Identification of epigenetic silencing of the SFRP2 gene in colorectal cancer as a clinical biomarker and molecular significance.

BACKGROUND: Several studies have suggested secreted frizzled-related protein 2 (SFRP2) gene as a potential clinical biomarker in colorectal cancer (CRC). However, its diagnostic role remains unclear. In this study, we aimed to investigate the significance of SFRP2 methylation levels in a large cohort of biological specimens (including blood, adipose and colonic tissues) from patients with CRC, thereby potentially identifying new biomarker utility. METHODS: We examined the expression (by qPCR) and methylation status (by 450 K DNA array and DNA pyrosequencing) of the SFRP2 gene in healthy participants (N = 110, aged as 53.7 (14.2), 48/62 males/females) and patients with CRC (N = 85, aged 67.7 (10.5), 61/24 males/females), across different biological tissues, and assessing its potential as a biomarker for CRC. Additionally, we investigated the effect of recombinant human SFRP2 (rhSFRP2) as a therapeutic target, on cell proliferation, migration, and the expression of key genes related to carcinogenesis and the Wnt pathway. RESULTS: Our findings revealed that SFRP2 promoter methylation in whole blood could predict cancer stage (I + II vs. III + IV) (AUC = 0.653), lymph node invasion (AUC = 0.692), and CRC recurrence (AUC = 0.699) in patients with CRC (all with p < 0.05). Furthermore, we observed a global hypomethylation of SFRP2 in tumors compared to the adjacent area (p < 0.001). This observation was validated in the TCGA-COAD and TCGA-READ cohorts, demonstrating overall hypermethylation (both with p < 0.001) and low expression (p < 0.001), as shown in publicly available scRNA-Seq data. Notably, neoadjuvant-treated CRC patients exhibited lower SFRP2 methylation levels compared to untreated patients (p < 0.05) and low promoter SFRP2 methylation in untreated patients was associated with poor overall survival (p < 0.05), when compared to high methylation. Finally, treatment with 5 µg of rhSFRP2 treatment in CRC cells (HCT116 cells) inhibited cell proliferation (p < 0.001) and migration (p < 0.05), and downregulated the expression of AXIN2 (p < 0.01), a gene involved in Wnt signaling pathway. CONCLUSIONS: These findings establish promoter methylation of the SFRP2 gene as a prognostic candidate in CRC when assessed in blood, and as a therapeutic prognostic candidate in tumors, potentially valuable in clinical practice. SFRP2 also emerges as a therapeutic option, providing new clinical and therapeutical avenues.

Effect of eccentricity and light level on the timing of light adaptation mechanisms.

We explored the complexity of the light adaptation process, assessing adaptation recovery (Ar) at different eccentricities and light levels. Luminance thresholds were obtained with transient background fields at mesopic and photopic light levels for temporal retinal eccentricities (0°-15°) with test/background stimulus size of 0.5°/1° using a staircase procedure in a two-channel Maxwellian view optical system. Ar was obtained in comparison with steady data [Vis. Res.125, 12 (2016)VISRAM0042-698910.1016/j.visres.2016.04.008]. Light level proportionally affects Ar only at fovea. Photopic extrafoveal thresholds were one log unit higher for transient conditions. Adaptation was equally fast at low light levels for different retinal locations with variations mainly affected by noise. These results evidence different timing in the mechanisms of adaptation involved.

Enantioselective Catalysis with Pyrrolidinyl Gold(I) Complexes: DFT and NEST Analysis of the Chiral Binding Pocket.

A new generation of chiral gold(I) catalysts based on variations of complexes with JohnPhos-type ligands with a remote C2-symmetric 2,5-diarylpyrrolidine have been synthesized with different substitutions at the top and bottom aryl rings: from replacing the phosphine by a N-heterocyclic carbene (NHC) to increasing the steric hindrance with bis- or tris-biphenylphosphine scaffolds, or by directly attaching the C2-chiral pyrrolidine in the ortho-position of the dialkylphenyl phosphine. The new chiral gold(I) catalysts have been tested in the intramolecular [4+2] cycloaddition of arylalkynes with alkenes and in the atroposelective synthesis of 2-arylindoles. Interestingly, simpler catalysts with the C2-chiral pyrrolidine in the ortho-position of the dialkylphenyl phosphine led to the formation of opposite enantiomers. The chiral binding pockets of the new catalysts have been analyzed by DFT calculations. As revealed by non-covalent interaction plots, attractive non-covalent interactions between substrates and catalysts direct specific enantioselective folding. Furthermore, we have introduced the open-source tool NEST, specifically designed to account for steric effects in cylindrical-shaped complexes, which allows predicting experimental enantioselectivities in our systems.

Temporal retinal sensitivity in mesopic adaptation.

PURPOSE: Night driving is a complex visual task with important ramifications for driver and pedestrian safety. It is usually performed under mesopic or scotopic conditions and frequently, in the presence of transient glare sources that can adapt parts of the central retina. The objective of this work was to analyze the time response of adaptation for the central 15° of the retina when part of it is exposed to transient or steady mesopic adapting fields. METHODS: Absolute visual thresholds and luminance thresholds when viewing steady and transient adaptation fields were measured for three observers, at temporal retinal eccentricities of 0°-14.5° in steps of 2.9° (subsequently described as 0°, 3°, 6°, 9°, 12° and 15°) using a two-channel Maxwellian view optical system. The adaptation field and stimulus subtended 1.05° and 0.45° respectively. The transient adaptation field was presented with a stimulus onset asymmetry (SOA) of 300 ms. Time course adaptation curves were also measured at 0°, 6° and 9°. RESULTS: The absolute dark adaptation threshold (threshold measured at dark adaptation conditions or L(a)(t) decreases in peripheral retina due to an increasing rod contribution. Luminance thresholds vs eccentricity curves for transient L(SOA300)(t) and steady L(LA)(t) mesopic adaptation fields intersect across the first 15° of the peripheral retina. CONCLUSIONS: While the fovea shows higher sensitivity than the areas of peripheral retina investigated in this study, the speed of adaptation, measured from the visibility loss, is greater for retinal regions between 6° and 9° than for the fovea or retinal eccentricities beyond 9°.

Quantitative and functional influence of surround luminance on the letter contrast sensitivity function.

AIM: To determine the influence of surround luminance on the letter contrast sensitivity function. METHOD: The binocular contrast sensitivities of 31 young and ocularly healthy individuals were measured with letters of sizes calculated to obtain the targeted fundamental frequencies of 3, 10, 20, and 30 c deg(-1), respectively; with surround luminances from 1 to 1000 cd m(-2), always with a test chart background luminance of 200 cd m(-2). RESULTS: The letter contrast sensitivity increased with surround luminance up to 100 cd m(-2) and decreased when surround luminance increased from 100 to 1000 cd m(-2). These increments are larger for higher fundamental spatial frequencies, while decrements are similar for all frequencies. To analyse pupil size influence, results were compared with theoretical predictions obtained by combining different ocular MTFs with a typical neural function, where pupil size decrease leads to letter contrast sensitivity increments and veiling luminance causes the observed decrements. Other possible optical or neural factors that influence these values have also been considered. CONCLUSIONS: Letter contrast sensitivity function depends on surround luminance and this influence should be considered in future standardized directives.

Thoracic thrust joint manipulation: An international survey of current practice and knowledge in IFOMPT member countries.

BACKGROUND: The perceived relative safety of thoracic thrust joint manipulation (TTJM) has contributed to a growth in evidence supporting use in practice. Yet adverse events (AE) have been documented following TTJM. Knowledge of current practice is therefore required to support further research. PURPOSE: To investigate TTJM knowledge and pre-TTJM examination across IFOMPT Member Organisations (MO) and Registered Interest Groups (RIG). METHODS: An e-survey was designed based on existing evidence and piloted. Eligibility criteria: physiotherapists from member countries of the International Federation of Orthopaedic Manipulative Physical Therapists (IFOMPT) who use TTJM. Recruitment was through IFOMPT networks (May 2018-March 2019). Data analyses included descriptive analyses and content analysis for free text data. RESULTS: Respondents (n = 363) from 20 countries. Pre-TTJM examination included patient history (22%, n = 81) and physical examination (69%, n = 248). Across presentations (>80% threshold of agreement) contraindications included osteomyelitis, fracture and metastatic disease. Spinal deformity, respiratory disease, serious joint disease and hypermobility achieved >60% agreement as precautions. Consent was obtained by 93% respondents (n = 250). Preferred technique was PA/AP thrust (61%, n = 144). Perception of primary effect was neurophysiological (52%, n = 134), biomechanical (42%, n = 109) and placebo (3%, n = 8). From those who reported AE (n = 100), these included fractures (36%, n = 42) and cord signs/symptoms (6%, n = 7). CONCLUSION: Pre-TTJM examination is common, although bias towards physical examination. Differential testing for upper versus lower thoracic spine is limited. Inconsistencies across knowledge of contraindications and precautions, and beliefs for biomechanical effect were found. Findings highlight the importance of high levels of clinical reasoning during patient history for TTJM.

Conjunctival mucin mRNA expression in contact lens wear.

PURPOSE: To investigate the influence of the water content in non-ionic hydrogel contact lenses (HCL) on the mRNA levels of human conjunctival mucin genes (MUCs). METHODS: Sixteen healthy subjects with no history of contact lenses wear were selected and randomized into two equal groups. Group 1 subjects wore low water content (38%, Soflens 38) non-ionic HCLs. Group 2 wore high water content (66%, Soflens 66) non-ionic HCLs. Conjunctival impression cytology was applied to the superior bulbar conjunctiva of both eyes before, 6 months, and 1 year after HCL fitting, and 15 days after discontinuation of wearing. Total RNA was isolated, retrotranscribed, and amplified by conventional polymerase chain reaction (PCR) and by quantitative real time PCR to study the mRNA levels of MUCs and to analyze variations during the study period. Time- and HCL-dependent variations in mRNA expression were analyzed using Student's test. RESULTS: From the known MUCs, transcripts from MUC1, MUC2, MUC4, MUC5AC, MUC7, MUC13, MUC15, MUC16, and MUC17 genes were detected in all subjects before HCL fitting. Except for MUC2, the expression of some MUC genes significantly increased whereas others significantly decreased at either the 6- and 12-month period. Statistically significant differences between both HCL groups (p < 0.001) were found in the MUC4, MUC13, and MUC15 mRNA expression after 1 year of wear and after the 15 days without HCL wear. However, these differences were not clearly related to the water content of the lenses. CONCLUSIONS: Low and high water content non-ionic HCLs induced different changes in the mRNA levels of several MUCs, but the water content was not related to the changes. Recovery to basal levels of conjunctival MUC mRNA expression after wearing HCL lenses for a year takes longer than 15 days for some MUCs.

Influence of age and spectral power distribution on mesopic visual sensitivity.

The ability of the human visual system to detect stimuli at low illumination levels provides awareness of potential risks. The influence of age and spectral power distribution on mesopic spectral sensitivity is analyzed. Two typical light sources are used, a high-pressure sodium lamp (HPS), with a higher content of long wavelengths, and a metal halide lamp (MH), with a higher content of short wavelengths. Two experiments were performed, using a two-channel Maxwellian-view optical system to measure contrast threshold under different experimental conditions. In Experiment 1, three age groups (young, middle-aged and old, n = 2 each), two retinal locations (on-axis and off-axis vision), four background luminances (0.01, 0.07, 0.45, and 3.2 cd/m2), and two photometry systems (photopic and the MES2 systems) were considered. In Experiment 2, contrast threshold measurement was performed with two age groups (young and old, n = 11 each), one retinal location (off-axis vision), one background luminance (0.01 cd/m2), and two photometry systems (photopic and the MES2 systems). In on-axis vision, neither age nor spectral power distribution have an effect on the contrast threshold. In off-axis vision, however, a significant interaction between age and spectral power distribution is obtained, albeit only at 0.01 cd/m2 with an MH lamp. Only at this lowest background luminance was the greater content of short wavelengths of this lamp responsible for higher rod stimulation in off-axis vision, with the subsequent improvement in detection performance in young subjects. However, the effect of diffused light inside the aged eye counteracted the benefits of increased rod sensitivity for the MH lamp.

Retinal toxic reactions following photopheresis.

BACKGROUND: Extracorporeal photochemotherapy (ECP), also known as photopheresis, is a generally well-tolerated therapeutic, immunomodulatory approach successfully used in cutaneous T-cell lymphoma and other diseases produced by T-lymphocytes such as graft vs host disease. OBSERVATIONS: On 2 separate occasions, a 54-year-old white man with Sézary syndrome developed cutaneous phototoxic reactions and chorioretinitis after being treated with ECP. A pharmacokinetic study showed therapeutic blood levels of 8-methoxypsoralen as long as 18 weeks after therapy had been terminated. However, the analysis of mutations in genes involved in the drug's disposition could not explain these abnormal levels. CONCLUSIONS: To our knowledge, there has been no previous description of ECP-related retinal toxic effects. This adverse effect was probably linked to impaired drug elimination. Further studies would be needed to determine the underlying mechanism.

Liquid chromatographic method for the quantification of zearalenone in baby food and animal feed: interlaboratory study.

An interlaboratory trial for determination of zearalenone (ZON) in baby food and animal feed was conducted. The study involved 39 participants in 16 European Union member states, as well as Turkey, Uruguay, and China, representing a cross-section of industry, and official food control and research institutes. The method is based on immunoaffinity column cleanup followed by high-performance liquid chromatography using fluorimetry (HPLC-FI). The test portion of the sample is extracted with methanol-water (75 + 25, v/v). The sample extract is filtered, diluted, and passed over an immunoaffinity column. ZON is eluted with methanol. The separation and determination of ZON is performed by reversed-phase HPLC-FI with an excitation wavelength of 274 nm and an emission wavelength of 446 nm. Test portions of the samples were spiked at levels of 20 and 30 microg/kg ZON in baby food and at levels of 100 and 150 microg/kg ZON in animal feed. Mean recoveries from each participant ranged from 78 to 119% with an average value of 92% for baby food and from 51 to 122% with an average value of 74% for animal feed. Based on results for spiked samples (blind duplicates at 2 levels), as well as naturally contaminated samples (blind duplicates at 3 levels), the relative standard deviation for repeatability (RSDr) in baby food ranged from 2.8 to 9.0%. For animal feed, this value ranged from 5.7 to 9.5%. The relative standard deviation for reproducibility (RSDR) in baby food ranged from 8.2 to 13.3%, and for animal feed this value ranged from 15.5 to 21.4%. The Horwitz ratio (HorRat) in baby food ranged from 0.3 to 0.4, and for animal feed this value ranged from 0.6 to 0.9. The method showed acceptable within- and between-laboratory precision for each matrix, as required by European legislation.

Liquid chromatographic determination of deoxynivalenol in baby food and animal feed: interlaboratory study.

An interlaboratory study was conducted for the determination of deoxynivalenol in baby food and animal feed by high-performance liquid chromatography with UV detection. The study included 14 participants representing a cross section of industry, official food control, and research facilities. Mean recoveries reported ranged from 89% (at 120 microg/kg) to 85% (at 240 microg/kg) for baby food and from 100% (at 200 microg/kg) to 93% (at 400 microg/kg) for animal feed. On the basis of the results for spiked samples (blind duplicates at 2 levels), as well as those for naturally contaminated samples (blind duplicates at 3 levels), the relative standard deviation for repeatability (RSDr) in analyses of baby food ranged from 6.4 to 14.0% and in analyses of animal feed, from 6.1 to 16.5%. The relative standard deviation for reproducibility (RSDR) in analyses of baby food ranged from 9.4 to 19.5% and in analyses of animal feed, from 10.5 to 25.2%. The HorRat values ranged from 0.4 to 1.0 and from 0.7 to 1.3, for baby food and animal feed, respectively. The method showed acceptable performance for within-laboratory and between-laboratory precision for each matrix, as required by European legislation.

Determination of aflatoxin B1 in medical herbs: interlaboratory study.

A method was developed for the determination of aflatoxin B1 in medical herbs (senna pods, botanical name Cassia angustifolia; devil's claw, botanical name Harpagophytum procumbens; and ginger roots, botanical name Zingiber officinale). The method, which was tested in a mini-collaborative study by 4 laboratories, is based on an immunoaffinity cleanup followed by reversed-phase high-performance liquid chromatography separation and fluorescence detection after post-column derivatization. It allows the quantitation of aflatoxin B1 at levels lower than 2 ng/g. A second extractant (acetone-water) was tested and compared to the proposed methanol-water extractant. Several post-column derivatization options (electrochemically generated bromine, photochemical reaction, and chemical bromination) as well as different integration modes (height versus area) were also investigated. No differences were found depending on the choice of derivatization system or the signal integration mode used. The method was tested for 3 different matrixes: senna pods, ginger root, and devil's claw. Performance characteristics were established from the results of the study and resulted in HorRat values ranging from 0.12 to 0.75 with mean recoveries from 78 to 91% for the extraction with methanol-water and HorRat values ranging from 0.10-1.03 with mean recoveries from 98 to 103% for the extraction with acetone-water. As a result, the method, with all tested variations, was found to be fit-for-purpose for the determination of aflatoxin B1 in medical herbs at levels of 1 microg/kg and above.

Influence of background size, luminance and eccentricity on different adaptation mechanisms.

Mechanisms of light adaptation have been traditionally explained with reference to psychophysical experimentation. However, the neural substrata involved in those mechanisms remain to be elucidated. Our study analyzed links between psychophysical measurements and retinal physiological evidence with consideration for the phenomena of rod-cone interactions, photon noise, and spatial summation. Threshold test luminances were obtained with steady background fields at mesopic and photopic light levels (i.e., 0.06-110cd/m(2)) for retinal eccentricities from 0° to 15° using three combinations of background/test field sizes (i.e., 10°/2°, 10°/0.45°, and 1°/0.45°). A two-channel Maxwellian view optical system was employed to eliminate pupil effects on the measured thresholds. A model based on visual mechanisms that were described in the literature was optimized to fit the measured luminance thresholds in all experimental conditions. Our results can be described by a combination of visual mechanisms. We determined how spatial summation changed with eccentricity and how subtractive adaptation changed with eccentricity and background field size. According to our model, photon noise plays a significant role to explain contrast detection thresholds measured with the 1/0.45° background/test size combination at mesopic luminances and at off-axis eccentricities. In these conditions, our data reflect the presence of rod-cone interaction for eccentricities between 6° and 9° and luminances between 0.6 and 5cd/m(2). In spite of the increasing noise effects with eccentricity, results also show that the visual system tends to maintain a constant signal-to-noise ratio in the off-axis detection task over the whole mesopic range.

Liquid chromatographic method for quantitation of patulin at 10 ng/mL in apple-based products intended for infants: interlaboratory study.

An interlaboratory trial for the determination of patulin in apple juice and fruit puree was conducted, involving 17 participants representing a cross section of industry, official food control, and research facilities. Mean recoveries reported ranged from 74 (10 ng/g) to 62% (25 ng/g) for apple juice and from 72 (25 ng/g) to 74% (10 ng/g) for fruit puree. Based on results for spiked samples (blind pairs at 2 levels), as well as naturally contaminated samples (blind pairs at 3 levels), the relative standard deviation for repeatability (RSDr) in juice ranged from 8.0 to 14.3% and in puree from 3.5 to 9.3%. The relative standard deviation for reproducibility (RSD(R)) in juice ranged from 19.8 to 39.5% and in puree from 12.5 to 35.2%, reflecting HORRAT values from 0.6 to 1.0 for juice and 0.4 to 0.9 for puree. The method showed acceptable within-laboratory and between-laboratory precision for each matrix, as required by current European legislation.

Valoración de la eficacia de una pauta más lenta y frecuente en la administración de hierro intravenoso en pacientes en hemodiálisis durante un año / Evaluation of the efficacy of a slower and more frequent administration of intravenous iron in haemodialysis patients during one year

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