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1.
Development ; 148(20)2021 10 15.
Artigo em Inglês | MEDLINE | ID: mdl-34541605

RESUMO

Programmed cell death (PCD) is a common cell fate in metazoan development. PCD effectors are extensively studied, but how they are temporally regulated is less understood. Here, we report a mechanism controlling tail-spike cell death onset during Caenorhabditis elegans development. We show that the zinc-finger transcription factor BLMP-1, which controls larval development timing, also regulates embryonic tail-spike cell death initiation. BLMP-1 functions upstream of CED-9 and in parallel to DRE-1, another CED-9 and tail-spike cell death regulator. BLMP-1 expression is detected in the tail-spike cell shortly after the cell is born, and blmp-1 mutations promote ced-9-dependent tail-spike cell survival. BLMP-1 binds ced-9 gene regulatory sequences, and inhibits ced-9 transcription just before cell-death onset. BLMP-1 and DRE-1 function together to regulate developmental timing, and their mammalian homologs regulate B-lymphocyte fate. Our results, therefore, identify roles for developmental timing genes in cell-death initiation, and suggest conservation of these functions.


Assuntos
Proteínas de Caenorhabditis elegans/genética , Caenorhabditis elegans/genética , Morte Celular/genética , Proteínas Repressoras/genética , Transcrição Gênica/genética , Animais , Apoptose/genética , Diferenciação Celular/genética , Regulação da Expressão Gênica no Desenvolvimento/genética
2.
Plant Biotechnol J ; 2024 Jun 11.
Artigo em Inglês | MEDLINE | ID: mdl-38861663

RESUMO

The length of hypocotyl affects the height of soybean and lodging resistance, thus determining the final grain yield. However, research on soybean hypocotyl length is scarce, and the regulatory mechanisms are not fully understood. Here, we identified a module controlling the transport of sucrose, where sucrose acts as a messenger moved from cotyledon to hypocotyl, regulating hypocotyl elongation. This module comprises four key genes, namely MYB33, SWEET11, SWEET21 and GA2ox8c in soybean. In cotyledon, MYB33 is responsive to sucrose and promotes the expression of SWEET11 and SWEET21, thereby facilitating sucrose transport from the cotyledon to the hypocotyl. Subsequently, sucrose transported from the cotyledon up-regulates the expression of GA2ox8c in the hypocotyl, which ultimately affects the length of the hypocotyl. During the domestication and improvement of soybean, an allele of MYB33 with enhanced abilities to promote SWEET11 and SWEET21 has gradually become enriched in landraces and cultivated varieties, SWEET11 and SWEET21 exhibit high conservation and have undergone a strong purified selection and GA2ox8c is under a strong artificial selection. Our findings identify a new molecular pathway in controlling soybean hypocotyl elongation and provide new insights into the molecular mechanism of sugar transport in soybean.

3.
Proc Natl Acad Sci U S A ; 118(39)2021 09 28.
Artigo em Inglês | MEDLINE | ID: mdl-34531300

RESUMO

The Down syndrome cell adhesion molecule (DSCAM) belongs to the immunoglobulin superfamily (IgSF) and plays important roles in neural development. It has a large ectodomain, including 10 Ig-like domains and 6 fibronectin III (FnIII) domains. Previous data have shown that DSCAM can mediate cell adhesion by forming homophilic dimers between cells and contributes to self-avoidance of neurites or neuronal tiling, which is important for neural network formation. However, the organization and assembly of DSCAM at cell adhesion interfaces has not been fully understood. Here we combine electron microscopy and other biophysical methods to characterize the structure of the DSCAM-mediated cell adhesion and generate three-dimensional views of the adhesion interfaces of DSCAM by electron tomography. The results show that mouse DSCAM forms a regular pattern at the adhesion interfaces. The Ig-like domains contribute to both trans homophilic interactions and cis assembly of the pattern, and the FnIII domains are crucial for the cis pattern formation as well as the interaction with the cell membrane. By contrast, no obvious assembly pattern is observed at the adhesion interfaces mediated by mouse DSCAML1 or Drosophila DSCAMs, suggesting the different structural roles and mechanisms of DSCAMs in mediating cell adhesion and neural network formation.


Assuntos
Moléculas de Adesão Celular/química , Adesão Celular , Síndrome de Down/patologia , Proteínas de Drosophila/química , Neurogênese , Animais , Moléculas de Adesão Celular/genética , Moléculas de Adesão Celular/metabolismo , Membrana Celular/metabolismo , Síndrome de Down/genética , Síndrome de Down/metabolismo , Drosophila , Proteínas de Drosophila/genética , Proteínas de Drosophila/metabolismo , Camundongos , Neuritos
4.
Mikrochim Acta ; 191(4): 188, 2024 Mar 08.
Artigo em Inglês | MEDLINE | ID: mdl-38457047

RESUMO

A solid-phase microextraction (SPME) Arrow and high-performance liquid chromatography-UV detector (HPLC-UV, detection at 225 nm) based method was developed for the selective determination of nine alkylphenols (APs) in milk. The functionalized mesoporous UiO-66 (4-meso-UiO-66) was utilized as the new coating material, which was synthesized by post-modification of pore-expanded UiO-66-NH2 by an esterification reaction with 4-pentylbenzoic acid. It was fully characterized by X-ray photoelectron spectroscopy (XPS), fourier transformation infrared spectrometry, nitrogen sorption-desorption test, scanning electron microscopy, transmission electron microscopy, and X-ray diffractometer. The characterization results showed the ester groups and benzene rings were introduced into the 4-meso-UiO-66, and the mesoporous structure was predominant in the 4-meso-UiO-66. The extraction mechanism of 4-meso-UiO-66 to APs is the synergistic effect of Zr-O electrostatic interaction and the size exclusion effect resulting from XPS, selectivity test, and nitrogen sorption-desorption test. The electrospinning technique was utilized to fabricate the 4-meso-UiO-66 coated SPME Arrow and polyacrylonitrile (PAN) was used as the adhesive. The mass rate of 4-meso-UiO-66 to PAN and the electrospinning time were evaluated. The extraction and desorption parameters were also studied. The linear range of this method was 0.2-1000 µg L-1 with a coefficient of determination greater than 0.9989 under the optimal conditions. The detection limits were 0.05-1 µg L-1, the inter-day and intra-day precision (RSD) were 2.8-11.5%, and the recovery was 83.6%-112%. The reusability study showed that the extraction performance of this new SPME Arrow could be maintained after 80 adsorption-desorption cycles. This method showed excellent applicability for the selective determination of APs in milk.

5.
Int Wound J ; 21(1): e14635, 2024 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-38272805

RESUMO

This study compared the ankle-brachial index (ABI) with transcutaneous oxygen pressure (TcPO2 ) in assessing peripheral vascular disease (PVD) prevalence in 100 diabetic foot ulcer (DFU) patients. Patients were categorized into vascular or nonvascular reconstruction groups and underwent both ABI and TcPO2 measurements four times over 6 months. Predictive validity for PVD diagnosis was analysed using the area under the receiver-operating characteristic curve (AUC). The study found TcPO2 to be a superior predictor of PVD than ABI. Among the DFU patients, 51 with abnormal TcPO2 values underwent vascular reconstruction. Only TcPO2 values showed significant pretreatment differences between the groups and increased post-reconstruction. These values declined over a 6-month follow-up, whereas ABI values rose. For those with end-stage renal disease (ESRD), TcPO2 values saw a sharp decrease within 3 months. Pre-reconstruction TcPO2 was notably lower in amputation patients versus limb salvage surgery patients. In conclusion, TcPO2 is more effective than ABI for evaluating ischemic limb perfusion and revascularization necessity. It should be prioritized as the primary follow-up tool, especially for ESRD patients.


Assuntos
Diabetes Mellitus , Pé Diabético , Falência Renal Crônica , Doenças Vasculares Periféricas , Humanos , Monitorização Transcutânea dos Gases Sanguíneos , Pé Diabético/cirurgia , Pé Diabético/complicações , Isquemia/diagnóstico , Isquemia/cirurgia , Oxigênio/uso terapêutico
6.
J Struct Biol ; 215(3): 107971, 2023 09.
Artigo em Inglês | MEDLINE | ID: mdl-37201639

RESUMO

The structural studies of macromolecules in their physiological context, particularly in tissue, is constrained by the bottleneck of sample preparation. In this study, we present a practical pipeline for preparing multicellular samples for cryo-electron tomography. The pipeline comprises sample isolation, vitrification, and lift-out-based lamella preparation using commercially available instruments. We demonstrate the efficacy of our pipeline by visualizing pancreatic ß cells from mouse islets at the molecular level. This pipeline enables the determination of the properties of insulin crystals in situ for the first time, using unperturbed samples.


Assuntos
Tomografia com Microscopia Eletrônica , Manejo de Espécimes , Animais , Camundongos , Tomografia com Microscopia Eletrônica/métodos , Microscopia Crioeletrônica/métodos , Substâncias Macromoleculares , Manejo de Espécimes/métodos , Vitrificação
7.
Dev Biol ; 486: 96-108, 2022 06.
Artigo em Inglês | MEDLINE | ID: mdl-35367457

RESUMO

Skin epidermis secretes apical extracellular matrix (aECM) as a protective barrier from the external environment. The aECM is highly dynamic and constantly undergoes remodeling during animal development. How aECM dynamics is temporally regulated during development, and whether and how its mis-regulation may impact epidermal cell morphology or function remains to be fully elucidated. Here, we report that the conserved Zn-finger transcription factor BLMP-1/Blimp1, which regulates epidermal development in C. elegans, controls apical cell shape of the epidermis by downregulation of aECM remodeling. Loss of blmp-1 causes upregulation of genes essential for molting, including bus-8 and mlt-8, in adult, leading to an abnormal shape in the apical region of adult epidermal cells. The apical epidermal morphological defect is suppressed by reduction of bus-8 or mlt-8. BUS-8 is a key mannosyltransferase, which functions in glycosylation of N-linked glycoproteins; MLT-8 has a ganglioside GM2 lipid-binding domain and is implicated in signaling during molting, a process where the old cuticle is shed and synthesized anew. Overexpression of bus-8 or mlt-8 induces an apical epidermal cell defect as observed in blmp-1 mutants. MLT-8::GFP fusion protein is localized to lysosomes and secreted to aECM. BUS-8 is important for MLT-8 stability and lysosomal targeting, which may be regulated by BUS-8-mediated glycosylation of MLT-8 and function as a molting signaling cue in aECM remodeling. We propose that BLMP-1 represses MLT-8 expression and glycosylation in the epidermis to prevent inappropriate aECM remodeling, which is essential for maintenance of apical epidermal cell morphology during larva-to-adult transition.


Assuntos
Proteínas de Caenorhabditis elegans , Caenorhabditis elegans , Animais , Caenorhabditis elegans/metabolismo , Proteínas de Caenorhabditis elegans/genética , Proteínas de Caenorhabditis elegans/metabolismo , Células Epidérmicas/metabolismo , Epiderme/metabolismo , Manosiltransferases/metabolismo , Muda/genética
8.
Environ Res ; 231(Pt 3): 116244, 2023 08 15.
Artigo em Inglês | MEDLINE | ID: mdl-37245567

RESUMO

The production and use of organophosphate esters (OPEs) as substitutes for traditional halogenated flame retardants is increasing, resulting in greater global concern related to their ecological risks to marine environments. In this study, polychlorinated biphenyls (PCBs) and OPEs, representing traditional halogenated and emerging flame retardants, respectively, were studied in multiple environmental matrices in the Beibu Gulf, a typical semi-closed bay in the South China Sea. We investigated the differences in PCB and OPE distributions, sources, risks, and bioremediation potentials. Overall, the concentrations of emerging OPEs were much higher than those of PCBs in both seawater and sediment samples. Sediment samples from the inner bay and bay mouth areas (L sites) accumulated more PCBs, with penta- and hexa-CBs as major homologs. Chlorinated OPEs were prevalent in both seawater and sediment samples from the L sites, whereas tri-phenyl phosphate (TPHP) and tri-n-butyl phosphate (TNBP) were predominant at the outer bay (B sites) sediment samples. Source identification via principal component analysis, land use regression statistics, and δ13C analysis indicate that PCBs were mainly sourced from the atmospheric deposition of sugarcane and waste incineration, whereas sewage inputs, aquaculture, and shipping activity were identified as sources of OPE pollution in the Beibu Gulf. A half-year sediment anaerobic culturing experiment was performed for PCBs and OPEs, and the results only exhibited satisfactory dechlorination for PCBs. However, compared with the low ecological risks of PCBs to marine organisms, OPEs (particularly trichloroethyl phosphate (TCEP) and TPHP) exhibited low to medium threats to algae and crustaceans at most sites. Given their increasing usage, high ecological risks, and low bioremediation potential in enrichment cultures, pollution by emerging OPEs warrants close attention.


Assuntos
Retardadores de Chama , Bifenilos Policlorados , Poluentes Químicos da Água , Bifenilos Policlorados/análise , Poluentes Químicos da Água/análise , Monitoramento Ambiental , Retardadores de Chama/análise , Biodegradação Ambiental , Organofosfatos/análise , Fosfatos/análise , China , Ésteres/análise
9.
Ann Plast Surg ; 90(1 Suppl 1): S60-S67, 2023 04 01.
Artigo em Inglês | MEDLINE | ID: mdl-37075295

RESUMO

BACKGROUND: Surgical site infection (SSI) after kidney transplantation can severely compromise graft function and prolong hospital stay. Organ/space SSI (osSSI) is a severe type of SSI associated with a significantly higher mortality rate. AIMS AND OBJECTIVES: This study aims to provide new strategies of managing (osSSI) after kidney transplant and other high-risk wound infections. METHOD: This is a single-center, retrospective study that analyzed the treatment outcomes of 4 patients who developed osSSI after kidney transplant at Shuang-Ho Hospital. The management strategy included real-time fluorescence imaging with MolecuLight, negative-pressure wound therapy (NPWT) with Si-Mesh, and incisional NPWT (iNPWT). RESULT: The average length of hospital stay was 18 days (range, 12-23 days). During hospitalization, all patients obtained high-quality debridement under real-time fluorescence image confirmation. The average duration of NPWT was 11.8 days (range, 7-17 days) and iNPWT was 7 days. All transplanted kidneys were preserved with normal function after 6 months of follow-up. CONCLUSIONS: Our strategies with real-time fluorescence imaging provide a novel and effective method that can be used in adjunct with the standard of care for managing osSSI after kidney transplantation. More studies are warranted to validate the efficacy of our approach.


Assuntos
Tratamento de Ferimentos com Pressão Negativa , Ferida Cirúrgica , Humanos , Infecção da Ferida Cirúrgica/terapia , Tratamento de Ferimentos com Pressão Negativa/métodos , Estudos Retrospectivos , Rim/diagnóstico por imagem
10.
Bull Environ Contam Toxicol ; 110(5): 96, 2023 May 15.
Artigo em Inglês | MEDLINE | ID: mdl-37188746

RESUMO

Microplastics (MPs) have become an important global issue in recent years. However, MPs in the soil have received far less attention than water. Effective and nondestructive extraction of MPs is important for studying MPs in agricultural soils. This study uses different floatation solutions as experiments and uses MgCl2 as the floatation solution of the density extraction method. Five types of standard MPs (PE, PP, PS, PVC, and PET) are used as the objects of this experiment. The recovery of the two particle sizes was between 90.82% and 109.69%. The extracted standard MPs were then subjected to IR and Raman spectroscopic analysis, and the results showed that Raman spectroscopy was more suitable for the identification of the extracted MPs. Finally, this method collected and verified a vast number of soil samples and further analyzed the abundance and characteristics of the collected MPs.


Assuntos
Microplásticos , Solo , Microplásticos/análise , Plásticos/análise , Cloreto de Sódio , China
11.
Comput Electr Eng ; 102: 108230, 2022 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-35965689

RESUMO

In 2019, a new strain of coronavirus pneumonia spread quickly worldwide. Viral propagation may be simulated using the Susceptible Infectious Removed (SIR) model. However, the SIR model fails to consider that separation of patients in the COVID-19 incubation stage entails difficulty and that these patients have high transmission potential. The model also ignores the positive effect of quarantine measures on the spread of the epidemic. To address the two flaws in the SIR model, this study proposes a new infectious disease model referred to as the Susceptible Quarantined Exposed Infective Removed (SQEIR) model. The proposed model uses the weighted least squares for the optimal estimation of important parameters in the infectious disease model. Based on these parameters, new differential equations were developed to describe the spread of the epidemic. The experimental results show that this model exhibits an accuracy 6.7% higher than that of traditional infectious disease models.

12.
J Biol Chem ; 295(46): 15727-15741, 2020 11 13.
Artigo em Inglês | MEDLINE | ID: mdl-32907880

RESUMO

Scavenger receptors are a superfamily of membrane-bound receptors that recognize both self and nonself targets. Scavenger receptor class A (SR-A) has five known members (SCARA1 to -5 or SR-A1 to -A5), which are type II transmembrane proteins that form homotrimers on the cell surface. SR-A members recognize various ligands and are involved in multiple biological pathways. Among them, SCARA5 can function as a ferritin receptor; however, the interaction between SCARA5 and ferritin has not been fully characterized. Here, we determine the crystal structures of the C-terminal scavenger receptor cysteine-rich (SRCR) domain of both human and mouse SCARA5 at 1.7 and 2.5 Å resolution, respectively, revealing three Ca2+-binding sites on the surface. Using biochemical assays, we show that the SRCR domain of SCARA5 recognizes ferritin in a Ca2+-dependent manner, and both L- and H-ferritin can be recognized by SCARA5 through the SRCR domain. Furthermore, the potential binding region of SCARA5 on the surface of ferritin is explored by mutagenesis studies. We also examine the interactions of ferritin with other SR-A members and find that SCARA1 (SR-A1, CD204) and MARCO (SR-A2, SCARA2), which are highly expressed on macrophages, also interact with ferritin. By contrast, SCARA3 and SCARA4, the two SR-A members without the SRCR domain, have no detectable binding with ferritin. Overall, these results provide a mechanistic view regarding the interactions between the SR-A members and ferritin that may help to understand the regulation of ferritin homeostasis by scavenger receptors.


Assuntos
Ferritinas/metabolismo , Receptores Depuradores Classe A/metabolismo , Animais , Sítios de Ligação , Cálcio/química , Cálcio/metabolismo , Cristalografia por Raios X , Humanos , Cinética , Macrófagos/citologia , Macrófagos/metabolismo , Camundongos , Simulação de Dinâmica Molecular , Mutagênese Sítio-Dirigida , Ligação Proteica , Domínios Proteicos , Estrutura Terciária de Proteína , Receptores Depuradores Classe A/química , Receptores Depuradores Classe A/genética
13.
Proc Natl Acad Sci U S A ; 115(37): 9246-9251, 2018 09 11.
Artigo em Inglês | MEDLINE | ID: mdl-30150416

RESUMO

Cell-cell adhesion is important for cell growth, tissue development, and neural network formation. Structures of cell adhesion molecules have been widely studied by crystallography, revealing the molecular details of adhesion interfaces. However, due to technical limitations, the overall structure and organization of adhesion molecules at cell adhesion interfaces has not been fully investigated. Here, we combine electron microscopy and other biophysical methods to characterize the structure of cell-cell adhesion mediated by the cell adhesion molecule Sidekick (Sidekick-1 and Sidekick-2) and obtain 3D views of the Sidekick-mediated adhesion interfaces as well as the organization of Sidekick molecules between cell membranes by electron tomography. The results suggest that the Ig-like domains and the fibronectin III (FnIII) domains of Sidekicks play different roles in cell adhesion. The Ig-like domains mediate the homophilic transinteractions bridging adjacent cells, while the FnIII domains interact with membranes, resulting in a tight adhesion interface between cells that may contribute to the specificity and plasticity of cell-cell contacts during cell growth and neural development.


Assuntos
Membrana Celular , Tomografia com Microscopia Eletrônica , Imunoglobulina G , Proteínas de Membrana , Animais , Adesão Celular/fisiologia , Membrana Celular/química , Membrana Celular/metabolismo , Membrana Celular/ultraestrutura , Células HEK293 , Humanos , Imunoglobulina G/química , Imunoglobulina G/metabolismo , Imunoglobulina G/ultraestrutura , Proteínas de Membrana/química , Proteínas de Membrana/metabolismo , Proteínas de Membrana/ultraestrutura , Camundongos , Domínios Proteicos
14.
Global Health ; 15(1): 18, 2019 02 28.
Artigo em Inglês | MEDLINE | ID: mdl-30819237

RESUMO

BACKGROUND: Although the worldwide incidence of tuberculosis (TB) has been slowly decreasing, the migrant workers remains an important gap for regional TB control. In Taiwan, the numbers of the migrant workers from countries with high TB incidence increase significantly in past decades and the impact on public health remains unknown. This study aimed to explore the difference of TB incidence between Taiwanese and the migrant workers. METHODS: The migrant workers are obligated to receive pre-arrival, post-arrival and regular chest X-ray screening during their stay in Taiwan. We retrospectively collected these data extracted from the Alien Workers Health Database in Centers for Disease Control, Taiwan from Jan. 1, 2004 to Dec. 31, 2013. Poisson regression models were used to compare the hazard ratios of TB between Taiwanese and the migrant workers after adjusting gender and age groups. RESULTS: The total migrant workers in Taiwan reached 314,034 persons in 2004 and 489,134 persons in 2013, accounting for 2% of Taiwan population. The TB incidence of migrant workers was similar to Taiwanese (53-73.7 per 105 vs 45.5-76.8 per 105). Comparing with Taiwanese, the TB risk was significantly lower in male migrant workers (HR: 0.76; 95% CI: 0.70-0.83, P < 0.001), but higher in female migrant workers (HR: 1.40; 95% CI: 1.35-1.46, P < 0.001). Besides, we found that the TB risk in migrant workers was 5.30-fold (95% CI: 4.83-5.83, P < 0.001) in youngest group (≤24 year-old) comparing with Taiwanese. CONCLUSIONS: Migrant workers in Taiwan have higher TB incidence than Taiwanese in young groups, especially in females. The mainstay young laborers with latent tuberculosis infection risk is an important vulnerability for public health. Further investigation and health screening are warranted.


Assuntos
Migrantes/estatística & dados numéricos , Tuberculose/epidemiologia , Adulto , Feminino , Humanos , Incidência , Masculino , Pessoa de Meia-Idade , Taiwan/epidemiologia , Adulto Jovem
15.
Global Health ; 14(1): 67, 2018 07 11.
Artigo em Inglês | MEDLINE | ID: mdl-29996856

RESUMO

BACKGROUND: With globalization, more and more people travel to countries where they are at risk of injuries and travel-related diseases. To protect travelers' health, it is crucial to understand whether travelers accurately perceive medical assistance resources before and during their trips. This study investigated the need, awareness, and previous usage of overseas emergency medical assistance services (EMAS) among people traveling abroad. METHODS: Anonymous questionnaires were distributed to patients (n = 500) at a travel clinic in Taipei, Taiwan. RESULTS: The results showed that EMAS were important, especially in the following categories: 24-h telephone medical consultation (91.8%), emergent medical repatriation (87.6%), and assistance with arranging hospital admission (87.4%). Patients were less aware of the following services: arrangement of appointments with doctors (70.7%) and monitoring of medical conditions during hospitalization (73.0%). Less than 5% of respondents had a previous experience with EMAS. CONCLUSIONS: EMAS are considered important to people who are traveling abroad. However, approximately 20-30% of travelers lack an awareness of EMAS, and the percentage of travelers who have previously received medical assistance through these services is extremely low. The discrepancy between the need and usage of EMAS emphasizes the necessity to adapt EMAS materials in pre-travel consultations to meet the needs of international travelers.


Assuntos
Conhecimentos, Atitudes e Prática em Saúde , Internacionalidade , Assistência Médica , Viagem , Adolescente , Adulto , Idoso , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Inquéritos e Questionários , Taiwan , Adulto Jovem
16.
PLoS Genet ; 10(8): e1004513, 2014 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-25144461

RESUMO

Programmed cell death (PCD) is the physiological death of a cell mediated by an intracellular suicide program. Although key components of the PCD execution pathway have been identified, how PCD is regulated during development is poorly understood. Here, we report that the epidermal growth factor (EGF)-like ligand LIN-3 acts as an extrinsic signal to promote the death of specific cells in Caenorhabditis elegans. The loss of LIN-3 or its receptor, LET-23, reduced the death of these cells, while excess LIN-3 or LET-23 signaling resulted in an increase in cell deaths. Our molecular and genetic data support the model that the LIN-3 signal is transduced through LET-23 to activate the LET-60/RAS-MPK-1/ERK MAPK pathway and the downstream ETS domain-containing transcription factor LIN-1. LIN-1 binds to, and activates transcription of, the key pro-apoptotic gene egl-1, which leads to the death of specific cells. Our results provide the first evidence that EGF induces PCD at the whole organism level and reveal the molecular basis for the death-promoting function of LIN-3/EGF. In addition, the level of LIN-3/EGF signaling is important for the precise fine-tuning of the life-versus-death fate. Our data and the previous cell culture studies that say EGF triggers apoptosis in some cell lines suggest that the EGF-mediated modulation of PCD is likely conserved in C. elegans and humans.


Assuntos
Proteínas de Caenorhabditis elegans/biossíntese , Proteínas de Caenorhabditis elegans/genética , Morte Celular/genética , Fator de Crescimento Epidérmico/genética , Proteínas Repressoras/biossíntese , Ativação Transcricional/genética , Animais , Caenorhabditis elegans , Proteínas de Caenorhabditis elegans/metabolismo , Linhagem da Célula/genética , Fator de Crescimento Epidérmico/metabolismo , Receptores ErbB/genética , Receptores ErbB/metabolismo , Regulação da Expressão Gênica no Desenvolvimento , Humanos , Sistema de Sinalização das MAP Quinases/genética , Proteínas Repressoras/genética , Fatores de Transcrição/genética , Fatores de Transcrição/metabolismo
17.
PLoS Genet ; 10(6): e1004428, 2014 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-24968003

RESUMO

Spatiotemporal regulation of cell migration is crucial for animal development and organogenesis. Compared to spatial signals, little is known about temporal signals and the mechanisms integrating the two. In the Caenorhabditis elegans hermaphrodite, the stereotyped migration pattern of two somatic distal tip cells (DTCs) is responsible for shaping the gonad. Guidance receptor UNC-5 is necessary for the dorsalward migration of DTCs. We found that BLMP-1, similar to the mammalian zinc finger transcription repressor Blimp-1/PRDI-BF1, prevents precocious dorsalward turning by inhibiting precocious unc-5 transcription and is only expressed in DTCs before they make the dorsalward turn. Constitutive expression of blmp-1 when BLMP-1 would normally disappear delays unc-5 transcription and causes turn retardation, demonstrating the functional significance of blmp-1 down-regulation. Correct timing of BLMP-1 down-regulation is redundantly regulated by heterochronic genes daf-12, lin-29, and dre-1, which regulate the temporal fates of various tissues. DAF-12, a steroid hormone receptor, and LIN-29, a zinc finger transcription factor, repress blmp-1 transcription, while DRE-1, the F-Box protein of an SCF ubiquitin ligase complex, binds to BLMP-1 and promotes its degradation. We have therefore identified a gene circuit that integrates the temporal and spatial signals and coordinates with overall development of the organism to direct cell migration during organogenesis. The tumor suppressor gene product FBXO11 (human DRE-1 ortholog) also binds to PRDI-BF1 in human cell cultures. Our data suggest evolutionary conservation of these interactions and underscore the importance of DRE-1/FBXO11-mediated BLMP-1/PRDI-BF1 degradation in cellular state transitions during metazoan development.


Assuntos
Proteínas de Caenorhabditis elegans/genética , Caenorhabditis elegans/genética , Proteínas F-Box/genética , Organogênese/genética , Proteínas Repressoras/genética , Animais , Caenorhabditis elegans/crescimento & desenvolvimento , Movimento Celular/genética , Evolução Molecular , Proteínas F-Box/metabolismo , Regulação da Expressão Gênica no Desenvolvimento , Gônadas/crescimento & desenvolvimento , Gônadas/metabolismo , Humanos , Fator 1 de Ligação ao Domínio I Regulador Positivo , Proteólise , Receptores de Superfície Celular/genética
18.
J Asian Nat Prod Res ; 18(3): 303-14, 2016.
Artigo em Inglês | MEDLINE | ID: mdl-26488075

RESUMO

Porcine reproductive and respiratory syndrome virus (PRRSV) is an important arterivirus that causes substantial economic losses to the swine industry. Current control strategies against PRRSV are still inadequate and there is an urgent need for new antiviral therapies. Tetrahydroaltersolanol C (TD-C) is a new anthraquinone derivative isolated from the marine-derived fungi. In the present study, we first demonstrated its anti-PRRSV activity in vitro through assessing the inhibition of TD-C on cytopathic effect, viral ORF7 gene and N protein expressions, progeny virions production by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay, relative-quantitative RT-PCR, Western blotting, and indirect immunofluorescence assay. Our experimental results showed that TD-C could significantly inhibit PRRSV replication in a dose-dependent manner. The 50% effective concentration, 50% cytotoxic concentration and the selectivity index were 12.11, 395.31 µM, and 32.64, respectively. Furthermore, the possible anti-PRRSV mechanism was explored by virucidal assay, virus adsorption inhibition assay, and the time-of-addition assay. The results showed that TD-C might inhibit the internalization and replication of PRRSV, but did not directly inactivate the virus or block its adsorption to cell surface. In conclusion, our findings indicated that TD-C possessed a significant anti-PRRSV activity, and provided a strong basis for further exploration of this compound as an antiviral agent against PRRSV.


Assuntos
Antraquinonas/farmacologia , Antivirais/farmacologia , Vírus da Síndrome Respiratória e Reprodutiva Suína/efeitos dos fármacos , Animais , Antraquinonas/química , Antivirais/química , Linhagem Celular , Estrutura Molecular , Reação em Cadeia da Polimerase , Suínos , Sais de Tetrazólio , Tiazóis , Replicação Viral/efeitos dos fármacos
19.
PLoS Genet ; 8(5): e1002663, 2012.
Artigo em Inglês | MEDLINE | ID: mdl-22615577

RESUMO

Clearance of apoptotic cells by engulfment plays an important role in the homeostasis and development of multicellular organisms. Despite the fact that the recognition of apoptotic cells by engulfment receptors is critical in inducing the engulfment process, the molecular mechanisms are still poorly understood. Here, we characterize a novel cell corpse engulfment pathway mediated by the integrin α subunit PAT-2 in Caenorhabditis elegans and show that it specifically functions in muscle-mediated engulfment during embryogenesis. Inactivation of pat-2 results in a defect in apoptotic cell internalization. The PAT-2 extracellular region binds to the surface of apoptotic cells in vivo, and the intracellular region may mediate signaling for engulfment. We identify essential roles of small GTPase CDC-42 and its activator UIG-1, a guanine-nucleotide exchange factor, in PAT-2-mediated cell corpse removal. PAT-2 and CDC-42 both function in muscle cells for apoptotic cell removal and are co-localized in growing muscle pseudopods around apoptotic cells. Our data suggest that PAT-2 functions through UIG-1 for CDC-42 activation, which in turn leads to cytoskeletal rearrangement and apoptotic cell internalization by muscle cells. Moreover, in contrast to PAT-2, the other integrin α subunit INA-1 and the engulfment receptor CED-1, which signal through the conserved signaling molecules CED-5 (DOCK180)/CED-12 (ELMO) or CED-6 (GULP) respectively, preferentially act in epithelial cells to mediate cell corpse removal during mid-embryogenesis. Our results show that different engulfing cells utilize distinct repertoires of receptors for engulfment at the whole organism level.


Assuntos
Benzenoacetamidas , Proteínas de Caenorhabditis elegans , Caenorhabditis elegans , Proteínas de Ciclo Celular , Desenvolvimento Embrionário , Proteínas de Ligação ao GTP , Músculo Esquelético/embriologia , Piridinas , Animais , Apoptose , Proteínas Reguladoras de Apoptose , Benzenoacetamidas/metabolismo , Caenorhabditis elegans/embriologia , Caenorhabditis elegans/genética , Proteínas de Caenorhabditis elegans/genética , Proteínas de Caenorhabditis elegans/metabolismo , Proteínas de Transporte/metabolismo , Proteínas de Ciclo Celular/genética , Proteínas de Ciclo Celular/metabolismo , Proteínas do Citoesqueleto/metabolismo , Citoesqueleto/genética , Citoesqueleto/metabolismo , Eptifibatida , Proteínas de Ligação ao GTP/genética , Proteínas de Ligação ao GTP/metabolismo , Fatores de Troca do Nucleotídeo Guanina/metabolismo , Integrinas/metabolismo , Proteínas de Membrana/metabolismo , Peptídeos/genética , Peptídeos/metabolismo , Fosfoproteínas/metabolismo , Piridinas/metabolismo , Transdução de Sinais
20.
Analyst ; 139(23): 6197-203, 2014 Dec 07.
Artigo em Inglês | MEDLINE | ID: mdl-25299249

RESUMO

In this work, a novel chemically modified electrode is constructed based on metal-organic frameworks and ß-cyclodextrin (Cu3(BTC)2/ß-CD, BTC = benzene-1,3,5-tricarboxylate) composite material. The electrode was used for simultaneous determination of guanine and adenine in a sequential injection lab-on-valve format and exhibited sensitive responses to guanine and adenine oxidation due to the π-π stacking interaction of Cu3(BTC)2 and the inclusion behavior of ß-CD. The analytical performance was assessed with respect to the supporting electrolyte and its pH, accumulation time and accumulation potential, and the fluid flow rates. Under optimal conditions, linear calibration ranges for both guanine and adenine were from 1.0 × 10(-7) to 1.0 × 10(-5) mol L(-1), and detection limits (S/N = 3) were found to be 5.2 × 10(-8) and 2.8 × 10(-8) mol L(-1), respectively. The proposed sensor showed advantages of high sensitivity, simple sample preparation protocol, enhanced throughput and good reproducibility. Finally, the practical application of the proposed sensor has been performed for the determination of guanine and adenine in real samples with satisfactory results.


Assuntos
Adenina/química , Técnicas Eletroquímicas/instrumentação , Guanina/química , Compostos Organometálicos/química , beta-Ciclodextrinas/química , Animais , Cerveja/análise , Sangue , Bovinos , Eletrodos , Análise de Alimentos/instrumentação , Análise de Alimentos/métodos , Carne/análise , Sensibilidade e Especificidade , Urina/química
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