Abstract: Fibromyalgia (FM) is a chronic disorder characterized by widespread pain, which can limit patients' physical function and daily activities. FM can be challenging to treat, and the treatment approach could benefit from a greater understanding of patients' perspectives on their condition and their care. Patients with FM participated in an online survey conducted in the USA that sought to identify the symptoms that had the greatest impact on patients' daily lives. The purpose of the survey was to facilitate efforts toward improving care of patients by nurse practitioners, primary care providers, and specialists, in addition to contributing to the development of new outcome measures in both clinical trials and general practice. A total of 1,228 patients with FM completed the survey, responding to specific questions pertaining to symptoms, impact of symptoms, management of FM, and the relationship with health care providers. Chronic pain was identified as the key FM symptom, affecting personal and professional relationships, and restricting physical activity, work, and social commitments. Patients felt that the severity of their condition was underestimated by family, friends, and health care providers. The results of this survey highlight the need for nurse practitioners, primary care providers, and specialists to provide understanding and support to patients as they work together to enable effective diagnosis and management of FM. Keywords: fibromyalgia, pain, survey, impact, support
Glycosyltransferases (GTs) are essential for the biosynthesis and diversification of many therapeutically important natural products. Of these, UDP-sugar: sterol glucosyltransferases (UGTs) (2.4.1.173) catalyse the synthesis of therapeutically important steryl glycosides (SGs). Guided by the sequence similarity with a previously characterised N-terminally truncated UGT from Saccharomyces cerevisiae (UGT51), this study reports the cloning of the gene fragment encoding the C-terminal catalytic domains from related yeasts and the expression and characterisation of their encoded products produced. N-terminally histidine tagged proteins were purified for in vitro assays against a panel of sterol and steroidal acceptors. Liquid chromatography-mass spectrometry (LC-MS) and kinetic analysis led to the successful characterisation of two novel UGTs from Pichia angusta and Kluyveromyces lactis. In addition, testosterone was shown to be utilized by all UGTs, including the previously characterised S. cerevisiae UGT51. Random mutagenesis of UGTs and homology modelling of the S. cerevisiae UGT revealed structural similarities with family 1 bacterial glycopeptide GTs. Given the structural and mechanistic similarities among GT family 1 UGTs, this approach may provide a template for genetic manipulation of novel UGTs from other members of the GT superfamily with a better understanding of catalytic domains and for broadening their scope in drug development. It may also aid the development of a generic process in the synthesis of SGs.
The main objectives of this work were to characterise a range of purified recombinant sterol 3β-glucosyltransferases and show that rational sampling of the diversity that exists within sterol 3β-glucosyltransferase sequence space can result in a range of enzyme selectivities. In our study the catalytically active domain of the Saccharomyces cerevisiae 3β-glucosyltransferase was used to mine putative sterol 3β-glucosyltransferases from the databases. Selected diverse sequences were expressed in and purified from Escherichia coli and shown to have different selectivities for the 3β-hydroxysteroids ergosterol and cholesterol. Surprisingly, three enzymes were also selective for testosterone, a 17β-hydroxysteroid. This study therefore reports for the first time sterol 3β-glucosyltransferases with selectivity for both 3β- and 17β-hydroxysteroids and is also the first report of recombinant 3β-glucosyltransferases with selectivity for steroids with a hydroxyl group at positions other than C-3. These enzymes could therefore find utility in the pharmaceutical industry for the green synthesis of a range of glycosylated compounds of medicinal interest.
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