Miracle-dz/UniFAR-FTUs huggingface dataset:
Confirmatory analysis of steroids in muscle using liquid chromatography–tandem mass spectrometry

A method is described for screening and confirmation of synthetic and endogenous steroids in muscle tissue. The method is sensitive, selective, and rapid and the consumption of organic solvents is low, compared to previously published methods. The procedure involves hydrolysis, defattening with heptane and final clean up with SPE using C18 cartridge. After filtration, the analytes are analysed by LC/MS/MS and quantification is performed using deuterated internal standards. Decision limits (CCα) varied from 0.02 to 0.33 µg kg−1 and the detection capabilities (CCβ) were <0.50 µg kg−1. The mean within-laboratory reproducibility ranged 5–22% (%RSDIR). Endogenous steroids (e.g. testosterone, epitestosterone and androstenedione) have been included in the method, to provide an insight into their levels, as the presence of these steroids was detected several times during analysis of imported meat.
Simultaneous detection of 12 mycotoxins in cereals using RP-HPLC-PDA-FLD with PHRED and a post-column derivatization system

A new method for the simultaneous quantification of 12 mycotoxins was developed and optimized using reverse phase high performance liquid chromatography (RP-HPLC) with a photodiode array (PDA) and fluorescence detector (FLD), a photochemical reactor for enhanced detection (PHRED) and post-column derivatization. The mycotoxins included aflatoxins (AFB1, AFB2, AFG1, and AFG2), ochratoxin A (OTA), zearalenone (ZEA), deoxynivalenol (DON), fumonisins (FB1, FB2, and FB3), T-2 and HT-2 toxins. A double sample extraction with a phosphate-buffered saline solution (PBS) and methanol was used for co-extraction of mycotoxins, and a multifunctional immunoaffinity column was used for cleanup. Optimum conditions for separation of the mycotoxins were obtained to separate 12 mycotoxins in FLD and PDA chromatograms with a high resolution. The method gave recoveries in the range 72–111% when applied to spiked corn samples. The limits of detection (LOD) were 0.025 ng/g for AFB1 and AFG1, 0.012 ng/g for AFB2 and AFG2, 0.2 ng/g for OTA, 1.5 ng/g for ZEA, 6.2 ng/g for FB1, FB3 and HT-2 toxin, 9.4 ng/g for FB2 and T-2 toxin, and 18.7 ng/g for DON. In addition, the limits of quantification (LOQ) ranged from 0.04 ng/g for AFB2 and AFG2 to 62 ng/g for DON. The method was successfully applied to the determination of these mycotoxins in 45 cereal samples obtained from the Malaysian market. The results indicated that the method can be applied for the multi-mycotoxin determination of cereals.
Methods for the determination of European Union-permitted added natural colours in foods: a review

Coupled to increasing consumer demand, food manufacturers have moved towards increased usage of approved natural colours. There is a legal requirement for governments to monitor the consumption of all food additives in the European Union to ensure the acceptable daily intakes (ADIs) are not exceeded, especially by young children. Validated analytical methods are needed to fulfil this requirement. The aim of this paper is to review the available literature on methods of extraction for approved natural colours in food and drink. Available analytical methods for the determination of European Union-permitted natural food colour additives in foods and beverages have been assessed for their fitness for purpose in terms of their key extraction and analysis procedures, selectivity and sensitivity, especially with regard to maximum permitted levels, and their applicability for use in surveillance and in an enforcement role. The advantages and disadvantages of available analytical methods for each of nine designated chemical classes (groups) of natural colours in different food and beverage matrices are given. Other important factors such as technical requirements, cost, transferability and applicability are given due consideration. Gaps in the knowledge and levels of validation are identified and recommendations made on further research to develop suitable methods. The nine designated natural colour classes covered are: 1. Curcumin (E100), 2. Riboflavins (E101i–ii), 3. Cochineal (E120), 4. Chlorophylls-including chlorophyllins and copper analogues (E140–141), 5. Caramel Classes I–IV (E150a–d), 6. Carotenoids (E160a–f, E161b, E161g), 7. Beetroot red (E162), 8. Anthocyanins (E163), and 9. Other colours – Vegetable carbon (E153), Calcium carbonate (E170), Titanium dioxide (E171) and Iron oxides and hydroxides (E172).
Genetic variants in TNF‐α promoter are predictors of recurrence in patients with squamous cell carcinoma of oropharynx after definitive radiotherapy

The promoter variants of TNF‐α, a major regulator of immune and inflammation responses, have been implicated in cancer development and prognosis. Thus, we investigated associations between four TNF‐α promoter variants and risk of recurrence of squamous cell carcinoma of the oropharynx (SCCOP). We evaluated associations of four TNF‐α polymorphisms with risk of recurrence in a cohort of 846 patients with SCCOP. Log‐rank test and multivariable Cox models were used to evaluate associations. Compared with patients with variant genotypes of the TNF‐α ‐308 and TNF‐α ‐863 polymorphisms, patients with common homozygous genotypes had worse disease‐free survival (log‐rank p = 0.0002 and p < 0.0001, respectively) and increased risk of SCCOP recurrence (HR, 1.9, 95% CI, 1.3–2.8 and HR, 1.9, 95% CI, 1.3–2.7, respectively) after multivariable adjustment. Furthermore, among patients with HPV16‐positive tumors, those with common homozygous genotypes of the TNF‐α ‐308 and ‐863 polymorphisms had worse disease‐free survival (log‐rank p = 0.005 and p = 0.007, respectively) and higher recurrence risk than patients with variant genotypes of these polymorphisms (HR, 5.1, 95% CI, 1.4–18.4 and HR, 3.7, 95% CI, 1.5–9.1, respectively), while no such significant associations were found for TNF‐α ‐857 or ‐1031 polymorphisms. Our findings suggest that TNF‐α ‐308 and ‐863 polymorphisms may modulate the risk of SCCOP recurrence in patients with HPV16‐positive tumors. However, larger studies are needed to validate these results.
Isotherm modeling of organic activated bentonite and humic acid polymer used as mycotoxin adsorbents

The aim of the current study was to evaluate and compare two representative samples of different classes of adsorbents intended for use as feed additives in the prevention or reduction of the adverse effects exerted by mycotoxins, specifically ochratoxin A (OTA) and zearalenone (ZEN). The adsorbents, an organically activated bentonite (OAB) and a humic acid polymer (HAP), were tested in a common in vitro model with a pH course comparing the maximum pH changes that can be expected in the digestive system of a monogastric animal, i.e. pH 7.4 for the oral cavity, pH 3.0 for the stomach, and pH 8.4 for the intestines. In the first experiment, the concentration-dependent adsorbent capacity of OAB and HAB were tested using a fixed concentration of either mycotoxin. Thereafter, adsorption was evaluated applying different isotherms models, such as Freundlich, Langmuir, Brunauer–Emmett–Teller (BET) and Redlich–Peterson, to characterize the adsorption process as being either homo- or heterogeneous and representing either mono- or multilayer binding. At the recommended statutory level for the mycotoxins of 0.1 mg kg−1 OTA and 0.5 mg kg−1 ZEN, OAB showed an adsorbed capacity of >96% towards both mycotoxins, regardless of the pH. The HAP product was also able to absorb >96% of both mycotoxins at pH 3.0, but extensive desorption occurred at pH 8.4. Based on χ-square (χ2) values, Langmuir and Redlich–Peterson equations proved to be the best models to predict monolayer equilibrium sorption of OTA and ZEN onto the organically activated bentonite and the humic acid polymer. The applied methodology has a sufficient robustness to facilitate further comparative studies with different mycotoxin-adsorbing agents.
Median Raphe Cysts of the Scrotum and Perineum Presenting with Recurrent Infection

Median raphe cysts of the perineum are rare congenital anomalies of the male genitalia, which form during embryological development, and can be found in the midline from the distal penis to the perineum. However, the incidence of median raphe cyst is likely under-reported and under-recognized. We report on the case of a median raphe cyst extending from the scrotum to the perineum with recurrent infection and purulent discharge in a 28-year-old man, which first developed at the age of 5 years. We believe it is important that urologists recognize median raphe cysts and have knowledge of their management in order to provide appropriate information to patients.
Detection of hazardous weight-loss substances in adulterated slimming formulations using ultra-high-pressure liquid chromatography with diode-array detection

The presence on the market of illegal products for slimming purposes or the treatment of overweight is a public health issue. These products may contain illicit chemicals in order to improve their effectiveness. Some of these weight-loss compounds are responsible for adverse events, including fatal outcomes. A general strategy for the analysis of any suspect formulation begins with a large screening for the general search of a wide range of compounds. A methodology for the qualitative and quantitative determination of 34 compounds in slimming preparations (such as dietary supplements or medicinal products) was used for the control of slimming formulations from the market, including over the Internet. The fast liquid chromatography system (ultra-high-pressure liquid chromatography) used a gradient of solvent (phosphate buffer and acetonitrile), a C18 endcapped column and a diode array detector. This system allows dual identification based on retention time and UV spectra. The analytical method is simple, fast and selective since 34 weight-loss compounds can be detected in a 15-min run time. Thus, 32 commercial slimming formulations were analysed using this method, allowing the detection and quantification of hazardous active substances: caffeine, clenbuterol, nicotinamide, phenolphthalein, rimonabant, sibutramine, didesmethylsibutramine, synephrine and yohimbine.
Waveband and Dose Dependency of Sunlight‐induced Immunomodulation and Cellular Changes †

Both the UVB and UVA wavebands within sunlight are immunosuppressive. This article reviews the relationship between wavebands and dose in UV‐induced immunosuppression mainly concentrating on responses in humans. It also contrasts the effects of UVB and UVA on cellular changes involved in immunosuppression. Over physiological sunlight doses to which humans can be exposed during routine daily living or recreational pursuits, both UVA and UVB suppress immunity. While there is a linear dose relationship with UVB commencing at doses less than half of what is required to cause sunburn, UVA has a bell‐shaped dose response over the range to which humans can be realistically exposed. At doses too low for either waveband to be suppressive, interactions between UVA and UVB augment each other, enabling immunosuppression to occur. At doses beyond where UVA is immunosuppressive, it still contributes to sunlight‐induced immunosuppression via this interaction with UVB. While there is little research comparing the mechanisms by which UVB, UVA and their interactions can cause immunosuppression, it is likely that different chromophores and early molecular events are involved. There is evidence that both wavebands disrupt antigen presentation and effect T cell responses. Different individuals are likely to have different immunomodulatory responses to sunlight.
Evaluation of counselling in primary care: How can research be made more useful for practitioners?

The paper presents a literature review of research on counselling in general practice critiqued from the perspective of the Practice Counsellor. It is noted that existing studies all focus primarily on outcome of psychological interventions in comparison to routine GP care. The contributions and limitations of these studies are discussed, and the methodological difficulties of conducting controlled trials in this context are explored. It is proposed that future research on counselling in primary care needs to draw on the paradigm shift in psychotherapy and a range of alternative research methodologies which will be more likely to produce research of relevance to Practice Counsellors.
Intertumor heterogeneity of non‐small‐cell lung carcinomas revealed by multiplexed mutation profiling and integrative genomics

Non‐small‐cell lung cancer (NSCLC) is a heterogeneous disease, with a burden of genomic alterations exceeding most other tumors. The goal of our study was to evaluate the frequencies of co‐occurring mutations and copy‐number aberrations (CNAs) within the same tumor and to evaluate their potential clinical impact. Mass‐spectrometry based mutation profiling using a customized lung cancer panel evaluating 214 mutations across 26 key NSCLC genes was performed on 230 nonsquamous NSCLC and integrated with genome‐wide CNAs and clinical variables. Among the 138 cases having at least one mutation, one‐third (41, 29.7%) showed two or more mutations, either in the same gene (double mutation) or in different genes (co‐mutations). In epidermal growth factor receptor (EGFR) mutant cancers, there was a double mutation in 18% and co‐mutations in the following genes: TP53 (10%), PIK3CA (8%), STK11 (6%) and MET (4%). Significant relationships were detected between EGFR mutation and 1p, 7p copy gains (harboring the EGFR gene) as well as 13q copy loss. KRAS mutation was significantly related with 1q gain and 3q loss. For Stage I, tumors harboring at least one mutation or PIK3CA mutation were significantly correlated with poor prognosis (p‐value = 0.02). When combining CNAs and mutational status, patients having both KRAS mutation and the highest related CNA (3q22.3 copy loss) showed a significant poorer prognosis (p‐value = 0.03). Our study highlights the clinical relevance of studying tumor complexity by integrative genomic analysis and the need for developing assays that broadly screen for both “actionable” mutations and copy‐number alterations to improve precision of stratified treatment approaches.
Increased Melanogenesis is a Risk Factor for Oxidative DNA Damage— Study on Cultured Melanocytes and Atypical Nevus Cells †

Melanin synthesis is an oxygen‐dependent process that acts as a potential source of reactive oxygen species (ROS) inside pigment‐forming cells. The synthesis of the lighter variant of melanin, pheomelanin, consumes cysteine and this may limit the capacity of the cellular antioxidative defense. We show that tyrosine‐induced melanogenesis in cultured normal human melanocytes (NHM) is accompanied by increased production of ROS and decreased concentration of intracellular glutathione. Clinical atypical (dysplastic) nevi (DN) regularly contain more melanin than do normal melanocytes (MC). We also show that in these cultured DN cells three out of four exhibit elevated synthesis of pheomelanin and this is accompanied by their early senescence. By using various redox‐sensitive molecular probes, we demonstrate that cultured DN cells produce significantly more ROS than do normal MC from the same donor. Our experiments employing single‐cell gel electrophoresis (comet assay) usually reveal higher fragmentation of DNA in DN cells than in normal MC. Even if in some cases the normal alkaline comet assay shows no differences in DNA fragmentation between DN cells and normal MC, the use of the comet assay with formamidopyrimidine DNA glycosylase can disclose that the DNA of the cultured DN cells harbor more oxidative damage than the DNA of normal MC from the same person.
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Surface Electromyographic Activity of the Upper Trapezius Before and After a Single Dry Needling Session in Female Office Workers With Trapezius Myalgia

Objective Myofascial pain can be accompanied by a disturbed surface electromyographic (sEMG) activity. Nevertheless, the effect of myofascial treatment techniques, such as dry needling (DN), on the sEMG activity is poorly investigated. Several DN studies also emphasize the importance of eliciting local twitch responses (LTRs) during treatment. However, studies investigating the added value of LTRs are scarce. Therefore, the aims of this study were first to evaluate the effect of DN on the sEMG activity of myalgic muscle tissue, compared with no intervention (rest), and secondly to identify whether this effect is dependent of eliciting LTRs during DN. Methods Twenty-four female office workers with work-related trapezius myalgia were included. After completion of a typing task, changes in sEMG activity were evaluated after a DN treatment of the upper trapezius, compared with rest. Results The sEMG activity increased after rest and after DN, but this increase was significantly smaller 10 minutes after DN, compared with rest. These differences were independent whether LTRs were elicited or not. Conclusions Dry needling leads to a significantly lower increase in sEMG activity of the upper trapezius, compared with no intervention, after a typing task. This difference was independent of eliciting LTRs.
Genetic association of gastric cancer with miRNA clusters including the cancer‐related genes MIR29, MIR25, MIR93 and MIR106: Results from the EPIC‐EURGAST study

MicroRNAs (miRNAs) are post‐transcriptional gene regulators involved in a wide range of biological processes including tumorigenesis. Deregulation of miRNA pathways has been associated with cancer but the contribution of their genetic variability to this disorder is poorly known. We analyzed the genetic association of gastric cancer (GC) and its anatomical and histological subtypes, with 133 single‐nucleotide polymorphisms (SNPs) tagging 15 isolated miRNAs and 24 miRNA clusters potentially involved in cancer, in 365 GC cases and 1,284 matched controls within the European Prospective Investigation into Cancer and Nutrition cohort. Various SNPs were associated with GC under the log‐additive model. Furthermore, several of these miRNAs passed the gene‐based permutation test when analyzed according to GC subtypes: three tagSNPs of the miR‐29a/miR‐29b‐1 cluster were associated with diffuse subtype (minimum p‐value = 1.7 × 10−4; odds ratio, OR = 1.72; 95% confidence interval, CI = 1.30–2.28), two tagSNPs of the miR‐25/miR‐93/miR‐106b cluster were associated with cardia GC (minimum p‐value = 5.38 × 10−3; OR = 0.56, 95% CI = 0.37–0.86) and one tagSNP of the miR‐363/miR‐92a‐2/miR‐19b‐2/miR‐20b/miR‐18b/miR‐106a cluster was associated with noncardia GC (minimum p‐value = 5.40 × 10−3; OR = 1.41, 95% CI = 1.12–1.78). Some functionally validated target genes of these miRNAs are implicated in cancer‐related processes such as methylation (DNMT3A, DNMT3B), cell cycle (E2F1, CDKN1A, CDKN1C), apoptosis (BCL2L11, MCL1), angiogenesis (VEGFA) and progression (PIK3R1, MYCN). Furthermore, we identified genetic interactions between variants tagging these miRNAs and variants in their validated target genes. Deregulation of the expression of these miRNAs in GC also supports our findings, altogether suggesting for the fist time that genetic variation in MIR29, MIR25, MIR93 and MIR106b may have a critical role in genetic susceptibility to GC and could contribute to the molecular mechanisms of gastric carcinogenesis.
Photo‐induced Reactive Oxygen Species Generation by Different Water‐soluble Fullerenes (C60) and Their Cytotoxicity in Human Keratinocytes

In this study we report the phototoxicity toward HaCaT keratinocytes that results from the photogeneration of superoxide and singlet oxygen (1O2) by four different “water‐soluble” fullerene (C60) preparations—monomeric (γ‐CyD)2/C60 (γ‐cyclodextrin bicapped C60) and three aggregated forms—THF/nC60 (prepared by solvent exchange from THF solution); Son/nC60 (prepared by sonication of a toluene/water mixture); and γ‐CyD/nC60 (prepared by heating the [γ‐CyD]2/C60 aqueous solution). Our results demonstrate that all four C60 preparations photogenerate 1O2 efficiently. However, the properties of C60‐generated 1O2, including its availability for reactions in solution, are markedly different for the monomeric and aggregated forms. 1O2 produced by monomeric (γ‐CyD)2/C60 is quenchable by NaN3 and its quantum yield in D2O, which is only weakly dependent on oxygen concentration, is as high as C60 in toluene. In contrast, 1O2 generated from aggregated C60 is not quenchable by NaN3, exhibits a solvent‐independent short‐lived lifetime (ca 2.9 μs), is highly sensitive to oxygen concentration while its phosphorescence is redshifted. All these features indicate that 1O2 is sequestered inside the C60 aggregates, which may explain why these preparations were not phototoxic toward HaCaT cells. Electron paramagnetic resonance studies demonstrated the generation of the C60 anion radical (C60•−) when (γ‐CyD)2/C60 was irradiated (λ > 300 nm) in the presence of a reducing agent (NADH); spin trapping experiments (λ > 400 nm) with 5,5‐dimethyl‐1‐pyrroline N‐oxide clearly showed the generation of superoxide resulting from the reaction of C60•− with oxygen. In vitro tests with HaCaT keratinocytes provided evidence that (γ‐CyD)2/C60 phototoxicity is mainly mediated by 1O2 (Type II mechanism) with only a minor contribution from free radicals (Type I mechanism).
A simple and reliable liquid chromatography-tandem mass spectrometry method for the determination of aflatoxin M1 in milk

A new chromatographic method is proposed for the analysis of aflatoxin M1 in milk. The method is based on liquid–liquid extraction followed by LC-MS/MS analysis. Liquid–liquid extraction (LLE) is performed on the defatted milk plus sodium chloride by using ethyl acetate as an extraction solvent. Accuracy and precision were evaluated at the LOQ (15 ng kg–1) spiked sample as well as with three other different naturally contaminated reference materials. The mean overall recovery (n = 24) was 95% with a confidence interval of 1.9% and a CV% of 4.5%. The performance of the proposed method was compared with that of the Official ISO Method based on the use of immunoaffinity chromatography columns (IAC): LLE protocol could be considered a valid alternative to the LC-IAC. In general it showed better accuracy with lower data dispersion. Moreover, the sample preparation is very simple and straightforward, potentially being applicable as a high-throughput method which, on account of its simplicity and low cost, may be applied to the analysis of a large number of samples in the occasion of outbreaks of large-scale contamination.