Abstract Background: Infections caused by multi-drug-resistant organisms are too common and represent a therapeutic challenge. Topical application of sevoflurane may be useful for the treatment of cutaneous infections, as this general anesthetic has antimicrobial properties in vitro. Case Report: A 43-year-old male patient received a liver transplant because of cirrhosis caused by hepatitis C. The surgical site was infected initially by multi-drug-resistant Pseudomonas aeruginosa that was susceptible to colistin, but parenteral administration of this antibiotic led to deterioration of renal function and was discontinued. The incision did not heal with daily lavage and debridement and was superinfected with Staphylococcus aureus. The off-label use of liquid sevoflurane in the form of site irrigations was followed by healing and closure of the site. This healing could be attributable to the mixed effect of three possible actions of sevoflurane: Direct antimicrobial effect, local analgesic effect that improved the quality of the dressings, and a direct vasodilator effect that, in theory, increased the supply of nutrients to the incision bed. Conclusions: The use of liquid sevoflurane on sites infected by microorganisms resistant to conventional antibiotics appears to be an attractive therapeutic option that deserves future research.
The effects of gold nanoparticles (AuNPs) incorporated in the hole transporting layer (HTL) of Poly[[4,8-bis[(2-ethylhexyl)oxy] benzo[1,2-b:4,5-b'] dithiophene-2, 6-diyl] [3-fluoro-2-[(2-ethylhexy)carbonyl]thieno[3,4-b]thiophened iyl]] (PTB7): [6,6]-Phenyl C71 butyric acid methyl ester (PC71BM) based solar cells are being systematically investigated in terms of the optical properties, electrical properties and photovoltaic performance. The impacts of AuNPs on the optical response of the devices are modeled by finite-difference time-domain (FDTD) simulation. The size of the AuNPs used in this work is around 50-70 nm, so that 10-20 nm penetrated from the HTL into the active layer. We found that the power conversion efficiencies (PCEs) of the devices with AuNPs are significantly enhanced from 7.5%, for the control device, to 8.0%, 8.1% and 8.2% for Au nanosphere-, nanorod- and nanocube-incorporated devices respectively. Among the photovoltaic parameters of the AuNP devices, the short circuit current density (JSC) exhibits the largest improvement, which can be attributed to the improved optical properties of the devices. Based on the calculation results, the scattering cross section for the samples in the presence of AuNPs can be enhanced by a factor of ~1010-1013 and Au nanocubes exhibit superior scattering cross section compared to the Au nanospheres and nanorods with the same linear dimension. From the experimental impedance spectroscopy results, we found that the addition of AuNPs had little effect on the electrical properties of the device. The device performance is also found to be sensitive to the concentration and morphology of the AuNPs.
Abstract Context: Miconazole (MIZ) and econazole (ECZ) are clinically used as antifungal drugs. Objective: The drug effect and binding property with transport protein human serum albumin of MIZ and ECZ were studied. Materials and methods: The antifungal efficiency was investigated by microdiluting drug solutions from 0 to 48??mol?L(-1) through microcalorimetry and voltammetry studies. Transmission electron microscopy was used for morphological observations of C. albicans. The interaction with HSA was studied by electrochemical methods, fluorescence spectrometry, electron microscopy, and molecular simulation. Results: IC50 of MIZ and ECZ for C. albicans were obtained as 19.72 and 29.90??mol?L(-1). Binding constants of MIZ and ECZ with HSA of 2.36?×?10(4) L?mol(-1) and 3.73?×?10(4) L?mol(-1) were obtained. After adding MIZ solution of 12 and 40??mol?L(-1), the peak currents increased to 4.887 and 6.024??A. The peak currents of C. albicans in the presence of 20 and 48??mol?L(-1) ECZ were 4.701 and 5.544??A. The docking scores for MIZ and ECZ of the best binding conformation in site I and site II were 5.60, 4.79, 5.63, and 5.85. Discussion and conclusion: Strong inhibition to the metabolism of C. albicans and destructive effect was proved for both drugs. The lower IC50, growth rate constant of C. albicans, and higher peak current, reveal stronger antifungal activity of MIZ. Both drugs show an efficient quenching effect to intrinsic fluorescence residues of protein. MIZ mainly binds on site I while ECZ on site II. Molecular modeling experiments give further insight of the binding mechanism.
Multiple myeloma (MM), a plasma cell malignancy, is the second most prevalent hematologic malignancy in the US. Although much effort has been made trying to understand the etiology and the complexities of this disease with the hope of developing effective therapies, MM remains incurable at this time. Because of their antiproliferative and proapoptotic activities, interferons (IFNs) have been used to treat various malignancies, including MM. Although some success has been observed, the inherent toxicities of IFNs limit their efficacy. To address this problem, we produced anti-CD138 antibody fusion proteins containing either IFN?2 or a mutant IFN?2 (IFN?2(YNS)) with the goal of targeting IFN to CD138-expressing cells, thereby achieving effective IFN concentrations at the site of the tumor in the absence of toxicity. The fusion proteins inhibited the proliferation and induced apoptosis of U266, ANBL-6, NCI-H929, and MM1-144 MM cell lines. The fusion proteins decreased the expression of IFN regulatory factor 4 (IRF4) in U266. In addition, the fusion proteins were effective against primary cells from MM patients, and treatment with fusion proteins prolonged survival in the U266 murine model of MM. These studies show that IFN? antibody fusion proteins can be effective novel therapeutics for the treatment of MM.
In this paper, the stabilisation times for all 24 h of the day are analysed for 12 measurement stations located in the Spanish town of Malaga and throughout 5 years (2007-2011) of sampling environmental noise levels. For the results to be generalised to sound level measurements made in other streets or cities where there have been no long-term measurements, this study was developed for different road types and urban shapes. This distinction was made according to the types of roads indicated in other studies in which a statistically significant relationship was found between noise levels and the road type. The final objective of the study is to determine the capacity to estimate and approximate the real equivalent hourly noise level (L Aeq,1h) from the integration of a number of consecutive minutes (short-term measurements) less than 60 (L Aeq,1h???L Aeq,T being T???60 min). Clearly, this strategy would save time and resources by making measurements of reduced duration. In summary and according to this analysis, a short-time measurement of 15 min is adequate to work with 90 % confidence levels and errors of ±2 dB, with 80 % confidence levels and errors of ±1 dB, and 50 % confidence levels and errors of ±0.5 dB. However, it is necessary to consider the measurement hour period to achieve these levels of confidence due to the high variability throughout the day.
Brucellosis is one of the major infectious and contagious bacterial diseases in Hinggan League of Inner Mongolia, China. The number of newly infected human brucellosis data in this area has increased dramatically in the last 10 years. In this study, in order to explore effective control and prevention measures we propose a deterministic model to investigate the transmission dynamics of brucellosis in Hinggan League. The model describes the spread of brucellosis among sheep and from sheep to humans. The model simulations agree with newly infected human brucellosis data from 2001 to 2011, and the trend of newly infected human brucellosis cases is given. We estimate that the control reproduction number Rc is about 1.9789 for the brucellosis transmission in Hinggan League and compare the effect of existing mixed cross infection between basic ewes and other sheep or not for newly infected human brucellosis cases. Our study demonstrates that combination of prohibiting mixed feeding between basic ewes and other sheep, vaccination, detection and elimination are useful strategies in controlling human brucellosis in Hinggan League.
The crystal structure of the [Ag62S12(SBu(t))32](2+) nanocluster (denoted as NC-I) has been successfully determined, and it shows a complete face-centered-cubic (FCC) Ag14 core structure with a Ag48(SBu(t))32 shell configuration interconnected by 12 sulfide ions, which is similar to the [Ag62S13(SBu(t))32](4+) structure (denoted as NC-II for short) reported by Wang. Interestingly, NC-I exhibits prominent differences in the optical properties in comparison with the case of the NC-II nanocluster. We employed femtosecond transient absorption spectroscopy to further identify the differences between the two nanoclusters. The results show that the quenching of photoluminescence in NC-I in comparison to that of NC-II is caused by the free valence electrons, which dramatically change the ligand to metal charge transfer (LMCT, S 3p ? Ag 5s). To get further insight into these, we carried out time-dependent density functional theory (TDDFT) calculations on the electronic structure and optical absorption spectra of NC-I and NC-II. These findings offer a new insight into the structure and property evolution of silver cluster materials.
Development of magnetic encapsulated microbubble agents that can integrate multiple diagnostic and therapeutic functions is a key focus in both biomedical engineering and nanotechnology and one which will have far-reaching impact on medical diagnosis and therapies. However, properly designing multifunctional agents that can satisfy particular diagnostic/therapeutic requirements has been recognized as rather challenging, because there is a lack of comprehensive understanding of how the integration of magnetic nanoparticles to microbubble encapsulating shells affects their mechanical properties and dynamic performance in ultrasound imaging and drug delivery. Here, a multifunctional imaging contrast and in-situ gene/drug delivery agent was synthesized by coupling super paramagnetic iron oxide nanoparticles (SPIOs) into albumin-shelled microbubbles. Systematical studies were performed to investigate the SPIO-concentration-dependence of microbubble mechanical properties, acoustic scattering response, inertial cavitation activity and ultrasound-facilitated gene transfection effect. These demonstrated that, with the increasing SPIO concentration, the microbubble mean diameter and shell stiffness increased and ultrasound scattering response and inertial cavitation activity could be significantly enhanced. However, an optimized ultrasound-facilitated vascular endothelial growth factor transfection outcome would be achieved by adopting magnetic albumin-shelled microbubbles with an appropriate SPIO concentration of 114.7?µg?ml(-1). The current results would provide helpful guidance for future development of multifunctional agents and further optimization of their diagnostic/therapeutic performance in clinic.
The crystal structure of selenolate-capped Au25(SePh)18(-) nanoclusters has been unambiguously determined for the first time, and provides a solid basis for a deeper understanding of the structure-property relationships. The selenolate-capped Au25 cluster shows noticeable differences from the previously reported Au25(SCH2CH2Ph)18(-) counterpart, albeit both share the icosahedral Au13 core and semi-ring Au2(SeR)3 or Au2(SR)3 motifs. Distinct differences in the electronic structure and optical, catalytic and electrochemical properties are revealed by the coupling experiments with density functional theory (TD-DFT) calculations. Overall, the successful determination of the Au25(SePh)18(-) structure removes any ambiguity about its structure, and comparison with the thiolated Au25 counterpart helps us to further understand how the ligands affect the properties of the nanocluster.
Two new xanthones, designated garcimangosxanthone F (1) and garcimangosxanthone G (2), were isolated from the EtOAc-soluble fraction of ethanolic extract from the pericarp of Garcinia mangostana. Their structures were established as 1,6,7-trihydroxy-5-(3-methylbut-2-enyl)-8-(3-hydroxy-3-methylbutyl)-6',6'-dimethylpyrano[2',3':3,2]xanthone and 1,6,7-trihydroxy-5-(3-methylbut-2-enyl)-8-(3-hydroxy-3-methylbutyl)-6',6'-dimethyl-4',5'-dihydropyrano[2',3':3,2]xanthone, respectively, on the basis of their 1D, 2D NMR and MS data interpretation.
Meningiomas have classically been considered to include benign and atypical/anaplastic tumors. Despite the availability of clinical and pathologic parameters for prognostic prediction prognosis, the behavior of each meningioma may be difficult to predict. Here, we used DNA flow-cytometric studies to predict biological tumor behaviors of intracranial meningiomas.
In this work, the anti-Escherichia coli activity of the bioactive substances produced by Bacillus amyloliquefaciens R3 was examined. A new and cheap medium for production of the anti-E. coli substances which contained 20.0 g L(-1) soybean powder, 20.0 g L(-1) wheat flour, pH 6.0 was developed. A crude surfactant concentration of 0.48 mg mL(-1) was obtained after 27 h of 10-L fermentation, and the diameter of the clear zone on the plate seeded with the pathogenic E. coli 2# was 23.3 mm. A preliminary characterization suggested that the anti-E. coli substances produced by B. amyloliquefaciens R3 were the biosurfactins (F1, F2, F3, F4, and F5) with amino acids (GLLVDLL) and hydroxy fatty acids (of 12-15 carbons in length). It was found that all the strains of the pathogenic E. coli showed resistance to several different antibiotics, suggesting that they were the multi-drug resistance and all the strains of the pathogenic E. coli were sensitive to the biosurfactins, indicating that the biosurfactins produced by B. amyloliquefaciens R3 had a broad spectrum of antibacterial activity against the pathogenic E. coli with multi-drug resistant profiles. After the treatment with the purified biosurfactin (F1), the cell membrane of both the whole cells and protoplasts of the E. coli 2# was damaged and the whole cells of the bacterium were broken.
Yunnan Province in China borders 3 countries (Vietnam, Laos, and Myanmar) in Southeast Asia. In the 1980s, a large-scale rabies epidemic occurred in this province, which subsided by the late 1990s. However, 3 human cases of rabies in 2000 indicated reemergence of the disease in 1 county. In 2012, rabies was detected in 77 counties; 663 persons died of rabies during this new epidemic. Fifty two rabies virus strains obtained during 2008-2012 were identified and analyzed phylogenetically by sequencing the nucleoprotein gene. Of the 4 clades identified, clades YN-A and YN-C were closely related to strains from neighboring provinces, and clade YN-B was closely related to strains from Southeast Asia, but formed a distinct branch. Rabies virus diversity might be attributed to dog movements among counties, provinces, and neighboring countries. These findings suggest that Yunnan Province is a focal point for spread of rabies between Southeast Asia and China.
Zinc finger E-box binding homeobox factor 1 (ZEB1), as a crucial mediator of "epithelial-mesenchymal transition," contributes to malignant progression of various epithelial tumors. However, its involvement in human esophageal squamous cell carcinoma (ESCC) remains unclear. In order to investigate the expression pattern of ZEB1 in ESCC tissues and evaluate its associations with tumor progression and patients' prognosis, 100 pairs of formalin-fixed and paraffin-embedded cancerous and adjacent noncancerous tissues from patients with ESCC were used to detect the expression pattern of ZEB1 by immunohistochemistry. Then, the association between ZEB1 expression, clinicopathological parameters, and prognosis of ESCC was examined. We also performed migration and invasion assays of small interfering RNA (siRNA)-targeted ZEB1-transfected cells in vitro. As a result, expression level of ZEB1 was significantly higher in ESCC tissues compared to that in adjacent noncancerous tissues (P?0.001). High expression of ZEB1 was observed in 55.00 % (55/100) of ESCCs. In addition, high ZEB1 expression was found to be closely correlated with advanced tumor stage (P?=?0.001), positive lymph node metastasis (P?=?0.001), great tumor depth (P?=?0.03), and high histologic grade (P?=?0.008). Moreover, multivariate analysis showed that the status of ZEB1 expression was an independent predictor for overall survival in ESCC. Furthermore, knockdown of ZEB1 by transfection of siRNA-ZEB1 abrogated the migration and invasion of ESCC cells in vitro. Taken together, our data offer the convincing evidence that ZEB1 may play a crucial role in promoting aggressive ESCC progression. ZEB1 may serve as an effective prognostic marker and a potential target for therapeutic intervention of ESCC.
We developed two efficient protocols for the synthesis of feruloyl and caffeoyl derivatives from commercial vanillin and veratraldehyde. Pharmacological activities were assessed against a panel of human cancer cell lines in vitro. Most synthesized compounds demonstrated attractive cytotoxicity. Several new compounds demonstrated significant antiproliferative and cytotoxic activities against HeLa and Bewo tumor cell lines. In particular, 5-nitro caffeic adamantyl ester showed broad spectrum of tumor inhibition in 10 cell lines, and reduced tumor weight by 36.7% in vivo when administered at a dose of 40 mg kg(-1).
The connection between an altered gut microbiota and metabolic disorders such as obesity, diabetes, and cardiovascular disease is well established. Defects in preserving the integrity of the mucosal barriers can result in systemic endotoxaemia that contributes to chronic low-grade inflammation, which further promotes the development of metabolic syndrome. Interleukin (IL)-22 exerts essential roles in eliciting antimicrobial immunity and maintaining mucosal barrier integrity within the intestine. Here we investigate the connection between IL-22 and metabolic disorders. We find that the induction of IL-22 from innate lymphoid cells and CD4(+) T cells is impaired in obese mice under various immune challenges, especially in the colon during infection with Citrobacter rodentium. While innate lymphoid cell populations are largely intact in obese mice, the upregulation of IL-23, a cytokine upstream of IL-22, is compromised during the infection. Consequently, these mice are susceptible to C. rodentium infection, and both exogenous IL-22 and IL-23 are able to restore the mucosal host defence. Importantly, we further unveil unexpected functions of IL-22 in regulating metabolism. Mice deficient in IL-22 receptor and fed with high-fat diet are prone to developing metabolic disorders. Strikingly, administration of exogenous IL-22 in genetically obese leptin-receptor-deficient (db/db) mice and mice fed with high-fat diet reverses many of the metabolic symptoms, including hyperglycaemia and insulin resistance. IL-22 shows diverse metabolic benefits, as it improves insulin sensitivity, preserves gut mucosal barrier and endocrine functions, decreases endotoxaemia and chronic inflammation, and regulates lipid metabolism in liver and adipose tissues. In summary, we identify the IL-22 pathway as a novel target for therapeutic intervention in metabolic diseases.
The present study investigated the temporal course of neural discriminations of acoustic cues of English lexical stress (i.e., pitch, intensity and duration) in Cantonese-speaking children. We used an event-related potential (ERP) measure with a multiple-deviant oddball paradigm to record auditory mismatch responses to four deviants, namely, a change in pitch, intensity, or duration, or a change in all three acoustic dimensions, of English lexical stress in familiar words. In the time window of 170-270ms, we found that the pitch deviant elicited significant positive mismatch responses (p-MMRs) and that the duration deviant elicited a mismatch negativity (MMN) response as compared with the standard. In the time window of 270-400ms, the intensity deviant elicited a significant p-MMR, whereas both the duration and the three-dimension changed deviants elicited significant MMNs. These results suggest that Cantonese-speaking children are sensitive to either single or convergent acoustic cues of English words, and that the relative weighting of pitch, intensity and duration in stress processing may correlate with different ERP components at different time windows in Cantonese second graders.
This study aimed to investigate the expression of metastasis suppressor 1 (MTSS1) in cervical intraepithelial neoplasia (CIN) and malignant cervical tissues, and the role of MTSS1 in carcinogenesis. MTSS1 expression was detected by immunohistochemistry in 147 cervical tissue specimens collected from 30 healthy individuals, 30 patients with cervical CIN I, 30 patients with CIN II-III and 57 patients with cervical cancer. The association between MTSS1 expression and clinicopathological factors was also examined. MTSS1 was found to be positively expressed in 43.33% CIN I cervical tissues, 100% CIN II-III cervical tissues and 100% malignant cervical tissues, but was weakly or negatively expressed in benign cervical tissues. The positive expression rates of MTSS1 were significantly higher in CIN II-III and malignant cervical tissues than in CIN I or normal cervical tissues (P<0.05). When examining MTSS1 expression and clinicopathological factors, the strong positive MTSS1 expression rates in early-stage versus middle- and advanced-stage cervical cancer tissues were 39.13% and 82.35%, respectively. Furthermore, the positive expression rates of MTSS1 were significantly higher in cervical tissues at an advanced clinical stage than those at an early clinical stage (P<0.05). The results suggest that the dysregulation of MTSS1 may be involved in cervical carcinogenesis, and thus MTSS1 may be a novel diagnostic biomarker or therapeutic target in cervical cancer patients.
Young adulthood is a high-risk life stage for weight gain. Evidence is needed to translate behavioural approaches into community practice to prevent weight gain in young adults. This systematic review assessed the effectiveness and reporting of external validity components in prevention interventions. The search was limited to randomized controlled trial (RCT) lifestyle interventions for the prevention of weight gain in young adults (18-35 years). Mean body weight and/or body mass index (BMI) change were the primary outcomes. External validity, quality assessment and risk of bias tools were applied to all studies. Twenty-one RCTs were identified through 14 major electronic databases. Over half of the studies were effective in the short term for significantly reducing body weight and/or BMI; however, few showed long-term maintenance. All studies lacked full reporting on external validity components. Description of the intervention components and participant attrition rates were reported by most studies. However, few studies reported the representativeness of participants, effectiveness of recruitment methods, process evaluation detail or costs. It is unclear from the information reported how to implement the interventions into community practice. Integrated reporting of intervention effectiveness and enhanced reporting of external validity components are needed for the translation and potential upscale of prevention strategies.
In this work, a combined strategy was developed to improve the production of glucose oxidase (GOD) (EC 22.214.171.124) in Pichia pastoris. One of the main challenges facing protein production by the high-density fermentation of P. pastoris is the high demand for oxygen. Another challenge is how to balance a reduction in oxygen consumption and its effects on protein production. Herein, a combined strategy involving mannitol co-feeding, two-stage methanol induction, and the co-expression of the transcriptional activator general control non-derepressible 4 (GCN4) from P. pastoris was used. A two-stage, co-feeding strategy, based on a mannitol/methanol mixture in a 3-L fermentor was used to enhance cell viability and protein production. This resulted in an increased GOD yield of 1208.2 U/mL compared with a control strain (427.6 U/mL). An increase in the copy number of the GCN4 gene enhanced the GOD yield (1634.7 U/mL) by 2.8-fold and the protein concentration (19.55 g/L) by 1.58-fold compared with the control (7.59 g/L). This strategy illustrates a way to overcome the high oxygen requirement during high-density fermentation of P. pastoris and balances the reduction of oxygen consumption and protein production. Moreover, the series of strategies presented in this work provide valuable and novel information for the industrial production of GOD.
Previous studies have shown that formaldehyde (FA) could cause immunotoxicity by changing the number of T lymphocytes and that cytokines play a pivotal role in the regulation of T lymphocytes. However, the previously used cytokine detection methods are difficult to use in the measurement of several cytokines in a small amount of sample for one test. Therefore, the cytometric bead array (CBA) technique was used. CBA showed better analytical efficiency and sensitivity than the previous methods. C57BL/6 mice were exposed to the control (normal saline), low FA concentration (0.5 mg/kg), and high FA concentration (2 mg/kg) for 1 week or 1 month. The contents of cytokines, including Th1-related cytokines (IL-2, IFN-?, and tumor necrosis factor), Th2-related cytokines (IL-4, IL-6, and IL-10), and Th17-related cytokines (IL-17A), were measured by using the BD FACS Canto II Flow Cytometer and analyzed by FCAP ArrayTM Software. Th1/Th2/Th17-related cytokines showed a slightly decreasing trend after low FA exposure. Conversely, a significantly increasing trend was found after high FA exposure. Th1/Th2/Th17-related cytokines all serve important functions in the immune reactions in mice after FA exposure.
To understand the effects of host stage on the fitness of the offspring Aenasius bambawalei Hayat, the sex ratio and body size of their offsprings parasitizing Phenacoccus solenopsis Tinsley of different stages (3rd instar nymph and adult) were determined under the laboratory conditions of (27 +/- 1) degree C, RH (70 +/- 5)% and 14L:10D. Sex ratio, body length and hind tibia length of the offsprings of A. bambawalei parasitizing the P. solenopsis adult females were significantly higher compared to those parasitizing the 3rd instar mealybugs. The hind tibia length of the offspring wasp was closely related to the body length. The body size of the parasitised 3rd instar mealybugs affected the size of the offspring wasp of A. bambawalei significantly, while such relationship was not found between the adult female of mealybugs and the offspring wasp.
Diffuse large B cell lymphoma (DLBCL) is the most common subtype of invasive non-Hodgkin's lymphoma (NHL), account for 30%-40% of NHL in adults, the 5-year survival rate is approximately 25%. Although there was a standard treatment to DLBCL today, approximately 50% of patients can not be cured. As a result, it is still a diligent direction of the researchers to understand the pathogenesis of DLBCL and to explore new effective treatment. Recently, the study of microRNA is a hot topic in biology, microRNAs are a class of small non-coding RNA that have emerged to regulate various of biological processes. MiR-155 is one of the most well-known oncogenic micro-RNA, miR-155 overexpression has been documented in a number of lymphoid neoplasms, extraordinarily in DLBCL. MiR-155 can promote the occurrence of lymphoma through various signaling pathways, such as the BMP/TGF-? and RhoA pathway, it is expected to become a new target for treatment of DLBCL. This article reviews the role and possible mechanisms of miR-155 in the pathogenesis of DLBCL.
In the present work, we constructed a new label-free "inter-sandwich" electrochemical aptasensor for thrombin (TB) detection by employing a cleavage-based hybridization chain reaction (HCR). The designed single-stranded DNA (defined as binding DNA), which contained the thrombin aptamer binding sequence, a DNAzyme cleavage site and a signal reporter sequence, was first immobilized on the electrode. In the absence of a target TB, the designed DNAzymes could combine with the thrombin aptamer binding sequence via complementary base pairing, and then Cu(2+) could cleave the binding DNA. In the presence of a target TB, TB could combine with the thrombin aptamer binding sequence to predominantly form an aptamer-protein complex, which blocked the DNAzyme cleavage site and prevented the binding DNA from being cleaved by Cu(2+)-dependent DNAzyme. As a result, the signal reporter sequence could leave the electrode surface to trigger HCR with the help of two auxiliary DNA single-strands, A1 and A2. Then, the electron mediator hexaammineruthenium (III) chloride ([Ru(NH3)6](3+)) was embedded into the double-stranded DNA (dsDNA) to produce a strong electrochemical signal for the quantitative measurement of TB. For further amplification of the electrochemical signal, graphene reduced by dopamine (PDA-rGO) was introduced as a platform in this work. With this strategy, the aptasensor displayed a wide linearity in the range of 0.0001 nM to 50 nM with a low detection limit of 0.05 pM. Moreover, the resulting aptasensor exhibited good specificity and acceptable reproducibility and stability. Because of these factors, the fabrication protocol proposed in this work may be extended to clinical application.
Lysobacter enzymogenes is a bacterial biological control agent emerging as a new source of antibiotic metabolites, such as heat-stable antifungal factor (HSAF) and the antibacterial factor WAP-8294A2. The regulatory mechanism(s) for antibiotic metabolite biosynthesis remains largely unknown in L. enzymogenes. Clp, a cyclic adenosine monophosphate (cAMP)-receptor-like protein, is shown to function as a global regulator in modulating biocontrol-associated traits in L. enzymogenes. However, the genetic basis of Clp signaling remains unclear. Here, we utilized transcriptome/microarray analysis to determine the Clp regulon in L. enzymogenes. We showed that Clp is a global regulator in gene expression, as the transcription of 775 genes belonging to 19 functional groups was differentially controlled by Clp signaling. Analysis of the Clp regulon detected previously characterized Clp-modulated functions as well as novel loci. These include novel loci involved in antibiotic metabolite biosynthesis and surface motility in L. enzymogenes. We further showed experimentally that Clp signaling played a positive role in regulating the biosynthesis of HSAF and WAP-8294A2, as well as surface motility which is a type-IV-pilus-dependent trait. The regulation by Clp signaling of antibiotic (HSAF and WAP-8294A2) biosynthesis and surface motility was found to be independent. Importantly, we identified a factor Lysobacter acetyltransferase (Lat), a homologue of histone acetyltransferase Hpa2, which was regulated by Clp and involved in HSAF biosynthesis, but not associated with WAP-8294A2 production and surface motility. Overall, our study provided new insights into the regulatory role and molecular mechanism of Clp signaling in L. enzymogenes.
The complete mitochondrial (mt) genome sequences of Neobenedenia melleni were determined and compared with those of Benedenia seriolae and B. hoshinai. This circular genome comprises 13,270 bp and includes all 36 typical mt genes found in flatworms. Total AT content of N. melleni is 75.9 %. ATG is the most common start codon, while nad4L is initiated by GTG. All protein-coding genes are predicted to terminate with TAG and TAA. N. melleni has the trnR with a TCG anticodon, which is the same to B. seriolae but different from B. hoshinai (ACG). The mt gene arrangement of N. melleni is similar to that of B. seriolae and B. hoshinai with the exception of three translocations (trnF, trnT and trnG). The overlapped region between nad4L and nad4 was found in the N. melleni mt genome, which was also reported for the published Gyrodactylus species, but it was not found in those of B. seriolae and B. hoshinai, which are non-coding regions instead. The present study provides useful molecular characters for species or strain identification and systematic studies of this parasite.
Paired Ig-like type 2 receptor (PILR)? inhibitory receptor and its counterpart PILR? activating receptor are coexpressed on myeloid cells. In this article, we report that PILR?, but not PILR?, is elevated in human rheumatoid arthritis synovial tissue and correlates with inflammatory cell infiltration. Pilr?(-/-) mice produce more pathogenic cytokines during inflammation and are prone to enhanced autoimmune arthritis. Correspondingly, engaging PILR? with anti-PILR? mAb ameliorates inflammation in mouse arthritis models and suppresses the production of proinflammatory cytokines. Our studies suggest that PILR? mediates an important inhibitory pathway that can dampen inflammatory responses.
Growing evidence demonstrates subtle left ventricular myocardial dysfunction in patients with metabolic syndrome (MetS), with central obesity, glucose intolerance and inflammation emerging as important contributors. Whether these results can be translated to the right ventricle (RV) is not yet fully elucidated. Furthermore, although lifestyle intervention favorably impacts MetS components and inflammatory biomarkers, its effect on RV myocardial function remains unknown today.
Formaldehyde (FA) is a ubiquitous compound used in a wide variety of industries, and is also a major indoor pollutant emitted from building materials, furniture, etc. Because FA is rapidly metabolized and endogenous to many materials, specific biomarkers for exposure have not been identified. In this study, we identified small metabolite biomarkers in urine that might be related FA exposure. Mice were allowed to inhale FA (0, 4, 8 mg/m3) 6 h per day for 7 consecutive days, and urine samples were collected on the 7th day of exposure. Liquid chromatography coupled with time of flight-mass spectrometry and principal component analysis (PCA) was applied to determine alterations of endogenous metabolites in urine. Additionally, immune toxicity studies were conducted to ensure that any resultant toxic effects could be attributed to inhalation of FA. The results showed a significant decrease in the relative rates of T lymphocyte production in the spleen and thymus of mice exposed to FA. Additionally, decreased superoxide dismutase activity and increased reactive oxygen species levels were found in the isolated spleen cells of exposed mice. A total of 12 small molecules were found to be altered in the urine, and PCA analysis showed that urine from the control and FA exposed groups could be distinguished from each other based on the altered molecules. Hippuric acid and cinnamoylglycine were identified in urine using exact mass and fragment ions. Our results suggest that the pattern of metabolites found in urine is significantly changed following FA inhalation, and hippuric acid and cinnamoylglycine might represent potential biomarker candidates for FA exposure.
The aim of this study was to investigate the acid tolerance response (ATR) in Lactobacillus casei by a combined physiological and proteomic analysis. To optimize the ATR induction, cells were acid adapted for 1 h at different pHs, and then acid challenged at pH 3.5. The result showed that acid adaptation improved acid tolerance, and the highest survival was observed in cells adapted at pH 4.5 for 1 h. Analysis of the physiological data showed that the acid-adapted cells exhibited higher intracellular pH (pHi), intracellular NH4 (+) content, and lower inner permeability compared with the cells without adaptation. Proteomic analysis was performed upon acid adaptation to different pHs (pH 6.5 vs. pH 4.5) using two-dimensional electrophoresis. A total of 24 proteins that exhibited at least 1.5-fold differential expression were identified. Four proteins (Pgk, LacD, Hpr, and Galm) involved in carbohydrate catabolism and five classic stress response proteins (GroEL, GrpE, Dnak, Hspl, and LCAZH_2811) were up-regulated after acid adaptation at pH 4.5 for 1 h. Validation of the proteomic data was performed by quantitative RT-PCR, and transcriptional regulation of all selected genes showed a positive correlation with the proteomic patterns of the identified proteins. Results presented in this study may be useful for further elucidating the acid tolerance mechanisms and may help in formulating new strategies to improve the industrial performance of this species during acid stress.
Using a multiple-deviant oddball paradigm, this study examined second graders' brain responses to Cantonese speech. We aimed to address the question of whether a change in a consonant or lexical tone could be automatically detected by children. We measured auditory mismatch responses to place of articulation and voice onset time (VOT), reflecting segmental perception, as well as Cantonese lexical tones including level tone and contour tone, reflecting suprasegmental perception. The data showed that robust mismatch negativities (MMNs) were elicited by all deviants in the time window of 300-500?ms in second graders. Moreover, relative to the standard stimuli, the VOT deviant elicited a robust positive mismatch response, and the level tone deviant elicited a significant MMN in the time window of 150-300?ms. The findings suggest that Hong Kong second graders were sensitive to neural discriminations of speech sounds both at the segmental and suprasegmental levels.
Safety and high efficacy of vectors are essential requirements for gene therapy. To address these challenges, poly(carboxy betaine methacrylate ethyl ester)-poly(carboxy betaine methacrylate) (PCBMAEE-PCBMA) diblock copolymers were synthesized to form core-shell vector for gene delivery. The hydrophobic PCBMAEE segment, a polyzwitterionic precursor, can condense plasmid DNA (pDNA) into a hydrophobic core, which improves pDNA protection from nuclease attack and maintains the condensed structure against dilution. Moreover, the hydrolysis of PCBMAEE in uptaken gene vectors can enhance the pDNA release and reduce the cytotoxicity caused by the cationic polymer accumulation in the host cells. The PCBMA segment, zwitterionic fouling resistant material, is utilized to stabilize the gene vector in the complex medium and reduce the interference from serum proteins without impeding the endocytosis of DNA vector like PEG protection layer. Results showed that the complex formed by PCBMAEE50-PCBMA14 with luciferase or pEGFP gene exhibit higher transfection efficacy of pDNA than that formed by PEI 25 kDa or Lipofectamine(®) 2000 in tested cell lines (COS-7, HepG-2, HeLa, and HUVEC), especially, in difficult-to-transfect ones, such as HeLa and HUVEC. The luciferase expression level infected by the vectors of PCBMAEE50-PCBMA14/pGL-4 at N/P = 20/1 is 27 times of the branched PEI 25 kDa in COS-7 cells and 16 times of Lipofectamine(®) 2000 in HUVEC. Furthermore, the complex formed by PCBMAEE50-PCBMA14 also show advantages in transfection rate, dosage effectiveness and preservation of transfecting activity in serum contained growth medium. The luciferase expression of the vectors of PCBMAEE50-PCBMA14/pGL-4 at N/P = 20/1 is 230 times higher than that of PEI complex at low vector dosage (the 5% standard dosage). And the transfection rate is 25 times higher than that of PEI complex in 10% serum contained growth medium. In short, all these results indicated that the polymeric gene vector, consisted of convertible hydrophobic polyzwitterionic precursor and fixed polyzwitterionic fouling resistant segment, is a promising candidate for high and stable gene transfection in complex growth medium.
Clinical adverse reactions to ampicillin sodium are closely related to its impurities and degradation products. Several unknown degradation products have been detected in the degradation samples of ampicillin sodium. Therefore, a sensitive and accurate method is required to rapidly identify unknown degradation products.
A first-principles calculation was carried out to investigate the stability and electronic properties of ultra-thin Cx(BN)y heteronanotubes which were composed by joining pure CNT and BNNT segments with different composition and configurations. We found that the stability of Cx(BN)y heteronanotubes is increased with the increasing number of B and N atoms. In addition, all armchair (3,3) Cx|(BN)y heteronanotubes were found to be semiconductors with tunable energy gaps between 0.45 to 1.62 eV. Whereas zigzag (5,0) Cx|(BN)y heteronanotubes can be metal (y?4) or semiconductor (y>4), and it is different from the relatively big zigzag Cx|(BN)y heteronanotubes which are always conductors. It indicates that the energy gap of (5,0) Cx|(BN)y heteronanotubes can be tuned by modifying the value of y. Further, zigzag (5,0) and armchair (3,3) C-BN heteronanotubes were found to be metal and semiconductor, respectively; but zigzag and armchair C-BN heteronanotubes with relatively big diameter are always semiconductor and conductor, respectively. Therefore, the electronic properties of ultra-thin Cx(BN)y heteronanotubes are abnormal when comparing with the relatively big ones. Highlights• The stability of Cx(BN)y heteronanotubes is increased with the increasing value of y.• Zigzag (5,0) Cx|(BN)y heteronanotubes can change from metal (y?4) to semiconductor (y>4) when the value of y increases from 1 to 7.• The band gaps of armchair (3,3) Cx|(BN)y heteronanotubes increase from 0.45 to 1.62 eV when the value of y increases from 1 to 7.• The electronic properties (energy gap) of Cx(BN)y heteronanotubes are tunable and different in comparison with the relatively big ones.
Stainless steel wire is a good substrate for jacket-free stir bar sorptive extraction (SBSE). In this work, we present the first example of chemical modification of graphene on stainless steel wire (SSW) for SBSE. Bio-inspired polydopamine was firstly modified on the SSW covalently; then graphene oxide was introduced and reacted with amino groups of polydopamine layer. The modification of polydopamine and graphene can be repeated by a layer-by-layer strategy, resulting in control of the thickness of graphene layer and increase of extraction capability. The prepared covalently immobilized graphene-stainless steel wire (G-SSW) exhibited good stability under stirring, ultrasonication and treatment with commonly used organic solvents, basic and acidic solutions. Application as stir bar for SBSE, G-SSW was found to possess good extraction efficiency towards pollutant polyaromatic hydrocarbons (PAHs), with enrichment factors from 10086 to 16896. After optimization of conditions, G-SSW based SBSE was coupled HPLC for determination of PAHs in environmental and food samples. The quantitative method has low limits of detection of 0.2-50pg/mL, which is better than some reported SBSE-HPLC methods. The method shows wide linear range (200-5000, 10-1000 and 1-1000pg/mL), good linearity (r?0.9950) and good reproducibility (RSD?4.97%). The method has been applied to soil and food samples, with good selectivity and good recoveries ranging from 88.5-113.6%.
Mandarin Chinese is a lexical tone language that has four tones, with a change in tone denoting a change in lexical meaning. There are few studies regarding lexical tone identification abilities in deafened children using either cochlear implants (CIs) or hearing aids (HAs). Furthermore, no study has compared the lexical tone identification abilities of deafened children with their hearing devices turned on and off. The present study aimed to investigate the lexical tone identification abilities of deafened children with CIs or HAs.
The aim of the present study was to investigate the protective effect of Asarum extract on rats with adjuvant arthritis (AA) and to determine the underlying mechanism. An AA model was established by injecting Freund's complete adjuvant into the rats. The degree of toe swelling, arthritis index, spleen index, and the expression levels of tumor necrosis factor (TNF)-?, interleukin (IL)-1? and IL-6 were measured. In addition, the underlying molecular mechanism was investigated using murine macrophage-derived RAW 264.7 cells. Asarum extract was found to significantly reduce the severity of arthritis by decreasing hind paw swelling, the arthritis index, the spleen index, and TNF-?, IL-1? and IL-6 expression levels in plasma. In vitro, Asarum extract inhibited the nuclear factor (NF)-?B and mitogen-activated protein kinase (MAPK) signaling pathways. These results indicate that Asarum extract may be a therapeutic agent for AA and may exert an anti-inflammatory effect by mediating the NF-?B and MAPK signaling pathways.
Up to one-third of patients who undergo cardiac resynchronisation therapy (CRT) are not responders. To identify potential responders to CRT may be sometimes difficult and time-consuming. Forty-five patients who had undergone CRT implantation for standard indications were evaluated. Electrical left ventricular (LV) lead location was assessed by left ventricular activation time (LVAT), LV lead electrical delay (LVLED), and RV-LV interlead electrical delay (RVsense-LVsense). Anatomic LV pacing location was assessed as basal or mid-ventricular between 3:00 to 5:00 (traditionally optimal site), and all the other positions (traditionally non-optimal site). CRT response was defined as a decrease in LV end-systolic volume (LVESV) exceeding 15% at six months. LVLED was larger in the responder group than that in the non-responder group (67.3 ± 8.5% vs. 55.3 ± 8.1%, P< 0.001). In the multivariate analysis, LVLED and cLBBB morphology were the two independent predictors of positive echocardiographic response to CRT (OR=1.180, P=0.003; OR=7.497, P=0.04, respectively). A cutoff value of LVLED> 54.82% predicted responders with 96.3% sensitivity and 75.2% specificity and the area under the receiver operating characteristic (ROC) curve was 0.844 for LVLED (P=0.002). No relationship was found between the anatomic LV pacing sites and response to CRT (P=0.188). The larger left ventricular lead electrical delay may predict response to cardiac resynchronisation therapy.
To introduce DNA into Streptomyces noursei xinao-4, which produces xinaomycins, we explored an intergeneric conjugal transfer system. High efficiency of conjugation (8×10(-3) exconjugants per recipient) was obtained when spores of S. noursei xinao-4 were heat-shocked at 50 °C for 10 min, mixed with Escherichia coli ET12567 (pUZ8002/pSET152) in the ratio of 1:100, plated on 2CMY medium containing 40 mmol/L MgCl2, and incubated at 30 °C for 22 h. With this protocol, the plasmids pKC1139 and pSET152 were successfully transferred from E. coli ET12567 (pUZ8002) with different frequencies. Among all parameters, the ratio of donor to recipient cell number had the strongest effect on the transformation efficiency. In order to validate the above intergeneric conjugal transfer system, a glycosyltransferase gene was cloned and efficiently knocked out in S. noursei xinao-4 using pSG5-based plasmid pKC1139.
Both blood stability and intelligent-responsiveness after reaching the drug-targeting site are very important features to make desirable nano-drug vehicles (NDVs). Here, a highly nonfouling cross-linked micelle based on a copolymer composed of carboxybetaine methacrylate (CBMA) as hydrophilic segment and 2-(methacryloyloxy)ethyl lipoate (MAEL) as hydrophobic and cross-linked segment is reported. Furthermore, a simple method to evaluate the hemocompatibility of NDVs through examining the activation of a blood-clotting protein (fibrinogen) was introduced. The micelles can encapsulate anticancer drug doxorubicin (DOX) conveniently and release DOX quickly in response to an intracellular reductive environment. With the advantages of excellent stability in fibrinogen (1 mg/mL) PBS solution and 50% fetal bovine serum (FBS), and accelerated intracellular drug release, the biocompatible zwitterionic micelles stabilized by reversible cross-linkage might be a promising drug carrier for cancer chemotherapy.
Cu/La2O3 catalysts with different Cu loadings were prepared by impregnation method and employed in the synthesis of glycerol carbonate and monoacetin from glycerol and CO2 in the presence of CH3CN. The interface between Cu and La2O3 and the structure of catalysts were characterized by XRD, TEM, H2-TPR, CO2-TPD, N2O chemisorptions, UV-vis DSR and XPS. Cu/La2O3 exhibited high catalytic activity for the conversion of glycerol to glycerol carbonate and monoacetin under mild reaction conditions. The effect of reaction parameters on the carbonylation of glycerol was also studied. CH3CN can break the thermodynamic limit of the reaction of glycerol with CO2 to glycerol carbonate through its hydrolysis. Over 2.3%Cu/La2O3 catalyst, the conversion of glycerol and the selectivity to glycerol carbonate and monoacetin were 33.4%, 45.4% and 52.9%, respectively (150°C, 7.0 MPa, 12 h). The size effect of Cu nanoparticles and the basic property of the supports were also examined. The characterization results suggested that the Cu sizes and basic sites on Cu/La2O3 had significant effect on the conversion and selectivity in the carbonylation of glycerol.
To review intermediate-term clinical outcomes of microwave ablation (MWA) compared with open radial nephrectomy (ORN) in small renal cell carcinoma (RCC) patients and to identify prognostic factors associated with two techniques.
Pollution associated to traffic can be considered as one of the most relevant pollution sources in our cities; noise is one of the major components of traffic pollution; thus, efforts are necessary to search adequate noise assessment methods and low pollution city designs. Different methods have been proposed for the evaluation of noise in cities, including the categorization method, which is based on the functionality concept. Until now, this method has only been studied (with encouraging results) for short-term, diurnal measurements, but nocturnal noise presents a behavior clearly different on respect to the diurnal one. In this work 45 continuous measurements of approximately one week each in duration are statistically analyzed to identify differences between the proposed categories. The results show that the five proposed categories highlight the noise stratification of the studied city in each period of the day (day, evening, and night). A comparison of the continuous measurements with previous short-term measurements indicates that the latter can be a good approximation of the former in diurnal period, reducing the resource expenditure for noise evaluation. Annoyance estimated from the measured noise levels was compared with the response of population obtained from a questionnaire with good agreement. The categorization method can yield good information about the distribution of a pollutant associated to traffic in our cities in each period of the day and, therefore, is a powerful tool for town planning and the design of pollution prevention policies.
Tissue inhomogeneity might have an important effect on the treatment accuracy of therapeutic ultrasound. Both computer simulation and measurement were performed to study the influence of tissue inhomogeneity on the temperature distribution and tissue lesion formation induced by focused ultrasound. The inhomogeneous tissue is considered a combination of a homogeneous medium and a phase aberration screen in this article. Temperature distributions and lesion dimensions were predicted using the combination of acoustic non-linear and bio-heat transfer equations. To verify the theoretical predictions, polyethylene plates with phase distributions of different correlation lengths and standard deviations were made to mimic inhomogeneous tissues such as human abdominal tissue, and a series of experiments were performed, including acoustic and thermal measurements. The results indicate that the tissue inhomogeneity caused phase aberration of the ultrasound beam. With increasing standard deviation and correlation length of phase aberration, the scattering level of the acoustic field increased, while ultrasound-induced peak temperature and lesion size decreased. This study provides a theoretical and experimental basis for future development of accurate treatment plans for high-intensity focused ultrasound.
This work investigated the suitability of lipid carriers as potential encapsulation method to improve the physical and chemical stability of microalgae oil high in docosahexaenoic acid (DHA). Lipid carriers with various oil contents were successfully prepared by a microfluidization method using stearic acid as solid lipid, microalgae oil as liquid lipid, and poloxamer 188 as surfactant. Results show that the mean particle diameter of the lipid carriers was in the range of 300 to 350 nm with the polydispersity index below 0.2. The lipid carriers were found to have spherical shape when examined under the transmission electron microscope. Data from the encapsulation efficiency and loading capacity indicate high distribution of microalgae oil throughout the lipid carriers and good physical stability as reflected by the particle size and size distribution during storage. Furthermore, the lower DPPH scavenging activity of lipid carriers compared with that of free microalgae oil suggests better chemical stability of microalgae oil encapsulated in lipid carriers. The addition of microalgae oil into lipid phase could disturb the crystalline order and form lattice defects to enable encapsulation of DHA as revealed by the results from differential scanning calorimetery. Current results suggest that this type of novel lipid carriers could be an efficient and promising carrier system for delivery of microalgae oil.
Iturin A is a potential lipopeptide antibiotic produced by Bacillus subtilis. Optimization of iturin A yield by adding various concentrations of asparagine (Asn), glutamic acid (Glu) and proline (Pro) during the fed-batch fermentation process was studied using an artificial neural network-genetic algorithm (ANN-GA) and uniform design (UD). Here, ANN-GA based on the UD data was used for the first time to analyze the fed-batch fermentation process. The ANN-GA and UD methodologies were compared based on their fitting ability, prediction and generalization capacity and sensitivity analysis.
Neuronal restricted progenitors (NRPs) represent a type of transitional intermediate cells that lie between multipotent neural progenitors and terminal differentiated neurons during neurogenesis. These NRPs have the ability to self-renew and differentiate into neurons, but not into glial cells, which is considered an advantage for cellular therapy of human neurodegenerative diseases. However, difficulty in the extraction of highly purified NRPs from normal nervous tissue prevents further studies and applications. In this study, we report the conversion of human fetal fibroblasts into human induced NRPs (hiNRPs) in 11 days by using just three defined factors: Sox2, c-Myc, and either Brn2 or Brn4. The hiNRPs exhibited distinct neuronal characteristics, including cell morphology, multiple neuronal marker expression, self-renewal capacity, and a genome-wide transcriptional profile. Moreover, hiNRPs were able to differentiate into various terminal neurons with functional membrane properties but not glial cells. Direct generation of hiNRPs from somatic cells will provide a new source of cells for cellular replacement therapy of human neurodegenerative diseases.
In traditional Chinese medicine, Ligusticum wallichii (Chuan Xiong) and its bioactive ingredient, tetramethylpyrazine (TMP), have been used to treat cardiovascular diseases and to relieve various neurological symptoms, such as those associated with ischemic injury. In the present study, we investigated whether ultrasound (US) exposure could enhance the protective effect of TMP against cerebral ischemia/reperfusion (I/R) injury. Glutamate-induced toxicity to pheochromocytoma (PC12) cells was used to model I/R injury. TMP was paired with US to examine whether this combination could alleviate glutamate-induced cytotoxicity. The administration of TMP effectively protected cells against glutamate-induced apoptosis, which could be further enhanced by US-mediated sonoporation. The anti-apoptotic effect of TMP was associated with the inhibition of oxidative stress and a change in the levels of apoptosis-related proteins, Bcl-2 and Bax. Furthermore, TMP reduced the expression of proinflammatory cytokines such as TNF-? and IL-8, which likely also contributes to its cytoprotective effects. Taken together, our findings suggest that ultrasound-enhanced TMP treatment might be a promising therapeutic strategy for ischemic stroke. Further study is required to optimize ultrasound treatment parameters.
Zinc-finger nucleases and transcription activator-like effector nucleases are novel gene-editing platforms contributing to redefine the boundaries of modern biological research. They are composed of a non-specific cleavage domain and a tailor made DNA-binding module, which enables a broad range of genetic modifications by inducing efficient DNA double-strand breaks at desired loci. Among other remarkable uses, these nucleases have been employed to produce gene knockouts in mid-size and large animals, such as rabbits and pigs, respectively. This approach is cost effective, relatively quick, and can produce invaluable models for human disease studies, biotechnology or agricultural purposes. Here we describe a protocol for the efficient generation of knockout rabbits using transcription activator-like effector nucleases, and a perspective of the field.
Brucellosis is a bacterial disease caused by brucella; mainly spread by direct contact transmission through the brucella carriers, or indirect contact transmission by the environment containing large quantities of bacteria discharged by the infected individuals. At the beginning of 21st century, the epidemic among dairy cows in Zhejiang province, began to come back and has become a localized prevalent epidemic. Combining the pathology of brucellosis, the reported positive data characteristics, and the feeding method in Zhejiang province, this paper establishes an [Formula: see text] dynamic model to excavate the internal transmission dynamics, fit the real disease situation, predict brucellosis tendency and assess control measures in dairy cows. By careful analysis, we give some quantitative results as follows. (1) The external input of dairy cows from northern areas may lead to high fluctuation of the number of the infectious cows in Zhejiang province that can reach several hundreds. In this case, the disease cannot be controlled and the infection situation cannot easily be predicted. Thus, this paper encourages cows farms to insist on self-supplying production of the dairy cows. (2) The effect of transmission rate of brucella in environment to dairy cattle on brucellosis spreading is greater than transmission rate of the infectious dairy cattle to susceptible cattle. The prevalence of the epidemic is mainly aroused by environment transmission. (3) Under certain circumstances, the epidemic will become a periodic phenomenon. (4) For Zhejiang province, besides measures that have already been adopted, sterilization times of the infected regions is suggested as twice a week, and should be combined with management of the birth rate of dairy cows to control brucellosis spread.
A 4-stage developmental model, in which auditory sensitivity is fully mediated by speech perception at both the segmental and suprasegmental levels, which are further related to word reading through their associations with phonological awareness, rapid automatized naming, verbal short-term memory and morphological awareness, was tested with concurrently collected data on 153 2nd- and 3rd-grade Hong Kong Chinese children. Nested model comparisons were conducted to test this model separately against alternatives in relation to both Chinese and English word reading using structural equation modeling. For Chinese word reading, the proposed 4-stage model was demonstrated to be the best model. Auditory sensitivity was associated with speech perception, which was related to Chinese word reading mainly through its relations to morphological awareness and rapid automatized naming. In contrast, for English word reading, the best model required an additional direct path from suprasegmental sensitivity (in Chinese) to English word reading. That is, in addition to phonological awareness, Chinese speech prosody was also directly associated with English word recognition. (PsycINFO Database Record (c) 2013 APA, all rights reserved).
The purpose of this research is to develop and evaluate Pluronic P123 (P123)-docetaxel (DTX) conjugate that could form core-shell-type micelles in aqueous solution. DTX was covalently conjugated to P123 via ester bond after the activation of DTX by succinic anhydride. The structure of P123-DTX conjugate was confirmed by 1H-NMR. The self-assembly behavior of the P123-DTX conjugate in aqueous solution was characterized by the measurement of critical micelle concentration (CMC). The CMC of P123-DTX conjugate was (1.34 +/- 0.05) x 10(-5) mol/L, which was lower than that of P123 ((1.91 +/- 0.07) x 10(-5) mol/L). The DTX content in the conjugate could reach 13.69% (wt.%). TEM and DLS analysis showed that the conjugate formed approximately spherical micelles with mean particle size of 85.3 +/- 1.59 nm. In vitro release of DTX from the conjugate micelles showed pH dependence, being faster at lower pH value than that at pH 7.4. In vitro cytotoxicity of P123-DTX conjugate micelles was evaluated by MTT method against HepG2, MCF-7 and B16 cell lines and the result indicated that P123-DTX conjugate micelles showed lower cytotoxicity than the DTX injection-Duopafei. Compared with Duopafei, in vivo antitumor activity of P123-DTX conjugate micelles in Kunming mice bearing B16 tumor was more effective and less toxic. These results indicated that the P123-DTX conjugate micelles prepared in this study may be considered as an alternative and promising DTX delivery system.
Glucose oxidase (GOD) is an important industrial enzyme with many potential applications. In order to increase the production and productivity of GOD by recombinant Pichia pastoris GS115, we investigated the feeding strategies of mixed carbon sources during induction phase, based on results of the optimization of initial cell and methanol concentration on GOD production. The optimal initial cell and methanol concentration were 100 g/L and 18 g/L. During induction phase, the mixed-carbon-sources strategies showed that glycerol, sorbitol or mannitol co-feeding with methanol could enhance GOD production. With mannitol co-feeding (20:1(W/W)), the maximum GOD production and maximum GOD productivity reached 711.3 U/mL and 4.60 U/(mL x h) after an induction period of 156 h. Compared to the control, the enhancements of GOD production and productivity were 66.3% and 67.9%, respectively. Meanwhile, we found an appropriate mannitol co-feeding strategy that would not inhibit the expression of promote. The activity of alcohol oxidase was 8.8 U/g, which was enhanced by 69.2% compared to the control (5.2 U/g). We can use the same optimization process to improve the production of other proteins from recombinant Pichia pastoris by changing the fermentation parameters.
A novel capillary with hydroxyapatite (HAP) as the stationary phase was prepared for open-tubular capillary electrochromatography (OT-CEC). To immobilize HAP, a mussel-inspired polydopamine method was utilized to modify the capillary firstly, generating a polydopamine layer; and then a layer of HAP would be formed on the polydopamine layer by a biomineralization process, to produce a HAP-modified capillary (HAP@capillary). Scanning electron microscopy (SEM), energy dispersive X-ray spectrometry (EDS) and X-ray photoelectron spectroscopy (XPS) provided evidence of nanostructured HAP grown on the surface of the capillary wall. The electroosmotic flow (EOF) characteristic of the HAP@capillary was investigated by varying the percentage of acetonitrile and pH value of the buffer with thiourea as a marker, and a pH-dependent EOF from anode to cathode was observed. The HAP@capillary exhibits high column efficiency for methylbenzene, up to 151?138 plates per meter. Different kinds of compounds including alkylbenzenes, phenols and amines have been successfully separated by the HAP@capillary in CEC mode. The HAP@capillary also possessed good separation ability in capillary liquid chromatography (CLC) mode because of the relatively large ratio of HAP in the capillary; however, the separation efficiency was not as good as that in CEC mode. The reproducibilities of the HAP@capillary were evaluated, and the relative standard deviations were found to be lower than 5%.
Recent evidence shows that the NMDAR-post-synaptic density-95 (PSD95), growth-associated protein 43 (GAP-43) and matrix metalloproteinase-9 (MMP-9) protein enhance neuroplasticity at the subacute stage of stroke. Here, we evaluated whether melatonin would modulate the PSD95, GAP-43 and MMP-9 proteins in cultured neurons exposed to glutamate excitotoxicity and in rats subjected to experimental stroke. Adult male Sprague-Dawley rats were treated with melatonin (5 mg/kg) or vehicle at reperfusion onset after transient occlusion of the right middle cerebral artery (tMCAO) for 90 min. Animals were euthanized for Western immunoblot analyses for the PSD-95 and GAP-43 proteins and gelatin zymography for the MMP-9 activity at 7-day post-insult. Another set of animals was sacrificed for histologic and Golgi-Cox-impregnated sections at 28 days post-insult. In cultured neurons exposed to glutamate excitotoxicity, melatonin significantly up-regulated the GAP-43 and PSD-95 expressions and improved dendritic aborizations (P < 0.05, respectively). Relative to controls, melatonin-treated stroke animals caused a significant improvement of GAP-43 and PSD-95 expressions as well as the MMP-9 activity in the ischemic brain (P < 0.05). Consequently, melatonin also significantly promoted the dendritic spine density and reduced infarction in the ischemic brain, and improved neurobehaviors as well at 28 days post-insult (P < 0.05, respectively). Together, melatonin upregulates GAP-43, PSD-95 and MMP-9 proteins, which likely accounts for its actions to improve neuroplasticity in cultured neurons exposed to glutamate excitotoxicity, and to enhance long-term neuroprotection, neuroplasticity and brain remodeling in stroke rats. This article is protected by copyright. All rights reserved.
The mechanical properties of the shell of ultrasonically synthesized lysozyme microbubbles, LSMBs, were evaluated by acoustic interrogation and nanoindentation techniques. The Youngs modulus of LSMBs was found to be 1.0 ± 0.3 MPa and 0.6 ± 0.1 MPa when analyzed by flow cytometry and AFM, respectively. The shell elasticity and Youngs modulus were not affected by the size of the microbubbles (MBs). The hydrogel-like protein shell of LSMBs offers a softer, more elastic and viscous interface compared to lipid-shelled MBs. We show that the acoustic interrogation technique is a real-time, fast, and high-throughput method to characterize the mechanical characteristics of air-filled microbubbles coated by a variety of materials.
Autoantibodies specific to the angiotensin II type I receptor (anti-AT1-AR) have been implicated in the pathology of congestive heart failure (CHF). Anti-AT1-AR may be associated with left ventricular function in CHF patients treated with perindopril.
A novel biocompatible polymer is developed for antimicrobial and nonstick coatings of wound dressing. The polymer is formed by copolymerization of carboxybetaine ester analogue methacrylate (CB-ester) and small partial poly(ethylene glycol) methacrylate (PEGMA) for cross-linking by hexamethylene diisocyanate (HDI), which is highly resistant to nonspecific protein adsorption and mammalian cell attachment after a quick hydrolysis. A small hydrophobic drug, aspirin, can be incorporated into the new polymer and slowly released to inhibit microorganism growth while the new polymer shows very low cytotoxicity. Moreover, the wound dressing, the new polymer coated medical gauze, shows good mechanic properties, such as flexibility and strength, for medical application. After all, this new nonfouling polymer offers great potential for an antimicrobial wound dressing and other applications.
The objective of the present study was to develop a new method for the simultaneous quantitation of imperatorin and its metabolite xanthotoxol in rat plasma. The samples were prepared with hollow fiber liquid phase microextraction (HF-LPME). The optimized extraction procedure was acquired by assessing extraction solvent, length of the fiber, agitation rate, extraction temperature and time. A comparison of sample pretreatment ways between HF-LPME and deproteinization with methanol was performed, which demonstrated less ion suppression and better sensitivity of HF-LPME. Analytes were separated on a C18 column with a gradient elution consisted of methanol and water containing 1mmol/L ammonium acetate. The detection was accomplished by electrospray ionization (ESI) source operating in the positive ionization mode. Selected-multiple-reaction monitoring (SMRM) scanning was employed, which guaranteed a higher sensitivity compared with MRM mode. Calibration curves were linear over investigated ranges with correlation coefficients greater than 0.9979. Precision varied from 0.26% to 14%, and the accuracy varied within ±5.5%. The developed method was successfully applied to the pharmacokinetic research of imperatorin and its metabolite xanthotoxol after oral administration of imperatorin to rats.
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