Comparative Analysis of Transcriptome and Proteome Revealed the Common Metabolic Pathways Induced by Prevalent ESBL Plasmids in Escherichia coli.

Antibiotic resistance has emerged as one of the most significant threats to global public health. Plasmids, which are highly efficient self-replicating genetic vehicles, play a critical role in the dissemination of drug-resistant genes. Previous studies have mainly focused on drug-resistant genes only, often neglecting the complete functional role of multidrug-resistant (MDR) plasmids in bacteria. In this study, we conducted a comprehensive investigation of the transcriptomes and proteomes of Escherichia coli J53 transconjugants harboring six major MDR plasmids of different incompatibility (Inc) groups, which were clinically isolated from patients. The RNA-seq analysis revealed that MDR plasmids influenced the gene expression in the bacterial host, in particular, the genes related to metabolic pathways. A proteomic analysis demonstrated the plasmid-induced regulation of several metabolic pathways including anaerobic respiration and the utilization of various carbon sources such as serine, threonine, sialic acid, and galactarate. These findings suggested that MDR plasmids confer a growth advantage to bacterial hosts in the gut, leading to the expansion of plasmid-carrying bacteria over competitors without plasmids. Moreover, this study provided insights into the versatility of prevalent MDR plasmids in moderating the cellular gene network of bacteria, which could potentially be utilized in therapeutics development for bacteria carrying MDR plasmids.

Socially Assistive Robots for People Living with Dementia in Long-Term Facilities: A Systematic Review and Meta-Analysis of Randomized Controlled Trials.

BACKGROUND AND OBJECTIVE: The purpose of our study was to explore the immediate and long-term effects of socially assistive robots (SARs) on neuropsychiatric symptoms (NPSs), behavioral and psychological symptoms of dementia (BPSD), positive emotional experiences, and social interaction in older people living with dementia. METHODS: We set keywords and used Boolean operators to search the CINAHL, Cochrane Library, EMBASE, IEEE Digital Library, MEDLINE, PsycINFO, PubMed, Web of Science, Scopus, and Chinese Electronic Periodical Service from inception to February 2022 for randomized controlled trials. The Cochrane Collaboration bias assessment tool was used to assess article quality, and RevMan 5.4.1 software was used to conduct the meta-analysis. RESULTS: A total of 14 studies were included in the meta-analysis. SARs can help people living with dementia reduce their NPS of depression and anxiety, provide happiness from positive emotional experiences, and improve their social interaction through conversation. However, there was no significant improvement in agitation behavior, overall BPSD, or quality of life in people living with dementia. In follow-up, it was found that the effect of SRT was limited. CONCLUSION: SARs can reduce depression and increase positive emotions in people living with dementia. They may also reduce the burden on healthcare workers during the COVID-19 pandemic. This research was registered on PROSPERO CRD42020169340.

Facile Fabrication of Self-Assembly Functionalized Polythiophene Hole Transporting Layer for High Performance Perovskite Solar Cells.

Crystallinity and crystal orientation have a predominant impact on a materials' semiconducting properties, thus it is essential to manipulate the microstructure arrangements for desired semiconducting device performance. Here, ultra-uniform hole-transporting material (HTM) by self-assembling COOH-functionalized P3HT (P3HT-COOH) is fabricated, on which near single crystal quality perovskite thin film can be grown. In particular, the self-assembly approach facilitates the P3HT-COOH molecules to form an ordered and homogeneous monolayer on top of the indium tin oxide (ITO) electrode facilitate the perovskite crystalline film growth with high quality and preferred orientations. After detailed spectroscopy and device characterizations, it is found that the carboxylic acid anchoring groups can down-shift the work function and passivate the ITO surface, retarding the interface carrier recombination. As a result, the device made with the self-assembled HTM show high open-circuit voltage over 1.10 V and extend the lifetime over 4,300 h when storing at 30% relative humidity. Moreover, the cell works efficiently under much reduced light power, making it useful as power source under dim-light conditions. The demonstration suggests a new facile way of fabricating monolayer HTM for high efficiency perovskite devices, as well as the interconnecting layer needed for tandem cell.

Laparoscopic metroplasty: reconstructive surgery for unicornuate uterus with noncommunicating, functional uterine horn.

OBJECTIVE: To demonstrate laparoscopic surgery for a patient with unicornuate uterus and a large hematometra in the noncommunicating uterine horn. DESIGN: Narrated video featuring the diagnostic tests and surgical management. SETTING: Academic tertiary hospital. PATIENT(S): A 13-year-old woman, gravida 0 with menarche at 9 years old, had severe dysmenorrhea during her recent menstrual cycles. The results of transrectal three-dimensional sonography, hysteroscopy, and magnetic resonance imaging (MRI) were consistent for a uterine didelphys composed of a right unicornuate uterus and a left hemiuterus with hematometra and no connection to the cervix. Left adnexal endometrioma and ipsilateral renal agenesis were also noted. INTERVENTION(S): Laparoscopic adhesiolysis and enucleation of the left ovarian endometrioma were first performed. Then, implementing the concepts of Strassman unification method, a horizontal incision was made on the medial side of each hemicorpus, starting from the left rudimentary horn and ending at the fundal area of the right hemiuterus. The incision was carried deep enough to reach the endometrium. The opposing, reverted myometrium was then reapproximated from the lower end of the uterus to the upper fundal part in two layers, using absorbable bidirectional monofilament barbed suture. The outer serosal layer was closed with 1-0 Monocryl via a running continuous suture. On postoperative day 2, the patient was discharged uneventfully. MAIN OUTCOME MEASURE(S): Symptomatic relief and restoration of normal and functional anatomy. RESULT(S): The patient has regular menstrual cycles with no recurrence of dysmenorrhea or ovarian endometrioma. At a 2-year postoperative follow-up evaluation, hysteroscopic examination revealed a single uterine cavity with a small fundal septum. The MRI scans showed an anteverted, symmetrical uterus. CONCLUSION(S): In comparison with the widely applied hemihysterectomy, reconstructive laparoscopic metroplasty not only effectively achieves symptomatic relief but also offers better uterine symmetry and a larger intrauterine cavity via unification of the uterus.

Simultaneous determination of nitrate and nitrite in vegetables by poly(vinylimidazole-co-ethylene dimethacrylate) monolithic capillary liquid chromatography with UV detection.

Simultaneous determination of nitrate (NO3‾) and nitrite (NO2‾) in vegetables was performed on a poly(1-vinylimidazole-co-ethylene dimethacrylate) (VIM-EDMA) monolithic column by capillary liquid chromatography (LC) with UV detection. Good linearity (0.5-100 µg mL-1 i.e. 12.5 -2500 µg g-1 in vegetables) was obtained with coefficient of determination > 0.996. Limits of detection (signal-to-noise ratio: S/N= 3) were estimated at 0.06 and 0.05 µg mL-1, which corresponded to 1.50 and 1.25 µg g-1 for NO2‾ and NO3‾, respectively, in vegetables. The limits of quantification (S/N= 10) were estimated at 0.17 and 0.16 µg mL-1 (4.25 and 4.00 µg g-1 in vegetables) for NO2‾ and NO3‾, respectively. Although the detection limits were relatively higher than other LC-UV techniques, this proposed method offered satisfactory sensitivity for complying the Acceptable Daily Intake (ADI) levels set by EU to monitor the occurrence of NO2‾ and NO3‾in vegetables. The intra-day/inter-day precision (0.14-3.35%/0.06-6.93%) and accuracy (90.33-103.32%/96.00-101.26%) were also examined for method validation. No obvious carry-over and decline of separation efficiency were observed for more than 200 analyses of real samples. The occurrence of NO2‾ and NO3‾in various vegetable samples was investigated to demonstrate the potential of utilizing the developed polymeric monolith-based capillary LC-UV method for food safety application.

Separation of inorganic anions by capillary ion chromatography with UV detection using poly(vinylimidazole-co-ethylene dimethacrylate) monolithic column.

Poly(1-vinylimidazole-co-ethylene dimethacrylate) (VIM-EDMA) monolithic stationary phase was applied for separation of inorganic anions by capillary ion chromatography (IC). The retention of inorganic anions on the VIM-EDMA column was investigated using various salt solutions as the eluent. Good separation of inorganic anions was obtained on VIM-EDMA monolithic column using NH4Cl as the eluent. Good mechanical stability and low swelling propensity values (0.12 and -0.02 for ACN and MeOH, respectively) were obtained. The column repeatability was also examined by determining the relative standard deviations (RSDs) of retention time and peak area of target anions. Relatively low RSDs (n = 7) of retention time (<2.3%) and peak area (<8.8%) were obtained on the VIM-EDMA column. The utilization of VIM-EDMA column for potential environmental application was also demonstrated by determining the occurrence of inorganic anions in various environmental water samples without sample preparation process.

History of Urinary Retention Is a Risk Factor for Infection after Prostate Biopsy: A Nationwide, Population-Based Cohort Study.

BACKGROUND AND AIM: Infection or bleeding after transrectal prostate biopsy remains a concern of both patients and urologists. We explored the risk of association of certain co-morbidities with both complications. PATIENTS AND METHODS: Using the Taiwan National Health Insurance Research Database, we identified patients undergoing prostate biopsy from 2000 to 2013. We used logistic multivariable regression to search for associations between post-biopsy hospitalization and the two co-morbidities within a year after biopsy. RESULTS: Among 3,601 prostate biopsies, 100 infections (3.77%) and 52 (1.44%) bleeding-related emergency room visits and hospitalizations were recorded within 30 days after biopsy. The group having the biopsy as an inpatient exhibited older age (p < 0.0001) and a higher percentage of having diabetes mellitus (p = 0.015) than patients without either complication. The logistic multivariable regression analysis showed that urinary retention, freedom from diabetes, and performance as an outpatient procedure were independent risk factors for infection-related hospitalization (odds ratios 1.81, 1.96, and 1.72; p values 0.031, 0.037, and 0.010, respectively). CONCLUSION: Patients with a recent history of urinary retention have a higher probability of infection-related hospitalization after prostate biopsy.

Enhanced catalytic activities and modified substrate preferences for taxoid 10ß-O-acetyl transferase mutants by engineering catalytic histidine residues.

OBJECTIVES: Taxoid 10ß-O-acetyl transferase (DBAT) was redesigned to enhance its catalytic activity and substrate preference for baccatin III and taxol biosynthesis. RESULTS: Residues H162, D166 and R363 were determined as potential sites within the catalytic pocket of DBAT for molecular docking and site-directed mutagenesis to modify the activity of DBAT. Enzymatic activity assays revealed that the kcat/KM values of mutant H162A/R363H, D166H, R363H, D166H/R363H acting on 10-deacetylbaccatin III were about 3, 15, 26 and 60 times higher than that of the wild type of DBAT, respectively. Substrate preference assays indicated that these mutants (H162A/R363H, D166H, R363H, D166H/R363H) could transfer acetyl group from unnatural acetyl donor (e.g. vinyl acetate, sec-butyl acetate, isobutyl acetate, amyl acetate and isoamyl acetate) to 10-deacetylbaccatin III. CONCLUSION: Taxoid 10ß-O-acetyl transferase mutants with redesigned active sites displayed increased catalytic activities and modified substrate preferences, indicating their possible application in the enzymatic synthesis of baccatin III and taxol.

Preparation and characterization of vinylimidazole-based polymer monolithic stationary phases for reversed-phase and hydrophilic interaction capillary liquid chromatography.

In this study, we applied 1-vinylimidazole (VIM) as the functional monomer to prepare a series of VIM-based monolithic stationary phases for both reversed-phase and hydrophilic interaction capillary liquid chromatography (LC) using various dimethacrylates (EDMA: ethylene dimethacrylate; HDDMA: 1,6-hexanediol dimethacrylate; DDDMA: 1,10-decanediol dimethacrylate) as cross-linkers. With a simple thermally initiated free-radical cross-linking polymerization process, VIM-based monolithic stationary phases have been successfully prepared. The porosity, permeability, and column efficiency of synthesized VIM-based monolithic stationary phases were characterized. With similar total porosity (85-90%), the VIM-HDDMA monoliths showed the lowest permeability among the three sets of VIM-based stationary phases. Various sets of non-polar (alkyl benzenes and polycyclic aromatic hydrocarbons) and polar analytes (phenol derivatives and amphenicol antibiotics) were applied as model compounds to further investigate the retention behavior of the VIM-based monolithic stationary phases for reversed-phase capillary LC analysis using selected VIM-based monolithic columns. While a mixture of organic acids was employed to perform HILIC analysis using the selected VIM-based monolithic columns. The separation selectivity and retention behavior of the VIM-based monolithic stationary phases were compared to those obtained using three previously prepared alkyl methacrylate-based monolithic columns. Strong retention and good resolution of polar analytes (such as phenol derivatives, amphenicol antibiotics, and organic acids) were observed using the selected VIM-based monolithic columns. The strong retention and good resolution might be attributed to the additional hydrogen-bonding between the hydrogen-donating analytes and the hydrogen-accepting imidazolium functionality on the VIM-based stationary phase. The applicability for both reversed-phase and HILIC capillary LC analysis has also been demonstrated using the selected VIM-based monolithic columns.

Bio-production of Baccatin III, an Important Precursor of Paclitaxel by a Cost-Effective Approach.

Natural production of anti-cancer drug taxol from Taxus has proved to be environmentally unsustainable and economically unfeasible. Currently, bioengineering the biosynthetic pathway of taxol is an attractive alternative production approach. 10-deacetylbaccatin III-10-O-acetyl transferase (DBAT) was previously characterized as an acyltransferase, using 10-deacetylbaccatin III (10-DAB) and acetyl CoA as natural substrates, to form baccatin III in the taxol biosynthesis. Here, we report that other than the natural acetyl CoA (Ac-CoA) substrate, DBAT can also utilize vinyl acetate (VA), which is commercially available at very low cost, acylate quickly and irreversibly, as acetyl donor in the acyl transfer reaction to produce baccatin III. Furthermore, mutants were prepared via a semi-rational design in this work. A double mutant, I43S/D390R was constructed to combine the positive effects of the different single mutations on catalytic activity, and its catalytic efficiency towards 10-DAB and VA was successfully improved by 3.30-fold, compared to that of wild-type DBAT, while 2.99-fold higher than the catalytic efficiency of WT DBAT towards 10-DAB and Ac-CoA. These findings can provide a promising economically and environmentally friendly method for exploring novel acyl donors to engineer natural product pathways.

Quantification of 7-aminoflunitrazepam in human urine by polymeric monolith-based capillary liquid chromatography coupled to tandem mass spectrometry.

Using a simple liquid-liquid extraction (LLE) procedure for sample pretreatment, 7-Aminoflunitrazepam (7-aminoFM2), a major metabolite of flunitrazepam (FM2), was determined in urine samples by polymeric monolith-based capillary liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS). The linearity was found in the range of 0.1-50ngmL-1 with a method detection limit (signal-to-noise ratio of 3) estimated at 0.05ngmL-1. Using the proposed method, good precision and recovery were also found in spiked urine samples at the levels of 0.5, 5.0, and 50ngmL-1 (intra-day/inter-day precision: 0.6-1.8% / 0.1-0.8%; post-spiked/pre-spiked recovery: 95.4-102.9% / 96.3-102.5%). In addition, acceptable relative differences (-24.2 - 0.8%) were observed by analyzing clinical urine samples using this monolith-based capillary LC-MS/MS method compared with the results obtained by the routine GC-MC method. Using the monolithic column, no noticeable deterioration of separation efficiency or carry-over was observed for more than 200 injections of urine samples. The applicability of the developed monolith-based capillary LC-MS/MS method was demonstrated by quantifying 7-aminoFM2 in various clinical urine samples. Based on these experimental results, the proposed LLE-monolith-based capillary LC-MS/MS method shows the potential for routine determination of drug metabolites in human urine for clinical and forensic applications.

Discovery of Novel 11-Triazole Substituted Benzofuro[3,2-b]quinolone Derivatives as c-myc G-Quadruplex Specific Stabilizers via Click Chemistry.

The specificity of nucleic acids' binders is crucial for developing this kind of drug, especially for novel G-quadruplexes' binders. Quindoline derivatives have been developed as G-quadruplex stabilizers with good interactive activities. In order to improve the selectivity and binding affinity of quindoline derivatives as c-myc G-quadruplex binding ligands, novel triazole containing benzofuroquinoline derivatives (T-BFQs) were designed and synthesized by using the 1,3-dipolar cycloaddition of a series of alkyne and azide building blocks. The selectivity toward c-myc G-quadruplex DNA of these novel T-BFQs was significantly improved, together with an obvious increase on binding affinity. Further cellular and in vivo experiments indicated that the T-BFQs showed inhibitory activity on tumor cells' proliferation, presumably through the down-regulation of transcription of c-myc gene. Our findings broadened the modification strategies of specific G-quadruplex stabilizers.

Discovery of Small Molecules for Repressing Cap-Independent Translation of Human Vascular Endothelial Growth Factor (hVEGF) as Novel Antitumor Agents.

Angiogenesis is important in tumorigenesis and tumor progression. Human vascular endothelial growth factor (hVEGF) is an angiogenic growth factor that plays a crucial role in tumor progression. The G-rich region within the 5'-untranslated regions (5'-UTR) of hVEGF-A mRNA can form a "switchable" RNA G-quadruplex structure that is essential for a cap-independent translation initiation. We screened our small-molecule library for binders of this G-tract. One novel quinazoline derivative, compound 1, showed a significant specific interaction with the G-tract and destabilized the G-quadruplex structure. The results of cellular experiments revealed that compound 1 down-regulated hVEGF-A translation and significantly impeded tumor cells migration. We also found that compound 1 exhibited tumor-inhibiting activity in MCF-7 xenograft tumors, which might be related to its ability to reduce hVEGF expression. These findings present a new strategy of hVEGF-A translational control in which small molecules interact with G-quadruplex structure in the 5'UTR.

Determination of sulfonamides in animal tissues by modified QuEChERS and liquid chromatography tandem mass spectrometry.

In this study, the salting-out solvent extraction and dispersive solid-phase extraction (dSPE) clean-up steps in QuEChERS (quick, easy, cheap, effective, rugged, and safe) method were optimized to reduce matrix effect and efficiently extract target sulfonamides from a variety of edible animal tissues. The extracted sulfonamides were then analyzed using liquid chromatography tandem mass spectrometry (LC-MS/MS). Good extraction recoveries (74.0-100.3% in five different sources of animal tissues; n=3) with acceptable matrix effect (<10%, except for liver samples) were obtained using the proposed method. For the first time, a commercial ND-lipids cartridge was used to remove hydrophobic matrix components from fat-rich animal tissues in the clean-up step of QuEChERS. In addition, good linearity (0.125-12.5ngg-1) was observed using matrix-matched calibration (in beef). Limits of detection (LODs) were estimated at 0.01-0.03ngg-1 in beef, pork, and chicken samples. For beef tripe and pig liver samples, the LODs were in the range of 0.02-0.04ngg-1. Good intra-day/inter-day precision (1.0-10.5%/0.4-8.0%) and accuracy (95.2-107.2%/97.8-102.1%) were also achieved using the modified QuEChERS for sample pretreatment. The applicability of the modified QuEChERS-LC-MS/MS method was demonstrated by determining the occurrence of target sulfonamides in various edible animal tissues for potential food safety analysis.

Chromatographic selectivity of poly(alkyl methacrylate-co-divinylbenzene) monolithic columns for polar aromatic compounds by pressure-driven capillary liquid chromatography.

In this study, divinylbenzene (DVB) was used as the cross-linker to prepare alkyl methacrylate (AlMA) monoliths for incorporating π-π interactions between the aromatic analytes and AlMA-DVB monolithic stationary phases in capillary LC analysis. Various AlMA/DVB ratios were investigated to prepare a series of 30% AlMA-DVB monolithic stationary phases in fused-silica capillaries (250-µm i.d.). The physical properties (such as porosity, permeability, and column efficiency) of the synthesized AlMA-DVB monolithic columns were investigated for characterization. Isocratic elution of phenol derivatives was first employed to evaluate the suitability of the prepared AlMA-DVB columns for small molecule separation. The run-to-run (0.16-1.20%, RSD; n = 3) and column-to-column (0.26-2.95%, RSD; n = 3) repeatabilities on retention times were also examined using the selected AlMA-DVB monolithic columns. The π-π interactions between the aromatic ring and the DVB-based stationary phase offered better recognition on polar analytes with aromatic moieties, which resulted in better separation resolution of aromatic analytes on the AlMA-DVB monolithic columns. In order to demonstrate the capability of potential environmental and/or food safety applications, eight phenylurea herbicides with single benzene ring and seven sulfonamide antibiotics with polyaromatic moieties were analyzed using the selected AlMA-DVB monolithic columns.