Pretreatment Protocol Research Articles

Targeted Lipidomics for Characterization of PUFAs and Eicosanoids in Extracellular Vesicles.

Lipids are increasingly recognized as bioactive mediators of extracellular vesicle (EV) functions. However, while EV proteins and nucleic acids are well described, EV lipids are insufficiently understood due to lack of adequate quantitative methods. We adapted an established targeted and quantitative mass spectrometry (LC-MS/MS) method originally developed for analysis of 94 eicosanoids and seven polyunsaturated fatty acids (PUFA) in human plasma. Additionally, the influence of freeze–thaw (FT) cycles, injection volume, and extraction solvent were investigated. The modified protocol was applied to lipidomic analysis of differently polarized macrophage-derived EVs. We successfully quantified three PUFAs and eight eicosanoids within EVs. Lipid extraction showed reproducible PUFA and eicosanoid patterns. We found a particularly high impact of FT cycles on EV lipid profiles, with significant reductions of up to 70%. Thus, repeated FT will markedly influence analytical results and may alter EV functions, emphasizing the importance of a standardized sample pretreatment protocol for the analysis of bioactive lipids in EVs. EV lipid profiles differed largely depending on the polarization of the originating macrophages. Particularly, we observed major changes in the arachidonic acid pathway. We emphasize the importance of a standardized sample pretreatment protocol for the analysis of bioactive lipids in EVs.

Electrical conductivity of foods and food components: The influence of formulation processes

AbstractDevelopment of safety protocols for solid–liquid mixtures processed by continuous flow ohmic heating systems requires the determination of the electrical conductivity of foods. In exploiting the advantages of ohmic heating, it is necessary to formulate the product to ensure uniform heating: toward this end, it is most desirable to match electrical conductivities of all phases as closely as possible. In this work, we investigate pretreatment protocols designed both to increase and decrease electrical conductivities of various solid particles, including components of chicken chow mein. Electrical conductivities of some particles, such as carrots, water chestnuts, and celery, could be increased or decreased by blanching in a medium of higher or lower conductivity. Products such as mushroom were more difficult to alter, requiring combinations of vacuum infusion and blanching. The electrical conductivity of chicken was most difficult to alter by blanching alone and required marination in conductive media for the appropriate lengths of time. Relationships were determined between treatment solution conductivities and the conductivity of the end product. A simulation of an incompletely equilibrated blanch process showed that subsequent thermal lag between fluid and solid during ohmic heating was reduced by increasing the blanch duration, and also by addition of an intermediate equilibration step postblanching. We also detail herein, treatments needed for formulating chicken alginate particles for use as surrogate inoculated particles.Practical applicationsIn this work, we investigate pretreatment protocols designed both to increase and decrease electrical conductivities of various solid particles. Electrical conductivities of some particles, such as carrots, water chestnuts, and celery, could be increased or decreased by blanching. Products such as mushroom were more difficult to alter, requiring combinations of vacuum infusion and blanching. The electrical conductivity of chicken was most difficult to alter by blanching alone and required marination.

Re-exposure to iodinated contrast media in two patients with prior severe hypersensitivity reaction

Most allergic reactions to iodinated contrast media can be managed using a pre-treatment protocol involving corticosteroids and antihistamines. However, under certain circumstances, patients may experience severe allergic symptoms despite pre-treatment. Two Chinese men with a history of severe contrast allergy and unstable angina underwent a desensitization protocol that allowed for successful percutaneous coronary intervention. Rapid desensitization is an effective and safe strategy that may allow other patients with similar allergies to successfully undergo angiography that requires the use of radiocontrast media.

Downregulation of topoisomerase 1 and 2 with acriflavine sensitizes bladder cancer cells to cisplatin-based chemotherapy.

Resistance to cisplatin is a major obstacle to effective treatment of bladder cancer (BC). The present study aimed to determine whether a combination of acriflavine (ACF) with cisplatin could potentiate the antitumor property of cisplatin against the BC cells. Furthermore, the molecular mechanism behind the anticancer action of ACF was considered. Two human BC cells (5637 and EJ138) contain mutated form of p53 was culture in standard condition. Cotreatment protocol (simultaneous combination of IC30 value of ACF + various dose of cisplatin for 72h) and pretreatment protocol (pretreatment with IC15 value of ACF for 24h + various dose of cisplatin for 48h) was used to determine the effect of ACF on the cells' sensitivity to main drug cisplatin. To assess the mechanism of action of ACF, real-time PCR was used to evaluate mRNA levels of hypoxia-inducible factor-1α (HIF-1α), Bax, Bcl-2, topoisomerase1 (TOP1) and topoisomerase 2 (TOP2A). Combination of ACF with cisplatin either as cotreatment or opretreatment protocol could significantly reduce the IC50 values of cisplatin as compared to the IC50 of cisplatin when use as a single drug. In addition, ACF could markedly decrease mRNA expression of TOP1 and TOP2A without changing the expression of HIF-1ɑ, Bax and Bcl-2. Our findings indicate that combination of cisplatin with ACF was able to significantly enhance the sensitivity of BC cells to cisplatin. The antitumor activity of ACF is exerted through the downregulation of TOP1 and TOP2A genes expression. ACF may serve as an adjuvant to boost cisplatin-based chemotherapy.

Comprehensive comparison of sample preparation workflows for proteomics.

Mass spectrometry-based proteomics experiments can be subject to a large variability, which forms an obstacle to obtaining deep and accurate protein identification. Here, to obtain an optimal sample preparation workflow for in-depth proteome identification in human tissues, we systematically compared typical procedures in the four key steps during sample preparation, including two lysis buffers (5% SDS and 7M urea/2M thiourea), acetone precipitation, two proteolytic enzyme methods (in-solution digestion and FASP), and two pre-fractionation methods (SDS-PAGE and hi-pH RPLC). Compared with other methods, the procedure, including urea/thiourea as the lysis buffer, in-solution digestion, followed by hi-pH RPLC, yields an increase in proteome coverage (+15%), matched peptides (+42.4%), and significantly high protein concentrations. We also used combinations of these sample preparation methods to demonstrate a high identification rate in the range of low molecular weight (LMW), and the performance of sample preparation workflows varied between different groups of proteins. Importantly, 3 proteins defined as missing proteins (MPs) following the Human Proteome Project (HPP) guidelines were found in our data set. Overall, our findings provide an optimal sample preparation workflow for highly efficient and unbiased global proteomic analysis in human tissues.

Impact of vitamin AD3E treatment before application of controlled intra-vaginal drug releaser on clinical and hematologic findings, ovarian hormones, and calving rates in she-camels (Camelus dromedarius).

The scope of using controlled intra-vaginal drug releasers (CIDR) in the camel industry was small relative to that of cow, sheep, and goat. The purpose of this study was to determine the effects of intramuscular injection of AD3E (25 ml/400 kg) as a premedication protocol before CIDR treatment on ovarian hormones profile and calving rate in dromedary she-camels (Camelus dromedarius) through measuring concentrations of progesterone (P4) and estradiol (E2) before, during, and after CIDR use. The study was conducted on she-camels (C. dromedarius) (n = 24) aged between 6 and 14 years. The animals were assigned into two equal groups: CIDR treated group (CIDRgr) and AD3E plus CIDR treated group (CIDR+ gr). Except for hemoglobin, the clinical and hematological findings showed no remarkable change either within each independent group or between CIDRgr and CIDR+ gr. Serum P4 and E2 concentrations differed significantly between the first 7 days (during CIDR use) and days from 9 to 15 (after CIDR use) in dromedary camels. Serum concentrations of P4 and E2 showed significant elevation at CIDR+ gr at day 0 compared with CIDRgr. This significant increase in serum of P4 and E2 at CIDR+ gr compared to those of CIDRgr was also observed either during CIDR use (days 3, 5, and 7) or after CIDR use (days 9, 11, 13, and 15). During CIDR use, serum concentrations of P4 were negatively correlated with those of E2 either in CIDRgr or CIDR+ gr, however, they were positively correlated after CIDR removal either in CIDRgror CIDR+ gr. Regarding the calving rate, it was highest in CIDR+ gr whereas CIDRgr showed the lowest one. It was concluded that the AD3E pretreatment protocol of she-camels before insertion of CIDR could be used successfully during the breeding season to improve fertility in she-camels whereas the serum ovarian hormones profile and calving rates were clearly changed due to pretreatment of she-camels with AD3E before CIDR insertion.

Extraction method development for nanoplastics from oyster and fish tissues

Nanoplastics are now found in some environmental media and consumer products. However, very limited data on nanoplastics are available for one of the main human consumption sources of microplastics: seafood. Unlike microplastics, a method for extracting nanoplastics from seafood is still lacking. Herein, a combination of common extraction techniques including enzymatic digestion, sequential membrane filtration, centrifugal concentration, and purification (dialysis and sodium dodecylsulfate (SDS) incubation), was developed to extract nanoplastics from oyster and fish tissues. Corolase with subsequent lipase treatment achieved the highest digestion efficiencies (88– 89%) for non-homogenized tissues compared to other proteases and additional cellulase or H2O2 treatment. With the exception of polyethylene terephthalate (PET), enzymatic digestion did not change the morphology or structure of polyvinyl chloride (PVC), polyethylene (PE), or polystyrene (PS) nanoplastic particles, and the subsequent extraction procedures had good recoveries of 71– 110% for fluorescence-labeled 76-nm PVC and 100- and 750-nm PS, as validated by a Nanoparticle Tracking Analysis (NTA). Few of the 1011 digested residual particles of 150– 300 nm in diameter per oyster or per serving of fish tissue were left in the method blank. Consequently, this efficient approach could be used as a pretreatment protocol for current potential nanoplastic detection methods.

Freeze-Thaw Pretreatment Can Improve Efficiency of Bacterial DNA Extraction From Meconium.

Although the presence of live microbes in utero remains under debate, newborn gastrointestinal bacteria are undoubtedly important to infant health. Measuring bacteria in meconium is an ideal strategy to understand this issue; however, the low efficiency of bacterial DNA extraction from meconium has limited its utilization. This study aims to improve the efficiency of bacterial DNA extraction from meconium, which generally has low levels of microflora but high levels of PCR inhibitors in the viscous matrix. The research was approved by the ethical committee of the Xiamen Maternity and Child Health Care Hospital, Xiamen, China. All the mothers delivered naturally, and their newborns were healthy. Meconium samples passed by the newborns within 24 h were collected. Each sample was scraped off of a sterile diaper, transferred to a 5-ml sterile tube, and stored at −80°C. For the assay, a freeze-thawing sample preparation protocol was designed, in which a meconium-InhibitEX buffer mixture was intentionally frozen 1–3 times at −20°C, −80°C, and (or) in liquid nitrogen. Then, DNA was extracted using a commercial kit and sequenced by 16S rDNA to verify the enhanced bacterial DNA extraction efficiency. Ultimately, we observed the following: (1) About 30 mg lyophilized meconium was the optimal amount for DNA extraction. (2) Freezing treatment for 6 h improved DNA extraction at −20°C. (3) DNA extraction efficiency was significantly higher with the immediate thaw strategy than with gradient thawing at −20°C, −80°C, and in liquid nitrogen. (4) Among the conditions of −20°C, −80°C, and liquid nitrogen, −20°C was the best freezing condition for both improving DNA extraction efficiency and preserving microbial species diversity in meconium, while liquid nitrogen was the worst condition. (5) Three freeze-thaw cycles could markedly enhance DNA extraction efficiency and preserve the species diversity of meconium microflora. We developed a feasible freeze-thaw pretreatment protocol to improve the extraction of microbial DNA from meconium, which may be beneficial for newborn bacterial colonization studies.

350 Effect of pre-hospital treatment for STEMI patients undergoing primary PCI

Abstract Aims The appropriate timing to administer antithrombotic therapies in ST-elevation myocardial infarction (STEMI) remains uncertain. This study aims to evaluate the role of antithrombotic therapy administration at first medical contact (FMC) compared to the administration in the Cathlab. Methods and results We conducted a ‘before-after’ observational study enrolling STEMI undergoing primary percutaneous coronary intervention (PCI). Outcomes were evaluated during two successive periods, before (control group: aspirin only at FMC) and after (pre-treated intervention group: heparin, aspirin plus ticagrelor at FMC) the introduction of a new regional pre-treatment protocol. 537 consecutive patients (300 in control vs. 237 in intervention group) were enrolled. The pre-treated compared to no pre-treated population showed better basal reperfusion, expressed as basal thrombolysis in myocardial Infarction (TIMI)-flow (p for trend P < 0.001). Pre-treated population showed lower frequency of TIMI 0 (56.5% vs. 73.7%, OR: 0.46, 95% CI: 0.32–0.67, P < 0.001) and higher frequency of TIMI 2–3 (33.3% vs. 19.7%; OR: 2.0; 95% CI: 1.38–2.00, P < 0.001) and TIMI 3 [14.3% vs. 9.7%, OR: 1.56, 95% CI: (0.92–2.65), P = 0.094]. Pre-treated compared to no pre-treated population showed reduced infarct size expressed as Troponin Peak [20 286 (8726–75027) vs. 48 676 (17229–113900), P = 0.001], and higher left ventricular ejection fraction at discharge [53% (44–59) vs. 50% (44–56), P = 0.027]. In-Hospital BARC ≥2 bleeding were similar (2.1% vs. 2.0%, P = 0.929, in pre-treated vs. no pre-treated population, respectively). Conclusions This study provides support for an early pre-treatment strategy in STEMI patients and confirmed the importance of an efficient organization of STEMI networks which allow initiation of antithrombotic treatment at FMC.

Bonding Universal Dental Adhesive to Developmentally Hypomineralised Enamel.

To investigate the effect of pretreatment protocols involving Papacarie Duo gel and Scotchbond Universal (SU) on the microshear bond strength (µSBS) of resin composite (RC) to hypomineralised enamel (HE). Specimens of normal enamel (NE) and HE were derived from extracted hypomineralised first permanent molars (FPMs). Based on the colour of demarcated opacities, HE specimens were classified as creamy/white (CW) or yellow/brown (YB). The specimens were randomly allocated into eight groups (n = 20). Each group involved pretreatment with Papacarie Duo gel or no pretreatment, and SU applied in etch-and-rinse (E&R) or self-etch (SE) mode. All specimens were bonded with RC and subjected to µSBS testing. Failure modes were analysed using an optical microscope and SEM. Comparing NE with HE, the following factors were found to be significant (p < 0.001): type of enamel substrate, deproteinising pretreatment, and etching mode. Comparing CW HE with YB HE, a significant interaction between "deproteinising pretreatment" and "etching mode" was demonstrated (p = 0.028). When subjected to the concurrent use of Papacarie Duo gel and phosphoric acid etching, HE specimens showed a significant increase in µSBS (p < 0.001). Deproteinising pretreatment using Papacarie Duo gel followed by the application of SU in E&R mode led to increased µSBS of resin composite to HE.

Rapid and multiplexed quantification of Salmonella, Escherichia coli O157:H7, and Shigella flexneri in ground beef using flow cytometry

Salmonella, Escherichia coli O157:H7 (E. coli O157:H7) and Shigella flexneri (S. flexneri) might contaminate similar types of meat products and cause deadly diseases in humans. In reality, ground beef samples may carry more than one pathogen and a rapid and accurate detection method for the simultaneous identification of multiple specific pathogenic strains in ground beef is crucial. In this study, a sample pretreatment protocol and a flow cytometry method were developed for rapid and multiplexed quantification of the three pathogens without cultural enrichment in ground beef. The whole process of sample pretreatment, staining, and instrument analysis can be accomplished within 1 h. The three bacteria upon sample pretreatment were demonstrated good recoveries (93.8%–101.2%). The quantitative detection range of the mothed was 103 to 108 cells/g for all three pathogens, and the detection limit for Salmonella, E. coli O157:H7 and S. flexneri in ground beef were 3.1 × 103 cells/g, 2.1 × 103 cells/g and 2.3 × 103 cells/g, respectively. Therefore, the as-developed approach is a rapid and quantitative method for multiplexed detection of Salmonella, E. coli O157:H7, and S. flexneri in ground beef.

Green sample pre-treatment technique coupled with UHPLC-MS/MS for the rapid biomonitoring of dietary poly-unsaturated (omega) fatty acids to predict health risks

Polyunsaturated fatty acids (PUFAs) consumption indicates beneficial effects on cardiovascular disease (CVD) and physiological processes in humans. However, the inappropriate ratio of omega-(ω)-PUFA levels in human blood is considered as raising the risk of CVD. Therefore, monitoring dietary ω-FAs in human serum is vital for early diagnosis for individuals to predict CVD risk. This work reports a fast green sample pre-treatment protocol for sensitive and simultaneous monitoring of ω-3-FAs and ω-6-FAs in serum by novel in-syringe-based ultrasonication-assisted alkaline hydrolysis coupled with vortex-induced liquid-liquid microextraction (IS-USAH-VI-LLME) technique connected with UHPLC–MS/MS analysis. Factors affecting extraction recoveries of ten ω-PUFAs by the presented method were well-studied. ω-3 and ω-6 PUFAs demonstrated excellent linearities between the concentrations between 0.1–10,000 ng mL−1 with good regression coefficients between 0.9910–0.9997. The detection and quantification limits were between 0.05–0.35 and 0.16–1.07 ng mL−1, demonstrating that the presented method is highly sensitive and versatile. The precision of the technique was <8.2% that deemed acceptable in clinical analysis. Further, the proposed method was applied for ω-PUFAs analysis in human blood samples, and spiked recoveries showed between 80.32–119.34% with <9.82% precision. Results proved that the developed method is green, sensitive, and reliable to simultaneously determine ten ω-PUFAs in human blood samples for clinical diagnosis applications for predicting health hazards.

Evaluation of GnRH antagonist pretreatment before ovarian stimulation in a GnRH antagonist protocol in normal ovulatory women undergoing IVF/ICSI: a randomized controlled trial

BackgroundSynchronization of follicles is key to improving ovulation stimulation with the gonadotropin-releasing hormone (GnRH) antagonist protocol. GnRH antagonist administration in the early follicular phase can quickly decrease gonadotrophin (Gn) levels and achieve downregulation before stimulation, which may improves synchronization. A previous small randomized controlled study (RCT) showed that pretreatment with a GnRH antagonist for 3 days before stimulation may increase oocyte retrieval but cannot increase the pregnancy rate. This study investigated whether the GnRH antagonist pretreatment protocol in ovulatory women can increase the synchronization of follicles and pregnancy outcomes compared with the conventional GnRH antagonist protocol.MethodsThis RCT included 136 normal ovulatory women undergoing in vitro fertilization (IVF)/intracytoplasmic sperm injection (ICSI). Both groups were treated with recombinant follicle-stimulating hormone (r-FSH) and a flexible GnRH antagonist protocol. The women were randomized into two equal groups with or without GnRH antagonist administration from day 2 of the menstrual cycle for 3 days before stimulation. Our primary outcome was the number of retrieved oocytes. Secondary outcomes included the pregnancy rate and live birth rate.ResultsBoth groups had similar baseline characteristics. The number of retrieved oocytes in the study group was comparable to that in the control group (9.5 [8.0–13.0] vs. 11.0 [7.0–14.8], P = 0.469). There was no significant difference in the follicle size. The fertilization rate, number of good-quality embryos, implantation rate, pregnancy rate, ongoing pregnancy rate, live birth rate per embryonic transfer cycle, and miscarriage rate were similar between the two groups.ConclusionThis large RCT analysed GnRH antagonist pretreatment with the GnRH antagonist protocol applied to normal ovulatory women undergoing IVF/ICSI. The number of retrieved oocytes and pregnancy outcomes did not significantly vary.Trial registrationChinese Clinical Trial Registry, ChiCTR1800019730. Registered 26 November 2018.

Effect of Diode Laser irradiation on the Bond Strength of Fiber Posts after Endodontic Retreatment Using Different Sealers.

Purpose: To investigate the effect of diode laser activated irrigation on bond strength between fiber post and radicular dentin after endodontic retreatment using two different sealers. Materials and Methods: Seventy-two extracted human maxillary central incisors were collected, decoronated, endodontically prepared then divided to three groups (n=24) according to sealer type: GP group: gutta-percha without sealer (control group); AD group: AD sealer; and BC group: Bioceramic sealer. Specimens were endodontically retreated after 30 days using the same sealers. 10 mm deep post spaces were prepared in each specimen. Each group was further sub-divided into three subgroups (n=8) according to post space treatment: irrigated with saline (Control), (EDTA) and (laser and EDTA). One tooth from each group was selected for SEM analysis. Posts were cemented, push out tests were performed. Failure modes were analyzed using a digital microscope. Results: With all dentin pretreatment protocols, AD group recorded the highest bond strength mean value followed by GP and BC group respectively, with significant difference (P 0.05). Predominant failure mode was the mixed type as detected by Chi square test. SEM revealed absence of smear layer and opened dentinal tubules after (laser and EDTA) treatment. Conclusions: laser activated irrigation of root dentin significantly improved the bond strength of fiber post after endodontic retreatment with both sealers.

Evaluation of developmentally hypomineralised enamel after surface pretreatment with Papacarie Duo gel and different etching modes: an in vitro SEM and AFM study.

This study aimed at investigating the surface morphology and nanotopography of normal enamel (NE) and developmentally hypomineralised enamel (HE) when subjected to various pretreatment protocols under scanning electron microscopy (SEM) and atomic force microscopy (AFM). Sixteen NE, 16 creamy/white (CW) HE and 16 yellow/brown (YB) HE specimens sectioned from extracted hypomineralised first permanent molars (FPMs) were included in this study. They were randomly distributed into 12 experimental groups (n = 4). Each group involved the following: (1) deproteinisation with Papacarie Duo® gel or no deproteinisation, and (2) the use of Scotchbond™ Universal Adhesive (Scotchbond) in self-etch (SE) mode or 37% phosphoric acid etchant. Subsequently, the surface morphology and nanotopography of pretreated enamel specimens were evaluated under SEM and AFM, respectively. SEM observation showed that deproteinisation with Papacarie Duo® gel before phosphoric acid etching led to favourable etching patterns. This was consistent across all groups irrespective of the type of enamel specimen and the severity of hypomineralisation. In contrast, AFM results identified three factors that influenced surface parameters: (1) type of enamel specimen, (2) severity of hypomineralisation and (3) etching mode. YB HE recorded higher surface roughness values than CW HE and NE when subjected to the same pretreatment protocol. Deproteinisation and the application of Scotchbond in SE mode led to minimal topographic changes; however, acid etching was associated with an increase in surface roughness. Deproteinisation with Papacarie Duo® gel followed by acid etching contributed to improved etching patterns on HE.

Preparation of activated carbon from pine sawdust with hydrothermal-pressure preconditioning

The aim of this paper has been to find a biomass pretreatment protocol for preparing activated carbons without using any of the commercially-applied chemical additives. Instead, we employed a hydrothermal-pressure preconditioning protocol (160 ℃, 300–390 psi) to induce rapid steam expansion within pine sawdust, followed by pyrolysis/activation. The pinewood sawdust was preconditioned via three alternative synthesis protocols: (a) High-Pressure Hydrothermal Eruption (HPE), (b) High-Pressure Hydrothermal (HP), and (c) non-pre-pressure Hydrothermal (HY) treatment. Activated carbons were characterized by BET, SEM, FTIR and XPS; and the results indicate that HP achieved the highest specific surface area (990 m2 g−1) and micropore volume (0.344 cm3 g−1). Adsorption tests with perfluorooctanoic acid (PFOA) and methylene blue (MB) show that the adsorption capacities of the prepared activated carbons were significantly better than a commercial activated carbon (CAC). The HP exhibited the highest adsorption capacity for PFOA (232 mg g−1) and MB (350 mg g−1). The PFOA adsorption rates were empirically described by pseudo-second-order and intra-particle diffusion models; and the equilibrium sorption of PFOA fit well to Langmuir isotherm model.

Insight into the Structural Evolution of the Cobalt Oxides Nanoparticles upon Reduction Process: An In Situ Transmission Electron Microscopy Study

AbstractAs a prerequisite elementary step, the reduction of cobalt (Co) oxides to form metallic Co is essential to display activity in heterogeneous catalysis. Nevertheless, the intrinsic principle of phase and morphology evolution for Co oxides and formation mechanism of the active Co site remains unclear. Here, we resolve this decades‐long question by using a state‐of‐the‐art in situ transmission electron microscopy (TEM) to elucidate the multi‐role of initial crystalline phase and size of Co oxides in modulation of reduction rate, phase and morphology evolution under H2 atmosphere, by excluding the possible interference from promoter and support. We anticipate that this study enriches current understanding on the surface structure and morphology evolution of Co oxides, and provides reference for rational design of Co‐based catalytic material as well as its pretreatment protocol.

The Use of Natural Fiber-Rich Food Product Is Safe and Reduces Aberrant Crypt Foci in a Pre-Clinical Model.

Background: Colorectal cancer is a highly prevalent disease, requiring effective strategies for prevention and treatment. The present research aimed to formulate a natural fiber-rich food product (NFRFP) and to evaluate its safety, toxicogenetics, and effects on aberrant crypt foci induced by 1,2-dimethyl-hydrazine in a preclinical model. Methods: A total of 78 male Wistar rats were distributed in six experimental groups: negative control, positive control (1,2-Dimethylhydrazine—40 mg/Kg), and four groups fed with 10% NFRFP: NFRFP, pre-treatment protocol, simultaneous treatment, and post-treatment protocol. Results: The NFRFP was shown to be a good source of fibers and did not change biometric, biochemical, hematological, and inflammatory parameters, and did not induce signs of toxicity and genotoxicity/carcinogenicity. NFRFP exhibited a chemopreventive effect, in all protocols, with damage reduction (% DR) of 75% in the comet test. NFRFP reduced the incidence of aberrant crypt outbreaks by 49.36% in the post-treatment protocol. Conclusions: The results suggest the applicability of NFRFP in the human diet due to potential production at an industrial scale and easy technological application in different products, since it could be incorporated in food without altering or causing small changes in final product sensory characteristics.

Pre-cementation procedures’ effect on dental zirconias with different yttria content

ObjectveSeveral pre-cementation procedures have been advocated to enhance adhesion between zirconia and resin-based cement. There is, however, limited documentation on how these pre-treatments affect the strength of zirconia crowns as most tests are performed on discs or bars.The aim was to assess the effect of pre-cementation procedures on fracture mode, fracture strength and cement retention on zirconia. MethodsTwo dental zirconia materials with different yttria content were assessed (<4 and>5 mol%). Both discs (n = 45) and crown-shaped specimens (n = 30) of the two materials were pretreated with either air-abrasion or hot-etching with KHF2 and compared with untreated controls with regards to surface roughness, crystallography, wettability, cement adhesion and fracture strength. Results and SignificanceAir-abrasion improves adhesion and strength of zirconia with moderate yttria content (<4 mol%). Acid etching with heated KHF2 showed the best effect on strength and cement retention on zirconia with higher yttria content (>5 mol%). Application of KHF2 was, however, complicated on crown-shaped specimens. Pre-treatment and cementation protocols should be optimized for different dental zirconias to improve both strength and retention.

Ameliorating hydroxychloroquine induced retinal toxicity through cerium oxide nanoparticle treatments.

The present study investigated the potential protective effects of cerium oxide nanoparticles (CNP) on human retinal pigment epithelium (ARPE-19) cells damaged by hydroxychloroquine (HCQ). Toxicity of HCQ on the ARPE-19 cells was explored with a dose response trial. CNP rescue both a pre-treatment protocol, where CNP were applied 24 hours prior to HCQ application and a simultaneous treatment protocol where both CNP and HCQ were applied together, were used. In the dose response trial, 250 µM HCQ showed 51.84% cell viability after 24 hours and 32.75% after 48 hours time period. This was selected as model HCQ dose for rescue trials. The simultaneous treatment trials did not show a significant increase in viability compared to model toxic dose. The CNP pre-treatment trials showed a significant increase in cellular viability compared to model toxic dose with 68.03% ± 3.27 viability (p = 4.56E-05) at 24 hours and 51.85% ± 4.96 (p = 1.18E-05) at 48 hours time period. CNP pre-treatment showed significant protection of cells from HCQ induced toxicity. The difference in efficacy of simultaneous and pre-treatment is hypothesized to lie in the cellular localization of CNP. Furthermore, including the reactive oxygen species (ROS) scavenging properties of CNP seems to be responsible for protection, the effect of CNP on autophagosome and lysosome colocalization are also hypothesized to play a significant role.