•Few studies have explored the phenolic variability in a high number of monovarietal olive oils.•Phenolic compounds were analyzed in 80 monovarietal oil samples by LC–MS/MS.•Secoiridoids were the ...most concentrated phenols in olive oil.•A high influence of the cultivar on the phenolic levels was detected.•Four groups of cultivars were found through their phenolic profiles by multivariate analysis.
Despite the evident influence of the cultivar on olive oil composition, few studies have been devoted to exploring the variability of phenols in a representative number of monovarietal olive oils. In this study, oil samples from 80 cultivars selected for their impact on worldwide oil production were analyzed to compare their phenolic composition by using a method based on LC–MS/MS. Secoiridoid derivatives were the most concentrated phenols in virgin olive oil, showing high variability that was significantly due to the cultivar. Multivariate analysis allowed discrimination between four groups of cultivars through their phenolic profiles: (i) richer in aglycon isomers of oleuropein and ligstroside; (ii) richer in oleocanthal and oleacein; (iii) richer in flavonoids; and (iv) oils with balanced but reduced phenolic concentrations. Additionally, correlation analysis showed no linkage among aglycon isomers and oleocanthal/oleacein, which can be explained by the enzymatic pathways involved in the metabolism of both oleuropein and ligstroside.
Objective
Systemic lupus erythematosus (SLE) is characterized by an unpredictable and fluctuating course. Although various methods have been developed to measure disease activity, there is still a ...lack of consensus about the optimal criteria for SLE remission. The principal aim of our study was to identify the number of lupus patients achieving a complete remission (implying that for 3 years there were no clinical or serologic features and no treatment with steroids and immunosuppressive drugs) in a single cohort of patients followed for a period of up to 32 years. In addition, we have identified patients in clinical but not serologic remission (known as serologically active, clinically quiescent disease SACQ) and vice versa. We were particularly interested to determine the factors associated with complete remission.
Methods
Eligible patients were followed up in the University College Hospital Lupus cohort from January 1978 until December 2010 for a period of at least 3 years. Complete remission was defined as a period of at least 3 years with clinical inactivity (British Isles Lupus Assessment Group scores of C, D, or E only) and laboratory remission (no antibodies to double‐stranded DNA and normal complement C3 levels), and being off‐treatment with corticosteroids and immunosuppressants. Antimalarial and nonsteroidal antiinflammatory drugs were allowed.
Results
Of 624 lupus patients at our hospital, a total of 532 patients met the strict inclusion criteria for the study. Of these 532 patients, 77 patients (14.5%) achieved complete remission for at least 3 years, and 23 (4.3%) achieved complete remission for a minimum period of 10 years. Ten of these 77 patients were subsequently lost to followup, and, interestingly, flares occurred subsequently in 15 of the 67 remaining patients (22.4%). Three patients relapsed after the tenth year of remission. Forty‐five patients (8.5%) fulfilled the requirement for SACQ, and 66 patients (12.4%) achieved only serologic remission.
Conclusion
Our study indicated that 14.5% of lupus patients achieved a complete remission for 3 years. However, flares may continue to occur beyond 10 years of remission. Long‐term followup of SLE is therefore mandatory.
Nerve growth factor (NGF) plays a central role in multiple chronic pain conditions. As such, anti‐NGF monoclonal antibodies (mAbs) that function by antagonizing NGF downstream signaling are leading ...drug candidates for non‐opioid pain relief. To evaluate anti‐canine NGF (cNGF) mAbs we sought a yeast surface display platform of cNGF. Both mature cNGF and pro‐cNGF displayed on the yeast surface but bound conformationally sensitive mAbs at most 2.5‐fold in mean fluorescence intensity above background, suggesting that cNGF was mostly misfolded. To improve the amount of folded, displayed cNGF, we used comprehensive mutagenesis, FACS, and deep sequencing to identify point mutants in the pro‐region of canine NGF that properly enhance the folded protein displayed on the yeast surface. Out of 1,737 tested single point mutants in the pro region, 49 increased the amount of NGF recognized by conformationally sensitive mAbs. These gain‐of‐function mutations cluster around residues A‐61–P‐26. Gain‐of‐function mutants were additive, and a construct containing three mutations increased amount of folded cNGF to 23‐fold above background. Using this new cNGF construct, fine conformational epitopes for tanezumab and three anti‐cNGF mAbs were evaluated. The epitope revealed by the yeast experiments largely overlapped with the tanezumab epitope previously determined by X‐ray crystallography. The other mAbs showed site‐specific differences with tanezumab. As the number of binding epitopes of functionally neutralizing anti‐NGF mAbs on NGF are limited, subtle differences in the individual interacting residues on NGF that bind each mAb contribute to the understanding of each antibody and variations in its neutralizing activity. These results demonstrate the potential of deep sequencing‐guided protein engineering to improve the production of folded surface‐displayed protein, and the resulting cNGF construct provides a platform to map conformational epitopes for other anti‐neurotrophin mAbs.
A deep sequencing‐guided protein engineering workflow combined with yeast display was developed to increase the production of folded canine nerve growth factor (NGF). Dozens of mutations within the pro‐region of NGF enhanced the display of mature NGF on the yeast surface. Combining a subset of these beneficial mutations enabled a NGF construct that was used for conformational epitope mapping of different anti‐NGF monoclonal antibodies.
We propose a model to generate electrocardiogram signals based on a discretized reaction-diffusion system to produce a set of three nonlinear oscillators that simulate the main pacemakers in the ...heart. The model reproduces electrocardiograms from healthy hearts and from patients suffering various well-known rhythm disorders. In particular, it is shown that under ventricular fibrillation, the electrocardiogram signal is chaotic and the transition from sinus rhythm to chaos is consistent with the Ruelle-Takens-Newhouse route to chaos, as experimental studies indicate. The proposed model constitutes a useful tool for research, medical education, and clinical testing purposes. An electronic device based on the model was built for these purposes.
This work presents a study on the application of wall open tubular column (WCOT) in liquid chromatography coupled with tandem mass spectrometry. Each process step reports the column preparation ...method in detail, subdivided into column pretreatment, silanization, stationary phase coating, and immobilization. Then, an evaluation of the parameters that can affect the efficiency of these columns was made. Atrazine, clomazone, and metolachlor were used as probes during this step. Factors such as stationary phase composition, length, internal diameter, stationary phase mass employed, and injection volume were investigated. In addition, with the help of Knox and Poppe graphs, the columns' performance was evaluated to determine the optimal flow rate and the speed-efficiency relationship, respectively. Based on the results, the best configurations for the WCOT column application to the LC system were defined: length-8 m; inner diameter-25 μm; mass of OV-210-2.5%
/
; and, injection volume-100 nL. Finally, the optimized WCOT column developed in this work was coupled with a commercially-packed trapping column in the nano liquid chromatography system (nanoLC). In this configuration, more significant results were obtained regarding separation resolution, with Rs = 5.9 achieved for the most retained pair of analytes (clomazone and metolachlor).
Hasta el presente se conocen 27 especies, pertenecientes a cinco géneros de Xylariales, creciendo sobre Ocotea porphyria “laurel del cerro”. Nuevos estudios permitieron acrecentar el conocimiento de ...la funga asociada a este hospedante citando, en esta oportunidad, a Hypoxylon cf. brevisporum e H. pulicicidum por primera vez para el país. Además se registra a Entonaema liquescens, Xylaria pseudoapiculata y se adiciona información sobre las características morfológicas de la fase teleomórfica de X. myosurus, lo cual fue posible debido al hallazgo de nuevas poblaciones.
Contents
Paraoxonase‐1 (PON1) is an enzyme found in serum and follicular fluid that protects cell membrane and circulating lipids against oxidative damage. The aims of this study were to measure the ...direct effects of recombinant PON1 (rPON1) on bovine oocyte maturation at the molecular level (gene expression) and to measure the carry‐over effects of PON1 on pre‐implantation embryo development in vitro. COCs were submitted to IVM with the addition of 0.0, 0.02, 0.04 and 0.08 mg ml−1 of rPON1, corresponding to an average PON1 arylesterase enzyme activity of 2.2 ± 0.4, 15.5 ± 1.5, 30.2 ± 3.0 and 57.9 ± 5.0 U ml−1, respectively. The results indicated that addition of rPON1 during IVM improved embryo development in a dose‐dependent manner as D7 embryo development was 22.2%, 29.4%, 32.2% and 37.0% for the treatment groups, respectively (p = 0.02). In conclusion, addition of PON1 enzyme during IVM exerted dose‐related positive effects on embryo development rates to blastocysts.
Refined edible oils (viz., oils from maize, soya, high-oleic sunflower, sunflower, olive, and rapeseed) enriched at two concentration levels (200 and 400 μg/mL total phenolic content) with phenolic ...extracts isolated from olive pomace and leaves have been characterized and compared with nonenriched oils and extra virgin olive oil (EVOO). Enriched oils were analyzed by LC–TOF/MS to generate representative fingerprints and compared with nonenriched oils and EVOO by unsupervised principal component analysis (PCA). The two raw materials reported enriched oils with profiles which were compared with those provided by EVOOs. Correlation analysis enabled us to establish the enriched oils with a composition more similar to EVOO. Discrimination according to the enrichment level depended on the raw material for extracts, and a global discussion about the enrichment on relevant phenolic compounds present in EVOO has reported quantitative results concerning the enrichment level for those significant compounds with known nutraceutical properties.
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