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1.
Heliyon ; 10(1): e22781, 2024 Jan 15.
Artículo en Inglés | MEDLINE | ID: mdl-38116191

RESUMEN

Getting employees to share their creative ideas has long been recognized as a vital source of organizational effectiveness. This research uses the conservation of resources theory to investigate how employee's creative idea sharing is affected by abusive supervision. Data for this research was collected from 209 employees and their immediate supervisors of generic nurses and medical dispensers of Southern Punjab public sector hospitals working under the Ministry of national health services regulation and Coordination. Data were then analyzed with the AMOS software package for simple regression and moderated mediation. This study found that with the increase in abusive supervision, employees develop cheating behavior, diminishing probability of sharing their creative ideas with coworkers. Along these lines, organizational justice moderates this relationship and attenuates the negative indirect effect of abusive supervision on creative idea sharing. The researchers recommended that organizations should develop training programs or coaching sessions for leaders to make them equip with essential interpersonal skills that can eradicate abusive supervision. Research implications, limitations, and future research directions are also discussed.

2.
Int J Gen Med ; 16: 5763-5777, 2023.
Artículo en Inglés | MEDLINE | ID: mdl-38089712

RESUMEN

Background: To evaluate the clinical value of metagenomic next-generation sequencing (mNGS) in screening of lower respiratory tract infections (LRTIs) and human tumors. Methods: Human samples included bronchoalveolar lavage fluid (BALF), sputum, lung biopsy tissue, and peripheral blood from 188 patients who were admitted to our hospital between January 2020 and September 2022 were analyzed using mNGS for simultaneous pathogen and chromosome copy number variation (CNV) detection. Traditional microbial culture and comprehensive microbial test (CMT) were also conducted. The diagnostic efficiencies of the three methods (mNGS, traditional culture, and CMT groups) were compared. Results: Among the 188 patients, 149 (79.3%) were in the LRTIs group and 39 (20.7%) were in the non-LRTIs group. The diagnostic sensitivity and accuracy of the mNGS group were higher than those of the traditional culture and CMT groups (P < 0.001; P < 0.001; P < 0.001; P < 0.001), and the specificity was higher than that of the CMT group (P = 0.039) but lower than that of the traditional culture group (P = 0.006). The positive predictive values of the mNGS and traditional culture groups were higher than that of the CMT group (P = 0.004; P = 0.011). The negative predictive value of the mNGS group was higher than that of the CMT group (P = 0.003). In addition, all samples were subjected to simultaneous chromosome CNV detection, and 8% (15/188) were positive for CNV. Of the 15 patients, 10 were initially misdiagnosed as non-neoplastic diseases, with a misdiagnosis rate of 66.7% (10/15). The BALF CNV test was performed on 13 patients diagnosed with primary or metastatic lung cancer, with a positivity rate of 38.5%. Conclusion: The sensitivity and accuracy of pathogen diagnosis using mNGS were better than those of traditional culture and CMT. CNV detection is an important auxiliary diagnostic tool for cancer, particularly for screening occult tumors.

3.
IEEE J Biomed Health Inform ; 27(10): 4938-4949, 2023 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-37471184

RESUMEN

The accurate diagnosis of significant liver fibrosis ( ≥ F2) in patients with chronic liver disease (CLD) is critical, as ≥ F2 is a crucial factor that should be considered in selecting an antiviral therapy for these patients. This article proposes a handcrafted-feature-assisted deep convolutional neural network (HFA-DCNN) that helps radiologists automatically and accurately diagnose significant liver fibrosis from ultrasound (US) brightness (B)-mode images. The HFA-DCNN model has three main branches: one for automatic region of interest (ROI) segmentation in the US images, another for attention deep feature learning from the segmented ROI, and the third for handcrafted feature extraction. The attention deep learning features and handcrafted features are fused in the back end of the model to enable more accurate diagnosis of significant liver fibrosis. The usefulness and effectiveness of the proposed model were validated on a dataset built upon 321 CLD patients with liver fibrosis stages confirmed by pathological evaluations. In a fivefold cross validation (FFCV), the proposed model achieves accuracy, sensitivity, specificity, and area under the receiver-operating-characteristic (ROC) curve (AUC) values of 0.863 (95% confidence interval (CI) 0.820-0.899), 0.879 (95% CI 0.823-0.920), 0.872 (95% CI 0.800-0.925), and 0.925 (95% CI 0.891-0.952), which are significantly better than those obtained by the comparative methods. Given its excellent performance, the proposed HFA-DCNN model can serve as a promising tool for the noninvasive and accurate diagnosis of significant liver fibrosis in CLD patients.

4.
Ginekol Pol ; 2023 May 10.
Artículo en Inglés | MEDLINE | ID: mdl-37162140

RESUMEN

OBJECTIVES: To investigate the prevalence and influencing factors of stress urinary incontinence (SUI) within 6-8 weeks postpartum in Jiangsu Province. MATERIAL AND METHODS: We designed a multi-center cross-sectional study involving seven hospitals in Jiangsu province, and enrolled women who underwent postpartum examination at 6-8 weeks in these hospitals between July 2019 and June 2021. According to the presence or absence of SUI, the enrolled patients were divided into two groups: the SUI group and the non-SUI group, respectively. We assessed the general health status, noted the details of delivery, and checked the pelvic floor electromyographic parameters of the postpartum women in both groups. RESULTS: Among 6,302 cases of postpartum women in Jiangsu province, there were 1,579 cases of SUI, with a prevalence of 25.06%. The prevalence of SUI increased significantly with age, BMI, increasing parity, coexisting constipation, organ prolapse, and diastasis recti abdominis. Compared to the non-SUI group, the SUI group had a lower mean value of the pre-baseline rest phase, shorter rise and fall times of fast muscle contractions, and a lower mean value of the endurance contraction phase. Multiple regression analysis revealed associations with weight (especially overweight and obesity), coexisting organ prolapse, constipation, parity, gestational week of delivery, mode of delivery, and mean value of endurance contraction phase. CONCLUSIONS: The prevalence of postpartum stress urinary incontinence in Jiangsu Province was 25.06%, and was linked to being overweight, parity > 2, coexisting organ prolapse, constipation, and a decrease in the mean value of the endurance contraction phase of the electromyograph. In this report, we offer a theoretical basis for the effective prevention of postpartum SUI clinically.

5.
Cell Res ; 33(4): 288-298, 2023 04.
Artículo en Inglés | MEDLINE | ID: mdl-36775821

RESUMEN

Intraflagellar transport (IFT) complexes, IFT-A and IFT-B, form bidirectional trains that move along the axonemal microtubules and are essential for assembling and maintaining cilia. Mutations in IFT subunits lead to numerous ciliopathies involving multiple tissues. However, how IFT complexes assemble and mediate cargo transport lacks mechanistic understanding due to missing high-resolution structural information of the holo-complexes. Here we report cryo-EM structures of human IFT-A complexes in the presence and absence of TULP3 at overall resolutions of 3.0-3.9 Å. IFT-A adopts a "lariat" shape with interconnected core and peripheral subunits linked by structurally vital zinc-binding domains. TULP3, the cargo adapter, interacts with IFT-A through its N-terminal region, and interface mutations disrupt cargo transport. We also determine the molecular impacts of disease mutations on complex formation and ciliary transport. Our work reveals IFT-A architecture, sheds light on ciliary transport and IFT train formation, and enables the rationalization of disease mutations in ciliopathies.


Asunto(s)
Cilios , Humanos , Cilios/metabolismo , Transporte Biológico , Transporte de Proteínas
6.
Nat Commun ; 14(1): 479, 2023 01 30.
Artículo en Inglés | MEDLINE | ID: mdl-36717564

RESUMEN

The transport of the CagA effector into gastric epithelial cells by the Cag Type IV secretion system (Cag T4SS) of Helicobacter pylori (H. pylori) is critical for pathogenesis. CagA is recruited to Cag T4SS by the Cagß ATPase. CagZ, a unique protein in H. pylori, regulates Cagß-mediated CagA transport, but the underlying mechanisms remain unclear. Here we report the crystal structure of the cytosolic region of Cagß, showing a typical ring-like hexameric assembly. The central channel of the ring is narrow, suggesting that CagA must unfold for transport through the channel. Our structure of CagZ in complex with the all-alpha domain (AAD) of Cagß shows that CagZ adopts an overall U-shape and tightly embraces Cagß. This binding mode of CagZ is incompatible with the formation of the Cagß hexamer essential for the ATPase activity. CagZ therefore inhibits Cagß by trapping it in the monomeric state. Based on these findings, we propose a refined model for the transport of CagA by Cagß.


Asunto(s)
Adenosina Trifosfatasas , Proteínas Bacterianas , Helicobacter pylori , Adenosina Trifosfatasas/metabolismo , Antígenos Bacterianos/metabolismo , Proteínas Bacterianas/genética , Proteínas Bacterianas/metabolismo , Helicobacter pylori/metabolismo , Sistemas de Secreción Tipo IV/metabolismo
7.
J Agric Food Chem ; 70(27): 8469-8480, 2022 Jul 13.
Artículo en Inglés | MEDLINE | ID: mdl-35771952

RESUMEN

The rice stripe virus (RSV) is responsible for devastating effects in East Asian rice-producing areas. The disease-specific protein (SP) level in rice plants determines the severity of RSV symptoms. Isothermal titration calorimetry (ITC) and bimolecular fluorescence complementation (BiFC) assays confirmed the interaction between an R3H domain-containing host factor, OsR3H3, and RSV SP in vitro and in vivo. This study determined the crystal structure of SP at 1.71 Å. It is a monomer with a clear shallow groove to accommodate host factors. Docking OsR3H3 into the groove generates an SP/OsR3H3 complex, which provides insights into the protein-binding mechanism of SP. Furthermore, SP's protein-binding properties and model-defined recognition residues were assessed using mutagenesis, ITC, and BiFC assays. This study revealed the structure and preliminary protein interaction mechanisms of RSV SP, shedding light on the molecular mechanism underlying the development of RSV infection symptoms.


Asunto(s)
Oryza , Tenuivirus , Oryza/metabolismo , Enfermedades de las Plantas , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Unión Proteica , Tenuivirus/genética , Tenuivirus/metabolismo
8.
Transbound Emerg Dis ; 69(5): e1923-e1935, 2022 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-35312168

RESUMEN

African Swine Fever (ASF) is a highly contagious viral haemorrhagic disease of swine, leading to enormous economic losses in the swine industry. However, vaccines and drugs to treat ASF have yet to be developed. African swine fever virus (ASFV) encodes more than 150 proteins, but 50% of them have unknown functions. Here, we present the crystal structure of the ASFV I73R protein at a resolution of 2.0 Å. Similar search tools based solely on amino acid sequence shows that it has no relationships to any proteins of known function. Interestingly, the overall structure of the I73R protein shares a winged helix-turn-helix fold, structural similarity with the Z-DNA binding domain (Zα). In accordance with this result, the I73R is capable of binding to a CpG repeats DNA duplex, which has a high propensity for forming Z-DNA during the DNA binding assays. In addition, the I73R protein was shown to be expressed at both early and late stages of ASFV post-infection in PAM cells as an 8.9 kDa protein. Immunofluorescence studies revealed that the I73R protein is expressed in the nucleus at early times post-infection and gradually translocated from the nucleus to the cytoplasm. Taken together, these data indicate that the I73R could be a member of Zα family that is important in host-pathogen interaction, which paves the way for the design of inhibitors to target this severe pathogen. Further exploring the biological role of I73R during ASFV infection in vitro and in vivo will provide new clues for development of new antiviral strategies.


Asunto(s)
Virus de la Fiebre Porcina Africana , Fiebre Porcina Africana , ADN de Forma Z , Enfermedades de los Porcinos , Virus de la Fiebre Porcina Africana/genética , Animales , Antivirales/farmacología , ADN , Proteínas de Unión al ADN/química , Proteínas de Unión al ADN/genética , Proteínas de Unión al ADN/metabolismo , Porcinos
9.
Plant J ; 103(3): 1215-1232, 2020 08.
Artículo en Inglés | MEDLINE | ID: mdl-32369638

RESUMEN

The Arabidopsis thaliana BON1 gene product is a member of the evolutionary conserved eukaryotic calcium-dependent membrane-binding protein family. The copine protein is composed of two C2 domains (C2A and C2B) followed by a vWA domain. The BON1 protein is localized on the plasma membrane, and is known to suppress the expression of immune receptor genes and to positively regulate stomatal closure. The first structure of this protein family has been determined to 2.5-Å resolution and shows the structural features of the three conserved domains C2A, C2B and vWA. The structure reveals the third Ca2+ -binding region in C2A domain is longer than classical C2 domains and a novel Ca2+ binding site in the vWA domain. The structure of BON1 bound to Mn2+ is also presented. The binding of the C2 domains to phospholipid (PSF) has been modeled and provides an insight into the lipid-binding mechanism of the copine proteins. Furthermore, the selectivity of the separate C2A and C2B domains and intact BON1 to bind to different phospholipids has been investigated, and we demonstrated that BON1 could mediate aggregation of liposomes in response to Ca2+ . These studies have formed the basis of further investigations into the important role that the copine proteins play in vivo.


Asunto(s)
Proteínas de Arabidopsis/química , Proteínas de Unión al Calcio/química , Proteínas Portadoras/metabolismo , Proteínas de la Membrana/química , Arabidopsis/genética , Arabidopsis/metabolismo , Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Calcio/metabolismo , Proteínas de Unión al Calcio/genética , Proteínas de Unión al Calcio/metabolismo , Membrana Celular/metabolismo , Liposomas/metabolismo , Manganeso/metabolismo , Proteínas de la Membrana/genética , Proteínas de la Membrana/metabolismo , Conformación Proteica , Alineación de Secuencia
10.
FEBS Lett ; 594(8): 1296-1306, 2020 04.
Artículo en Inglés | MEDLINE | ID: mdl-31898812

RESUMEN

The Teosinte branched 1/Cycloidea/Proliferating cell factor (TCP) domain is an evolutionarily conserved DNA binding domain unique to the plant kingdom. To date, the functions of TCPs have been well studied, but the three-dimensional structure of the TCP domain is lacking. Here, we have determined the crystal structure of the TCP domain from OsPCF6. The structure reveals that the TCP domain adopts three short ß-strands followed by a helix-loop-helix structure, distinct from the canonical basic helix-loop-helix structure. This folded domain shows high structural similarity to the ribbon-helix-helix (RHH) transcriptional repressors, a family of DNA binding proteins with a conserved 3D structural motif (RHH fold), indicating that TCPs could be reclassified as RHH proteins. Our work will provide insight toward a better understanding of the mechanisms underlying TCP protein function.


Asunto(s)
Oryza/química , Proteínas de Plantas/química , Sitios de Unión , Cristalografía por Rayos X , ADN de Plantas/química , ADN de Plantas/metabolismo , Secuencias Hélice-Asa-Hélice , Modelos Moleculares , Simulación del Acoplamiento Molecular , Proteínas de Plantas/metabolismo , Dominios Proteicos , Pliegue de Proteína , Homología de Secuencia de Aminoácido
11.
Front Psychol ; 10: 2612, 2019.
Artículo en Inglés | MEDLINE | ID: mdl-31849746

RESUMEN

Organizations are confronted with increasing social responsibility to contribute to environmental sustainability. Employee organizational citizenship behavior toward the environment (OCBE) is considered essential to organizational environment performance. Drawing upon the theory of psychological ownership, the study investigated the effects of empowering leadership on employee OCBE by a sample of 374 employees in China. With the use of the bootstrapping technique in SPSS 25 to test our proposed moderated mediation model, results demonstrated a positive relationship between empowering leadership and OCBE through the mechanism of employee psychological ownership. Further, we found that the indirect effect is stronger when employees hold high rather than low future time perspectives. The theoretical implications for sustainability literature and practical implications for organizations striving for environmental sustainability are discussed.

12.
FEBS J ; 286(21): 4294-4309, 2019 11.
Artículo en Inglés | MEDLINE | ID: mdl-31230405

RESUMEN

The VirB/D type IV secretion system (T4SS) plays an essential role in materials transport between host cells and pathogenic Helicobacter pylori and is considered the major pathogenic mediator of H. pylori-associated gastric disease. VirB8, an inner membrane protein that interacts with many other proteins, is a crucial component for secretory function. Here, we present a crystal structure of the periplasmic domain of CagV, the VirB8 counterpart in the H. pylori Cag-T4SS. The structure reveals a fold similar to that of other VirB8 members except for the absence of the α5 helix, a discontinuous ß1 strand, a larger angle between the α2 and α3 helices, a more hydrophobic surface groove, but exhibits a different dimer interface. Whether the dimerization occurs in solution was proved by mutagenesis, size-exclusion chromatography and cross-linking assays. Unlike the classical dimerization mode, the interface of the CagV dimer is principally formed by several hydrogen bonds, which indicates instability of dimerization. The structure here demonstrates the difference in dimerization among VirB8 homologues and indicates the considerable compositional and functional diversity of them in T4SS. DATABASE: Coordinates and structure factors have been deposited in the Protein Data Bank under accession codes 6IQT.


Asunto(s)
Infecciones por Helicobacter/microbiología , Helicobacter pylori/química , Proteínas de la Membrana/ultraestructura , Conformación Proteica , Cristalografía por Rayos X , Helicobacter pylori/patogenicidad , Helicobacter pylori/ultraestructura , Interacciones Huésped-Patógeno/genética , Humanos , Proteínas de la Membrana/química , Periplasma/química , Periplasma/ultraestructura , Unión Proteica , Pliegue de Proteína , Multimerización de Proteína/genética , Sistemas de Secreción Tipo IV/química , Sistemas de Secreción Tipo IV/genética
13.
FEBS J ; 286(14): 2809-2821, 2019 07.
Artículo en Inglés | MEDLINE | ID: mdl-30974511

RESUMEN

The phosphate starvation response 1 (PHR1) protein has a central role in mediating the response to phosphate starvation in plants. PHR1 is composed of a number of domains including a MYB domain involved with DNA binding and a coiled-coil domain proposed to be involved with dimer formation. PHR1 binds to the promoter of phosphate starvation-induced genes to control the levels of phosphate required for nutrition. Previous studies have shown that both the MYB domain and the coiled-coil domain of PHR1 are required for binding the target DNA. Here, we describe the crystal structure of the PHR1 MYB domain and two structures of its complex with the PHR1-binding DNA sequence (P1BS). Structural and isothermal titration calorimetry has been carried out showing that the MYB domain of PHR1 alone is sufficient for target DNA recognition and binding. Two copies of the PHR1 MYB domain bind to the same major groove of the P1BS DNA with few direct interactions between the individual MYB domains. In addition, the PHR1 MYB-P1BS DNA complex structures reveal amino acid residues involved in DNA recognition and binding. Mutagenesis of these residues results in lost or impaired ability of PHR1 MYB to bind to its target DNA. The results presented reveal the structural basis for DNA recognition by the PHR1 MYB domain and demonstrate that two PHR1 MYB domains attach to their P1BS DNA targeting sequence. DATABASE: Coordinates and structure factors have been deposited in the Protein Data Bank under accession codes 6J4K (PHR1 MYB), 6J4R (PHR1 MYB-R-P1BS), 6J5B (MYB-CC-R2-P1BS).


Asunto(s)
Proteínas de Arabidopsis/química , ADN/química , Factores de Transcripción/química , Sitios de Unión , Dominios Proteicos , Estructura Cuaternaria de Proteína
14.
Sheng Wu Gong Cheng Xue Bao ; 32(8): 1133-1144, 2016 Aug 25.
Artículo en Chino | MEDLINE | ID: mdl-29022314

RESUMEN

The expression of dentin sialophosphoprotein (DSPP) is the marker for cells differentiated into odontoblasts. This study attempted to analyze the DSPP promoter and build the reporter LacZ expression system driven by this promoter, which allows convenient and quick detection of odontoblast cells. First, we separated the human dental mesenchymal cells in which the expression of DSPP can be effectively induced by dexamethasone. Second, four 5' flanking regions of human DSPP gene (- 4 000-+54, -2 500-+54, -1 447-+54 and -1 027-+54) were analyzed, the results showed that the highest promoter activity lied in the -2 500-+54 region. The promoter activity is reduced when the 5' flanking region was extended from -2 500 to -4 000 bp upstream from the transcription start site; The promoter activity are also decreased when the 5' flanking regions were shorted from -2 500 to -1 447 bp and from -1 447 to -1 027 bp, indicating that potential suppresser elements are lied in the region between -4 000 and -2 500 bp and potential activator elements are lied in the region between -2 500 and -1 027 bp. Then we constructed the lentiviral report vector phDSPP-LacZ containing the -2 500-+ 54 promoter region in front of the LacZ gene. The expression of LacZ was detected using X-Gal staining in both human dental mesenchymal cells and immortalized human dental mesenchymal cells infected with phDSPP-LacZ. The phDSPP-LacZ lentiviral vector may provide a more convenient method to detect the expression of DSPP in human odontogenic differentiation, tooth development and tooth regeneration studies.


Asunto(s)
Proteínas de la Matriz Extracelular/genética , Operón Lac , Odontoblastos/citología , Fosfoproteínas/genética , Regiones Promotoras Genéticas , Sialoglicoproteínas/genética , Diferenciación Celular , Genes Reporteros , Humanos
15.
J Dent ; 43(5): 576-82, 2015 May.
Artículo en Inglés | MEDLINE | ID: mdl-25687752

RESUMEN

OBJECTIVES: Due to the rarity of human embryonic samples and limited proliferating capability of primary human dental mesenchymal cells, it is valuable to create an immortalized human dental mesenchymal cell line for studying dental mesenchymal cell differentiation and signalling pathways during dentinogenesis in humans. METHODS: In this study, dental mesenchymal cells from human molar tooth germs at 19-week gestation were isolated and immortalized with pSV40. Single cell colonies were then selected by 96-well plate dilution. The immortalized cell line was characterized using immunofluorescent microscopy, RT-PCR and Western blot for the expression of SV40 large T antigen and five genes specific for the mesenchymal stage during tooth development. The differentiation and mineralization activities of the immortalized and primary cells were compared using adipogenic and calcifying induction. RESULTS: The immortalized dental mesenchymal cell line displayed a higher proliferation rate, expressed several tooth-specific markers including Msx1, Pax9, Lhx6, Barx1, and Runx2, and maintained the ability to differentiate and form mineralized nodules. CONCLUSIONS: Our results demonstrated that the immortalized human mesenchymal cell line retained the characteristics similar to primary human dental mesenchymal cells and can be used for studying the mechanisms of human dental mesenchymal cell differentiation and signalling pathways involved in human odontogenesis.


Asunto(s)
Células Madre Mesenquimatosas/citología , Mesodermo/citología , Germen Dentario/citología , Feto Abortado/citología , Calcificación Fisiológica , Diferenciación Celular/fisiología , Línea Celular , Proliferación Celular/fisiología , Células Cultivadas , Humanos , Diente Molar/citología , Germen Dentario/metabolismo
16.
Biotechnol Appl Biochem ; 62(1): 48-54, 2015.
Artículo en Inglés | MEDLINE | ID: mdl-24825511

RESUMEN

Inducible co-expression of multiple genes is often needed in research. Here we describe a single-vector-based Tet-On inducible system for co-expression of two transgenes. The two transgenes (DsRed1 and eGFP as model genes) and reverse tetracycline-controlled transactivator were separated by internal ribosomal entry sites and 2A sequences, and their transcription was controlled by the same tetracycline responsive element. Two novel vectors with different internal ribosomal entry sites and 2A positions on the vectors were constructed. The DsRed1 and eGFP in cells transduced with both vectors are undetectable in the absence of doxycycline and can be efficiently induced in the presence of doxycycline in vitro and in vivo. These two vectors can be useful tools when regulated co-expression of two ecotopic genes is needed.


Asunto(s)
Regulación de la Expresión Génica/genética , Vectores Genéticos/genética , Lentivirus/genética , Transducción Genética/métodos , Transgenes/genética , Doxiciclina/farmacología , Virus de la Encefalomiocarditis/genética , Virus de la Fiebre Aftosa/genética , Regulación de la Expresión Génica/efectos de los fármacos , Proteínas Fluorescentes Verdes/genética , Células HEK293 , Humanos , Proteínas Luminiscentes/genética , ARN Mensajero/genética
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