RESUMEN
Aquaculture has suffered significant financial losses as a result of the infection of zoonotic Aeromonas hydrophila, which has a high level of resistance to classic antibiotics. In this study, we isolated an A. hydrophila strain B3 from diseased soft-shelled turtle (Pelodiscus sinensis), which is one of the most commercially significant freshwater farmed reptiles in East Asia, and found that A. hydrophila was its dominant pathogen. To better understand the inhibition effect and action mechanism of Chinese herbs on A. hydrophila, we conducted Chinese herbs screening and found that Lonicera japonica had a significant antibacterial effect on A. hydrophila B3. Experimental therapeutics of L. japonica on soft-shelled turtle showed that the supplement of 1% L. japonica to diet could significantly upregulate the immunity-related gene expression of soft-shelled turtle and protect soft-shelled turtle against A. hydrophila infection. Histopathological section results validated the protective effect of L. japonica. As the major effective component of L. japonica, chlorogenic acid demonstrated significant inhibitory effect on the growth of A. hydrophila with MIC at 6.4 mg/mL. The in vitro assay suggested that chlorogenic acid could inhibit the hemolysin/protease production and biofilm formation of A. hydrophila and significantly decrease the expression of quorum sensing, biofilm formation, and hemolysin-related genes in A. hydrophila. Our results showed that the Chinese herb L. japonica would be a promising candidate for the treatment of A. hydrophila infections in aquaculture, and it not only improves the immune response of aquatic animals but also inhibits the virulence factor (such as biofilm formation) expression of A. hydrophila. KEY POINTS: ⢠A. hydrophila was the dominant pathogen of the diseased soft-shelled turtle. ⢠L. japonica can protect soft-shelled turtle against A. hydrophila infection. ⢠Chlorogenic acid inhibits the growth and biofilm formation of A. hydrophila.
Asunto(s)
Lonicera , Animales , Aeromonas hydrophila/genética , Ácido Clorogénico , Proteínas Hemolisinas , Reptiles , Antibacterianos/farmacología , BiopelículasRESUMEN
AIMS AND OBJECTIVES: To examine the relationships among nursing students' perceived nursing image, caring behaviours and gender-friendliness barriers to determine whether gender-friendliness barriers affect nursing image and caring behaviour among male nursing students. BACKGROUND: Because caring is typically seen as a feminine trait, male nurses face gender-role strains in the current nursing environment. Gender-friendliness barriers may have an impact on the vital relationship between professional nursing image and caring behaviour. DESIGN: This study used a quantitative and cross-sectional research design. METHODS: Participants were 141 male students who had obtained at least 1 month of clinical practice experience. We collected data using three instruments: The Caring Assessment Report Evaluation Q-sort (CARE-Q), Gender-Friendliness Barriers in Nursing Programs (GFB-NP), and Nursing Image-as a Profession Questionnaire (NIPQ). Data were collected from August 2016-July 2017. Partial least squares structural equation modelling (PLS-SEM) with bootstrapping was used to test the hypothesis model. RESULTS: The full model results indicated a direct positive and significant path from professional nursing image to caring behaviour (ß = 0.47, 95% CI = 0.32 to 0.61, t = 6.19, p < 0.001). Gender-friendliness barriers had a direct and significant negative relationship between professional nursing image (ß = -0.31, 95% CI = -0.49 to -0.12, t = 3.17, p < 0.01) and caring behaviour (ß = -0.18, 95% CI = -0.35 to -0.02, t = 2.18, p < 0.05). In addition, the variable of student-perceived barriers to gender-friendliness was indirectly and significantly negatively related to caring behaviour (ß = -0.15, 95% CI = -0.27 to -0.05, t = 2.57, p < 0.05) through professional nursing image. CONCLUSION: Male nursing students with a higher nursing image engage in greater caring behaviour. Gender-friendliness barriers, however, decrease students' nursing image and caring behaviour. RELEVANCE TO CLINICAL PRACTICE: As applied to nursing education, the goal should be to improve male nursing students' caring behaviours and professional nursing image and decrease gender-friendliness barriers.
Asunto(s)
Empatía , Enfermeros/psicología , Estudiantes de Enfermería/psicología , Adulto , Estudios Transversales , Femenino , Humanos , Masculino , Encuestas y CuestionariosRESUMEN
AIM: This study presents a workshop on the Situation Model Nursing Education Action program, examines how to promote gender awareness and decrease barriers related to differences in gender friendliness through the implementation of this program and tracks the trends of the relevant variables. DESIGN: Pre- and post-observations and a cohort study were conducted. METHODS: A total of 58 nursing students (42 females and 16 males) were included. Nursing students were offered a gender-care intervention as two 90-minute workshops and surveys that measure gender awareness and gender friendliness were administered. RESULTS: The results concerned gender-bias awareness and gender-friendliness barriers scores across the four weeks of the program intervention, with the generalized estimated difference score compared with the Week 1 baseline. Mean student scores showed that gender-bias awareness was significantly decreased at Week 2 (p < .001) and Week 4 (p < .001) and that mean gender-friendliness barrier scores significantly declined at Week 4 (p < .001). CONCLUSIONS: The findings indicate that a faculty professional learning community workshop that facilitates intentional behavioral change can help faculty to become aware of gender bias, which can improve students' gender awareness through clinical case discussion and lead to a decline in barriers to students' gender friendliness.
Asunto(s)
Educación en Enfermería , Estudiantes de Enfermería , Concienciación , Estudios de Cohortes , Femenino , Humanos , Masculino , Sexismo , Encuestas y CuestionariosRESUMEN
PURPOSE: Phosphorylation is an important post-translational modification for the cellular regulation of various biosignaling pathways. We have identified in vivo phosphorylation sites of various lens proteins including especially the major structural proteins of the crystallin family from porcine eye lenses by means of two-dimensional gel electrophoresis (2-DE) or immobilized metal affinity chromatography (IMAC) followed by liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS). METHODS: For the identification of phosphorylated residues in various lens proteins of porcine lens extracts, we have adapted two complementary proteomic approaches, i.e., pre-fractionation of protein samples with 2-DE or enrichment of phosphopeptides with IMAC followed by LC-MS/MS analysis and database search. The results were compared and validated with those in phosphoproteomics databases. RESULTS: Two subunits of alpha-crystallin, alphaA-crystallin and alphaB-crystallin, as well as other lens crystallins and non-crystallin cellular proteins, such as beta-enolase, heat shock protein beta-1 (HSP27), and glucose-6-phosphate isomerase (GPI) were found to be phosphorylated in vivo at specific sites. Moreover, alphaA- and alphaB-crystallins were found to be the most abundantly phosphorylated proteins in porcine lenses, being extensively phosphorylated on serine or threonine, but not on tyrosine residues. CONCLUSIONS: The complementary gel-based and gel-free proteomic strategies have been compared and evaluated for the study of crystallin phosphorylation from whole tissue extracts of porcine eye lenses. Technically, the IMAC method facilitates direct site-specific identification of phosphorylation residues in lens proteins, which does not necessitate the pre-MS/MS 2-DE separation of protein samples. Moreover, the improved strategy using gel-free phosphoproteomics analysis affords a more effective and simplistic method for the determination of in vivo phosphorylation sites than the conventional 2-DE pre-separation of protein mixture. This study should form a firm basis for the comprehensive analysis of post-translational modification of lens proteins in terms of aging or various diseased states.
Asunto(s)
Cristalinas/química , Cristalinas/metabolismo , Cristalino/metabolismo , Fosfoproteínas/química , Fosfoproteínas/metabolismo , Proteoma/metabolismo , Proteómica/métodos , Secuencia de Aminoácidos , Animales , Cromatografía Liquida , Electroforesis en Gel Bidimensional , Glucosa-6-Fosfato Isomerasa/química , Glucosa-6-Fosfato Isomerasa/metabolismo , Proteínas de Choque Térmico HSP27/química , Proteínas de Choque Térmico HSP27/metabolismo , Datos de Secuencia Molecular , Fosfopiruvato Hidratasa/química , Fosfopiruvato Hidratasa/metabolismo , Fosforilación , Sus scrofa , Espectrometría de Masas en Tándem , Extractos de Tejidos , Cadena B de alfa-Cristalina/química , Cadena B de alfa-Cristalina/metabolismoRESUMEN
AIM: We explored the impact of gender role orientation (masculinity and femininity) on student nurses' caring behaviour and critical thinking. BACKGROUND: Caring and critical thinking are at the core of professional nursing education. Previous studies revealed inconsistent findings regarding the impact of gender roles on caring behaviour and critical thinking. DESIGN AND METHODS: We employed a quantitative correlational study. Nursing students (N = 449; female = 310, male = 139) who had at least had one month of clinical practice experience were recruited from four universities in Taiwan. Students' ages ranged from 19 to 29 years (Mean age = 21.24 years, SD = 1.28). Data were collected from August 2016 to July 2017, using three questionnaires: Taiwan Critical Thinking Disposition Inventory (CTDI), Caring Assessment Report Evaluation Q-sort Scale (CARE-Q), and Bem Sex Role Inventory (BSRI). Partial least squares structural equation modelling and generalized linear models were conducted to test the research model and hypotheses. RESULTS: Findings indicated that students who reported higher caring and masculinity presented greater critical thinking (ß = .37 and ß = 0.24, respectively; ps < .001). Students' gender, age, femininity, or clinical practice experience, however, were not significantly associated with critical thinking (ß = -0.01, ß = 0.09, ß = .10, and ß = 0.01, respectively; ps > .05). In addition, students who reported higher masculinity and femininity presented greater caring behaviour (ß = .22 and ß = 0.38, respectively; ps < .001). Students' gender, age or clinical practice experience were not significantly associated with caring behaviour (ß = .04, ß = .03, and ß = -0.05, respectively; ps > 0.05). The findings confirmed a direct influence of caring and masculinity on critical thinking. Masculinity indirectly affected critical thinking via caring behaviour. Caring and masculinity accounted for 34.4% of the variance in critical thinking, and masculinity and femininity accounted for 29.1% of the variance in caring behaviour. CONCLUSION: Our study confirms the effect of age, gender role, and caring behaviour on critical thinking. We recommend that the cultivation of nursing care behaviour focus on students' gender role orientation. In addition, clinical nurse educators, when working with male students on patient caring, should consider their gender role orientation and support male nursing students' ways of presenting caring behaviours.
Asunto(s)
Empatía , Identidad de Género , Estudiantes de Enfermería/psicología , Pensamiento , Adulto , Educación en Enfermería , Femenino , Humanos , Masculino , Relaciones Enfermero-Paciente , Taiwán , Adulto JovenRESUMEN
Multidrug resistance (MDR) is a phenomenon in which cells become resistant to structurally and mechanistically unrelated drugs, and it is one of the emerging problems in cancer therapy today. The relation between overexpression of the ABC transporter subfamily B member 1 (ABCB1/P-glycoprotein) and resistant cancers has been well characterized. In the present study, we successfully synthesized 52 novel benzodipyranone analogs and evaluated for their P-gp inhibitory activity in a P-gp transfected cell line, ABCB1/Flp-In™-293. Among these derivatives, 5a bearing on the 3-methylphenyl substituent, displayed the most potent P-gp inhibitory activity, which can enable the increase of the intracellular accumulation of P-gp substrate Calcein-AM. 5a exhibited more potency on promoted anticancer drugs cytotoxicity by reversing P-gp-mediated drug resistance in both ABCB1/Flp-In™-293 and KBvin cell lines. In particular, the compound 5a sensitized ABCB1/Flp-In™-293 cells toward paclitaxel, vincristine, and doxorubicin by 16.1, 21.0, and 1.6-fold at 10 µM, respectively. Further, 5a dramatically sensitized the resistant cell line KBvin toward paclitaxel and vincristine by 23.1 and 29.7-fold at 10 µM, respectively. It's possible that its mechanism of MDR inhibition can restore the intracellular accumulation of drugs and eventually chemosensitize cancer cells to anticancer drugs and reduce ABCB1 mRNA expression level.
Asunto(s)
Subfamilia B de Transportador de Casetes de Unión a ATP/antagonistas & inhibidores , Diseño de Fármacos , Resistencia a Múltiples Medicamentos/efectos de los fármacos , Resistencia a Antineoplásicos/efectos de los fármacos , Piranos/síntesis química , Piranos/farmacología , Subfamilia B de Transportador de Casetes de Unión a ATP/química , Subfamilia B de Transportador de Casetes de Unión a ATP/metabolismo , Miembro 1 de la Subfamilia B de Casetes de Unión a ATP/genética , Línea Celular Tumoral , Técnicas de Química Sintética , Regulación Neoplásica de la Expresión Génica/efectos de los fármacos , Humanos , Conformación Proteica , Piranos/química , ARN Mensajero/genética , ARN Mensajero/metabolismoRESUMEN
It has been observed that a modified peptide and its non-modified counterpart, when analyzed with reverse phase liquid chromatography, usually share a very similar elution property [1-3]. Inasmuch as this property is common to many different types of protein modifications, we propose an informatics-based approach, featuring the generation of segmental average mass spectra ((sa)MS), that is capable of locating different types of modified peptides in two-dimensional liquid chromatography-mass spectrometric (LC-MS) data collected for regular protease digests from proteins in gels or solutions. To enable the localization of these peptides in the LC-MS map, we have implemented a set of computer programs, or the (sa)MS package, that perform the needed functions, including generating a complete set of segmental average mass spectra, compiling the peptide inventory from the Sequest/TurboSequest results, searching modified peptide candidates and annotating a tandem mass spectrum for final verification. Using ROCK2 as an example, our programs were applied to identify multiple types of modified peptides, such as phosphorylated and hexosylated ones, which particularly include those peptides that could have been ignored due to their peculiar fragmentation patterns and consequent low search scores. Hence, we demonstrate that, when complemented with peptide search algorithms, our approach and the entailed computer programs can add the sequence information needed for bolstering the confidence of data interpretation by the present analytical platforms and facilitate the mining of protein modification information out of complicated LC-MS/MS data.
Asunto(s)
Cromatografía Líquida de Alta Presión , Espectrometría de Masas en Tándem , Quinasas Asociadas a rho/química , Secuencia de Aminoácidos , Anticuerpos/inmunología , Células HEK293 , Humanos , Inmunoprecipitación , Datos de Secuencia Molecular , Fosfopéptidos/análisis , Procesamiento Proteico-Postraduccional , Quinasas Asociadas a rho/inmunología , Quinasas Asociadas a rho/metabolismoRESUMEN
The expression of heat shock proteins (Hsps) induced by nonlethal heat treatment confers acquired thermotolerance (AT) to organisms against subsequent challenges of otherwise lethal temperature. After the stress signal is removed, AT gradually decays, with decreased Hsps during recovery. AT of sufficient duration is critical for sessile organisms such as plants to survive repeated heat stress in their environment, but little is known regarding its regulation. To identify potential regulatory components, we took a reverse genetics approach by screening for Arabidopsis (Arabidopsis thaliana) T-DNA insertion mutants that show decreased thermotolerance after a long recovery (2 d) under nonstress conditions following an acclimation heat treatment. Among the tested mutants corresponding to 48 heat-induced genes, only the heat shock transcription factor HsfA2 knockout mutant showed an obvious phenotype. Following pretreatment at 37 degrees C, the mutant line was more sensitive to severe heat stress than the wild type after long but not short recovery periods, and this could be complemented by the introduction of a wild-type copy of the HsfA2 gene. Quantitative hypocotyl elongation assay also revealed that AT decayed faster in the absence of HsfA2. Significant reduction in the transcript levels of several highly heat-inducible genes was observed in HsfA2 knockout plants after 4 h recovery or 2 h prolonged heat stress. Immunoblot analysis showed that Hsa32 and class I small Hsp were less abundant in the mutant than in the wild type after long recovery. Our results suggest that HsfA2 as a heat-inducible transactivator sustains the expression of Hsp genes and extends the duration of AT in Arabidopsis.
Asunto(s)
Aclimatación/genética , Proteínas de Arabidopsis/fisiología , Arabidopsis/fisiología , Proteínas de Unión al ADN/fisiología , Proteínas de Choque Térmico/fisiología , Calor , Proteínas de Plantas/fisiología , Factores de Transcripción/fisiología , Arabidopsis/genética , Arabidopsis/metabolismo , Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Proteínas de Unión al ADN/genética , Proteínas de Unión al ADN/metabolismo , Regulación de la Expresión Génica de las Plantas , Factores de Transcripción del Choque Térmico , Proteínas de Choque Térmico/genética , Proteínas de Choque Térmico/metabolismo , Hipocótilo/genética , Hipocótilo/crecimiento & desarrollo , Hipocótilo/fisiología , Immunoblotting , Mutagénesis Insercional , Fenotipo , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , ARN Mensajero/metabolismo , Factores de Transcripción/genética , Factores de Transcripción/metabolismoRESUMEN
Plants and animals share similar mechanisms in the heat shock (HS) response, such as synthesis of the conserved HS proteins (Hsps). However, because plants are confined to a growing environment, in general they require unique features to cope with heat stress. Here, we report on the analysis of the function of a novel Hsp, heat-stress-associated 32-kD protein (Hsa32), which is highly conserved in land plants but absent in most other organisms. The gene responds to HS at the transcriptional level in moss (Physcomitrella patens), Arabidopsis (Arabidopsis thaliana), and rice (Oryza sativa). Like other Hsps, Hsa32 protein accumulates greatly in Arabidopsis seedlings after HS treatment. Disruption of Hsa32 by T-DNA insertion does not affect growth and development under normal conditions. However, the acquired thermotolerance in the knockout line was compromised following a long recovery period (>24 h) after acclimation HS treatment, when a severe HS challenge killed the mutant but not the wild-type plants, but no significant difference was observed if they were challenged within a short recovery period. Quantitative hypocotyl elongation assay also revealed that thermotolerance decayed faster in the absence of Hsa32 after a long recovery. Similar results were obtained in Arabidopsis transgenic plants with Hsa32 expression suppressed by RNA interference. Microarray analysis of the knockout mutant indicates that only the expression of Hsa32 was significantly altered in HS response. Taken together, our results suggest that Hsa32 is required not for induction but rather maintenance of acquired thermotolerance, a feature that could be important to plants.