation followed by intracavitary brachytherapy. OUTCOMES: She showed no evidence of disease after treatment but, it recurred after 7 months of last treatment. LESSONS: Different approaches to diagnosis and treatment of this rare disease are needed and molecular pathological studies related to the onset of the disease are required.


Assuntos
Carcinoma de Células em Anel de Sinete , Colo do Útero , Quimiorradioterapia/métodos , Cisplatino/administração & dosagem , Neoplasias do Colo do Útero , Esfregaço Vaginal/métodos , Adulto , Antineoplásicos/administração & dosagem , Biópsia/métodos , Braquiterapia/métodos , Carcinoma de Células em Anel de Sinete/patologia , Carcinoma de Células em Anel de Sinete/fisiopatologia , Carcinoma de Células em Anel de Sinete/terapia , Colo do Útero/diagnóstico por imagem , Colo do Útero/patologia , Evolução Fatal , Feminino , Humanos , Recidiva Local de Neoplasia/patologia , Papillomaviridae/isolamento & purificação , Retratamento/métodos , Neoplasias do Colo do Útero/patologia , Neoplasias do Colo do Útero/fisiopatologia , Neoplasias do Colo do Útero/terapia , Hemorragia Uterina/diagnóstico , Hemorragia Uterina/etiologia
2.
Reprod Sci ; 28(11): 3137-3146, 2021 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-34106437

RESUMO

The role and mechanisms of progesterone in preterm premature rupture of membranes (PPROM) remains unclear. This study aims to investigate the molecular mechanisms of action of progesterone in pre-labor full-term fetal amniotic membrane cells with and without stimulation by microbial, pro-inflammatory, or thrombogenic agents. Fetal amniotic membranes were collected from 30 women with a normal singleton pregnancy undergoing elective cesarean section at term prior to the onset of labor. The human amniotic epithelial cells isolated were pretreated with and without medroxyprogesterone acetate for 24 h. Then, cells were treated with and without TLR/NLR agonists, pro-inflammatory cytokines, or thrombin for 48 h. Semi-quantitative RT-PCR, Western blot, and caspase-3 activity measurement were performed. Progesterone stimulation decreased the expression of TLR2, TLR5, and Nod2 genes (alone and/or in combination with TLR/NLR agonists) and decreased the expression of IL-1ß and IL-8 genes increased by stimulation with specific agonists for TLR2, TLR4, TLR5, Nod1, and Nod2. Moreover, progesterone decreased thrombin-induced IL-8 gene expression. Progesterone also decreased expression of Bax and Bid proteins (pro-apoptotic factors) increased by stimulation with pro-inflammatory cytokines (TNF-α, NGAL, IL-18, and IL-1ß) and thrombin. Progesterone stimulation alone as well as co-stimulation with TNF-α, NGAL, IL-18, IL-1ß, or thrombin with progesterone either increased, decreased, or did not change the expression of Bcl-2, Bcl-XL, or XIAP genes (anti-apoptotic factors). These data suggest progesterone plays protective roles against PPROM through anti-microbial, anti-inflammatory, and anti-thrombogenic actions on human-term fetal amniotic membrane cells. Progesterone alters pro-inflammatory cytokine- and thrombin-induced apoptosis by controlling the expression of pro-apoptotic and anti-apoptotic factors.

3.
Antibiotics (Basel) ; 10(5)2021 May 10.
Artigo em Inglês | MEDLINE | ID: mdl-34068711

RESUMO

Pseudomonas syringae pv. actinidiae (Psa) is a Gram-negative bacterium that causes bacterial canker disease in kiwifruit. Copper or antibiotics have been used in orchards to control this disease, but the recent emergence of antibiotic-resistant Psa has called for the development of a new control agent. We previously reported that the bacteriophage (or phage) PPPL-1 showed antibacterial activity for both biovar 2 and 3 of Psa. To investigate the possibility of PPPL-1 to control bacterial canker in kiwifruit, we further tested the efficacy of PPPL-1 and its phage cocktail with two other phages on suppressing disease development under greenhouse conditions using 6 weeks old kiwifruit plants. Our results showed that the disease control efficacy of PPPL-1 treatment was statistically similar to those of phage cocktail treatment or AgrimycinTM, which contains streptomycin and oxytetracycline antibiotics as active ingredients. Moreover, PPPL-1 could successfully kill streptomycin-resistant Psa isolates, of which the treatment of BuramycinTM carrying only streptomycin as an active ingredient had no effect in vitro. The phage PPPL-1 was further characterized, and stability assays showed that the phage was stable in the field soil and at low temperature of 0 ± 2 °C. In addition, the phage could be scaled up quickly up to 1010 pfu/mL at 12 h later from initial multiplicity of infection of 0.000005. Our results indicate that PPPL-1 phage is a useful candidate as a biocontrol agent and could be a tool to control the bacterial canker in kiwifruit by Psa infection in the field conditions.

4.
Microorganisms ; 9(4)2021 Apr 08.
Artigo em Inglês | MEDLINE | ID: mdl-33917817

RESUMO

Pectobacterium odoriferum has recently emerged as a widely infective and destructive pathogen causing soft-rot disease in various vegetables. Bacteriophage phiPccP-1 isolated from Pyeongchang, South Korea, showed lytic activity against P. odoriferum Pco14 and two other Pectobacterium species. The transmission electron microscopy and genome phylograms revealed that phiPccP-1 belongs to the Unyawovirus genus, Studiervirinae subfamily of the Autographivirinae family. Genome comparison showed that its 40,487 bp double-stranded DNA genome shares significant similarity with Pectobacterium phage DU_PP_II with the identity reaching 98% of the genome. The phiPccP-1 application significantly inhibited the development of soft-rot disease in the mature leaves of the harvested Kimchi cabbage up to 48 h after Pco14 inoculation compared to the untreated leaves, suggesting that phiPccP-1 can protect Kimchi cabbage from soft-rot disease after harvest. Remarkably, bioassays with phiPccP-1 in Kimchi cabbage seedlings grown in the growth chamber successfully demonstrated its prophylactic and therapeutic potential in the control of bacterial soft-rot disease in Kimchi cabbage. These results indicate that bacteriophage phiPccP-1 can be used as a potential biological agent for controlling soft rot disease in Kimchi cabbage.

5.
Plant Pathol J ; 36(5): 450-458, 2020 Oct 01.
Artigo em Inglês | MEDLINE | ID: mdl-33082729

RESUMO

Pepper and tomato plants infected with two Clavibacter species, C. capsici and C. michiganensis have shown different patterns of disease development depending on their virulence. Here, we investigated how pepper and tomato plants respond to infection by the high-virulent or low-virulent Clavibacter strains. For this, we chose two strains of each Clavibacter species to show different virulence level in the host plants. Although low-virulent strains showed less disease symptoms, they grew almost the same level as the high-virulent strains in both plants. To further examine the response of host plants to Clavibacter infection, we analyzed the expression patterns of plant defense-related genes in the leaves inoculated with different strains of C. capsici and C. michiganensis. Pepper plants infected with high-virulent C. capsici strain highly induced the expression of CaPR1, CaDEF, CaPR4b, CaPR10, and CaLOX1 at 5 days after inoculation (dai), but their expression was much less in low-virulent Clavibacter infection. Expression of CaSAR8.2 was induced at 2 dai, regardless of virulence level. Expression of GluA, Pin2, and PR2 in tomato plants infected with high-virulent C. michiganensis were much higher at 5 dai, compared with mock or low-virulent strain. Expression of PR1a, Osmotin-like, Chitinase, and Chitinase class 2 was increased, regardless of virulence level. Expression of LoxA gene was not affected by Clavibacter inoculation. These results suggested that Clavibacter infection promotes induction of certain defense-related genes in host plants and that differential expression of those genes by low-virulent Clavibacter infection might be affected by their endophytic lifestyle in plants.

6.
Mol Plant Pathol ; 21(6): 808-819, 2020 06.
Artigo em Inglês | MEDLINE | ID: mdl-32196887

RESUMO

The gram-positive bacterial species Clavibacter capsici causes necrosis and canker in pepper plants. Genomic and functional analyses of C. capsici type strain PF008 have shown that multiple virulence genes exist in its two plasmids. We aimed to identify the key determinants that control the virulence of C. capsici. Pepper leaves inoculated with 54 natural isolates exhibited significant variation in the necrosis. Six isolates showed very low virulence, but their population titres in plants were not significantly different from those of the highly virulent isolates. All six isolates lacked the pCM1Cc plasmid that carries chpG, which has been shown to be required for virulence and encodes a putative serine protease, but two of them, isolates 1,106 and 1,207, had the intact chpG elsewhere in the genome. Genomic analysis of these two isolates revealed that chpG was located in the pCM2Cc plasmid, and two highly homologous regions were present next to the chpG locus. The chpG expression in isolate 1,106 was not induced in plants. Introduction of chpG of the PF008 strain into the six low-virulence isolates restored their virulence to that of PF008. Our findings indicate that there are at least three different variant groups of C. capsici and that the plasmid composition and the chpG gene are critical for determining the virulence level. Moreover, our findings also indicate that the virulence level of C. capsici does not directly correlate with bacterial titres in plants.


Assuntos
Capsicum/microbiologia , Clavibacter/patogenicidade , Doenças das Plantas/microbiologia , Plasmídeos/genética , Fatores de Virulência/genética , Clavibacter/genética , Loci Gênicos , Folhas de Planta/microbiologia , Virulência/genética
7.
Plant Pathol J ; 36(1): 29-42, 2020 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-32089659

RESUMO

Acidovorax citrulli causes bacterial fruit blotch in Cucurbitaceae, including watermelon. Although A. citrulli is a seed-borne pathogen, it can cause diverse symptoms in other plant organs like leaves, stems and fruits. To determine the infection routes of A. citrulli, we examined the virulence of six isolates (Ac0, Ac1, Ac2, Ac4, Ac8, and Ac11) on watermelon using several inoculation methods. Among six isolates, DNA polymorphism reveals that three isolates Ac0, Ac1, and Ac4 belong to Clonal Complex (CC) group II and the others do CC group I. Ac0, Ac4, and Ac8 isolates efficiently infected seeds during germination in soil, and Ac0 and Ac4 also infected the roots of watermelon seedlings wounded prior to inoculation. Infection through leaves was successful only by three isolates belonging to CC group II, and two of these also infected the mature watermelon fruits. Ac2 did not cause the disease in all assays. Interestingly, three putative type III effectors (Aave_2166, Aave_2708, and Aave_3062) with intact forms were only found in CC group II. Overall, our results indicate that A. citrulli can infect watermelons through diverse routes, and the CC grouping of A. citrulli was only correlated with virulence in leaf infection assays.

8.
Mol Plant Microbe Interact ; 32(4): 491-501, 2019 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-30345870

RESUMO

Diverse plant pathogens secrete cellulases to degrade plant cell walls. Previously, the plasmid-borne cellulase gene celA was shown to be important for the virulence of the gram-positive bacterium Clavibacter michiganensis in tomato. However, details of the contribution of cellulases to the development of wilting in tomato have not been well-determined. To better understand the contribution of cellulases to the virulence of C. michiganensis in tomato, a mutant lacking cellulase activity was generated and complemented with truncated forms of certain cellulase genes, and virulence of those strain was examined. A celA mutant of the C. michiganensis type strain LMG7333 lost its cellulase activity and almost all its ability to cause wilting in tomato. The cellulase catalytic domain and cellulose-binding domain of CelA together were sufficient for both cellulase activity and the development of wilting in tomato. However, the expansin domain did not affect virulence or cellulase activity. The celA ortholog of Clavibacter sepedonicus restored the full virulence of the celA mutant of C. michiganensis. Another cellulase gene, celB, located in the chromosome, carries a single-base deletion in most C. michiganensis strains but does not carry a functional signal peptide in its N terminus. Nevertheless, an experimentally modified CelB protein with a CelA signal peptide was secreted and able to cause wilting in tomato. These results indicate that cellulases are major virulence factors of C. michiganensis that causes wilting in tomato. Furthermore, there are natural variations among cellulase genes directly affecting their function.


Assuntos
Actinobacteria , Celulase , Lycopersicon esculentum , Doenças das Plantas , Actinobacteria/enzimologia , Actinobacteria/genética , Celulase/genética , Celulase/metabolismo , Lycopersicon esculentum/microbiologia , Doenças das Plantas/microbiologia , Sinais Direcionadores de Proteínas/genética , Sinais Direcionadores de Proteínas/fisiologia , Virulência/genética
9.
Theor Appl Genet ; 131(5): 1017-1030, 2018 May.
Artigo em Inglês | MEDLINE | ID: mdl-29352323

RESUMO

KEY MESSAGE: Genotyping of disease resistance to bacterial wilt in tomato by a genome-wide SNP analysis Bacterial wilt caused by Ralstonia pseudosolanacearum is one of the destructive diseases in tomato. The previous studies have identified Bwr-6 (chromosome 6) and Bwr-12 (chromosome 12) loci as the major quantitative trait loci (QTLs) contributing to resistance against bacterial wilt in tomato cultivar 'Hawaii7996'. However, the genetic identities of two QTLs have not been uncovered yet. In this study, using whole-genome resequencing, we analyzed genome-wide single-nucleotide polymorphisms (SNPs) that can distinguish a resistant group, including seven tomato varieties resistant to bacterial wilt, from a susceptible group, including two susceptible to the same disease. In total, 5259 non-synonymous SNPs were found between the two groups. Among them, only 265 SNPs were located in the coding DNA sequences, and the majority of these SNPs were located on chromosomes 6 and 12. The genes that both carry SNP(s) and are near Bwr-6 and Bwr-12 were selected. In particular, four genes in chromosome 12 encode putative leucine-rich repeat (LRR) receptor-like proteins. SNPs within these four genes were used to develop SNP markers, and each SNP marker was validated by a high-resolution melting method. Consequently, one SNP marker, including a functional SNP in a gene, Solyc12g009690.1, could efficiently distinguish tomato varieties resistant to bacterial wilt from susceptible varieties. These results indicate that Solyc12g009690.1, the gene encoding a putative LRR receptor-like protein, might be tightly linked to Bwr-12, and the SNP marker developed in this study will be useful for selection of tomato cultivars resistant to bacterial wilt.


Assuntos
Resistência à Doença/genética , Lycopersicon esculentum/genética , Doenças das Plantas/genética , Locos de Características Quantitativas , Mapeamento Cromossômico , Genes de Plantas , Lycopersicon esculentum/microbiologia , Fenótipo , Doenças das Plantas/microbiologia , Polimorfismo de Nucleotídeo Único , Ralstonia
10.
New Phytol ; 217(3): 1177-1189, 2018 02.
Artigo em Inglês | MEDLINE | ID: mdl-29134663

RESUMO

Clavibacter michiganensis ssp. capsici is a Gram-positive plant-pathogenic bacterium causing bacterial canker disease in pepper. Virulence genes and mechanisms of C. michiganensis ssp. capsici in pepper have not yet been studied. To identify virulence genes of C. michiganensis ssp. capsici, comparative genome analyses with C. michiganensis ssp. capsici and its related C. michiganensis subspecies, and functional analysis of its putative virulence genes during infection were performed. The C. michiganensis ssp. capsici type strain PF008 carries one chromosome (3.056 Mb) and two plasmids (39 kb pCM1Cmc and 145 kb pCM2Cmc ). The genome analyses showed that this bacterium lacks a chromosomal pathogenicity island and celA gene that are important for disease development by C. michiganensis ssp. michiganensis in tomato, but carries most putative virulence genes in both plasmids. Virulence of pCM1Cmc -cured C. michiganensis ssp. capsici was greatly reduced compared with the wild-type strain in pepper. The complementation analysis with pCM1Cmc -located putative virulence genes showed that at least five genes, chpE, chpG, ppaA1, ppaB1 and pelA1, encoding serine proteases or pectate lyase contribute to disease development in pepper. In conclusion, C. michiganensis ssp. capsici has a unique genome structure, and its multiple plasmid-borne genes play critical roles in virulence in pepper, either separately or together.


Assuntos
Capsicum/microbiologia , Genes Bacterianos , Micrococcaceae/genética , Micrococcaceae/patogenicidade , Doenças das Plantas/microbiologia , Plasmídeos/genética , Perfilação da Expressão Gênica , Regulação Bacteriana da Expressão Gênica , Micrococcaceae/crescimento & desenvolvimento , Mapeamento Físico do Cromossomo , Doenças das Plantas/genética , Virulência/genética
11.
Planta ; 246(3): 351-364, 2017 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-28508261

RESUMO

MAIN CONCLUSION: Pepper leucine-rich repeat protein (CaLRR1) interacts with defense response proteins to regulate plant cell death and immunity. This review highlights the current understanding of the molecular functions of CaLRR1 and its interactor proteins. Plant cell death and immune responses to microbial pathogens are controlled by complex and tightly regulated molecular signaling networks. Xanthomonas campestris pv. vesicatoria (Xcv)-inducible pepper (Capsicum annuum) leucine-rich repeat protein 1 (CaLRR1) serves as a molecular marker for plant cell death and immunity signaling. In this review, we discuss recent advances in elucidating the functional roles of CaLRR1 and its interacting plant proteins, and understanding how they are involved in the cell death and defense responses. CaLRR1 physically interacts with pepper pathogenesis-related proteins (CaPR10 and CaPR4b) and hypersensitive-induced reaction protein (CaHIR1) to regulate plant cell death and defense responses. CaLRR1 is produced in the cytoplasm and trafficked to the extracellular matrix. CaLRR1 binds to CaPR10 in the cytoplasm and CaPR4b and CaHIR1 at the plasma membrane. CaLRR1 synergistically accelerates CaPR10-triggered hypersensitive cell death, but negatively regulates CaPR4b- and CaHIR1-triggered cell death. CaHIR1 interacts with Xcv filamentous hemagglutinin (Fha1) to trigger disease-associated cell death. The subcellular localization and cellular function of these CaLRR1 interactors during plant cell death and defense responses were elucidated by Agrobacterium-mediated transient expression, virus-induced gene silencing, and transgenic overexpression studies. CaPR10, CaPR4b, and CaHIR1 positively regulate defense signaling mediated by salicylic acid and reactive oxygen species, thereby activating hypersensitive cell death and disease resistance. A comprehensive understanding of the molecular functions of CaLRR1 and its interacting protein partners in cell death and defense responses will provide valuable information for the molecular genetics of plant disease resistance, which could be exploited as a sustainable disease management strategy.


Assuntos
Capsicum/genética , Morte Celular/genética , Imunidade Vegetal/genética , Proteínas de Plantas/genética , Proteínas/genética , Capsicum/metabolismo , Doenças das Plantas , Proteínas de Plantas/fisiologia , Proteínas/fisiologia
12.
Plant Pathol J ; 33(2): 163-169, 2017 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-28381963

RESUMO

SlMAPKKKα, a tomato (Solanum lycopersicum) mitogen-activated protein kinase kinase kinase, is a positive regulator of Pto-mediated effector-triggered immunity, which elicits programmed cell death (PCD) in plants. In this study, we examined whether putative phosphorylation sites in the conserved activation segment of the SlMAPKKKα kinase domain are critical for eliciting PCD. Three amino acids, threonine353, serine360 (Ser360), or serine364 (Ser364), in the conserved activation segment of SlMAPKKKα kinase domain were substituted to alanine (T353A, S360A, or S364A), and these variants were transiently expressed in tomato and Nicotiana benthamiana plants. Two alanine substitutions, S360A and S364A, completely abolished SlMAPKKKα PCD-eliciting activity in both plants, while T353A substitution did not affect its PCD-eliciting activity. SlMAPKKKα wild type and variant proteins accumulated to similar levels in plant leaves. However, SlMAPKKKα protein with the largest size was missed when either S360A or S364A substitutions were expressed, whereas proteins with the smaller masses were more accumulated than those of full-length of SIMAPKKKα and T353A. These results suggest that phosphorylation of SlMAPKKKα at Ser360 and Ser364 is critical for PCD elicitation in plants.

13.
Int J Syst Evol Microbiol ; 66(10): 4065-4070, 2016 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-27412674

RESUMO

Clavibacter michiganensis is a Gram-stain-positive bacterium with eight subspecies. One of these subspecies is C. michiganensis subsp. michiganensis, which causes bacterial canker disease in tomato. Bacterial strains showing very similar canker disease symptoms to those of a strain originally classified as C. michiganensis have been isolated from pepper. In this paper, we reclassified strains isolated from pepper. On the basis of phylogenetic analysis with 16S rRNA gene sequences, the strains isolated from pepper were grouped in a separate clade from other subspecies of C. michiganensis. Biochemical, physiological and genetic characteristics of strain PF008T, which is the representative strain of the isolates from pepper, were examined in this study. Based on multi-locus sequence typing and other biochemical and physiological features including colony color, utilization of carbon sources and enzyme activities, strain PF008T was categorically differentiated from eight subspecies of C. michiganensis. Moreover, genome analysis showed that the DNA G+C content of strain PF008T is 73.2 %. These results indicate that PF008T is distinct from other known subspecies of C. michiganensis. Therefore, we propose a novel subspecies, C. michiganensis subsp. capsici, causing bacterial canker disease in pepper, with a type strain of PF008T (=KACC 18448T=LMG 29047T).


Assuntos
Capsicum/microbiologia , Micrococcaceae/classificação , Filogenia , Doenças das Plantas/microbiologia , Técnicas de Tipagem Bacteriana , Composição de Bases , DNA Bacteriano/genética , Micrococcaceae/genética , Micrococcaceae/isolamento & purificação , Tipagem de Sequências Multilocus , RNA Ribossômico 16S/genética , República da Coreia , Análise de Sequência de DNA
14.
Plant Pathol J ; 32(3): 173-81, 2016 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-27298592

RESUMO

Gene disruption by homologous recombination is widely used to investigate and analyze the function of genes in Fusarium fujikuroi, a fungus that causes bakanae disease and root rot symptoms in rice. To generate gene deletion constructs, the use of conventional cloning methods, which rely on restriction enzymes and ligases, has had limited success due to a lack of unique restriction enzyme sites. Although strategies that avoid the use of restriction enzymes have been employed to overcome this issue, these methods require complicated PCR steps or are frequently inefficient. Here, we introduce a cloning system that utilizes multi-fragment assembly by In-Fusion to generate a gene disruption construct. This method utilizes DNA fragment fusion and requires only one PCR step and one reaction for construction. Using this strategy, a gene disruption construct for Fusarium cyclin C1 (FCC1 ), which is associated with fumonisin B1 biosynthesis, was successfully created and used for fungal transformation. In vivo and in vitro experiments using confirmed fcc1 mutants suggest that fumonisin production is closely related to disease symptoms exhibited by F. fujikuroi strain B14. Taken together, this multi-fragment assembly method represents a simpler and a more convenient process for targeted gene disruption in fungi.

15.
Artigo em Inglês | MEDLINE | ID: mdl-25263347

RESUMO

A number of 188 food and dietary supplement samples were collected from 2009 to the first half of 2013 in Korean online and offline stores. A method to identify phosphodiesterase-5 (PDE-5) inhibitors and their analogues using liquid chromatography-electrospray ionisation-mass spectrometry/mass spectrometry (LC-ESI-MS/MS) was validated. Limit of detection and limit of quantitation of liquid-type and solid-type negative samples ranged from 0.05 to 3.33 ng/mL or ng/g and from 0.15 to 10.00 ng/mL or ng/g, respectively. Recoveries ranged from 83% to 112%. Nineteen PDE-5 inhibitors and their analogues were detected, with tadalafil group compounds being the most frequently observed (53.0%), followed by the sildenafil group (42.5%). Tadalafil concentrations ranged from 0.08 to 138.69 mg/g. Compounds were most frequently detected in capsules (in 40 of 80 adulterated samples). To protect public health and food safety, appropriate monitoring of PDE-5 inhibitors and their analogues in foods and dietary supplements is recommended.


Assuntos
Cromatografia Líquida/métodos , Suplementos Nutricionais/análise , Análise de Alimentos/métodos , Inibidores da Fosfodiesterase 5/química , Espectrometria de Massas por Ionização por Electrospray/métodos , Espectrometria de Massas em Tandem/métodos , Contaminação de Alimentos , Humanos , Limite de Detecção , República da Coreia
16.
J Forensic Sci ; 61(1): 226-9, 2016 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-26346959

RESUMO

The objective of this study was to determine the presence of corticosteroids in illegal herbal medicines using ultra-high-performance liquid chromatography-tandem mass spectrometry. We collected 212 herbal medicine samples that were advertised as being effective for treatment of joint pain and bone aches. Samples were from the Korean commercial market during a span of four years (2010-2013), and the method was validated. The limits of quantification ranged from 0.47 to 15.0 ng/mL, and recoveries ranged from 80.6% to 119.5%. The intra- and interday precision ranged from 0.18% to 8.82% and from 0.09% to 8.96%, respectively. Among the samples, three samples (1.4%) were identified as adulterants. Dexamethasone was the only compound detected in the adulterated products. As the corticosteroid-adulteration of herbal medicines may become a major problem and lead to side effects, the continued development of screening procedures for herbal medicines is critical.


Assuntos
Contaminação de Medicamentos , Glucocorticoides/análise , Medicina Tradicional Coreana , Extratos Vegetais/química , Cromatografia Líquida de Alta Pressão , Dexametasona/análise , Humanos , Espectrometria de Massas , República da Coreia
17.
Am J Vet Res ; 76(9): 814-21, 2015 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-26309110

RESUMO

OBJECTIVE: To compare the characteristics and multipotential and in vivo cartilage formation capabilities of porcine adipose-derived stem cells (pASCs) with those of porcine skin-derived stem cell-like cells (pSSCs). ANIMALS: Three 6-month-old female pigs and four 6-week-old female athymic mice. PROCEDURES: Adipose and skin tissue specimens were obtained from each pig following slaughter and digested to obtain pASCs and pSSCs. For each cell type, flow cytometry and reverse transcription PCR assays were performed to characterize the expression of cell surface and mesenchymal stem cell markers, and in vitro cell cultures were performed to determine the adipogenic, osteogenic, and chondrogenic capabilities. Each cell type was then implanted into athymic mice to determine the extent of in vivo cartilage formation after 6 weeks. RESULTS: The cell surface and mesenchymal stem cell marker expression patterns, multipotential capability, and extent of in vivo cartilage formation did not differ significantly between pASCs and pSSCs. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggested that pSSCs may be a viable alternative to pASCs as a source of progenitor cells for tissue engineering in regenerative medicine.


Assuntos
Cartilagem/citologia , Diferenciação Celular/fisiologia , Condrogênese/fisiologia , Células-Tronco/fisiologia , Suínos , Tecido Adiposo/citologia , Animais , Células Cultivadas , Feminino , Citometria de Fluxo , Regulação da Expressão Gênica/fisiologia , Células-Tronco Mesenquimais/citologia , Camundongos , Células-Tronco/citologia , Engenharia Tecidual
18.
Plant Cell Physiol ; 56(5): 930-42, 2015 May.
Artigo em Inglês | MEDLINE | ID: mdl-25657344

RESUMO

In plants, lipoxygenases (LOXs) are involved in various physiological processes, including defense responses to biotic and abiotic stresses. Our previous study had shown that the pepper 9-LOX gene, CaLOX1, plays a crucial role in cell death due to pathogen infection. Here, the function of CaLOX1 in response to osmotic, drought and high salinity stress was examined using CaLOX1-overexpressing (CaLOX1-OX) Arabidopsis plants. Changes in the temporal expression pattern of the CaLOX1 gene were observed when pepper leaves were treated with drought and high salinity, but not when treated with ABA, the primary hormone in response to drought stress. During seed germination and seedling development, CaLOX1-OX plants were more tolerant to ABA, mannitol and high salinity than wild-type plants. In contrast, expression of the ABA-responsive marker genes RAB18 and RD29B was higher in CaLOX1-OX Arabidopsis plants than in wild-type plants. In response to high salinity, CaLOX1-OX plants exhibited enhanced tolerance, compared with the wild type, which was accompanied by decreased accumulation of H2O2 and high levels of RD20, RD29A, RD29B and P5CS gene expression. Similarly, CaLOX1-OX plants were also more tolerant than wild-type plants to severe drought stress. H2O2 production and the relative increase in lipid peroxidation were lower, and the expression of COR15A, DREB2A, RD20, RD29A and RD29B was higher in CaLOX1-OX plants, relative to wild-type plants. Taken together, our results indicate that CaLOX1 plays a crucial role in plant stress responses by modulating the expression of ABA- and stress-responsive marker genes, lipid peroxidation and H2O2 production.


Assuntos
Capsicum/enzimologia , Capsicum/fisiologia , Secas , Lipoxigenase/metabolismo , Pressão Osmótica , Salinidade , Estresse Fisiológico , Ácido Abscísico/farmacologia , Adaptação Fisiológica/efeitos dos fármacos , Adaptação Fisiológica/genética , Arabidopsis/efeitos dos fármacos , Arabidopsis/genética , Arabidopsis/fisiologia , Capsicum/efeitos dos fármacos , Capsicum/genética , Regulação da Expressão Gênica de Plantas , Germinação/efeitos dos fármacos , Folhas de Planta/efeitos dos fármacos , Folhas de Planta/genética , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Plantas Geneticamente Modificadas , Tolerância ao Sal/efeitos dos fármacos , Tolerância ao Sal/genética , Sementes/efeitos dos fármacos , Sementes/crescimento & desenvolvimento , Cloreto de Sódio/farmacologia , Estresse Fisiológico/efeitos dos fármacos , Estresse Fisiológico/genética
19.
Reprod Fertil Dev ; 27(7): 1088-96, 2015 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-24718263

RESUMO

We tried to prevent the mitochondrial and DNA damage caused by mechanical stress-associated reactive oxygen species (ROS), and to improve the reprogramming of bovine somatic cell nuclear transfer (SCNT) embryos by antioxidant treatment during the manipulation procedures of SCNT. Bovine recipient oocytes and reconstituted oocytes were treated with antioxidants during manipulation procedures. The H2O2 level, mitochondrial morphology, membrane potential and apoptosis at the one-cell stage, and in vitro development and DNA methylation status of blastocysts were evaluated. Antioxidant treatment during manipulation procedures reduced the H2O2 level of SCNT embryos. Antioxidant-treated SCNT embryos normally formed mitochondrial clumps, similar to IVF embryos, and showed higher mitochondrial membrane potential versus the SCNT control (P<0.05). Apoptosis and DNA fragmentation were reduced by antioxidant treatment. The development rate to the blastocyst stage was higher (P<0.05) in the antioxidant treatment groups (30.5±2.5 to 30.6±1.6%) versus the control (23.0±1.9%). The DNA methylation status of blastocysts in the antioxidant treatment groups was lower (P<0.05) than that of the control and similar to that of IVF embryos. These results indicate that antioxidant treatment during manipulation procedures can prevent cellular damage that may be caused by mechanical stress-associated ROS, and improve nuclear reprogramming.


Assuntos
Antioxidantes/farmacologia , Ácido Ascórbico/farmacologia , Blastocisto/efeitos dos fármacos , Reprogramação Celular/efeitos dos fármacos , Dano ao DNA/efeitos dos fármacos , Mercaptoetanol/farmacologia , Mitocôndrias/efeitos dos fármacos , Animais , Apoptose/efeitos dos fármacos , Blastocisto/metabolismo , Caspase 3/metabolismo , Bovinos , Fragmentação do DNA/efeitos dos fármacos , Metilação de DNA/efeitos dos fármacos , Feminino , Peróxido de Hidrogênio/metabolismo , Potencial da Membrana Mitocondrial/efeitos dos fármacos , Mitocôndrias/metabolismo , Técnicas de Transferência Nuclear , Espécies Reativas de Oxigênio/metabolismo
20.
Plant J ; 77(4): 521-33, 2014 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-24304389

RESUMO

To control defense and cell-death signaling, plants contain an abundance of pathogen recognition receptors such as leucine-rich repeat (LRR) proteins. Here we show that pepper (Capsicum annuum) LRR1 interacts with the pepper pathogenesis-related (PR) protein 4b, PR4b, in yeast and in planta. PR4b is synthesized in the endoplasmic reticulum, interacts with LRR1 in the plasma membrane, and is secreted to the apoplast via the plasma membrane. Binding of PR4b to LRR1 requires the chitin-binding domain of PR4b. Purified PR4b protein inhibits spore germination and mycelial growth of plant fungal pathogens. Transient expression of PR4b triggers hypersensitive cell death. This cell death is compromised by co-expression of LRR1 as a negative regulator in Nicotiana benthamiana leaves. LRR1/PR4b silencing in pepper and PR4b over-expression in Arabidopsis thaliana demonstrated that LRR1 and PR4b are necessary for defense responses to Pseudomonas syringae pv. tomato and Hyaloperonospora arabidopsidis (Hpa) infection. The mutant of the PR4b Arabidopsis ortholog, pr4, showed enhanced susceptibility to Hpa infection. Together, our results suggest that PR4b functions as a positive modulator of plant cell death and defense responses. However, the activity of PR4b is suppressed by interaction with LRR1.


Assuntos
Capsicum/fisiologia , Regulação da Expressão Gênica de Plantas , Doenças das Plantas/imunologia , Proteínas de Plantas/metabolismo , Proteínas/metabolismo , Arabidopsis/citologia , Arabidopsis/genética , Arabidopsis/imunologia , Arabidopsis/fisiologia , Capsicum/citologia , Capsicum/genética , Capsicum/imunologia , Morte Celular , Membrana Celular/metabolismo , Resistência à Doença , Interações Hospedeiro-Patógeno , Peróxido de Hidrogênio/metabolismo , Mutação , Óxido Nítrico/metabolismo , Oomicetos/patogenicidade , Oomicetos/fisiologia , Doenças das Plantas/microbiologia , Folhas de Planta/citologia , Folhas de Planta/genética , Folhas de Planta/imunologia , Folhas de Planta/fisiologia , Proteínas de Plantas/genética , Plantas Geneticamente Modificadas , Transporte Proteico , Proteínas/genética , Pseudomonas syringae/patogenicidade , Transdução de Sinais , Tabaco/citologia , Tabaco/genética , Tabaco/imunologia , Tabaco/fisiologia , Xanthomonas campestris/patogenicidade
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