Chlorogenic acids (CGAs) are a large class of esters formed between quinic acid and hydroxycinnamic acids. They are present in coffee as a complex mixture of positional and geometric isomers, where caffeoylquinic acids (CQAs) are the most abundant, followed by dicaffeoylquinic acids (diCQAs), feruloylquinic acids (FQAs), and p-coumaroylquinic acids (p-CoQAs). The aim of this work is to develop a new reliable and fast liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for simultaneous identification and quantification of total amount of 11 CGAs in roasted coffee. Regarding sample preparation step, aqueous methanol and 100% aqueous ultrasonic extractions were evaluated. For the filtration Step 4, different membranes were tested, in order to fill the void of complete evaluation of extractables recovery when using different membranes, highlighting an incomplete recovery when using nylon filters. An LC/electrospray ionization (ESI)-MS/MS method was developed and validated following the European rules in terms of specificity, linearity, concentration range, limit of detection (LOD) and limit of quantification (LOQ), precision, and trueness, in order to obtain a useful quality control tool for roasted coffee. The method was applied for quantification of CGAs of a roasted coffee sample previously characterized by an interlaboratory circuit (LVU), and results were compared with the quantitation of CGAs via UHPLC-DAD. The quantitative results were expressed as 5-CQA equivalents, and the validation of this approach opens the way to reliable, cheap, and environmental-friendly tools for quality control purposes.CDX2 plays a crucial role in the formation and maintenance of the trophectoderm epithelium in preimplantation embryos. Follistatin supplementation during the first 72 hr of in vitro culture triggers a significant increase in blastocyst rates, CDX2 expression, and trophectoderm cell numbers. However, the underlying epigenetic mechanisms by which follistatin upregulates CDX2 expression are not known. Here, we investigated whether stimulatory effects of follistatin are linked to alterations in DNA methylation within key regulatory regions of the CDX2 gene. In vitro-fertilized (IVF) zygotes were cultured with or without 10 ng/ml of recombinant human follistatin for 72 hr, then cultured without follistatin until Day 7. The bisulfite-sequencing analysis revealed differential methylation (DM) at specific CpG sites within the CDX2 promoter and intron 1 following follistatin treatment. These DM CpG sites include five hypomethylated sites at positions -1384, -1283, -297, -163, and -23, and four hypermethylated sites at positions -1501, -250, -243, and +20 in the promoter region. There were five hypomethylated sites at positions +3060, +3105, +3219, +3270, and +3545 in intron 1. Analysis of transcription factor binding sites using MatInspector combined with a literature search revealed a potential association between differentially methylated CpG sites and putative binding sites for key transcription factors involved in regulating CDX2 expression. The hypomethylated sites are putative binding sites for FXR, STAF, OCT1, KLF, AP2 family, and P53 protein, whereas the hypermethylated sites are putative binding sites for NRSF. Collectively, our results suggest that follistatin may increase CDX2 expression in early bovine embryos, at least in part, by modulating DNA methylation at key regulatory regions.COVID-19 remains a global pandemic with more than 10 million cases and half a million deaths worldwide. The disease manifestations in patients with chronic kidney disease and especially those on haemodialysis are still being understood, with only a few overseas case series, and small observational trials thus far. It appears that the disease is more severe in this patient cohort. Part of the pathophysiology of severe COVID-19 is related to accompanying cytokine release syndrome (CRS). Tocilizumab, an interleukin-6 inhibitor, has been trialled for treatment of CRS in COVID-19, but not yet approved. We present a case of an Australian patient on long-term haemodialysis with severe COVID-19 who was successfully treated with Tocilizumab. The peak of her illness was on day 7, with a C-reactive protein of 624 mg/L (reference  less then  5 mg/L), ferritin of 5293 ng/mL (reference 30-500 ng/mL), and interleukin-6 level 1959.7 pg/mL, consistent with CRS. She was severely hypoxic on a ventilator, with rising inotropic requirements. With the use of Tocilizumab, there was a significant and immediate response in her inflammatory markers, and she made a steady recovery. https://www.selleckchem.com/products/ABT-869.html The patient was discharged home 6 weeks after presentation. The dedicated education unit (DEU) is an innovative clinical model that prepares preceptors for success in clinical settings with nursing students. Though the DEU is mostly used in acute-care settings, this project explores the implementation of a DEU in a public health setting. Better preparation of public health nurses and social workers as clinical preceptors for nursing students with the implementation of a DEU in a public health setting. IRB approved, pre/post survey with participant comments. Clinical Nurse Teacher Survey was assessed pre/post intervention with registered nurses and social work staff (n=13). Paired t-tests analysis was used to determine significance. The Clinical Learning Environment and Nurse Teacher (CLES+T) scale completed postimplementation by nursing students (n=8) after the clinical rotation. Clinical Nurse Teacher Survey mean scores preintervention was 4.56 and increased postintervention to 4.89, though not statistically significant (p-value .11). CLES+T showing 100% fully agree or agree that the Public Health DEU is an effective learning environment. The DEU model in a public health setting is an opportunity to improve lived clinical experiences of preceptors and nursing students, which may increase nursing students' positive perceptions of, and increase interest in serving as a public health nurse after graduation. The DEU model in a public health setting is an opportunity to improve lived clinical experiences of preceptors and nursing students, which may increase nursing students' positive perceptions of, and increase interest in serving as a public health nurse after graduation.