RESUMEN
Horizontal gene transfer has been demonstrated to be an important driver for the emergency of multidrug-resistant pathogens. Recently, a transferable gene cluster tmexCD1-toprJ1 of the resistance-nodulation-division (RND) superfamily was identified in the plasmids of animal-derived Klebsiella pneumoniae strains, with a higher efflux capacity for various drugs than the Escherichia coli AcrAB-TolC homolog system. In this study, we focused on the differences in the inner membrane pump of these two systems and identified some key residues that contribute to the robust efflux activity of the TMexCD1 system. With the aid of homologous modeling and molecular docking, eight residues from the proximal binding pocket (PBP) and nine from the distal binding pocket (DBP) were selected and subjected to site-directed mutagenesis. Several of them, such as S134, I139, D181, and A290, were shown to be important for substrate binding in the DBP region, and all residues in PBP and DBP showed certain substrate preferences. Apart from the conservative switch loop (L613-623TMexD1) previously identified in the E. coli AcrB (EcAcrB), a relatively unconservative loop (L665-675TMexD1) at the bottom of PBP was proposed as a critical element for the robust activity of TMexD1, due to variations at sites E669, G670, N673, and S674 compared to EcAcrAB, and the significantly altered efflux activity due to their mutations. The conservation and flexibility of these key factors can contribute to the evolution of the RND efflux pumps and thus serve as potential targets for developing inhibitors to block the widespread of the TMexCD1 system.
Asunto(s)
Proteínas de Escherichia coli , Escherichia coli , Animales , Escherichia coli/genética , Escherichia coli/metabolismo , Proteínas de Escherichia coli/metabolismo , Antibacterianos/química , Simulación del Acoplamiento Molecular , Farmacorresistencia Bacteriana Múltiple/genética , Proteínas Asociadas a Resistencia a Múltiples Medicamentos/genética , Pruebas de Sensibilidad MicrobianaRESUMEN
BACKGROUND: Biological-derived hydroxyapatite is widely used as a bone substitute for addressing bone defects, but its limited osteoconductive properties necessitate further improvement. The osteo-immunomodulatory properties hold crucial promise in maintaining bone homeostasis, and precise modulation of macrophage polarization is essential in this process. Metabolism serves as a guiding force for immunity, and fluoride modification represents a promising strategy for modulating the osteoimmunological environment by regulating immunometabolism. In this context, we synthesized fluorinated porcine hydroxyapatite (FPHA), and has demonstrated its enhanced biological properties and osteogenic capacity. However, it remains unknown whether and how FPHA affects the immune microenvironment of the bone defects. METHODS: FPHA was synthesized and its composition and structural properties were confirmed. Macrophages were cultured with FPHA extract to investigate the effects of FPHA on their polarization and the related osteo-immune microenvironment. Furthermore, total RNA of these macrophages was extracted, and RNA-seq analysis was performed to explore the underlying mechanisms associated with the observed changes in macrophages. The metabolic states were evaluated with a Seahorse analyzer. Additionally, immunohistochemical staining was performed to evaluate the macrophages response after implantation of the novel bone substitutes in critical size calvarial defects in SD rats. RESULTS: The incorporation of fluoride ions in FPHA was validated. FPHA promoted macrophage proliferation and enhanced the expression of M2 markers while suppressing the expression of M1 markers. Additionally, FPHA inhibited the expression of inflammatory factors and upregulated the expression of osteogenic factors, thereby enhancing the osteogenic differentiation capacity of the rBMSCs. RNA-seq analysis suggested that the polarization-regulating function of FPHA may be related to changes in cellular metabolism. Further experiments confirmed that FPHA enhanced mitochondrial function and promoted the metabolic shift of macrophages from glycolysis to oxidative phosphorylation. Moreover, in vivo experiments validated the above results in the calvarial defect model in SD rats. CONCLUSION: In summary, our study reveals that FPHA induces a metabolic shift in macrophages from glycolysis to oxidative phosphorylation. This shift leads to an increased tendency toward M2 polarization in macrophages, consequently creating a favorable osteo-immune microenvironment. These findings provide valuable insights into the impact of incorporating an appropriate concentration of fluoride on immunometabolism and macrophage mitochondrial function, which have important implications for the development of fluoride-modified immunometabolism-based bone regenerative biomaterials and the clinical application of FPHA or other fluoride-containing materials.
Asunto(s)
Durapatita , Glucólisis , Macrófagos , Fosforilación Oxidativa , Ratas Sprague-Dawley , Animales , Durapatita/química , Macrófagos/metabolismo , Macrófagos/efectos de los fármacos , Fosforilación Oxidativa/efectos de los fármacos , Glucólisis/efectos de los fármacos , Ratas , Porcinos , Proliferación Celular/efectos de los fármacos , Masculino , Osteogénesis/efectos de los fármacos , Cráneo/patología , Cráneo/efectos de los fármacos , Ratones , Microambiente Celular/efectos de los fármacos , Células RAW 264.7 , Huesos/metabolismo , Huesos/efectos de los fármacosRESUMEN
BACKGROUND: Studies examining whether diet sugar intake increases the risk of depression have produced inconsistent results. Therefore, we investigated this relationship, using the US' National Health and Nutrition Examination Survey (NHANES) database. METHODS: This cross-sectional study included 18,439 adults (aged ≥ 20 years) from NHANES (2011-2018). Depressive symptoms were assessed using the nine-item version of the Patient Health Questionnaire (PHQ-9). Covariates, including age, sex, race/ethnicity, poverty-income ratio, education, marital status, hypertension, diabetes mellitus, cardiovascular disease, alcohol intake, smoking status, physical activity, and dietary energy intake, were adjusted in multivariate logistic regression models. Subgroup and threshold saturation effect analyses were performed. RESULTS: After adjusting for potential confounders, we found that a 100 g/day increase in dietary sugar intake correlated with a 28% higher prevalence of depression (odds ratio = 1.28, 95% confidence interval = 1.17-1.40, P < 0.001). CONCLUSION: Dietary sugar intake is positively associated with depression in US adults.
Asunto(s)
Depresión , Dieta , Humanos , Adulto , Encuestas Nutricionales , Estudios Transversales , Depresión/epidemiología , Depresión/etiología , Azúcares de la Dieta/efectos adversosRESUMEN
BACKGROUND: Endometriosis is a common chronic gynecological condition characterized by the presence of endometrial tissue outside the uterine cavity, leading to chronic inflammation, pelvic nodules and masses, pelvic pain, and infertility. Acupuncture has been shown to improve pain associated with endometriosis by modulating abnormal levels of prostaglandins, ß-endorphins, dynorphins, electrolytes, and substance P. This review aims to evaluate the clinical efficacy of acupuncture in treating endometriosis, specifically focusing on its efficacy in relieving pain associated with endometriosis. METHODS: A comprehensive search was conducted in eight databases (PubMed, EMBASE, Cochrane, Web of Science, China National Knowledge Infrastructure (CNKI), the China Biology Medicine (CBM), Wanfang, and Weipu database) to identify randomized controlled trials (RCTs) published from database inception to December 16, 2022, which investigated the use of acupuncture for endometriosis-related pain. Two researchers independently screened articles, extracted data, and assessed methodological quality using the Cochrane Collaboration's risk of bias tool. Meta-analysis was performed using Stata statistical software. RESULTS: A total of 1991 articles were identified, and ultimately, 14 studies involving 793 patients (387 in the acupuncture group and 359 in the control group) were included. The control interventions in the included studies included placebo, traditional Chinese medicine (TCM), and Western medicine treatments. Meta-analysis results showed that compared to the control group, acupuncture treatment for pain associated with endometriosis demonstrated significant reductions in pain severity [SMD = - 1.10, 95% CI (- 1.45, - 0.75), P < 0.001], improved response rate [RR = 1.25, 95% CI (1.09, 1.44), P = 0.02], and decreased serum CA-125 levels [SMD = - 0.62, 95% CI (- 1.15, - 0.08), P = 0.024]. Furthermore, subgroup analysis revealed that electroacupuncture and auricular acupuncture were superior to the control group in reducing pain severity, while auricular acupuncture and warm needling showed greater clinical efficacy compared to the control group. However, there were no significant differences between electroacupuncture or fire needling and the control group in terms of pain relief. The findings suggest that acupuncture is effective in improving pain associated with endometriosis. CONCLUSIONS: In conclusion, acupuncture is effective in alleviating dysmenorrhea and pelvic pain associated with endometriosis, reducing serum CA-125 levels, decreasing the size of nodules, improving patients' quality of life, and lowering the recurrence rate. However, it should be noted that the current evidence is limited by the design and quality flaws of the original studies, as well as a lack of research specifically focusing on subtypes of acupuncture. Therefore, caution should be exercised when interpreting the results.
RESUMEN
Sodium-proton (Na+ /H+ ) antiporters from the ion transporter (IT) superfamily play a vital role in controlling the pH and electrolyte homeostasis. However, very limited information regarding their structural functions is available to date. In this study, the structural model of the NhaD antiporter was proposed as a typical hairpin structure of IT proteins, with two symmetrically conserved scaffold domains that frame the core substrate-binding sites, and four motifs were identified. Furthermore, 25 conserved sites involving these domains were subjected to site-directed mutagenesis, and all mutations resulted in an impact on transport abilities. In particular, as candidates for Na+ -binding sites, D166 and D405 mutations at hairpin discontinuities were detrimental to transport activities and were found to induce pronounced conformational changes using fluorescence resonance energy transfer (FRET) assays. In addition, as observed in the NhaA structure, some charged residues, for example, E64, E65, R454, and R464, are predicted to be involved in the net charge switch of NhaD activation, by collectively form a "pH sensor" at the entrance of the cytoplasmic funnel. Mutations encompassing these residues were detrimental to the transport activity of NhaD or lost the capacity to respond to pH signals and trigger conformational changes for Na+ translocation.
Asunto(s)
Antiportadores , Proteínas de Escherichia coli , Secuencia de Aminoácidos , Antiportadores/metabolismo , Proteínas Bacterianas/metabolismo , Proteínas de Escherichia coli/metabolismo , Concentración de Iones de Hidrógeno , Protones , Sodio/metabolismo , Intercambiadores de Sodio-Hidrógeno/química , Intercambiadores de Sodio-Hidrógeno/genética , Intercambiadores de Sodio-Hidrógeno/metabolismoRESUMEN
IFAP syndrome is a rare genetic disorder characterized by ichthyosis follicularis, atrichia, and photophobia. Previous research found that mutations in MBTPS2, encoding site-2-protease (S2P), underlie X-linked IFAP syndrome. The present report describes the identification via whole-exome sequencing of three heterozygous mutations in SREBF1 in 11 unrelated, ethnically diverse individuals with autosomal-dominant IFAP syndrome. SREBF1 encodes sterol regulatory element-binding protein 1 (SREBP1), which promotes the transcription of lipogenes involved in the biosynthesis of fatty acids and cholesterols. This process requires cleavage of SREBP1 by site-1-protease (S1P) and S2P and subsequent translocation into the nucleus where it binds to sterol regulatory elements (SRE). The three detected SREBF1 mutations caused substitution or deletion of residues 527, 528, and 530, which are crucial for S1P cleavage. In vitro investigation of SREBP1 variants demonstrated impaired S1P cleavage, which prohibited nuclear translocation of the transcriptionally active form of SREBP1. As a result, SREBP1 variants exhibited significantly lower transcriptional activity compared to the wild-type, as demonstrated via luciferase reporter assay. RNA sequencing of the scalp skin from IFAP-affected individuals revealed a dramatic reduction in transcript levels of low-density lipoprotein receptor (LDLR) and of keratin genes known to be expressed in the outer root sheath of hair follicles. An increased rate of in situ keratinocyte apoptosis, which might contribute to skin hyperkeratosis and hypotrichosis, was also detected in scalp samples from affected individuals. Together with previous research, the present findings suggest that SREBP signaling plays an essential role in epidermal differentiation, skin barrier formation, hair growth, and eye function.
Asunto(s)
Artrogriposis/genética , Mutación/genética , Proteína 1 de Unión a los Elementos Reguladores de Esteroles/genética , Adolescente , Adulto , Niño , Preescolar , Femenino , Regulación de la Expresión Génica/genética , Humanos , Queratosis/genética , Masculino , Persona de Mediana Edad , Linaje , Fenotipo , Adulto JovenRESUMEN
The arsenical resistance-3 (ACR3) family constitutes the most common pathway that confers high-level resistance to toxic metalloids in various microorganisms and lower plants. Based on the structural model constructed by AlphaFold2, the Acr3 antiporter from Bacillus subtilis (Acr3Bs ) exhibits a typical NhaA structure fold, with two discontinuous helices of transmembrane (TM) segments, TM4 and TM9, interacting with each other and forming an X-shaped structure. As the structural information available for these important arsenite-efflux pumps is limited, we investigated the evolutionary conservation among 300 homolog sequences and identified three conserved motifs in both the discontinuous helices and TM5. Through site-directed mutagenesis, microscale thermophoresis (MST), and fluorescence resonance energy transfer (FRET) analyses, the identified Motif C in TM9 was found to be a critical element for substrate binding, in which N292 and E295 are involved in substrate coordination, while R118 in TM4 and E322 in TM10 is responsible for structural stabilization. In addition, the highly conserved residues on Motif B of TM5 are potentially key factors in the protonation/deprotonation process. These consensus motifs and residues are essential for metalloid compound translocation of Acr3 antiporters, by framing the core domain and the typical X-shaped of NhaA fold.
Asunto(s)
Antiportadores , Arsenitos , Antiportadores/genética , Antiportadores/metabolismo , Arsenitos/metabolismo , Proteínas de Transporte de Membrana/metabolismo , Estructura Secundaria de ProteínaRESUMEN
BACKGROUND: Generalized pustular psoriasis (GPP) is a rare and life-threatening autoinflammatory dermatological disease. IL36RN was reported to be the main pathogenetic basis for GPP. Only a few studies have reported on the correlation analysis of IL36RN variants and the phenotype of pediatric-onset GPP. METHODS: IL36RN was screened in 60 children diagnosed with GPP from January 2013 to January 2020, and their detailed clinical profiles were obtained. RESULTS: Forty-six out of 60 (76.67%) patients harbored IL36RN variants, and six IL36RN variants were found, of which two were novel variants that were reported for the first time. The frequency of IL36RN variants was significantly different among the subtypes of GPP (GPP with acrodermatitis continua of Hallopeau group (ACH), 100%; GPP without plaque psoriasis (PV) and ACH, 78.05%; GPP with PV group, 44.44%) (p = 0.018), while the percentage of IL36RN variants in the GPP with ACH group was higher than that in the GPP with PV group (p < 0.05). IL36RN variants were associated with a lower percentage of PV, longer length of hospitalization, and longer time to reach normal body temperature after treatment (p < 0.05). After treatment, marked responses, moderate responses, and no responses were recorded in 75.00%, 8.33%, and 16.67% of patients, respectively. No significant difference was observed during efficacy assessment in patients with or without IL36RN variants (χ2 = 1.122, p > 0.05). CONCLUSIONS: IL36RN variants are associated with GPP with ACH subtypes, an absence of concurrent PV, and a greater extent of severe inflammation. Acitretin was an effective treatment for patients in our study and mostly resulted in a marked response in our cohort.
Asunto(s)
Psoriasis , Enfermedades Cutáneas Vesiculoampollosas , Humanos , Acitretina/uso terapéutico , Enfermedad Aguda , Enfermedad Crónica , Interleucinas/genética , Psoriasis/tratamiento farmacológico , Psoriasis/genética , Psoriasis/patología , Enfermedades Cutáneas Vesiculoampollosas/genética , Resultado del TratamientoRESUMEN
BACKGROUND: Despite the close relationship between sleep-wake cycles and depression symptoms, the relationship between sleep midpoint and depression symptoms in adults remains understudied. METHODS: In this cross-sectional study, 18280 adults aged ≥ 18 years from the National Health and Nutrition Examination Survey (NHANES) 2015-2020 were analyzed. Covariates included age, sex, race/ethnicity, education level, marital status, family income, body mass index, smoking status, drinking status, physical activity, comorbid condition, sleep duration, and sleep disturbance were adjusted in multivariate regression models. RESULTS: Weighted restricted cubic spline based on the complex sampling design of NHANES showed that in participants with a sleep midpoint from 2:18 AM to 6:30 AM, the prevalence of depression symptoms increased by 0.2 times (adjusted odds ratio [OR] = 1.20, 95% confidence interval [CI]: 1.08-1.33) per 1-h increment in sleep midpoint compared to the reference point of 2:18 AM. For participants with a sleep midpoint after 6:30 AM and before 2:18 AM the next day, the relationship between sleep midpoint and depression symptoms was not significant after adjusting for all covariates (adjusted OR = 1.01, 95% CI: 0.99-1.03). CONCLUSIONS: The findings indicate a significant nonlinear association between sleep midpoint and depression symptoms in a nationally representative sample of adults.
Asunto(s)
Depresión , Sueño , Humanos , Adulto , Estudios Transversales , Depresión/epidemiología , Encuestas Nutricionales , Duración del SueñoRESUMEN
BACKGROUND: As the most abundant fatty acid in plasma, oleic acid has been found to be associated with multiple neurological diseases; however, results from studies of the relationship between oleic acid and depression are inconsistent. METHODS: This cross-sectional study analyzed 4,459 adults from the National Health and Nutrition Examination Survey 2011-2014. The following covariates were adjusted in multivariable logistic regression models: age, sex, race/ethnicity, education level, marital status, body mass index, physical activity, smoking status, alcohol status, metabolic syndrome, omega-3 polyunsaturated fatty acids, and total cholesterol. RESULTS: Serum oleic acid levels were positively associated with depression. After adjusting for all covariates, for every 1 mmol/L increase in oleic acid levels, the prevalence of depression increased by 40% (unadjusted OR: 1.35, 95%CI: 1.16-1.57; adjusted OR: 1.40, 95% CI: 1.03-1.90). CONCLUSIONS: Our study suggests that oleic acid may play a role in depression. Further research is needed to investigate the potential benefits of changing oleic acid levels for the treatment and prevention of depression.
Asunto(s)
Depresión , Ácidos Grasos Omega-3 , Adulto , Humanos , Estados Unidos/epidemiología , Depresión/epidemiología , Ácido Oléico , Estudios Transversales , Encuestas NutricionalesRESUMEN
BACKGROUND: The scientific evidence of methotrexate (MTX) in children with severe plaque psoriasis is scarce. OBJECTIVES: To retrospectively evaluate the efficacy and safety of oral MTX in children with severe plaque psoriasis in a single center in China. METHODS: We enrolled 42 children with severe plaque psoriasis who were administrated MTX. Efficacy was evaluated by the psoriasis area and severity index (PASI) score, physician global assessment (PGA) score, and body surface area (BSA) score. The Children's Dermatology Life Quality Index (CDLQI) score and safety data were recorded. RESULTS: Among 42 children (22 males, 20 females), the mean age was 11.2 years old. The initial weight-based dosage of oral MTX ranged from 0.1 to 0.3 mg/kg weekly. Overall, 80.6 and 47.2% of patients achieved PASI75 (at least 75% improvement from baseline in PASI score) and PASI90 (at least 90% improvement from baseline in PASI score) at week 12, respectively. 72.2% of patients achieved PGA 0/1 at week 12. BSA and PGA scores significantly decreased from baseline from week 4, accompanied by CDLQI score improvement from week 8. The steady effect of MTX could be reached at week 16. Elevated liver enzymes (28.6%) and infections (28.6%) were the most common side effects. Relapse was recorded in 9 (30.0%) of 30 patients, and the mean posttherapy disease-free interval was 7.2 months. CONCLUSIONS: MTX is an effective and safe option for children with severe plaque psoriasis with adequate monitoring.
Asunto(s)
Metotrexato , Psoriasis , Niño , Femenino , Humanos , Masculino , Superficie Corporal , Metotrexato/efectos adversos , Metotrexato/uso terapéutico , Psoriasis/tratamiento farmacológico , Estudios Retrospectivos , Índice de Severidad de la Enfermedad , Resultado del TratamientoRESUMEN
STATEMENT OF PROBLEM: Both the placement accuracy and primary stability of implants are important to implant therapy in the esthetic zone. The effect of dynamic and static computer-assisted navigation on the primary stability of implants in the esthetic zone remains uncertain. PURPOSE: The purpose of this case-control study was to investigate the effect of dynamic and static computer-assisted navigation on the placement accuracy and primary stability of implants in the esthetic zone. MATERIAL AND METHODS: Partially edentulous participants who received at least 1 implant in the anterior maxilla using either fully guided static or dynamic computer-assisted implant surgery (s-CAIS, d-CAIS) from January 2020 to February 2022 were screened. Participant demographic information, timing of implant placement, primary stability represented by the insertion torque value (ITV) in Ncm, and implant survival were collected from the treatment record. Bone quality at the implant sites was determined according to the Lekholm and Zarb classification. The accuracy of implant placement represented by the linear (platform: Dpl, mm; apex: Dap, mm) and angular deviations (axis: Dan, degree) between the planned and placed implants was evaluated based on the preoperative surgical plan and postoperative cone beam computed tomography (CBCT) data. A statistical analysis of the data was completed by using the chi-square, Fisher exact, Student t, and Mann-Whitney U tests (α=.05). RESULTS: A total of 32 study participants (38 implants) were included. The groups of s-CAIS (16 participants, 18 implants) and d-CAIS (16 participants, 20 implants) were statistically comparable in sex (P=.072), age (P=.548), bone quality (P=.671), and timing of implant placement (P=.719). All implants survived during an average follow-up period of 13 months. The d-CAIS group showed close linear deviations (Dpl 1.07 ±0.57 mm, Dap 1.26 ±0.53 mm) but lower angular deviation (Dan 2.14 ±1.20 degrees) and primary stability (ITV 25.25 ±7.52 Ncm) than the s-CAIS group (Dpl 0.92 ±0.46 mm, Dap 1.31 ±0.43 mm, Dan 3.31 ±1.61 degrees, ITV 30.56 ±11.23 Ncm, PDpl=.613, PDap=.743, PDan=.016, PITV=.028). CONCLUSIONS: Comparable linear positioning accuracy and higher angular deviation were found for implants placed in the esthetic zone by using s-CAIS than when using d-CAIS. Higher primary stability of implants may be achieved by using s-CAIS, as s-CAIS seemed to have higher osteotomy accuracy than d-CAIS.
RESUMEN
BACKGROUND: Lotus seedpods are an agricultural by-product of lotus (Nelumbo nucifera Gaertn.), which is widely cultivated in Southeast Asia and Australia. Most lotus seedpods are considered waste and are abandoned or incinerated, resulting in significant waste of resources and heavy environmental pollution. For recycling lotus seedpods, the extraction optimization, physicochemical properties, antioxidant activity, and α-glucosidase inhibitory effect of the polysaccharides contained therein were investigated in this study. RESULTS: Hot water extraction of lotus seedpod polysaccharides was optimized by using a response surface methodology combined with a Box-Behnken design, with the optimum conditions being as follows: a liquid/solid ratio of 25.0 mL g-1 , an extraction temperature of 98.0 °C, and an extraction time of 138.0 min. Under these conditions, an experimental yield of 5.88 ± 0.06% was obtained. Physicochemical analyses suggested that lotus seedpod polysaccharides belong to acidic heteropolysaccharides and are principally composed of rhamnose, arabinose, galactose, glucose, mannose, and galacturonic acid. The polysaccharides content has a broad molecular weight distribution (2.15 × 105 to 1.77 × 107 Da), an α-configuration, and mainly possesses smooth and sheet-like structures. Biological evaluations showed that the polysaccharides possessed good scavenging activity on 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt, 1,1-diphenyl-2-picryl-hydrozyl, and hydroxyl radicals, and exerted an obvious inhibitory effect on α-glucosidase activity. Moreover, the polysaccharides content was determined to be a mixed-type noncompetitive inhibitor of α-glucosidase. CONCLUSION: The results indicate that lotus seedpod polysaccharides have potential as natural antioxidants and hypoglycaemic substitutes. This study provides the theoretical bases for the exploitation and application of polysaccharides from lotus seedpod by-product resources. © 2022 Society of Chemical Industry.
Asunto(s)
Antioxidantes , Lotus , Antioxidantes/química , Antioxidantes/farmacología , Polisacáridos/química , Polisacáridos/farmacología , Semillas , alfa-Glucosidasas/químicaRESUMEN
OBJECTIVES: The NorA efflux pump in Staphylococcus aureus mediates resistance to many fluoroquinolone (FQ) antibiotics. Three norA alleles with high sequence similarity are found in various S. aureus strains exhibiting different FQ resistance profiles. This study aimed to elucidate the underlying molecular basis for the varying efflux activity of these three allelic variations. METHODS: The norA genotypes of 20 S. aureus isolates were analysed. Multiple alignments and conservative analyses were conducted to explore the evolutionary variations. After heterologous expression in Escherichia coli, seven mutants were constructed for MIC tests, efflux activity and conformational change measurements. RESULTS: Three NorA alleles were identified that displayed different FQ MICs and varying efflux activity for ethidium bromide, with the NorAII protein showing the strongest activity. A total of 29 single amino acid polymorphisms were identified by conservative analysis within three allelic peptides, with seven sites densely distributed in the 277-297 region. Mutations of these seven residues in NorAII all significantly impaired drug resistance and efflux activity, and three key mutants showed conformational changes in fluorescence resonance energy transfer (FRET) analysis. CONCLUSIONS: Evolutionary variations of the 277-297 region could be a major explanation for the functional difference of three norA alleles and serve as a potential target for the development of novel NorA inhibitors.
Asunto(s)
Proteínas Asociadas a Resistencia a Múltiples Medicamentos , Staphylococcus aureus , Alelos , Antibacterianos/farmacología , Proteínas Bacterianas/genética , Proteínas Bacterianas/metabolismo , Pruebas de Sensibilidad Microbiana , Proteínas Asociadas a Resistencia a Múltiples Medicamentos/genética , Staphylococcus aureus/genética , Staphylococcus aureus/metabolismoRESUMEN
OBJECTIVES: For the stress from fermenters, downstream processing equipment, and wastewater treatment to be alleviated, lowering salt-dependence in the ectoine synthesis process is of great significance in the moderately halotolerant Halomonas hydrothermalis Y2. RESULTS: In H. hydrothermalis Y2, the σ70- and σ38-controlled promoters of ectA are predicted to be involved in the osmotic regulation of ectoine synthesis. By substituting the ectA promoter with a promoter P265 that identified in the outer membrane pore protein E of H. hydrothermalis Y2, the salt dependence of ectoine synthesis was significantly decreased. In the 500-ml flask containing various NaCl contents, the engineered strain (p/Y2/â³ectD/â³doeA) showed a remarkably enhanced ability in ectoine synthesis, especially under lower saline stress. After a 36-h fed-batch fermentation in the 1-l fermenter, p/Y2/â³ectD/â³doeA synthesized 11.5 g ectoine l-1 in the presence of 60 g NaCl-1 l, with a high 0.32 g ectoine l-1 h-1 productivity, a specific productivity of 512.2 mg ectoine per g cell dry weight (CDW)-1, and an excretion ratio of 67 % ectoine. CONCLUSIONS: As no impaired growth was observed in strain p/Y2/â³ectD/â³doeA while ectoine synthesis was increased, this promoter engineering strategy provides a practical protocol for lowering the salt-dependence of ectoine synthesis in this moderately halotolerant strain.
Asunto(s)
Aminoácidos Diaminos/biosíntesis , Proteínas Bacterianas/genética , Técnicas de Cultivo Celular por Lotes/métodos , Halomonas/crecimiento & desarrollo , Proteínas de la Membrana Bacteriana Externa/genética , Reactores Biológicos/microbiología , Medios de Cultivo/química , Fermentación , Perfilación de la Expresión Génica , Regulación Bacteriana de la Expresión Génica , Ingeniería Genética , Halomonas/metabolismo , Regiones Promotoras Genéticas , Cloruro de Sodio/químicaRESUMEN
BACKGROUND: The accuracy of digital impressions for fully edentulous cases is currently insufficient for routinely clinical application. To overcome the challenge, a modified scan body was introduced, which demonstrated satisfactory accuracy in vitro. The aim of this study was to evaluate the accuracy of digital impressions using the modified scan bodies with extensional structure versus scan bodies without extensional structure in mandible with two implants in beagle dogs. METHODS: The unilateral mandibular second premolar to second molar were extracted in four beagle dogs. Twelve weeks later, two implants were placed. Five repeated digital impressions were performed with an intraoral scanner on each dog using each of the two different scan bodies: Group I-scan body without extensional structure (SB); Group II-scan body with extensional structure (SBE). The scans were exported to Standard Tessellation Language (STL) files to serve as test data. The dogs were sacrificed and the dissected mandibles were digitalized with a lab scanner to provide reference data. Linear and angular deviations were calculated in an inspection software for accuracy assessment. Statistical analysis was performed with two-way ANOVA. The level of significance was set at α = 0.05. RESULTS: For trueness assessment, the mean of absolute linear/angular deviations were 119.53 µm/0.75 degrees in Group I and 68.89 µm/0.36 degrees in Group II. SBE was more accurate than SB regarding both linear (p = 0.008) and angular (p = 0.049) deviations. For precision assessment, the mean of absolute linear/angular deviations were 63.01 µm/0.47 degrees in Group I and 38.38 µm/0.24 degrees in Group II. No significant difference was found. CONCLUSIONS: The application of SBE significantly improved the trueness of digital impressions in mandible with two implants compared to SB. No significant difference was found in terms of precision.
Asunto(s)
Implantes Dentales , Técnica de Impresión Dental , Animales , Diseño Asistido por Computadora , Perros , Imagenología Tridimensional , Modelos DentalesRESUMEN
OBJECTIVES: To compare the accuracy of an original and two newly designed CAD/CAM scan bodies used in digital impressions with one another as well as conventional implant impressions. MATERIAL AND METHODS: A reference model containing four implants was fabricated. Digital impressions were taken using an intraoral scanner with different scan bodies: original scan bodies for Group I (DO), CAD/CAM scan bodies without extensional structure for Group II (DC), and CAD/CAM scan bodies with extensional structure for Group III (DCE). For Group IV, conventional splinted open-tray impressions (CI) were taken. The reference model and conventional stone casts were digitalized with a laboratory reference scanner. The Standard Tessellation Language datasets were imported into an inspection software for trueness and precision assessment. Statistical analysis was performed with a Kruskal-Wallis test and Dunn-Bonferroni test. The level of significance was set at α = .05. RESULTS: The median of trueness was 35.85, 38.50, 28.45, and 25.55 µm for Group I, II, III, and IV, respectively. CI was more accurate than DO (p = .015) and DC (p = .002). The median of precision was 48.40, 48.90, 27.30, and 19.00 for Group I, II, III, and IV, respectively. CI was more accurate than DO (p < .001), DC (p < .001), and DCE (p = .007). DCE was more accurate than DC (p < .001) and DO (p < .001). CONCLUSIONS: The design of the extensional structure could significantly improve scanning accuracy. Conventional splinted open-tray impressions were more accurate than digital impressions for full-arch implant rehabilitation.
Asunto(s)
Implantes Dentales , Técnica de Impresión Dental , Diseño Asistido por Computadora , Materiales de Impresión Dental , Imagenología Tridimensional , Modelos Dentales , CintigrafíaAsunto(s)
Anticuerpos Monoclonales Humanizados , Proteínas Adaptadoras de Señalización CARD , Guanilato Ciclasa , Proteínas de la Membrana , Mutación , Nevo Sebáceo de Jadassohn , Humanos , Anticuerpos Monoclonales Humanizados/uso terapéutico , Proteínas Adaptadoras de Señalización CARD/genética , Guanilato Ciclasa/genética , Nevo Sebáceo de Jadassohn/genética , Nevo Sebáceo de Jadassohn/patología , Nevo Sebáceo de Jadassohn/tratamiento farmacológico , Proteínas de la Membrana/genética , Femenino , MasculinoRESUMEN
BACKGROUND AND OBJECTIVES: Café-au-lait spots, also known as café-au-lait macules (CALMs), are a common pigmentary disorder. Although various laser modalities have been used to treat CALMs, the efficacy of laser treatment in children differs from that in adults. We investigated the efficacy, safety, and clinical factors of the treatment of CALMs using Q-switched alexandrite laser (755 nm) therapy in children. METHODS: In total, 471 children with CALMs underwent Q-switched alexandrite laser therapy at a treatment interval of 3-12 months. The safety and efficacy of the laser treatment were evaluated by reviewing clinical records and photographs before and after treatments. RESULTS: Of the 471 patients, 140 (29.72%) were cured completely, 124 (26.33%) showed substantial improvement, 110 (23.35%) showed improvement, and 97 (20.60%) showed no improvement after one to nine treatments. The overall treatment success rate was 79.41%, and the treatment efficacy was positively correlated with the number of laser treatments (rs = 0.26, P < 0.0001). Sex and the interval of laser treatments were also associated with significant differences in treatment outcomes (P < 0.05). No obvious adverse effects were observed. Multivariate logistic regression analysis showed that the number of treatments influenced the treatment efficacy (odds ratio, 2.130; 95% confidence interval, 1.561-2.908). CONCLUSIONS: Q-switched alexandrite laser (755 nm) therapy is safe and highly effective for CALMs in children, and the number of treatments affects the treatment efficacy. Lasers Surg. Med. © 2019 The Authors. Lasers in Surgery and Medicine Published by Wiley Periodicals, Inc.