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1.
Antimicrob Agents Chemother ; : e0034124, 2024 May 14.
Artigo em Inglês | MEDLINE | ID: mdl-38742905

RESUMO

Cell culture-based screening of a chemical library identified diphenoxylate as an antiviral agent against severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). The observed 50% effective concentrations ranged between 1.4 and 4.9 µM against the original wild-type strain and its variants. Time-of-addition experiments indicated that diphenoxylate is an entry blocker targeting a host factor involved in viral infection. Fluorescence microscopic analysis visualized that diphenoxylate prevented SARS-CoV-2 particles from penetrating the cell membrane and also impaired endo-lysosomal acidification. Diphenoxylate exhibited a synergistic inhibitory effect on SARS-CoV-2 infection in human lung epithelial Calu-3 cells when combined with a transmembrane serine protease 2 (TMPRSS2) inhibitor, nafamostat. This synergy suggested that efficient antiviral activity is achieved by blocking both TMPRSS2-mediated early and endosome-mediated late SARS-CoV-2 entry pathways. The antiviral efficacy of diphenoxylate against SARS-CoV-2 was reproducible in a human tonsil organoids system. In a transgenic mouse model expressing the obligate SARS-CoV-2 receptor, human angiotensin-converting enzyme 2, intranasal administration of diphenoxylate (10 mg/kg/day) significantly reduced the viral RNA copy number in the lungs by 70% on day 3. This study underscores that diphenoxylate represents a promising core scaffold, warranting further exploration for chemical modifications aimed at developing a new class of clinically effective antiviral drugs against SARS-CoV-2.

2.
J Org Chem ; 87(11): 7574-7580, 2022 06 03.
Artigo em Inglês | MEDLINE | ID: mdl-35549260

RESUMO

Pd-Catalyzed intramolecular allylic C-H amination of 1,1-disubstituted alkenyl amines with various allylic tethers (X = O, NMs, CH2) was developed. This process allows for the divergent synthesis of 1,3-X,N-heterocycles through a regioselective allylic C-H cleavage and π-allylpalladium formation. Particularly noteworthy is the use of substrates containing a labile allylic moiety and new simple catalytic systems capable of promoting highly chemo- and regioselective allylic C-H amination by overcoming significant challenges.


Assuntos
Aminas , Paládio , Aminação , Aminas/química , Catálise , Paládio/química
3.
Neth Heart J ; 30(12): 541-545, 2022 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-35360895

RESUMO

Due to population ageing, the number of older and frail patients with cardiovascular disease is increasing. In the current guidelines of the European Society of Cardiology specific recommendations for this older population are missing or scarce, probably due to limited evidence concerning diagnosis and treatment of cardiovascular disease in older patients. Moreover, recommendations on shared decision making, palliative care and advanced care planning are also essential in these guidelines. In this article we evaluate the current European of Society of Cardiology guidelines (2013-2020) to determine whether specific recommendations for older patients have been included.

4.
Age Ageing ; 50(6): 2105-2115, 2021 11 10.
Artigo em Inglês | MEDLINE | ID: mdl-34304264

RESUMO

BACKGROUND: after hospitalisation for cardiac disease, older patients are at high risk of readmission and death. OBJECTIVE: the cardiac care bridge (CCB) transitional care programme evaluated the impact of combining case management, disease management and home-based cardiac rehabilitation (CR) on hospital readmission and mortality. DESIGN: single-blind, randomised clinical trial. SETTING: the trial was conducted in six hospitals in the Netherlands between June 2017 and March 2020. Community-based nurses and physical therapists continued care post-discharge. SUBJECTS: cardiac patients ≥ 70 years were eligible if they were at high risk of functional loss or if they had had an unplanned hospital admission in the previous 6 months. METHODS: the intervention group received a comprehensive geriatric assessment-based integrated care plan, a face-to-face handover with the community nurse before discharge and follow-up home visits. The community nurse collaborated with a pharmacist and participants received home-based CR from a physical therapist. The primary composite outcome was first all-cause unplanned readmission or mortality at 6 months. RESULTS: in total, 306 participants were included. Mean age was 82.4 (standard deviation 6.3), 58% had heart failure and 92% were acutely hospitalised. 67% of the intervention key-elements were delivered. The composite outcome incidence was 54.2% (83/153) in the intervention group and 47.7% (73/153) in the control group (risk differences 6.5% [95% confidence intervals, CI -4.7 to 18%], risk ratios 1.14 [95% CI 0.91-1.42], P = 0.253). The study was discontinued prematurely due to implementation activities in usual care. CONCLUSION: in high-risk older cardiac patients, the CCB programme did not reduce hospital readmission or mortality within 6 months. TRIAL REGISTRATION: Netherlands Trial Register 6,316, https://www.trialregister.nl/trial/6169.


Assuntos
Cuidado Transicional , Assistência ao Convalescente , Idoso , Idoso de 80 Anos ou mais , Humanos , Alta do Paciente , Readmissão do Paciente , Método Simples-Cego
5.
J Appl Clin Med Phys ; 22(1): 184-190, 2021 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-33340391

RESUMO

PURPOSE: The purpose of this study was to develop automated planning for whole-brain radiation therapy (WBRT) using a U-net-based deep-learning model for predicting the multileaf collimator (MLC) shape bypassing the contouring processes. METHODS: A dataset of 55 cases, including 40 training sets, five validation sets, and 10 test sets, was used to predict the static MLC shape. The digitally reconstructed radiograph (DRR) reconstructed from planning CT images as an input layer and the MLC shape as an output layer are connected one-to-one via the U-net modeling. The Dice similarity coefficient (DSC) was used as the loss function in the training and ninefold cross-validation. Dose-volume-histogram (DVH) curves were constructed for assessing the automatic MLC shaping performance. Deep-learning (DL) and manually optimized (MO) approaches were compared based on the DVH curves and dose distributions. RESULTS: The ninefold cross-validation ensemble test results were consistent with DSC values of 94.6 ± 0.4 and 94.7 ± 0.9 in training and validation learnings, respectively. The dose coverages of 95% target volume were (98.0 ± 0.7)% and (98.3 ± 0.8)%, and the maximum doses for the lens as critical organ-at-risk were 2.9 Gy and 3.9 Gy for DL and MO, respectively. The DL technique shows the consistent results in terms of the DVH parameter except for MLC shaping prediction for dose saving of small organs such as lens. CONCLUSIONS: Comparable with the MO plan result, the WBRT plan quality obtained using the DL approach is clinically acceptable. Moreover, the DL approach enables WBRT auto-planning without the time-consuming manual MLC shaping and target contouring.


Assuntos
Neoplasias Encefálicas , Aprendizado Profundo , Encéfalo/diagnóstico por imagem , Neoplasias Encefálicas/diagnóstico por imagem , Neoplasias Encefálicas/radioterapia , Irradiação Craniana , Estudos de Viabilidade , Humanos , Planejamento da Radioterapia Assistida por Computador
6.
Pract Neurol ; 20(6): 486-488, 2020 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-32554439

RESUMO

Acute intermittent porphyria is a rare genetic condition in which disrupted haem synthesis causes overproduction of porphyrin precursors. Occasionally, it is associated with posterior reversible encephalopathy syndrome (PRES), presenting with headache, confusion, seizures and visual disturbance. We describe a patient with acute intermittent porphyria who presented with seizures and PRES, and who had previous unexplained severe abdominal pain. Acute intermittent porphyria should be considered as a possible cause of PRES, especially in those with unexplained abdominal pain, since delays in its diagnosis can result in permanent complications.


Assuntos
Porfiria Aguda Intermitente , Síndrome da Leucoencefalopatia Posterior , Dor Abdominal/etiologia , Humanos , Imageamento por Ressonância Magnética , Porfiria Aguda Intermitente/complicações , Porfiria Aguda Intermitente/diagnóstico , Síndrome da Leucoencefalopatia Posterior/complicações , Síndrome da Leucoencefalopatia Posterior/diagnóstico por imagem , Convulsões
7.
Microbiology (Reading) ; 163(3): 410-420, 2017 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-28100297

RESUMO

Microbotryum lychnidis-dioicae is an obligate biotrophic parasite of the wildflower species Silene latifolia. This dikaryotic fungus, commonly known as an anther smut, requires that haploid, yeast-like sporidia of opposite mating types fuse and differentiate into dikaryotic hyphae that penetrate host tissue as part of the fungal life cycle. Mating occurs under conditions of cool temperatures and limited nutrients. Further development requires host cues or chemical mimics, including a variety of lipids, e.g. phytols. To identify global changes in transcription associated with developmental shifts, RNA-Seq was conducted at several in vitro stages of fungal propagation, i.e. haploid cells grown independently on rich and nutrient-limited media, mated cells on nutrient-limited media as well as a time course of such mated cells exposed to phytol. Comparison of haploid cells grown under rich and nutrient-limited conditions identified classes of genes probably associated with general nutrient availability, including components of the RNAi machinery. Some gene enrichment patterns comparing the nutrient-limited and mated transcriptomes suggested gene expression changes associated with the mating programme (e.g. homeodomain binding proteins, secreted proteins, proteins unique to M. lychnidis-dioicae¸ multicopper oxidases and RhoGEFs). Analysis for phytol treatment compared with mated cells alone allowed identification of genes likely to be involved in the dikaryotic switch (e.g. oligopeptide transporters). Gene categories of particular note in all three conditions included those in the major facilitator superfamily, proteins containing PFAM domains of the secretory lipase family as well as proteins predicted to be secreted, many of which have the hallmarks of fungal effectors with potential roles in pathogenicity.

8.
Int J Mol Sci ; 18(11)2017 Nov 22.
Artigo em Inglês | MEDLINE | ID: mdl-29165363

RESUMO

(1) Background: Plant pathogenic fungi often display high levels of host specificity and biotrophic fungi; in particular, they must manipulate their hosts to avoid detection and to complete their obligate pathogenic lifecycles. One important strategy of such fungi is the secretion of small proteins that serve as effectors in this process. Microbotryum violaceum is a species complex whose members infect members of the Caryophyllaceae; M. lychnidis-dioicae, a parasite on Silene latifolia, is one of the best studied interactions. We are interested in identifying and characterizing effectors of the fungus and possible corresponding host targets; (2) Methods: In silico analysis of the M. lychnidis-dioicae genome and transcriptomes allowed us to predict a pool of small secreted proteins (SSPs) with the hallmarks of effectors, including a lack of conserved protein family (PFAM) domains and also localized regions of disorder. Putative SSPs were tested for secretion using a yeast secretion trap method. We then used yeast two-hybrid analyses for candidate-secreted effectors to probe a cDNA library from a range of growth conditions of the fungus, including infected plants; (3) Results: Roughly 50 SSPs were identified by in silico analysis. Of these, 4 were studied further and shown to be secreted, as well as examined for potential host interactors. One of the putative effectors, MVLG_01732, was found to interact with Arabidopsis thaliana calcium-dependent lipid binding protein (AtCLB) and with cellulose synthase interactive protein 1 orthologues; and (4) Conclusions: The identification of a pool of putative effectors provides a resource for functional characterization of fungal proteins that mediate the delicate interaction between pathogen and host. The candidate targets of effectors, e.g., AtCLB, involved in pollen germination suggest tantalizing insights that could drive future studies.


Assuntos
Fungos/fisiologia , Interações Hospedeiro-Patógeno , Doenças das Plantas/microbiologia , Proteínas de Plantas/metabolismo , Proteínas Fúngicas/metabolismo , Fungos/patogenicidade , Genes Reporter , Ligação Proteica , Mapeamento de Interação de Proteínas , Técnicas do Sistema de Duplo-Híbrido
9.
J Neurosci ; 35(15): 6209-20, 2015 Apr 15.
Artigo em Inglês | MEDLINE | ID: mdl-25878291

RESUMO

Methyl-CpG binding protein 2 (MeCP2) is a chromatin regulator highly expressed in mature neurons. Mutations of MECP2 gene cause >90% cases of Rett syndrome, a neurodevelopmental disorder featured by striking psychomotor dysfunction. In Mecp2-null mice, the motor deficits are associated with reduction of dopamine content in the striatum, the input nucleus of basal ganglia mostly composed of GABAergic neurons. Here we investigated the causal role of MeCP2 in modulation of striatal dopamine content and psychomotor function. We found that mice with selective removal of MeCP2 in forebrain GABAergic neurons, predominantly in the striatum, phenocopied Mecp2-null mice in dopamine deregulation and motor dysfunction. Selective expression of MeCP2 in the striatum preserved dopamine content and psychomotor function in both males and females. Notably, the dopamine deregulation was primarily confined to the rostral striatum, and focal deletion or reactivation of MeCP2 expression in the rostral striatum through adeno-associated virus effectively disrupted or restored dopamine content and locomotor activity, respectively. Together, these findings demonstrate that striatal MeCP2 maintains local dopamine content in a non-cell autonomous manner in the rostral striatum and that is critical for psychomotor control.


Assuntos
Córtex Cerebral/metabolismo , Corpo Estriado/metabolismo , Dopamina/metabolismo , Proteína 2 de Ligação a Metil-CpG/metabolismo , Desempenho Psicomotor/fisiologia , Análise de Variância , Animais , Cromatografia Líquida de Alta Pressão , Comportamento Exploratório/fisiologia , Proteínas de Homeodomínio/genética , Proteínas de Homeodomínio/metabolismo , Masculino , Aprendizagem em Labirinto/fisiologia , Proteína 2 de Ligação a Metil-CpG/genética , Camundongos , Camundongos Transgênicos , Atividade Motora/genética , Mutação/genética , Transdução Genética
10.
Mol Biol Evol ; 32(4): 928-43, 2015 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-25534033

RESUMO

Dimorphic mating-type chromosomes in fungi are excellent models for understanding the genomic consequences of recombination suppression. Their suppressed recombination and reduced effective population size are expected to limit the efficacy of natural selection, leading to genomic degeneration. Our aim was to identify the sequences of the mating-type chromosomes (a1 and a2) of the anther-smut fungi and to investigate degeneration in their nonrecombining regions. We used the haploid a1 Microbotryum lychnidis-dioicae reference genome sequence. The a1 and a2 mating-type chromosomes were both isolated electrophoretically and sequenced. Integration with restriction-digest optical maps identified regions of recombination and nonrecombination in the mating-type chromosomes. Genome sequence data were also obtained for 12 other Microbotryum species. We found strong evidence of degeneration across the genus in the nonrecombining regions of the mating-type chromosomes, with significantly higher rates of nonsynonymous substitution (dN/dS) than in nonmating-type chromosomes or in recombining regions of the mating-type chromosomes. The nonrecombining regions of the mating-type chromosomes also showed high transposable element content, weak gene expression, and gene losses. The levels of degeneration did not differ between the a1 and a2 mating-type chromosomes, consistent with the lack of homogametic/heterogametic asymmetry between them, and contrasting with X/Y or Z/W sex chromosomes.


Assuntos
Basidiomycota/genética , Genes Fúngicos Tipo Acasalamento , Recombinação Genética , Cromossomos Sexuais , Sequência de Bases , Elementos de DNA Transponíveis , Deleção de Genes , Dados de Sequência Molecular , Filogenia , Análise de Sequência de DNA
11.
Arch Microbiol ; 198(8): 813-25, 2016 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-27215216

RESUMO

Microbotryum lychnidis-dioicae is a member of a species complex infecting host plants in the Caryophyllaceae. It is used as a model system in many areas of research, but attempts to make this organism tractable for reverse genetic approaches have not been fruitful. Here, we exploited the recently obtained genome sequence and transcriptome analysis to inform our design of constructs for use in Agrobacterium-mediated transformation techniques currently available for other fungi. Reproducible transformation was demonstrated at the genomic, transcriptional and functional levels. Moreover, these initial proof-of-principle experiments provide evidence that supports the findings from initial global transcriptome analysis regarding expression from the respective promoters under different growth conditions of the fungus. The technique thus provides for the first time the ability to stably introduce transgenes and over-express target M. lychnidis-dioicae genes.


Assuntos
Basidiomycota/genética , Técnicas de Transferência de Genes , Genoma Fúngico/genética , Doenças das Plantas/microbiologia , Plantas/microbiologia , Transformação Genética/genética , Mapeamento Cromossômico , Perfilação da Expressão Gênica , Genômica , Transcriptoma/genética
12.
Org Biomol Chem ; 14(7): 2200-4, 2016 Feb 21.
Artigo em Inglês | MEDLINE | ID: mdl-26804765

RESUMO

A highly regioselective synthesis of 3-arylindoles by using the cooperative FeCl3/DDQ system has been developed. This new protocol represents an attractive route for the synthesis of 3-arylindoles from readily accessible non-indole precursors, ß-aryl-substituted 2-styrylanilines, using an inexpensive catalyst and oxidant. Noteworthy is the unique synergetic and synergistic effect of FeCl3 and DDQ on the 1,2-aryl migratory process.

13.
Occup Environ Med ; 73(10): 694-700, 2016 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-27466611

RESUMO

BACKGROUND: Industrial radiography is known to be one of the most vulnerable lines of work among the range of different radiation work. According to the relevant law in Korea, every worker registered in this work should check their blood cell counts every year in addition to their thermoluminescent dosimeter (TLD) doses. Since the law was enacted, however, few follow-up studies have been carried out based on the obtained results. OBJECTIVES: To ascertain the clinical usefulness of complete blood cell count (CBC) results and suggest a proper protocol of health protection for radiation workers. METHODS: After reviewing all the consecutive results of CBC and TLD doses from radiation workers registered nationwide, we selected two groups of high-risk radiation workers, CBC-high risk (CBC-HR) and TLD-high risk (TLD-HR) groups. A control group of unexposed healthy adults was also included. We compared the absorbed doses calculated by cytogenetic biodosimetry among those three groups, and examined possible confounding factors for each group. RESULTS: Both groups of high-risk radiation workers, CBC-HR and TLD-HR, showed higher chromosome aberrations than the control group. In the control group, previous medical history of a CT scan increased the frequency of chromosome aberrations. In contrast, the frequency of chromosome aberrations in the high-risk radiation workers was affected not by the previous CT history but only by the duration of their work. CONCLUSIONS: We ascertain that reviewing consecutive results of blood cell counts and cytogenetic biodosimetry are useful complementary tools to TLD doses for health protection regulation. Several confounding factors including work duration and previous medical history need to be considered for the interpretation of biodosimetry results.


Assuntos
Aberrações Cromossômicas/efeitos da radiação , Doenças Profissionais/induzido quimicamente , Doenças Profissionais/epidemiologia , Exposição Ocupacional/efeitos adversos , Lesões por Radiação/epidemiologia , Radiografia/efeitos adversos , Adulto , Idoso , Análise de Variância , Contagem de Células Sanguíneas , Aberrações Cromossômicas/induzido quimicamente , Citogenética , Relação Dose-Resposta à Radiação , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Doenças Profissionais/sangue , Exposição Ocupacional/análise , Radiação , Doses de Radiação , Lesões por Radiação/sangue , República da Coreia , Estudos Retrospectivos , Fumar/epidemiologia , Dosimetria Termoluminescente , Adulto Jovem
14.
Phytopathology ; 106(11): 1244-1254, 2016 11.
Artigo em Inglês | MEDLINE | ID: mdl-27111800

RESUMO

The smut fungi form a large, diverse, and nonmonophyletic group of plant pathogens that have long served as both important pests of human agriculture and, also, as fertile organisms of scientific investigation. As modern techniques of molecular genetic analysis became available, many previously studied species that proved refractive to these techniques fell by the wayside and were neglected. Now, as the advent of rapid and affordable next-generation sequencing provides genomic and transcriptomic resources for even these "forgotten" fungi, several species are making a comeback and retaking prominent places in phytopathogenic research. In this review, we highlight several of these smut fungi, with special emphasis on Microbotryum lychnidis-dioicae, an anther smut whose molecular genetic tools have finally begun to catch up with its historical importance in classical genetics and now provide mechanistic insights for ecological studies, evolution of host-pathogen interaction, and investigations of emerging infectious disease.


Assuntos
Basidiomycota/genética , Perfilação da Expressão Gênica , Genômica , Doenças das Plantas/microbiologia , Basidiomycota/fisiologia , Ecologia , Interações Hospedeiro-Patógeno
15.
BMC Genomics ; 16: 461, 2015 Jun 16.
Artigo em Inglês | MEDLINE | ID: mdl-26076695

RESUMO

BACKGROUND: The genus Microbotryum includes plant pathogenic fungi afflicting a wide variety of hosts with anther smut disease. Microbotryum lychnidis-dioicae infects Silene latifolia and replaces host pollen with fungal spores, exhibiting biotrophy and necrosis associated with altering plant development. RESULTS: We determined the haploid genome sequence for M. lychnidis-dioicae and analyzed whole transcriptome data from plant infections and other stages of the fungal lifecycle, revealing the inventory and expression level of genes that facilitate pathogenic growth. Compared to related fungi, an expanded number of major facilitator superfamily transporters and secretory lipases were detected; lipase gene expression was found to be altered by exposure to lipid compounds, which signaled a switch to dikaryotic, pathogenic growth. In addition, while enzymes to digest cellulose, xylan, xyloglucan, and highly substituted forms of pectin were absent, along with depletion of peroxidases and superoxide dismutases that protect the fungus from oxidative stress, the repertoire of glycosyltransferases and of enzymes that could manipulate host development has expanded. A total of 14% of the genome was categorized as repetitive sequences. Transposable elements have accumulated in mating-type chromosomal regions and were also associated across the genome with gene clusters of small secreted proteins, which may mediate host interactions. CONCLUSIONS: The unique absence of enzyme classes for plant cell wall degradation and maintenance of enzymes that break down components of pollen tubes and flowers provides a striking example of biotrophic host adaptation.


Assuntos
Fungos/genética , Genoma Fúngico/genética , Parasitos/genética , Doenças das Plantas/microbiologia , Plantas/microbiologia , Silene/microbiologia , Transcriptoma/genética , Animais , Mapeamento Cromossômico/métodos , Perfilação da Expressão Gênica/métodos , Interações Hospedeiro-Parasita/genética , Lipase/genética , Peroxidases/genética , Superóxido Dismutase/genética
16.
J Biomed Sci ; 22: 102, 2015 Nov 17.
Artigo em Inglês | MEDLINE | ID: mdl-26572230

RESUMO

BACKGROUND: Mutations in mitogen-activated protein kinase (MAPK) kinase 1 (MEK1) that occur during cell proliferation and tumor formation are well described. Information on the roles of MEK2 in these effects is still limited. We established a constitutive MEK2 transgenic zebrafish, Tg(krt14:MEK2S219D-GFP), to elucidate the role of MEK2 in skin tumor formation. RESULTS: We found that both constitutive MEK2 and MEK1 are able to phosphorylate the extracellular signal-regulated kinase 1 (ERK1) protein. Transient expression of constitutive MEK2 and MEK1 in the zebrafish epidermis induced papillary formation at 48 h post-fertilization, but no effects were observed due to the expression of MEK1, MEK2, or the dominant negative form of MEK2. The transgenic zebrafish, Tg(krt14:MEK2S219D-GFP), developed skin papillomas in the epidermis within 6 days post-fertilization (dpf). The phospho-ERK signal was detected in section of skin papillomas in an immunohistochemical experiment. Treatment with 50 µM of the MEK inhibitor, U0126, had significantly decreased the skin papilloma formation in Tg(krt14:MEK2S219D-GFP) zebrafish by 6 dpf. In vitro and in vivo proliferation assay in COS-1 cells and in Tg(krt14:MEK2S219D-GFP) transgenic fish show significantly increased cell number and Ki-67 signaling. CONCLUSION: Our data indicate that MEK2 is sufficient to induce epidermal papilloma formation through MAPK signaling in zebrafish, and this transgenic model can be used as a new platform for drug screening.


Assuntos
MAP Quinase Quinase 2/metabolismo , Papiloma/metabolismo , Neoplasias Cutâneas/metabolismo , Proteínas de Peixe-Zebra/metabolismo , Peixe-Zebra/metabolismo , Animais , Animais Geneticamente Modificados/genética , Animais Geneticamente Modificados/metabolismo , Ativação Enzimática/genética , MAP Quinase Quinase 2/genética , Papiloma/genética , Neoplasias Cutâneas/genética , Peixe-Zebra/genética , Proteínas de Peixe-Zebra/genética
17.
Int J Mol Sci ; 16(9): 22027-45, 2015 Sep 11.
Artigo em Inglês | MEDLINE | ID: mdl-26378529

RESUMO

Dioecious plants in the Caryophyllaceae family are susceptible to infection by members of the anthericolous smut fungi. In our studies of the Silene latifolia/Microbotryum lychnidis-dioicae pathosystem, we were interested in characterizing the plant-pathogen interaction at the molecular level before and during teliosporogenesis. This takes place during floral bud development, and we hoped to capture the interaction by Illumina Next-Gen RNA-Sequencing. Using previous literature that documented the stages of the floral buds for S. latifolia, we examined the floral buds from plants grown and infected under growth chamber conditions, using the disserting microscope to determine the stage of floral buds based on the morphology. We compiled the information and determined the size of floral buds that correspond to the desired stages of development for tissue collection, for the purpose of RNA-sequencing. This offers a practical approach for researchers who require a large number of floral buds/tissue categorized by stages of development, ascertaining whether infected/uninfected buds are at comparable stages of development and whether this also holds true for male vs. female buds. We also document our experience in infecting the plants and some of the unusual morphologies we observed after infection.


Assuntos
Regulação da Expressão Gênica de Plantas , Silene/genética , Transcriptoma , Flores/genética , Flores/microbiologia , Perfilação da Expressão Gênica , Silene/microbiologia
18.
MMWR Morb Mortal Wkly Rep ; 63(8): 178, 2014 Feb 28.
Artigo em Inglês | MEDLINE | ID: mdl-24572614

RESUMO

Dog-to-dog transmission of rabies in Taiwan was eliminated in 1961; the island was considered canine rabies-free for 52 years. On July 16, 2013, three ferret-badgers (Melogale moschata) tested positive for rabies by fluorescent antibody testing at the Animal Health Research Institute, Council of Agriculture of Taiwan. This was the first time wild animals other than bats were tested. During 1999-2012, a total of 6,841 clinically healthy dogs and five apparently normal cats from shelters were tested and found negative for rabies. During 2009-2012, a total of 322 bats were tested and found negative for rabies.


Assuntos
Animais Selvagens/virologia , Vírus da Raiva/isolamento & purificação , Raiva/veterinária , Animais , Erradicação de Doenças/estatística & dados numéricos , Doenças do Cão/epidemiologia , Cães , Raiva/epidemiologia , Raiva/prevenção & controle , Vacina Antirrábica/administração & dosagem , Taiwan/epidemiologia
19.
Tzu Chi Med J ; 36(2): 152-165, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-38645788

RESUMO

Objectives: The protective effects and related mechanisms of Jing-Si herbal tea (JSHT) were investigated in cellular damage mediated by pro-inflammatory cytokines, including interleukin (IL)-1ß, IL-6, and tumor necrosis factor-α, on normal human lung fibroblast by multiomic platform analysis. Materials and Methods: The in silico high-throughput target was analyzed using pharmacophore models by BIOVIA Discovery Studio 2022 with ingenuity pathway analysis software. To assess cell viability, the study utilized the MTT assay technique. In addition, the IncuCyte S3 ZOOM System was implemented for the continuous monitoring of cell confluence of JSHT-treated cytokine-injured HEL 299 cells. Cytokine concentrations were determined using a Quantibody Human Inflammation Array. Gene expression and signaling pathways were determined using next-generation sequencing. Results: In silico high-throughput target analysis of JSHT revealed ingenuity in canonical pathways and their networks. Glucocorticoid receptor signaling is a potential signaling of JSHT. The results revealed protective effects against the inflammatory cytokines on JSHT-treated HEL 299 cells. Transcriptome and network analyses revealed that induction of helper T lymphocytes, TNFSF12, NFKB1-mediated relaxin signaling, and G-protein coupled receptor signaling play important roles in immune regulatory on JSHT-treated cytokine-injured HEL 299 cells. Conclusion: The findings from our research indicate that JSHT holds promise as a therapeutic agent, potentially offering advantageous outcomes in treating virus infections through various mechanisms. Furthermore, the primary bioactive components in JSHT justify extended research in antiviral drug development, especially in the context of addressing coronavirus.

20.
Int J Radiat Biol ; 100(5): 709-714, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-38394348

RESUMO

PURPOSE: Networking with other biodosimetry laboratories is necessary to assess the radiation exposure of many individuals in large-scale radiological accidents. The Korea biodosimetry network, K-BioDos, prepared harmonized scoring guidelines for dicentric chromosome assay to obtain homogeneous results within the network and investigated the efficiency of the guidelines. MATERIALS AND METHODS: Three laboratories in K-BioDos harmonized the scoring guidelines for dicentric chromosome assay. The results of scoring dicentric chromosomes using the harmonized scoring guidelines were compared with the laboratories' results using their own methods. Feedback was collected from the scorers following the three intercomparison exercises in 3 consecutive years. RESULTS: K-BioDos members showed comparable capacity to score dicentrics in the three exercises. However, the results of the K-BioDos guidelines showed no significant improvement over those of the scorers' own methods. According to the scorers, our harmonized guidelines led to more rejected metaphases and ultimately decreased the number of scorable metaphases compared with their own methods. Moreover, the scoring time was sometimes longer with the K-BioDos protocol because some scorers were not yet familiar with the guidelines, though most scorers reported that the time decreased or was unchanged. These challenges may cause low adherence to the guidelines. Most scorers expressed willingness to use the guidelines to select scorable metaphases or identify dicentrics for other biodosimetry works, whereas one did not want to use it due to the difference from their calibration curves. CONCLUSIONS: We identified potential resistance to following the harmonized guidelines and received requests for more detailed methods. Our findings suggest that the harmonized criteria should be continually updated, and education and training should be provided for all scorers. These changes could allow members within the biodosimetry network to successfully collaborate and support each other in large-scale radiological accidents.


Assuntos
Aberrações Cromossômicas , República da Coreia , Humanos , Cromossomos Humanos/genética , Cromossomos Humanos/efeitos da radiação
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