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Stacking two-dimensional van der Waals (vdW) materials rotated with respect to each other show versatility for studying exotic quantum phenomena. In particular, anisotropic layered materials have great potential for such twistronics applications, providing high tunability. Here, we report anisotropic superconducting order parameters in twisted Bi2Sr2CaCu2O8+x (Bi-2212) vdW junctions with an atomically clean vdW interface, achieved using the microcleave-and-stack technique. The vdW junctions with twist angles of 0° and 90° showed the maximum Josephson coupling, comparable to that of intrinsic Josephson junctions. As the twist angle approaches 45°, Josephson coupling is suppressed, and eventually disappears at 45°. The observed twist angle dependence of the Josephson coupling can be explained quantitatively by theoretical calculation with the d-wave superconducting order parameter of Bi-2212 and finite tunneling incoherence of the junction. Our results revealed the anisotropic nature of Bi-2212 and provided a novel fabrication technique for vdW-based twistronics platforms compatible with air-sensitive vdW materials.
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Eriobotrya japonica leaf is included in the Chinese Pharmacopoeia, and is widely used as a medicinal material in traditional medicine. The present study investigated the potential genotoxic effects of E. japonica leaf extract (EJE) using three standard battery systems. Genotoxicity tests were conducted following the test guidelines of the Organisation for Economic Cooperation and Development (OECD) and Ministry of Food and Drug Safety (MFDS), with application of Good Laboratory Practice. The bacterial reverse mutation test was conducted using the pre-incubation method in the presence or absence of the metabolic activation system (S9 mixture). The in vitro chromosome aberration test was performed using cultured Chinese hamster lung cell line in the presence or absence of the S9 mixture. The in vivo micronucleus test was performed using ICR mice. The bacterial reverse mutation test with Salmonella typhimurium strains TA98, TA100, TA1535, and TA1537 and Escherichia coli strain WP2uvrA showed that EJE did not induce gene mutations at any dose level in all the strains tested. EJE also did not show any chromosomal aberrations in the in vitro chromosomal aberration test and in the in vivo micronucleus test. These results showed that EJE did not induce mutagenicity or clastogenicity in either in vitro or in vivo systems.
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Eriobotrya/toxicidade , Extratos Vegetais/toxicidade , Folhas de Planta/toxicidade , Animais , Linhagem Celular , Aberrações Cromossômicas/efeitos dos fármacos , Cricetinae , Cricetulus , Relação Dose-Resposta a Droga , Escherichia coli/efeitos dos fármacos , Masculino , Camundongos , Camundongos Endogâmicos ICR , Testes para Micronúcleos/métodos , Testes de Mutagenicidade/métodos , Mutagênicos/toxicidade , Mutação/efeitos dos fármacos , Salmonella typhimurium/efeitos dos fármacosRESUMO
A layered two-dimensional superconducting material 2H-NbSe2 is used to build a van der Waals heterostructure, where a proximity-coupled superconducting order can be induced in the interfacing materials. Vertically stacked NbSe2-graphene-NbSe2 is fabricated using van der Waals interlayer coupling, producing defect-free contacts with a high interfacial transparency. The atomically thin graphene layer allows the formation of a highly coherent proximity Josephson coupling between the two NbSe2 flakes. The temperature dependence of the junction critical current (Ic) reveals short and ballistic Josephson coupling characteristics that agree with theoretical prediction. The strong Josephson coupling is confirmed by a large junction critical current density of 1.6 × 104 A/cm2, multiple Andreev reflections in the subgap structure of the differential conductance, and a magnetic-field modulation of Ic. This is the first demonstration of strongly proximity-coupled Josephson junctions with extremely clean interfaces in a dry-transfer-stacked van der Waals heterostructure.
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PURPOSE: The purpose of the present study was to describe the use of a novel hybrid surgical technique-arthroscopic-assisted plate fixation-and evaluate its clinical and anatomical outcomes in the management of large, displaced greater tuberosity (GT) fractures with comminution. METHODS: From 2009 to 2011, this novel technique was performed in 11 patients [2 men and 9 women; median age, 64 years (range 41-83 years)] with large, comminuted GT fractures, with fragment displacements of >5 mm. The preoperative mean posterior and superior migration of the fractured fragment, as measured on computed tomography (CT), was 19.5 and 5.5 mm, respectively. Two patients had shoulder fracture-dislocation, and three had associated undisplaced surgical neck fracture. The mean duration between injury and surgery was 4 days. The mean follow-up duration was 26 months. RESULTS: At the final follow-up, the mean postoperative ASES, UCLA and SST scores were 84, 29, and 8, respectively. The mean range of motion was as follows: forward flexion, 138°; abduction, 135°; external rotation at the side, 19°; and internal rotation, up to the L2 level. The mean posterior and superior displacements of fracture fragments on postoperative CT scan [0.7 ± 0.8 mm (range 0-2.1 mm) and 2.8 ± 0.5 mm (range 3.4-5.3 mm), respectively] were significantly improved (p < 0.05). On arthroscopy, a partial articular-side supraspinatus tendon avulsion lesion was identified in 10 of 11 patients (91 %), and 1 of these patients had a partial tear of the biceps and 1 had a partial subscapularis tear, respectively (9 %). Intraoperatively, 1 anchor pullout and 1 anchor protrusion through the humeral head were noted and corrected. Postoperatively, the loss of reduction in the fracture fragment was noted in 1 patient at 4 weeks, after corrective reduction and fixation surgery. CONCLUSIONS: The novel arthroscopic-assisted anatomical plate fixation technique was found to be effective in reducing large-sized, displaced, comminuted GT fractures and in allowing concurrent management of intra-articular pathologies and early functional rehabilitation. Compared with the conventional plate fixation or arthroscopic suture anchor fixation technique, arthroscopic-assisted plate fixation enabled accurate restoration of the medial footprint of the GT fracture and provided an effective buttress to the large-sized GT fracture fragments. LEVEL OF EVIDENCE: Retrospective clinical study, Level IV.
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Artroscopia/métodos , Placas Ósseas , Fixação Interna de Fraturas/métodos , Fraturas Cominutivas/cirurgia , Fraturas do Ombro/cirurgia , Âncoras de Sutura , Adulto , Idoso , Idoso de 80 Anos ou mais , Feminino , Fraturas Cominutivas/diagnóstico por imagem , Humanos , Masculino , Pessoa de Meia-Idade , Amplitude de Movimento Articular , Recuperação de Função Fisiológica , Estudos Retrospectivos , Fraturas do Ombro/diagnóstico por imagem , Técnicas de Sutura , Tomografia Computadorizada por Raios X , Resultado do TratamentoRESUMO
This study compared the overall postoperative recovery of patients who underwent total intravenous anesthesia with remimazolam or propofol, using the Quality of Recovery-15 questionnaire (QoR-15). Seventy-two patients who underwent spine surgery with intraoperative neurophysiological monitoring (IONM) were randomly categorized into the remimazolam group (group R) or propofol group (group P). On the first postoperative day, the QoR-15 scores for groups P and R were 114 and 112, respectively, indicating no significant difference (p = 0.691). Similarly, group-time interaction effects on QoR-15 scores were not significantly different. In the post-anesthesia care unit, the pain intensity at rest was notably higher in group P than in group R (3.0 [0.0] vs. 2.8 [0.5], respectively, p = 0.009). Although the intraoperative consumption of remifentanil was higher in group R (1452.4 µg vs. 2066.8 µg, respectively, p < 0.001), the intraoperative use of vasopressors was lower in group R (1705.6 µg vs. 286.1 µg, respectively, p < 0.001) compared to group P. Group R exhibited significantly lower variability in mean blood pressure over time compared to group P. Remimazolam was viewed as a promising intravenous agent for general anesthesia, showing potential to replace propofol in spine surgery with IONM, considering both recovery quality and intraoperative hemodynamic stability.
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BACKGROUND: Myocardial injury after non-cardiac surgery (MINS) has been known to be associated with mortality in various surgical patients; however, its prognostic role in abdominal aortic aneurysm (AAA) open repair remains underexplored. This study aimed to investigate the role of MINS as a predictor of mortality in patients who underwent AAA open repair. METHODS: This retrospective study investigated 352 patients who underwent open repair for non-ruptured AAA. The predictors of 30-day and 1-year mortalities were investigated using logistic regression analysis. RESULTS: MINS was diagnosed in 41% of the patients after AAA open repair in this study. MINS was an independent risk factor of 30-day mortality (odds ratio [OR]: 10.440, 95% confidence interval [CI]: 1.278-85.274, p = 0.029) and 1-year mortality (OR: 5.189, 95% CI: 1.357-19.844, p = 0.016). Kaplan-Meier survival curves demonstrated significantly lower overall survival rates in patients with MINS compared to those without MINS (p = 0.003). CONCLUSION: This study revealed that MINS is a common complication after AAA open repair and is an independent risk factor of 30-day and 1-year mortalities. Patients with MINS have lower overall survival rates than those without MINS.
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INTRODUCTION: The present study aimed to compare the correlation and agreement of epidural depth estimation using ultrasound in the paramedian sagittal oblique (PSO) versus the transverse median (TM) plane relative to the actual epidural depth observed during midline epidural punctures in children with scoliosis. METHODS: In this prospective observational study, we enrolled 55 children aged 3-14 years with thoracolumbar scoliosis (Cobb angle >10°) requesting postoperative epidural analgesia. Ultrasound imaging was performed to estimate the distance from the skin to the epidural space in the bilateral PSO and TM planes. An anesthesiologist performed midline epidural puncture and measured the actual epidural depth from the skin to the epidural space. The correlation and degree of agreement between the ultrasound-estimated and actual epidural depths were investigated using Pearson's and concordance correlation coefficients. The image quality of the ligamentum flavum and posterior dura mater was compared. RESULTS: In the PSO view, where the larger of the two measurements from both sides was used, both Pearson's and concordance correlation coefficients for comparing the actual epidural and ultrasound-estimated depths were significantly higher than those in the TM view (0.964 vs 0.930, p value=0.002; 0.952 vs 0.892, p value=0.004, respectively). The ligamentum flavum-posterior dura mater unit was more easily distinguished in the PSO view than in the TM view (72.7% vs 38.2%, p value<0.001). CONCLUSIONS: The PSO view can be a reliable guide to facilitate epidural puncture in children with scoliosis with better visualization. TRIAL REGISTRATION NUMBER: NCT04877964.
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We evaluated the potential genotoxic effects of the nutrient supplement SUNACTIVE Zn-P240 in vitro and in vivo. Genotoxicity tests were performed at the Korea Testing and Research Institute, a GLP certification institution. A bacterial reverse mutation test was performed using the pre-incubation method, while the in vitro chromosome aberration test was performed using a cultured Chinese hamster lung cell line in the presence or absence of metabolic activation. The in vivo micronucleus test was performed using ICR mice. The bacterial reverse mutation test revealed that SUNACTIVE Zn-P240 did not induce genetic mutations at the tested doses in Salmonella typhimurium (TA98, TA100, TA1535, and TA1537) and Escherichia coli (WP2uvrA) tester strains. Meanwhile, the results of the in vitro chromosomal aberration and in vivo micronucleus tests revealed that SUNACTIVE Zn-P240 did not induce chromosomal aberrations. These results suggest that SUNACTIVE Zn-P240 did not exhibit mutagenic or clastogenic properties in vitro and in vivo.
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Granulocyte-colony stimulating factor (G-CSF), a pleiotropic cytokine, belongs to the hematopoietic growth factor family. Recent studies have reported that G-CSF is a predictive biomarker of oocyte and embryo developmental competence in humans. The aim of our study was to determine whether CSF3 and its receptor (CSF3R) were expressed in porcine maternal reproductive tissues (oviduct and uterus), cumulus cells, and embryos and to investigate the effects of human recombinant G-CSF (hrG-CSF) supplementation during in vitro culture (IVC) on the developmental competence of pre-implantation embryos. To do this, we first performed reverse-transcription polymerase chain reaction (RT-PCR). Second, we performed parthenogenetic activation (PA), in vitro fertilization (IVF), and somatic cell nuclear transfer (SCNT) to evaluate the embryonic developmental potential after hrG-CSF supplementation based on various concentrations (0 ng/mL, 10 ng/mL, 50 ng/mL, and 100 ng/mL) and durations (Un-treated, Days 0-3, Days 4-7, and Days 0-7) of IVC. Finally, we examined transcriptional levels of several marker genes in blastocysts. The results of our study showed that CSF3 transcript was present in all samples we assessed. CSF3-R was also detected, except in cumulus cells and blastocysts from PA. Furthermore, 10 ng/mL and Days 0-7 were the optimal concentration and duration for the viability of in vitro embryonic development, especially for SCNT-derived embryos. The rate of blastocyst formation and the total cell number of blastocysts were significantly enhanced, while the number and index of apoptotic nuclei were significantly decreased in optimal condition groups compared to others. Moreover, the transcriptional levels of anti-apoptotis- (BCL2), proliferation- (PCNA), and pluripotency- (POU5F1) related genes were dramatically upregulated. In conclusion, for the first time, we demonstrated that CSF3 and CSF3R were expressed in porcine reproductive organs, cells, and embryos. Additionally, we determined that hrG-CSF treatment improved porcine embryonic development capacity in vitro.
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Desenvolvimento Embrionário/efeitos dos fármacos , Fator Estimulador de Colônias de Granulócitos/farmacologia , Proteínas Recombinantes/farmacologia , Animais , Apoptose/efeitos dos fármacos , Blastocisto/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Células do Cúmulo/efeitos dos fármacos , Técnicas de Cultura Embrionária/métodos , Feminino , Fertilização in vitro/métodos , Regulação da Expressão Gênica no Desenvolvimento/efeitos dos fármacos , Humanos , Técnicas de Transferência Nuclear , Oócitos/efeitos dos fármacos , Gravidez , SuínosRESUMO
In the present study, we investigated the effect of melatonin on the preimplantation development of porcine parthenogenetic and somatic cell nuclear transfer (SCNT) embryos. Parthenogenetic embryos were cultured in mNCSU-23 supplemented with various concentrations of melatonin for 7 days. The results revealed that 100 pM was the optimal concentration, which resulted in significantly increased cleavage and blastocyst formation rates. Additionally, 100 pM melatonin provided the highest increase in total cell number of blastocysts. Therefore, the subsequent experiments were performed with 100 pM melatonin. ROS level in 2-8 cell stage embryos in the presence or absence of melatonin was evaluated. Embryos cultured with melatonin showed significantly decreased ROS. Blastocysts cultured with melatonin for 7 days were analyzed by the TUNEL assay. It was observed that melatonin not only increased (P < 0.05) the total cell number but also decreased (P < 0.05) the rate of apoptotic nuclei. Blastocysts cultured with melatonin were assessed for the expression of apoptosis-related genes Bcl-xl and Bax, and of pluripotency marker gene Oct-4 by real-time quantitative PCR. Analysis of data showed that the expression of Bcl-xl was higher (1.7-fold) compared to the control while the expression of Bax was significantly decreased relative to the control (0.7-fold) (P < 0.05). Moreover, the expression of Oct-4 was 1.7-fold higher than the control. These results indicated that melatonin had beneficial effects on the development of porcine parthenogenetic embryos. Based on the findings of parthenogenetic embryos, we investigated the effect of melatonin on the development of porcine SCNT embryos. The results also demonstrated increased cleavage and blastocyst formation rates, and the total cell numbers in blastocysts were significantly higher when the embryos were cultured with melatonin. Therefore, these data suggested that melatonin may have important implications for improving porcine preimplantation SCNT embryo development.
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Apoptose/efeitos dos fármacos , Blastocisto/fisiologia , Desenvolvimento Embrionário/efeitos dos fármacos , Melatonina/farmacologia , Partenogênese/fisiologia , Animais , Blastocisto/citologia , Blastocisto/efeitos dos fármacos , Feminino , Marcação In Situ das Extremidades Cortadas , Técnicas de Transferência Nuclear , Partenogênese/efeitos dos fármacos , Reação em Cadeia da Polimerase , Gravidez , SuínosRESUMO
BACKGROUND AND AIM: The extrastriate cortex is a visual processing structure beyond the striate cortex. This region contains cells that differ in selectivity for different features of stimuli such as color, motion and stereopsis. We studied stereopsis and color vision in patients with right extrastriate cerebral lesions. METHODS: Patients with right extrastriate cerebral lesions determined by MRI were recruited among consecutive inpatients. Patients with cognitive impairment (Mini Mental Status Examination score <24) were excluded. An ophthalmologist performed cortical visual testing: visual acuity, stereopsis, color vision and strabismus within 1 week after the insult. RESULTS: Cortical visual testing was performed in 12 patients. Among the patients studied, 11 demonstrated abnormalities: reduced stereoacuity (8/12), color vision abnormalities (7/12). Seven patients showed complex cortical visual abnormalities. However, only 2 patients complained of subjective clinical symptoms. The MRI lesions in patients with disturbance of stereopsis were located in the dorsal occipitoparietal area and the ventral occipitotemporal area. Color vision abnormalities corresponded to lesions in the ventral occipitotemporal area. CONCLUSIONS: Patients with lesions in the right extrastriate cortex tend to have disturbance of stereopsis and color vision abnormalities, even in the absence of visual complaints.
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Lesões Encefálicas/complicações , Defeitos da Visão Cromática/etiologia , Percepção de Profundidade/fisiologia , Lobo Occipital/lesões , Transtornos da Percepção/etiologia , Adulto , Idoso , Lesões Encefálicas/patologia , Defeitos da Visão Cromática/patologia , Feminino , Humanos , Imageamento por Ressonância Magnética , Masculino , Pessoa de Meia-Idade , Lobo Occipital/patologia , Transtornos da Percepção/patologiaRESUMO
Burkholderia gladioli is a causal agent of bacterial panicle blight and sheath/grain browning in rice in many countries. Many strains produce the yellow pigment toxoflavin, which is highly toxic to plants, fungi, animals and microorganisms. Although there have been several studies on the toxoflavin biosynthesis system of B. glumae, it is still unclear how B. gladioli activates toxoflavin biosynthesis. In this study, we explored the genomic organization of the toxoflavin system of B. gladioli and its biological functions using comparative genomic analysis between toxoflavin-producing strains (B. glumaeâ BGR1 and B. gladioliâ BSR3) and a strain not producing toxoflavin (B. gladioliâ KACC11889). The latter exhibits normal physiological characteristics similar to other B. gladioli strains. Burkholderia gladioliâ KACC11889 possesses all the genes involved in toxoflavin biosynthesis, but lacks the quorum-sensing (QS) system that functions as an on/off switch for toxoflavin biosynthesis. These data suggest that B. gladioli has evolved to use the QS signalling cascade of toxoflavin production (TofI/TofR of QS â ToxJ or ToxR â tox operons) similar to that in B. glumae. However, some strains may have evolved to eliminate toxoflavin production through deletion of the QS genes. In addition, we demonstrate that the toxoflavin biosynthetic system enhances the virulence of B. gladioli. These findings provide another line of evidence supporting the differential regulation of the toxoflavin system in Burkholderia strains.
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Burkholderia gladioli/metabolismo , Burkholderia gladioli/patogenicidade , Pirimidinonas/metabolismo , Triazinas/metabolismo , Vias Biossintéticas/genética , Burkholderia gladioli/genética , Genes Bacterianos , Teste de Complementação Genética , Movimento , Cebolas/microbiologia , Óperon/genética , Reprodutibilidade dos Testes , Virulência/genéticaRESUMO
Tomato yellow leaf curl virus (TYLCV), a member of the genus Begomovirus, is responsible for one of the most devastating viral diseases in tomato-growing countries and is becoming a serious problem in many subtropical and tropical countries. The climate in Korea is getting warmer and developing subtropical features in response to global warming. These changes are being accompanied by TYLCV, which is now becoming a large problem in the Korean tomato industry. The most effective way to reduce damage caused by TYLCV is to breed resistant varieties of tomatoes. To accomplish this, it is necessary to establish a simple inoculation technique for the efficient evaluation of resistance to TYLCV. Here, we present the rolling circle amplification (RCA) method, which employs a bacteriophage using phi-29 DNA polymerase for construction of infectious TYLCV clones. The RCA method is simple, does not require sequence information for cloning, and is less expensive and time consuming than conventional PCR based-methods. Furthermore, RCA-based construction of an infectious clone can be very useful to other emerging and unknown geminiviruses in Korea.
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Burkholderia glumae is a causal agent of rice grain and sheath rot. Similar to other phytopathogens, B. glumae adapts well to the host environment and controls its biology to induce diseases in the host plant; however, its molecular mechanisms are not yet fully understood. To gain a better understating of the actual physiological changes that occur in B. glumae during infection, we analyzed B. glumae transcriptome from infected rice tissues using an RNA-seq technique. To accomplish this, we analyzed differentially expressed genes (DEGs) and identified 2653 transcripts that were significantly altered. We then performed KEGG pathway and module enrichment of the DEGs. Interestingly, most genes involved bacterial chemotaxis-mediated motility, ascorbate and trehalose metabolisms, and sugar transporters including l-arabinose and d-xylose were found to be highly enriched. The in vivo transcriptional profiling of pathogenic B. glumae will facilitate elucidation of unknown plant-pathogenic bacteria interactions, as well as the overall infection processes.
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Burkholderia/metabolismo , Oryza/microbiologia , Transdução de Sinais , Transcriptoma , Burkholderia/genética , Burkholderia/patogenicidade , Genes Bacterianos , Oryza/genética , Reação em Cadeia da Polimerase em Tempo Real , Análise de Sequência de RNARESUMO
PURPOSE: The widespread use of nanoparticles (NPs) in industrial and biomedical applications has prompted growing concern regarding their potential toxicity and impact on human health. This study therefore investigated the subchronic, systemic oral toxicity and no-observed-adverse-effect level (NOAEL) of 20 nm, negatively charged zinc oxide (ZnO(SM20(-))) NPs in Sprague Dawley rats for 90 days. METHODS: The high-dose NP level was set at 500 mg/kg of bodyweight, and the mid- and low-dose levels were set at 250 and 125 mg/kg, respectively. The rats were observed during a 14-day recovery period after the last NP administration for the persistence or reduction of any adverse effects. Toxicokinetic and distribution studies were also conducted to determine the systemic distribution of the NPs. RESULTS: No rats died during the test period. However, ZnO(SM20(-)) NPs (500 mg/kg) induced changes in the levels of anemia-related factors, prompted acinar cell apoptosis and ductular hyperplasia, stimulated periductular lymphoid cell infiltration and excessive salivation, and increased the numbers of regenerative acinar cells in the pancreas. In addition, stomach lesions were seen at 125, 250, and 500 mg/kg, and retinal atrophy was observed at 250 and 500 mg/kg. The Zn concentration was dose-dependently increased in the liver, kidney, intestines, and plasma, but not in other organs investigated. CONCLUSION: A ZnO(SM20(-)) NP NOAEL could not be established from the current results, but the lowest-observed-adverse-effect level was 125 mg/kg. Furthermore, the NPs were associated with a number of undesirable systemic actions. Thus, their use in humans must be approached with caution.
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Nanopartículas Metálicas , Óxido de Zinco , Administração Oral , Animais , Ânions , Apoptose/efeitos dos fármacos , Nanopartículas Metálicas/administração & dosagem , Nanopartículas Metálicas/química , Nanopartículas Metálicas/toxicidade , Pâncreas/efeitos dos fármacos , Tamanho da Partícula , Ratos , Ratos Sprague-Dawley , Distribuição Tecidual , Testes de Toxicidade Subcrônica , Óxido de Zinco/administração & dosagem , Óxido de Zinco/química , Óxido de Zinco/farmacocinética , Óxido de Zinco/toxicidadeRESUMO
Burkholderia glumae causes rice grain rot and sheath rot by producing toxoflavin, the expression of which is regulated by quorum sensing (QS). The QS systems of B. glumae rely on N-octanoyl homoserine lactone, synthesized by TofI and its cognate receptor TofR, to activate the genes for toxoflavin biosynthesis and an IclR-type transcriptional regulator gene, qsmR. To understand genome-wide transcriptional profiling of QS signaling, we employed RNAseq of the wild-type B. glumae BGR1 with QS-defective mutant, BGS2 (BGR1 tofI::Ω) and QS-dependent transcriptional regulator mutant, BGS9 (BGR1 qsmR::Ω). A comparison of gene expression profiling among the wild-type BGR1 and the two mutants before and after QS onset as well as gene ontology (GO) enrichment analysis from differential expressed genes (DEGs) revealed that genes involved in motility were highly enriched in TofI-dependent DEGs, whereas genes for transport and DNA polymerase were highly enriched in QsmR-dependent DEGs. Further, a combination of pathways with these DEGs and phenotype analysis of mutants pointed to a couple of metabolic processes, which are dependent on QS in B. glumae, that were directly or indirectly related with bacterial motility. The consistency of observed bacterial phenotypes with GOs or metabolic pathways in QS-regulated genes implied that integration RNAseq with GO enrichment or pathways would be useful to study bacterial physiology and phenotypes.
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Human interferon α (IFN-α) and erythropoietin (EPO) have been used for a variety of purposes in clinical medicine. Human IFN-α has been used to treat several types of viral infection and cancer, as well as renal anemia, via stimulation of erythrocyte formation in the bone marrow. Transgenic cattle are excellent candidates for pharmaceutical production for humans due to their ability to produce recombinant proteins in milk. The purpose of the present study was to generate bovine transgenic fibroblasts capable of producing recombinant human IFN-α and EPO proteins in transgenic cattle milk. First, we analyzed the promoter activities of various bovine milk protein genes in HC11 mouse mammary epithelial cells. The bovine milk protein gene promoters were cloned into the Luc gene in a promoter-less pGL3-Basic vector. Presence of the αS1-casein promoter (-175 to +796 nt) resulted in an up to 16-fold increase in luciferase activity compared with that of the promoter-less construct. In addition, the human IFN-α and EPO genes were identified as significantly overexpressed in HC11 cells compared with the promoter-less construct. Together, the present results demonstrate that the construct with the αS1-casein promoter may induce secretion of recombinant human IFN-α and EPO into bovine milk. Furthermore, we generated transgenic fibroblasts expressing human IFN-α and EPO cDNA controlled by the αS1-casein promoter and two screening markers, enhanced green fluorescent protein and neomycin resistance. These transgenic fibroblasts may be a source of somatic cells for generating transgenic cattle that produce recombinant human IFN-α and EPO proteins during lactation.