High Incorporation Rate Research Articles

Decorating phenylalanine side-chains with triple labeled 13C/19F/2H isotope patterns.

We present an economic and straightforward method to introduce 13C-19F spin systems into the deuterated aromatic side chains of phenylalanine as reporters for various protein NMR applications. The method is based on the synthesis of [4-13C, 2,3,5,6-2H4] 4-fluorophenylalanine from the commercially available isotope sources [2-13C] acetone and deuterium oxide. This compound is readily metabolized by standard Escherichia coli overexpression in a glyphosate-containing minimal medium, which results in high incorporation rates in the corresponding target proteins.

Nonionic polymer and amino acid-assisted synthesis of ZSM-5 nanocrystals and their catalytic application in the alkylation of 2-methylnaphthalene.

The synthesis of nanosized ZSM-5 zeolites with high crystallinity and suitable acidity is very significant for their great potential in various catalytic applications. Herein, a series of zeolite ZSM-5 crystals with different particle sizes and SiO2/Al2O3 ratios (10-30) were synthesized by a temperature-varying two-step crystallization method in a concentrated gel system containing L-lysine and/or polyvinylpyrrolidone (PVP) additives. By optimizing the addition amounts of the two additives, the crystal size of the ZSM-5 zeolite could be reduced to less than 100 nm. Meanwhile, relatively high crystallinity and framework Al incorporation rates could be achieved, resulting in the generation of high-quality ZSM-5 nanocrystals. The nanosized H-form ZSM-5 zeolite with a SiO2/Al2O3 ratio of 20 showed enhanced catalytic efficiency and stability for the alkylation of 2-methylnaphthalene (2-MN) with methanol to produce an important intermediate, 2,6-dimethylnaphthalene (2,6-DMN). A relatively high and steady yield of 2,6-DMN (above 7.2%) could be achieved during 20 h time-on-stream at 400 °C. The smaller crystal size, higher crystallinity and framework Al content could provide more accessible Brønsted acid sites in the 10-membered ring channel of the ZSM-5 nanocrystals, which are the main active sites responsible for the shape-selectivity of the targeted product of 2,6-DMN. As a result, the formation of other side products like 1-MN and poly-MN could be effectively inhibited, thus leading to an improved 2,6-DMN yield and coke resistance over the nanosized ZSM-5 catalyst.

Polyethyleneimine-mediated gene transfection in porcine fetal fibroblasts.

Polyethylenimine (PEI) has been explored as an efficient non-viral system for delivering genes to cells; however, there were no protocols for its use in porcine fetal fibroblasts (PFF). Therefore, we compared different concentrations of FITC-PEI (0.625, 1.25, 2.5, 5, 10, 20, 40, or 80 µg/mL) and incubation times (30 min, 1 h, or 2 h). It was observed that the incubation time did not affect the internalization of the PEI-FITC and that 30 min was sufficient to capture the complex. The concentrations higher than 10 µg/mL could reach many marked PFF (>90%). Then, two PEI concentrations were tested, 10 or 40 µg/mL, combined with an N/P of 2 with the pmhyGENIE-5 for 30 min. The percentage of PFF-GFP positive was similar between the PEI concentrations in the evaluation time points (24 h, 48 h, and 72 h). However, 40 µg/mL caused higher membrane damage rates. Thus, it can be concluded that concentrations between 10 - 80 µg/ml of PEI promote high incorporation rates, even in periods as short as 30 minutes. Furthermore, it can be stated that the transfection condition used in Polyplexes 1 (10 µg/mL of PEI and 37.5 µg/mL of pmhyGENIE-5 for 30 min) efficiently produces genetically edited porcine fetal fibroblasts with low cell damage.

Could high RAP mixtures be multi-recycled? Validation through long-term performance assessment

Reclaimed Asphalt Pavement (RAP) multi-recycling is a step forward in the circular economy by the reuse of materials in successive phases of the life cycle of infrastructures without downgrading their functionality. RAP application continues to be faced with some challenges and barriers that are yet to be overcome to enable its widespread use mainly for rates up to 100 %. With the increment of RAP mixtures use, the multi-recycling capacity will be a near future problem. This paper assessed the multi-recycling capacity of RAP in high incorporation rate (75 %) mixture for wearing courses, before and after ageing, using two recycling cycles. Both the short- and long-term performances of RAP mixtures were evaluated by mechanical and functional tests. In general, the second recycled RAP mixture showed similar or higher performance than the virgin asphalt mixture. It showed a 23 % lower stiffness modulus, a 37 % higher fatigue resistance and 62 % lower permanent deformation than virgin asphalt mixture. Moreover, it presented non-sensitivity to water damage and good functional performance. The RAP multi-recycling capacity was validated. Further studies can be developed for other types of asphalt mixtures.

Both angled bony-increased offset and metal-augmented baseplates provide satisfactory bone incorporation to the glenoid in reverse total shoulder arthroplasty: a radiographic evaluation using tomosynthesis

Angled bony-increased offset and metal-augmented baseplate have recently been used to achieve neutral to inferior inclination of the glenoid implant. Nonetheless, bone incorporation is difficult to evaluate using computed tomography or other conventional methods owing to the presence of metal artifacts; therefore, whether bone incorporation between the grafted bone and glenoid or between the graft and baseplate implant can be achieved remains unclear. Several studies have reported the effectiveness of tomosynthesis in reducing metal artifacts for the evaluation of implant loosening, bone resorption, and spot welds. We aimed to evaluate and compare the bone incorporation rates between angled bony-increased offset and metal-augmented implants using tomosynthesis with metal artifact reduction technology. We hypothesized that a high bone incorporation rate would be obtained with angled bony-increased offset and a metal-augmented baseplate. A total of 52 patients who underwent reverse total shoulder arthroplasty (TSA) with angled bony-increased offset and 42 patients who underwent reverse total shoulder arthroplasty with metal-augmented baseplate were assessed and followed up for a minimum of 2years. The bone incorporation and implant loosening rates were compared between the 2 groups, and the sites of spot welds and trabeculation were recorded according to zones. Bone incorporation between the bone and prosthesis was defined as a confirmation of spot welds connecting the porous area and bone in more than three zones. Bone incorporation between the native bone and grafted bone was defined as an observation of trabeculation. Glenoid loosening was defined as the presence of at least 1mm radiolucency around the prosthesis in more than 2 zones. Both the angled bony-increased offset and metal-augmented baseplate groups achieved sufficient bone incorporation rates (98% [51/52 cases] vs. 100% [42/42 cases], P=1.0) and low implant loosening rates (2% [1/52 cases] vs. 0% [0/42 cases], P=1.0). Spot welds and trabeculation were likely to be confirmed in the lower parts of the glenoid. The two groups did not show any significant differences regarding bone incorporation rates. Considering the complexity of performing the procedure with angled bony-increased offset, the use of a metal-augmented baseplate can serve as an alternative treatment to avoid superior inclination in reverse total shoulder arthroplasty.

Selenomethionine mis-incorporation and redox-dependent voltage-gated sodium channel gain of function.

Selenomethionine (SeMet) readily replaces methionine (Met) residues in proteins during translation. Long-term dietary SeMet intake results in the accumulation of the amino acid in tissue proteins. Despite the high rates of SeMet incorporation in proteins and its stronger susceptibility to oxidation compared to Met, little is known about the effect of SeMet mis-incorporation on electrical excitability and ion channels. Fast inactivation of voltage-gated sodium (NaV ) channels is essential for exact action potential shaping with even minute impairment of inactivation resulting in a plethora of adverse phenotypes. Met oxidation of the NaV channel inactivation motif (Ile-Phe-Met) and further Met residues causes a marked loss of inactivation. Here, we examined the impact of SeMet mis-incorporation on the function of NaV channels. While extensive SeMet incorporation into recombinant rat NaV 1.4 channels preserved their normal function, it greatly sensitized the channels to mild oxidative stress, resulting in loss of inactivation and diminished maximal current, both reversible by dithiothreitol-induced reduction. SeMet incorporation similarly affected human NaV 1.4, NaV 1.2, NaV 1.5, and NaV 1.7. In mouse dorsal root ganglia (DRG) neurons, 1 day of SeMet exposure exacerbated the oxidation-mediated broadening of action potentials. SeMet-treated DRGs also exhibited a stronger increase in the persistent NaV current in response to oxidation. SeMet incorporation in NaV proteins coinciding with oxidative insults may therefore result in hyperexcitability pathologies, such as cardiac arrhythmias and neuropathies, like congenital NaV channel gain-of-function mutations.

Femoral head allograft for glenoid bone loss in primary reverse shoulder arthroplasty: functional and radiologic outcomes

Several techniques have been adopted during primary reverse shoulder arthroplasty (RSA) to manage glenoid bone defect. Among bone grafts, humeral head autograft is currently the mainstream option. However, autologous humeral heads may be unavailable or inadequate and allografts may be a viable alternative. The aim of the present study was to evaluate the functional and radiological outcomes of femoral head allografts for glenoid bone defects in primary RSA. We conducted a retrospective study with prospective data collection enrolling 20 consecutive patients who underwent RSA with femoral head allografts for glenoid bone defects. Indications for surgery were eccentric cuff tear arthropathy in 10 (50%) cases, concentric osteoarthritis in 9 (45%) cases, and fracture sequelae in one (5%) case. Each patient was evaluated preoperatively and at follow-up by X-ray and computed tomography (CT) and by assessing the range of motion (ROM) and the Constant-Murley score (CMS). A CT-based software, a patient-specific 3D model of the scapula, and patient specific instrumentation were used to shape the graft and to assess the position of K-wire for central peg. Postoperatively, CT scans were used to identify graft incorporation and resorption. After a median follow-up of 26.5 months (24-38), ROM and CMS showed a statistically significant improvement (all p=0.001). The median measures of the graft were as follows: 28 mm (28-29) for diameter, 22° (10°-31°) for angle, 4 mm (2-8 mm) for minimum thickness, and 15 mm (11-21 mm) for maximum thickness. Before the surgery, the median glenoid version was 21.8° (16.5°-33.5°) for the retroverted glenoids and -13.5° (-23° to -12°) for the anteverted glenoids. At the follow-up, the median postoperative baseplate retroversion was 5.7° (2.2°- 1.5°) (p=0.001), and this value was close to the 4° retroversion planned on the preoperative CT-based software. Postoperative major complications were noted in four patients: two dislocations, one baseplate failure following a high-energy trauma, and one septic baseplate failure. Partial graft resorption without glenoid component failure was observed in three cases that did not require revision surgery. The femoral head allograft for glenoid bone loss in primary RSA restores shoulder function with CMS values comparable to those of sex- and age-matched healthy individuals. A high rate of incorporation of the graft and satisfactory correction of the glenoid version can be expected after surgery. The management of glenoid bone defects remains a challenging procedure, and a 15% risk of major complication must be considered.

Impact of Cannabis Seed Incorporation in Layer Diet on Productive Performance and Egg Quality Traits.

The production of nonindustrial cannabis is highly developed in the Moroccan Rif region; however, local farmers consider hemp seeds which are rich in omega 3 and tocopherols, only as by-products of cannabis cultivation with low market value. The local ecotype is considered to be a plant with a cannabinoid content of more than 0.4%. So, the objective of this research is to investigate how the incorporation of this local hemp seed affects productive performance and egg quality traits. The experiment is conducted to evaluate the effects of hemp seed (HS) incorporation on hen laying performance and physical egg quality at three levels: 10% (HS-10% group), 20% (HS-20% group), and 30% (HS-30% group). Ninety-six Lohmann Brown classic laying hens were randomly assigned to a control group and three feed treatments. The sampling was taken after the 28-week rearing period (peak egg laying). Throughout the experiment, low-rate HS inclusion (HS-10%) showed no significant differences in egg-laying performance (p > 0.05). However, the high incorporation rates of HS (20% and 30%) negatively affected the egg-laying performance (84-94% and 80-86%, respectively). The albumen quality was also improved by the HS inclusion, where the highest values of the Haugh unit were recorded, ranging between 68.69 and 73.91 for the HS-30% groups. The results also show that HS inclusion and duration influence significantly the yolk color (p < 0.001). The yellow intensity decreases with HS incorporation and aging, from a dark yellow (b ∗ = 38.63 for the control group) to a very pale yellow (b ∗ = 26.29 for HS 30% group). Based on these findings, we can conclude that the incorporation of nonindustrial Moroccan cannabis seeds (ecotype Beldiya) at low rate in the diet of laying hens does not alter the laying performance or the quality of the egg; therefore, they could be used in poultry feeding as an alternative constituent to partially replace high-cost imported ingredients, such as corn and soybeans.

Antibacterial Electrodeposited Copper-Doped Calcium Phosphate Coatings for Dental Implants.

Dental implants provide a good solution for the replacement of tooth roots. However, the full restoration of tooth functions relies on the bone-healing period before positioning the abutment and the crown on the implant, with the associated risk of post-operative infection. This study aimed at developing a homogeneous and adherent thin calcium phosphate antibacterial coating on titanium dental implants by electrodeposition to favor both implant osseointegration and to limit peri-implantitis. By combining global (XRD, FTIR-ATR, elemental titration) and local (SEM, Raman spectroscopy on the coating surface and thickness) characterization techniques, we determined the effect of electrodeposition time on the characteristics and phases content of the coating and the associated mechanism of its formation. The 1-min-electrodeposited CaP coating (thickness: 2 ± 1 μm) was mainly composed of nano-needles of octacalcium phosphate. We demonstrated its mechanical stability after screwing and unscrewing the dental implant in an artificial jawbone. Then, we showed that we can reach a high copper incorporation rate (up to a 27% Cu/(Cu+Ca) molar ratio) in this CaP coating by using an ionic exchange post-treatment with copper nitrate solution at different concentrations. The biological properties (antibiofilm activity and cytotoxicity) were tested in vitro using a model of mixed bacteria biofilm mimicking peri-implantitis and the EN 10993-5 standard (direct contact), respectively. An efficient copper-doping dose was determined, providing an antibiofilm property to the coating without cytotoxic side effects. By combining the electrodeposition and copper ionic exchange processes, we can develop an antibiofilm calcium phosphate coating on dental implants with a tunable thickness and phases content.

Assessment of the long-term photocatalytic performance of cementitious systems incorporating different mineral admixtures

This work aims to perform an experimental study to examine the long-term photocatalytic performance of TiO2-incorporated cementitious systems under carbonation exposure and the effect of the incorporation of different mineral admixtures (metakaolin or calcite) on the time-dependent photocatalytic performance stability of the cementitious systems. To do this, TiO2-containing mixtures were produced by replacing cement with different mineral admixtures (metakaolin or calcite) at a mass percentage of 15 and 30 % of total powder material. Produced specimens were kept under two different environmental conditions [95 ± 5 % relative humidity (RH) and 20 ± 2 °C (Curing Condition I), and 65 ± 5 % RH, 50 ± 5 °C and %4 CO2 (Curing Condition II)]. Photocatalytic activity of the specimens was assessed by determining NOx degradation rate (%), NO degradation rate (%), NO2 formation rate (%) and system selectivity (%) via photocatalytic degradation tests. To assess the microstructural, physical and chemical properties of the specimens, mercury intrusion porosimeter (MIP) and thermogravimetry (TG/DTG) analyses were used. The results indicated that metakaolin incorporation into the cement-based systems enhanced the long-term photocatalytic performance of the specimens. For Curing Condition II, a better time-dependent performance stability was achieved from the mixture with higher metakaolin incorporation rate. Calcite incorporation into the cement-based systems also improved the time-dependent photocatalytic performance of the mixtures, which was more pronounced for the high calcite incorporation rate. While the carbonation mechanism determined the time-dependent photocatalytic performance of the metakaolin-incorporated mixtures, the presence of defects in the matrix because of the calcite incorporation determined that of the calcite-incorporated mixtures.

447 Early Experience With Ct Evaluation Of Valve Incorporation Prior To Tavr Explant

Introduction: An increasing number of transcatheter aortic valve replacement (TAVR) recipients require reintervention after index TAVR failure. Those with unfavorable anatomy for redo TAVR, endocarditis and/or concurrent cardiac pathologies require surgical TAVR explantation (TAVR-explant). Early results of TAVR-explant have shown a high rate of incorporation of the TAVR valve into surrounding tissues, sometimes requiring unplanned repair/replacement and adding to procedural complexity. We aimed to assess if pre-operative computed tomography angiography (CTA) could accurately assess TAVR incorporation.

A phase 1 study of NTX-301, an oral DNMT1 inhibitor, in patients with MDS and AML (trial in progress).

TPS7077 Background: Patients with relapsed and/or refractory (r/r) myelodysplastic syndrome (MDS), chronic myelomonocytic leukemia (CMML), and acute myeloid leukemia (AML) who are not candidates for stem-cell transplantation have dismal outcomes. Novel therapies are needed. The nucleoside analogue NTX-301 (5-aza-4’-thio-2’-deoxycytidine or Aza-TdC) is incorporated into DNA, where it engages the active site of DNA methyltransferase I (DNMT1), a maintenance methyltransferase that contributes to the hypermethylation and silencing of tumor suppressor genes. DNMT1 can become trapped in a covalent complex with DNA, thus depleting free enzyme and inhibiting the normal maintenance methylation of CpG sites, resulting in re-activation of tumor suppressor genes. Preclinical data suggest a correlation between NTX-301 activity in leukemia xenograft models and decreased levels of DNMT1. NTX-301 offers an improvement over traditional DNA methyltransferase inhibitors (azacitidine, decitabine) by virtue of a higher incorporation rate into DNA at lower levels of cytotoxicity. Methods: This is a phase 1, open-label, multi-center, dose-escalation study to assess the safety, tolerability, and recommended myeloid monotherapy dose of NTX-301 in patients with r/r myeloid neoplasms. Patients ≥ 18 years with r/r higher-risk MDS (intermediate, high, very-high risk by IPSS-R), high-risk CMML (int-2/high by CPSS or CPSS-mol), or AML (marrow blasts ≤ 30% or WBC &lt; 20,000 cells/µL without leukoreduction) are eligible. Other key eligibility criteria: Eastern Cooperative Oncology Group performance status 0–2; adequate cardiac, renal, and liver function; and resolved acute effects of any prior therapy. Successive cohorts of patients will receive escalating doses of NTX-301 starting from 2 mg QD. Each cycle is 21 days in duration (treatment for 5 days/week x 2 weeks and 1 week off). The trial incorporates accelerated titration design for dose level 1 and 2 followed by traditional 3+3 dose escalation design (3-6 patients per cohort) from dose level 3. Intrapatient dose escalation is allowed. NTX-301 treatment will continue until disease progression or unacceptable toxicity, whichever occurs first. Primary endpoints: incidence and severity of adverse events, dose-limiting toxicities (DLTs), maximum tolerated dose (MTD), biologically effective dose (BED). Secondary endpoints: PK parameters of NTX-301, PD assessment through global hypomethylation assay and other markers in blood and marrow, and clinical efficacy [overall response rate (ORR), complete remission (CR), marrow CR (mCR), partial remission (PR), stable disease (SD), hematologic improvement (HI) per mIWG criteria]. After the MTD/BED is identified, the safety and efficacy of NTX-301 will be explored further in selected myeloid malignancies. The study began enrolling patients in January 2021 and is still recruiting. Clinical trial information: NCT04167917.

The influence of 4-thiouridine labeling on pre-mRNA splicing outcomes.

Metabolic labeling is a widely used tool to investigate different aspects of pre-mRNA splicing and RNA turnover. The labeling technology takes advantage of native cellular machineries where a nucleotide analog is readily taken up and incorporated into nascent RNA. One such analog is 4-thiouridine (4sU). Previous studies demonstrated that the uptake of 4sU at elevated concentrations (>50μM) and extended exposure led to inhibition of rRNA synthesis and processing, presumably induced by changes in RNA secondary structure. Thus, it is possible that 4sU incorporation may also interfere with splicing efficiency. To test this hypothesis, we carried out splicing analyses of pre-mRNA substrates with varying levels of 4sU incorporation (0-100%). We demonstrate that increased incorporation of 4sU into pre-mRNAs decreased splicing efficiency. The overall impact of 4sU labeling on pre-mRNA splicing efficiency negatively correlates with the strength of splice site signals such as the 3' and the 5' splice sites. Introns with weaker splice sites are more affected by the presence of 4sU. We also show that transcription by T7 polymerase and pre-mRNA degradation kinetics were impacted at the highest levels of 4sU incorporation. Increased incorporation of 4sU caused elevated levels of abortive transcripts, and fully labeled pre-mRNA is more stable than its uridine-only counterpart. Cell culture experiments show that a small number of alternative splicing events were modestly, but statistically significantly influenced by metabolic labeling with 4sU at concentrations considered to be tolerable (40 μM). We conclude that at high 4sU incorporation rates small, but noticeable changes in pre-mRNA splicing can be detected when splice sites deviate from consensus. Given these potential 4sU artifacts, we suggest that appropriate controls for metabolic labeling experiments need to be included in future labeling experiments.

Phase I trial of 5-aza-4’-thio-2’-deoxycytidine (Aza-TdC) in patients with advanced solid tumors.

3088 Background: The nucleoside analog Aza-TdC inhibits DNA methyltransferase 1 (DNMT1), which regulates methylation-mediated silencing of tumor suppressor genes. Aza-TdC offers an improvement over traditional DNA methyltransferase inhibitors by virtue of a higher incorporation rate into DNA at lower levels of cytotoxicity. Aza-TdC has also shown improved preclinical antitumor activity compared to other hypomethylating agents in some solid tumor xenograft models. In an ongoing phase I trial, we evaluate the safety and activity of Aza-TdC in patients (pts) with advanced solid tumors. Methods: Adult pts with solid tumors whose disease has progressed on standard therapy or for which there is no standard therapy were treated with Aza-TdC administered orally once a day for 5 days of each week for 2 weeks in 21-day cycles. The study followed Simon accelerated titration design 3, with 1 pt per dose level at 100% dose increments. Accelerated titration continued until 1 pt experienced a dose-limiting toxicity (DLT) or 2 pts experience drug-related grade 2 toxicity at any dose level, after which, a 3 + 3 dose escalation design was used. Intrapatient dose escalation was allowed. Correlative studies included pharmacokinetic assays and pharmacodynamic assays in circulating tumor cells. Results: As of January 2021, a total of 18 pts have been enrolled on study. Median pt age is 61.5 years (range 35-84). Tumor types included colorectal adenocarcinoma (5 pts), sarcoma (3), breast carcinoma (2), and ovarian carcinoma (2). The DLTs at 48 mg were grade 3 rash and grade 3 acute kidney injury in one pt and &lt; 75% of dosing completed in another pt due to grade 3 myelosuppression. Among the 10 pts treated at 32 mg, 1 pt experienced a DLT: grade 4 neutropenia. The maximum tolerated dose (MTD) is 32 mg. Grade 3 or 4 toxicities across all cycles possibly attributable to study drug were leukopenia (6), lymphopenia (6), neutropenia (4), rash (2), febrile neutropenia (1), anemia (1), thrombocytopenia (1), acute kidney injury (1), elevated AST (1), elevated ALT (1), diarrhea (1), and dehydration (1). Of the 14 pts evaluable for response, 11 had a best response of stable disease, and 3 had a best response of progressive disease. Median cycles on study is 4 (range 1-10+). A pt with clear cell ovarian carcinoma has been on study for &gt; 10 cycles with stable disease. Conclusions: At the MTD of 32 mg, Aza-TdC is safe and well tolerated with a toxicity profile similar to currently approved hypomethylating agents. Global DNA methylation profiling, RNAseq, and DNMT immunohistochemical analyses of tumor biopsies are planned for the currently accruing dose expansion cohort. Funded by NCI Contract No. HHSN261200800001E. Clinical trial information: NCT03366116.

Performance Assessment of Reclaimed Asphalt Pavement (RAP) in Road Surface Mixtures

Considerable amounts of Reclaimed Asphalt Pavement (RAP) are produced every year, as the road network requires maintenance to ensure the safety and comfort of its users. RAP is a 100% recyclable material and a useful fit to be re-introduced into another cycle without downgrading its functionality. Despite the current knowledge about the benefits associated with RAP use, it is not yet largely applied in several countries. This paper aims to validate, on the basis of both short- and long-term mechanical behaviours, the application of a bituminous mixture with a high RAP incorporation rate (75%) in road pavement wearing courses. A crude tall oil rejuvenator was used. Both short- and long-term oven ageing procedures were employed to simulate the ageing that occurs during mixture production and in-service life, respectively. The tests for validating the RAP mixture as an alternative solution comprised stiffness, resistance to fatigue, permanent deformation, and determination of the water sensitivity. Furthermore, the RAP bitumen mobilisation degree was evaluated and a mixing protocol was established. In comparison with virgin bituminous mixtures, it was found that, in general, the high RAP mixtures presented similar or better behaviour. The ageing process had a hardening effect namely in terms of stiffness and resistance to permanent deformation, without significant effects on the resistance to fatigue and water damage.

Bicelles as a carrier for bioactive compounds in beverages: application to nerolidol, an active sesquiterpene alcohol.

Nerolidol is a natural sesquiterpene alcohol with promising but limited application in food and pharmaceutical fields due to several factors including low photostability and low aqueous solubility. Recently, several carriers loading nerolidol were prepared and tested in fresh orange juice. Lipid vesicles loading nerolidol did not exhibit satisfactory organoleptic properties in this beverage. Hence, DMPC/DHPC bicelles were prepared as a new phospholipid-based carrier for nerolidol at different molar ratios. The bicelle suspensions were characterized in terms of homogeneity, particles size, and morphology. The optimal formulation (phospholipid:nerolidol molar ratio 100:1) was selected based on transparent appearance, homogeneity, and particle size (~ 45nm). Besides, it showed a high encapsulation efficiency of nerolidol and a high incorporation rate of phospholipids. Transmission electron microscopy analysis demonstrated the formation of bicelles. The bicelles membrane fluidity was assessed by 1,6-diphenyl-1,3,5-hexatriene fluorescence anisotropy and differential scanning calorimetry analysis. The membrane fluidity of bicelles appeared to increase in the presence of nerolidol in a concentration dependent manner. To our knowledge this is the first study dealing with the encapsulation of an essential oil component in bicelles.

Mesoporous iron oxide nanoparticles loaded with ciprofloxacin as a potential biocompatible antibacterial system

Nowadays, infectious diseases are still a great challenge for human health. Bacterial infections are one of the primary causes of worldwide mortality. Due to the development of resistance processes, the search for therapeutic alternatives has focused on many researchers of biomedical areas. In this sense, antimicrobial drugs' encapsulation has emerged as a relevant alternative, capable of improving the therapeutic effectiveness, while a significant reduction in the side effects can be observed. Following in this same direction, in the present work, mesoporous iron oxide nanoparticles (MIONs), loaded with ciprofloxacin, were prepared, aiming to evaluate the system's potential application against bacterial infections. MIONs and silanized MIONs (MIONs@APTES) were prepared, presenting an average size below 100 nm and a superficial area of 258.27 and 186.27 m 2 g −1 , respectively. Both samples allowed to load relevant amounts of the drug, reaching a high incorporation rate and a controlled release profile, with different kinetics behavior, when incubated with simulated body fluid (SBF). The in vitro evaluation revealed that fabricated systems display biocompatible profiles over normal human cell lineage, where no significant cytotoxic profile was observed. However, it presented relevant antibiofilm (S aureus strain) properties when compared to controls constituted by free ciprofloxacin and nanoparticles without the drug. From all Results obtained in this work, the produced nanosystems can be considered a potential system to be applied in bacterial infection treatment. • CIPRO-MIONs and CIPRO-MIONS@APTES can be considered as potential new antibacterial systems. • MIONS and silanized MIONs (MIONs@APTES) can load high amounts of Ciprofloxacin drug (CIPRO). • MIONs and MIONs-APTES matrices allowed a controlled release profile of ciprofloxacin. • CIPRO-MIONs and CIPRO-MIONs@APTES displayed a high biocompatible profile over human cells. • CIPRO-MIONs and CIPRO-MIONs@APTES have relevant antibiofilm profiles.

Nomophobia in Lebanon: Scale validation and association with psychological aspects.

ObjectivesNomophobia, an abbreviation of “No mobile phone phobia”, is characterized by the illogical fear of being detached from the mobile phone or unable to use it. Research have provided evidence of an association between increased cellular phone use and multiple health issues, such as anxiety, depression, insomnia, and others. To our knowledge, there are no Lebanese studies about nomophobia, despite the high incorporation rate of mobile phones in Lebanon and the likelihood of suffering from anxiety, depression, and other conditions due to nomophobic attitudes. The study objectives were to validate and confirm psychometric properties of the Nomophobia Questionnaire (NMP-Q) and examine the associations between particular psychological conditions (anxiety, depression, stress, insomnia and impulsivity) and nomophobia among a representative sample of Lebanese people.MethodsThis cross-sectional study was carried out between January and July 2019. It enrolled 2260 residents of the community randomly selected from Lebanon’s Mohafazat. Two villages per sub-district and households from each village were chosen using a random sampling technique. A questionnaire was distributed randomly to the households. SPSS version 25 was used to perform the statistical analysis. A multinomial regression was computed taking the nomophobia categories as the dependent variable (and taking the absence of nomophobia as the reference category) and all variables that showed a significant association in the bivariate analysis as independent variables.ResultsA total of 2260 (80.71%) out of 2800 questionnaires distributed was collected back. The mean age of the participants was 27.98 ± 9.66 years (58.8% females). Moreover, the mean nomophobia score was 71.56 ± 26.92 (median = 71; minimum = 14; maximum = 140). The results showed that 46 (2.0%) had no nomophobia, 769 (34.1%) mild nomophobia [95% CI 0.322–0.361], 1089 (48.3%) moderate nomophobia [95% CI 0.463–0.504] and 349 (15.5%) severe nomophobia [95% CI 0.140–0.170]. Items of the nomophobia scale converged over a solution of three factors that had an Eigenvalue over 1 (Factor 1 = emotions associated to losing connectedness, Factor 2 = not being able to communicate, Factor 3 = not being able to access information; total variance explained = 66.65%, and Cronbach’s alpha = 0.948). The results of a multinomial regression, taking the nomophobia score as the dependent variable, showed that higher age was significantly associated with lower odds of having mild (aOR = 0.97), moderate (aOR = 0.93) and severe (aOR = 0.97) nomophobia respectively. Higher anxiety (aOR = 1.09) and higher insomnia (aOR = 1.04) were significantly associated with higher odds of having severe nomophobia.ConclusionThe results suggest a positive correlation between nomophobia and psychological conditions. There is a need for longitudinal and prospective studies that furnish information with regards of the impact of time on the variables measured, in order to better understand the nature, causes, and attributes of nomophobia.

Riboflavin-Sensitized Photooxidation of Low-Density-Lipoprotein (LDL) Cholesterol: A Culprit in the Development of Cardiovascular Diseases (CVDs).

The oxidation of human low-density-lipoprotein (LDL) particles is responsible for the development of cardiovascular diseases (CVDs). In the present study, the occurrence of riboflavin-sensitized photooxidation of LDL particles was examined in an in vitro system. The presence of light, oxygen, and photosensitizer (50 μM riboflavin) caused the riboflavin-sensitized photooxidation of human LDL particles thereby increasing in the conjugated dienes (CDs) by 32.5 ± 4.8% (p < 0.05), indicating that this could serve as a major culprit in the development of CVDs. A 1 h radiation caused a 63.6 ± 0.3% degradation of the riboflavin content, and this indicates the extremely fast reaction of the riboflavin-sensitized photooxidation. The singlet oxygen quenching capacity of β-carotene was determined at three different concentrations (10, 50, and 100 μM), exhibiting both antioxidant and prooxidant effects, depending on the concentrations used. In addition, ascorbic acid displayed a high incorporation rate into the LDL particles, implying its potential in preventing riboflavin photosensitization of LDL particles. To the best of our knowledge, this is the first report on the riboflavin-sensitized photooxidation of LDL particles in an in vitro system, proposing a new possible mechanism in the development of CVDs.

Identification of permissive amber suppression sites for efficient non-canonical amino acid incorporation in mammalian cells.

The genetic code of mammalian cells can be expanded to allow the incorporation of non-canonical amino acids (ncAAs) by suppressing in-frame amber stop codons (UAG) with an orthogonal pyrrolysyl-tRNA synthetase (PylRS)/tRNAPylCUA (PylT) pair. However, the feasibility of this approach is substantially hampered by unpredictable variations in incorporation efficiencies at different stop codon positions within target proteins. Here, we apply a proteomics-based approach to quantify ncAA incorporation rates at hundreds of endogenous amber stop codons in mammalian cells. With these data, we compute iPASS (Identification of Permissive Amber Sites for Suppression; available at www.bultmannlab.eu/tools/iPASS), a linear regression model to predict relative ncAA incorporation efficiencies depending on the surrounding sequence context. To verify iPASS, we develop a dual-fluorescence reporter for high-throughput flow-cytometry analysis that reproducibly yields context-specific ncAA incorporation efficiencies. We show that nucleotides up- and downstream of UAG synergistically influence ncAA incorporation efficiency independent of cell line and ncAA identity. Additionally, we demonstrate iPASS-guided optimization of ncAA incorporation rates by synonymous exchange of codons flanking the amber stop codon. This combination of in silico analysis followed by validation in living mammalian cells substantially simplifies identification as well as adaptation of sites within a target protein to confer high ncAA incorporation rates.