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Nucleic Acids Research Feb 2019The assembly of DNA fragments with homologous arms is becoming popular in routine cloning. For an in vitro assembly reaction, a DNA polymerase is often used either alone...
The assembly of DNA fragments with homologous arms is becoming popular in routine cloning. For an in vitro assembly reaction, a DNA polymerase is often used either alone for its 3'-5' exonuclease activity or together with a 5'-3' exonuclease for its DNA polymerase activity. Here, we present a 'T5 exonuclease DNA assembly' (TEDA) method that only uses a 5'-3' exonuclease. DNA fragments with short homologous ends were treated by T5 exonuclease and then transformed into Escherichia coli to produce clone colonies. The cloning efficiency was similar to that of the commercial In-Fusion method employing a proprietary DNA polymerase, but higher than that of the Gibson method utilizing T5 exonuclease, Phusion DNA polymerase, and DNA ligase. It also assembled multiple DNA fragments and did simultaneous site-directed mutagenesis at multiple sites. The reaction mixture was simple, and each reaction used 0.04 U of T5 exonuclease that cost 0.25 US cents. The simplicity, cost effectiveness, and cloning efficiency should promote its routine use, especially for labs with a budget constraint. TEDA may trigger further development of DNA assembly methods that employ single exonucleases.
Topics: Cloning, Molecular; Escherichia coli; Exodeoxyribonucleases; Genetic Vectors; Mutagenesis, Site-Directed; Polyethylene Glycols; Tromethamine
PubMed: 30462336
DOI: 10.1093/nar/gky1169 -
Bioanalysis Jul 2016
Topics: Acetates; Edetic Acid; Enzyme-Linked Immunosorbent Assay; Hydrogen-Ion Concentration; Nucleic Acid Hybridization; Oligonucleotides; Tromethamine
PubMed: 27269964
DOI: 10.4155/bio.11.18a1 -
Dental Materials : Official Publication... Nov 2022The aim of this study was to investigate the degradation of inert glass fillers which are commonly used in conventional resin-based composites to provide radiopacity,...
OBJECTIVES
The aim of this study was to investigate the degradation of inert glass fillers which are commonly used in conventional resin-based composites to provide radiopacity, reduce the polymerization shrinkage and improve the mechanical properties.
METHODS
75 mg of five different glass powders (1 µm) was immersed separately into 50 mL of acetic acid (pH 4) and tris buffer (pH 7.4) for up to 4 weeks. At each time point the glass powder was filtered and dried for characterization using ATR-FTIR and XRD to assess the degradation behavior and crystallization. ICP-OES, ISE and pH measurements were performed on the supernatant solutions to monitor the pH and ion release.
RESULTS
Although FTIR and XRD analysis showed no significant glass degradation or crystallization upon immersion, there was a substantial release of ions from the inert fillers, especially from BABFG and CDL. Barium release for these fillers were 270 and 165 ppm respectively. G018-373 glass presented the lowest ion release followed by GM27884 and BABG. The ion release was more pronounced in acidic conditions compared to neutral conditions apart from the fluoride release.
SIGNIFICANCE
Inert glasses are not as inert as previously thought. This may result in leaching of ions, potentially causing toxicity, reduction in mechanical properties, increased wear and subsequent failure of the composite material. The ions released from the inert glass may interfere with other glass fillers such as bioactive glass fillers, inhibiting degradation of the bioactive glass, beneficial ion release from the bioactive glass, pH neutralization and apatite formation.
Topics: Apatites; Barium; Fluorides; Glass; Materials Testing; Powders; Tromethamine
PubMed: 36154969
DOI: 10.1016/j.dental.2022.09.004 -
British Medical Journal Jan 1972
Topics: Acidosis; Airway Obstruction; Aniline Compounds; Benzamides; Bronchitis; Chronic Disease; Expectorants; Humans; Male; Middle Aged; Nikethamide; Oxygen Inhalation Therapy; Respiratory Insufficiency; Respiratory Tract Infections; Smoking Prevention; Toluene; Tromethamine
PubMed: 5066640
DOI: No ID Found -
Biomedicine & Pharmacotherapy =... May 2022Dexketoprofen is an enantiomer of ketoprofen (S+) that belongs to nonsteroidal anti-inflammatory drugs and has analgesic, anti-inflammatory, and antipyretic properties.... (Review)
Review
Dexketoprofen is an enantiomer of ketoprofen (S+) that belongs to nonsteroidal anti-inflammatory drugs and has analgesic, anti-inflammatory, and antipyretic properties. Dexketoprofen has a stronger effect than ketoprofen, which makes it a readily used preparation. The review aims to find in recent original publications data about dexketoprofen and its comparison with other painkilling medications. The systematic literature review was conducted in November 2021 (2018 onwards). We selected 12 articles from PubMed, Google Scholar, Medline Complete databases. In the last 4 years, there have been many publications that shed a new light on dexketoprofen. The article is a comparative analysis of dexketoprofen's action vs other nonsteroidal anti-inflammatory drugs and the combination of dexketoprofen with tramadol vs paracetamol with tramadol. The findings of the review confirm that dexketoprofen is a very good pain reliever more potent than paracetamol. Dexketoprofen produces similar effects to lidocaine and dexmedetomidine. Complex preparations containing dexketoprofen and tramadol are very effective painkilling tandem and are more effective than tramadol and paracetamol therapy in the treatment of acute pain.
Topics: Acetaminophen; Anti-Inflammatory Agents, Non-Steroidal; Ketoprofen; Tramadol; Tromethamine
PubMed: 35299123
DOI: 10.1016/j.biopha.2022.112819 -
Drug Design, Development and Therapy 2016Migraine is a common neurovascular disorder, affecting millions of people worldwide. Current guidelines recommend triptans as first-line treatment for moderate-to-severe... (Review)
Review
Migraine is a common neurovascular disorder, affecting millions of people worldwide. Current guidelines recommend triptans as first-line treatment for moderate-to-severe migraine attacks. Frovatriptan is a second-generation triptan with a longer terminal elimination half-life in blood than other triptans (~26 hours). Three double-blind, randomized crossover preference studies have been recently conducted, assessing efficacy and safety of frovatriptan versus rizatriptan, zolmitriptan, and almotriptan, respectively. Frovatriptan showed favorable tolerability and sustained effect, with a significantly lower rate of relapse over 48 hours versus the other triptans. These findings were confirmed in a series of analyses of patient subsets from the three studies, including patients with menstrually related and oral contraceptive-induced migraine, hypertension, obesity, weekend migraine, as well as patients with migraine with aura. In all patient subsets analyzed, lower headache recurrence rates were observed versus the comparator triptans, indicating a more sustained pain-relieving effect on migraine symptoms. A further randomized, double-blind study demonstrated that frovatriptan given in combination with the fast-acting cyclooxygenase inhibitor dexketoprofen provided improved migraine pain-free activity at 2 hours, and gave more sustained pain-free activity at 24 hours, versus frovatriptan alone. These benefits were observed both when the combination was administered early (<1 hour after symptom onset) or late (>1 hour after onset). Different pharmacokinetic, but synergistic, properties between frovatriptan and dexketoprofen may make the combination of these agents particularly effective in migraine treatment, with rapid onset of action and sustained effect over 48 hours. These benefits, together with potential cost-effectiveness advantages versus other triptans could drive selection of the most appropriate treatment for acute migraine attacks.
Topics: Anti-Inflammatory Agents, Non-Steroidal; Carbazoles; Cross-Over Studies; Double-Blind Method; Drug Therapy, Combination; Female; Humans; Italy; Ketoprofen; Male; Migraine Disorders; Randomized Controlled Trials as Topic; Triazoles; Tromethamine; Tryptamines
PubMed: 27757013
DOI: 10.2147/DDDT.S105932 -
The Oncologist Aug 2021MET overexpression is uncommon, and positive MET immunohistochemistry (1+/2+) was an independent positive prognostic factor for response rate and progression-free...
LESSONS LEARNED
MET overexpression is uncommon, and positive MET immunohistochemistry (1+/2+) was an independent positive prognostic factor for response rate and progression-free survival. Whether MET overexpression can be considered a potential predictive biomarker and be used as an inclusion criterion is worth investigating in a future study.
BACKGROUND
Metatinib tromethamine tablet (metatinib) is a small molecule receptor kinase inhibitor targeting both c-MET and vascular endothelial growth factor receptor 2. This phase I trial aimed to determine the dose-limiting toxicity (DLT) and maximum tolerated dose (MTD), pharmacokinetics, safety, and efficacy of metatinib in patients with advanced solid tumors.
METHODS
Eligible patients received a single dose of metatinib in a 3 + 3 dose-escalation design with dose levels of 25-800 mg/day, after a single dose on day 1, then 2 days off, and then a multidose schedule of once-daily doses for 25 consecutive days (days 4-28). Primary endpoints were MTD and safety; secondary and exploratory endpoints included pharmacokinetics (PK), efficacy, and biomarkers.
RESULTS
Eighteen patients (including nine patients with hepatocellular carcinoma [HCC]) received at least one dose of study drug (one patient quit the study without continuous multiple-dose administration after receiving a single dose of metatinib). Hand-foot skin reaction, diarrhea, and liver dysfunction were the DLTs, and 200 mg/day was the MTD. The most common treatment-related adverse events (TRAEs) were skin toxicity (50%), diarrhea (33.3%), and liver dysfunction (27.8%). Three patients (only one of six in the 200 mg/day cohort; the other two in the 300 mg/day cohort) experienced severe TRAEs: one patient with severe liver dysfunction and two patients with severe liver dysfunction and skin toxicity, respectively. Pharmacokinetics assessment indicated that metatinib was rapidly absorbed and metabolized to the formation of reactive metabolite, SCR-1510, after single-dose administration. The mean time taken to achieve maximum concentration and terminal elimination half-life of SCR-1510 was approximately 2.0-3.0 hours and ranged from 8 to 14 hours. Two patients had partial responses. The objective response rate and disease control rate (DCR) were 11.1% and 61.1%, respectively. The median progression-free survival (PFS) was 2.75 months.
CONCLUSION
Metatinib administration of 200 mg/day was well tolerated, safe, and effective. The MTD was 200 mg/day, which should be recommended in further investigations.
Topics: Carcinoma, Hepatocellular; Humans; Liver Neoplasms; Tablets; Tromethamine; Vascular Endothelial Growth Factor A
PubMed: 33749934
DOI: 10.1002/onco.13760 -
Antimicrobial Agents and Chemotherapy Jun 1988The pharmacokinetics of two oral forms of fosfomycin, tromethamine (trometamol) salt and calcium salt, were studied in five young (age, 29 +/- 3 [standard deviation]... (Comparative Study)
Comparative Study
The pharmacokinetics of two oral forms of fosfomycin, tromethamine (trometamol) salt and calcium salt, were studied in five young (age, 29 +/- 3 [standard deviation] years) and eight elderly (age, 72 +/- 6 years) adults. The subjects received a single 40-mg/kg (body weight) (approximately equal to 3-g) calcium fosfomycin dose and a 25-mg/kg (approximately equal to 2-g) tromethamine fosfomycin dose in fosfomycin acid form. Blood and urine samples were collected for 24 h. Antibiotic concentrations in serum and urine were measured by microbiological assay. In all subjects, the peak levels of the calcium salt in serum were two- to fourfold lower than those of the tromethamine salt (6 to 7 and 18 to 22 micrograms/ml, respectively), indicating poor intestinal absorption of the calcium form. The elimination half-life of the two oral forms was about 5 h in young adults, and the half-life was only moderately longer in elderly subjects, with large individual variations: 8.28 +/- 5.51 h for tromethamine fosfomycin and 11.80 +/- 6.86 h for calcium fosfomycin. In elderly subjects, absorption of the tromethamine salt form was not modified, but the time to peak level was delayed for the calcium salt (2.58 +/- 0.54 h versus 1.41 +/- 0.67 h in young adults). Pharmacokinetic elimination of the two forms of fosfomycin was only moderately affected in elderly subjects; we observed lower urinary elimination, about 58 versus 28% of the dose in 24-h urines for the tromethamine salt and decreased renal clearance of both forms. However, the dosages of tromethamine and calcium fosfomycin need not be adjusted for elderly subjects who have endogenous creatinine clearances above 50 ml/min per 1.73 m2.
Topics: Adult; Aged; Aged, 80 and over; Aging; Female; Fosfomycin; Half-Life; Humans; Male; Middle Aged; Tromethamine
PubMed: 3415215
DOI: 10.1128/AAC.32.6.938 -
The Journal of Antimicrobial... Jul 2023Fosfomycin is a potentially attractive option as step-down therapy for bacteraemic urinary tract infections (BUTI), but available data are scarce. Our objective was to... (Randomized Controlled Trial)
Randomized Controlled Trial
Effectiveness of fosfomycin trometamol as oral step-down therapy for bacteraemic urinary tract infections due to MDR Escherichia coli: a post hoc analysis of the FOREST randomized trial.
BACKGROUND
Fosfomycin is a potentially attractive option as step-down therapy for bacteraemic urinary tract infections (BUTI), but available data are scarce. Our objective was to compare the effectiveness and safety of fosfomycin trometamol and other oral drugs as step-down therapy in patients with BUTI due to MDR Escherichia coli (MDR-Ec).
METHODS
Participants in the FOREST trial (comparing IV fosfomycin with ceftriaxone or meropenem for BUTI caused by MDR-Ec in 22 Spanish hospitals from June 2014 to December 2018) who were stepped-down to oral fosfomycin (3 g q48h) or other drugs were included. The primary endpoint was clinical and microbiological cure (CMC) 5-7 days after finalization of treatment. A multivariate analysis was performed using logistic regression to estimate the association of oral step-down with fosfomycin with CMC adjusted for confounders.
RESULTS
Overall, 61 patients switched to oral fosfomycin trometamol and 47 to other drugs (cefuroxime axetil, 28; amoxicillin/clavulanic acid and trimethoprim/sulfamethoxazole, 7 each; ciprofloxacin, 5) were included. CMC was reached by 48/61 patients (78.7%) treated with fosfomycin trometamol and 38/47 (80.9%) with other drugs (difference, -2.2; 95% CI: -17.5 to 13.1; P = 0.38). Subgroup analyses provided similar results. Relapses occurred in 9/61 (15.0%) and 2/47 (4.3%) of patients, respectively (P = 0.03). The adjusted OR for CMC was 1.11 (95% CI: 0.42-3.29, P = 0.75). No relevant differences in adverse events were seen.
CONCLUSIONS
Fosfomycin trometamol might be a reasonable option as step-down therapy in patients with BUTI due to MDR-Ec but the higher rate of relapses would need further assessment.
Topics: Humans; Fosfomycin; Tromethamine; Anti-Bacterial Agents; Escherichia coli; Urinary Tract Infections; Escherichia coli Infections; Recurrence
PubMed: 37260299
DOI: 10.1093/jac/dkad147 -
Journal of Dentistry Oct 2022To investigate the degradation, fluorapatite formation, biological safety and cutting efficiency on dentine of the mixed fluoride- and chloride-containing bioactive...
OBJECTIVES
To investigate the degradation, fluorapatite formation, biological safety and cutting efficiency on dentine of the mixed fluoride- and chloride-containing bioactive glasses (BGs).
METHODS
Two series of mixed fluoride- and chloride-containing glasses (GPFCl and GPF2.3Cl series) were synthesized using a melt-quench method. Glass transition temperature (T) and the bioactivity in term of glass degradation and fluorapatite formation were evaluated in Tris buffer solution. The cutting efficiency of the powdered BGs (GPF2.3Cl series) on dentine via air abrasion was investigated using white light profilometry and scanning electron microscope. The cytotoxicity of GPF2.3Cl series on human periodontal ligament stem cells (hPLSCs) and oral fibroblasts (OFB) were examined by MTT.
RESULTS
These BGs are highly degradable and able to form fluorapatite within 3h of immersion. The formation of CaF was also found in the high fluoride-containing BGs. The faster glass degradation was evidenced in the BGs with higher chloride. A significant reduction of T from 790°C to 463°C was seen with increasing in calcium halide content. Air abrasion on dentine using the low and intermediate chloride-containing glasses demonstrates clear depressions, while no depression was found using the high chloride-containing glass. Moreover, the studied BGs showed no cytotoxicity to hPLSCs and OFB.
CONCLUSIONS
The glasses with mixed fluoride and chloride integrate the benefits from the presence of both, showing rapid glass degradation, fast fluorapatite formation, excellent biocompatibility and controllable hardness to provide a selective cutting efficiency on dentine.
CLINICAL SIGNIFICANCE
The developed BGs air abrasive with tunable hardness by varying chloride content can selectively cut different dental tissues. In clinic, a relatively hard BG is of great interest for caries preparation, while a soft glass is attractive for tooth cleaning.
Topics: Air Abrasion, Dental; Apatites; Calcium; Chlorides; Fluorides; Glass; Humans; Surface Properties; Tromethamine
PubMed: 36030643
DOI: 10.1016/j.jdent.2022.104252