An environmental metabolomics investigation of the effects of chiral pharmaceuticals and environmental pollutants on microorganisms

Active pharmaceutical ingredients (APIs) and their metabolites are ubiquitous in the environment and their occurrence in the aquatic environment is of growing concern. However, despite the fact that these may cause harmful effects in organisms found within this niche, little is currently known about the effects of APIs in the aquatic environment. Chiral pharmaceuticals are of particular concern as the enantiomers may be metabolised differently, with the potential for the production of an array of harmful compounds. There are many racemic APIs for treating human and animal conditions, and even in these target organisms the pharmacodynamic effects of the enantiomers are not always known. Within recent years the importance of the interactions of these compounds within the aquatic environment has been realised and information regarding the fate and biodegradation of such environmental pollutants is of great importance. The advent of post-genomic technologies has proved advantageous in the study of the effects of these environmental pollutants. In this thesis, the effects of a range of chiral APIs, and other environmental pollutants, on environmentally relevant microorganisms were investigated at the metabolome level. The effects of chiral APIs were investigated in a number of prokaryotic and eukaryotic systems in order to provide a comprehensive study of the effects of the APIs in the aquatic environment. FT-IR spectroscopy was employed for metabolic fingerprinting of some environmentally relevant bacteria and GC-MS was subsequently employed for metabolite profiling of two pseudomonads that had shown differential chiral effects with Propranolol. In addition, FT-IR microspectroscopy was employed for the investigation of the phenotypic and localised effects of chiral APIs in a eukaryotic system. Furthermore, the effects of a range of environmental pollutants on a complex bacterial community were investigated with the use of FT-IR spectroscopy and multivariate analysis. Initial results indicated a large phenotypic response in relation to phenol, and this was further explored with a range of ageing experiments and metabolic fingerprinting. An FT-IR peak was found to be characteristic of the phenotypic changes in the actively degrading communities and this was likely to be a degradation product of phenol, and armed with this knowledge the activated sludge community was monitored during the active degradation of phenol with the use of GC-MS.The work presented in this thesis has shown for the first time that metabolomics allows subtle phenotypes in microorganisms to be revealed when they are exposed to chiral forms of APIs which are commonly found in the aquatic environment. Despite these APIs not being designed for any interaction with bacteria and aquatic life in general these are significant findings and may have implications as more and more APIs become detectable and concentrated in the environment due to continued use in man and indeed animals or aquaculture.EThOS - Electronic Theses Online ServiceGBUnited Kingdo

A woman with diabetes presenting with pyomyoma and treated with subtotal hysterectomy: a case report

<p>Abstract</p> <p>Introduction</p> <p>Pyomyoma (suppurative leiomyoma of the uterus) is a rare condition resulting from infarction and infection of a leiomyoma. It is more usual in pregnant women or postmenopausal women who have vascular disease. The condition is usually fatal unless treated with appropriate antibiotics and surgical intervention.</p> <p>Case presentation</p> <p>We report a case of a 44-year-old Afro-Caribbean woman with diabetes who presented with recurrent episodes of abdominal pain and fever over a period of five months. Her problem proved to be a diagnostic dilemma mimicking cholecystitis, pyelonephritis and ovarian cancer. Her blood cultures were positive on one occasion for methicillin-resistant <it>Staphylococcus epidermidis</it>. An ultrasound scan suggested uterine fibroids but a computed tomography scan suggested an ovarian malignancy because the mass appeared heterogeneous with fluid filled areas. She was treated with several courses of antibiotics and eventually at laparotomy, she was found to have a large pyomyoma which was successfully removed by subtotal hysterectomy with immediate and complete resolution of her symptoms.</p> <p>Conclusion</p> <p>The diagnosis of pyomyoma should be considered in perimenopausal women with large fibroids and pyrexia of unknown origin.</p

Biochemical characterization of the initial steps of the Kennedy pathway in Trypanosoma brucei:the ethanolamine and choline kinases

Note related output below contains correction of this paper.Ethanolamine and choline are major components of the trypanosome membrane phospholipids, in the form of GPEtn (glycero-phosphoethanolamine) and GPCho (glycerophosphocholine). Ethanolamine is also found as an integral component of the GPI (glycosylpliosphatidylinositol) anchor that is required for membrane attachment of cell-surface proteins, most notably the variant-surface glycoproteins. The de novo synthesis of GPEtn and GPCho starts with the generation of phosphoethanolamine and phosphocholine by ethanolamine and choline kinases via the Kennedy pathway. Database mining revealed two putative C/EKs (choline/ethanolamine kinases) in the Trypanosoma brucei genome, which were cloned, overexpressed, purified and characterized. TbEK 1 (T brucei ethanolamine kinase 1) was shown to be catalytically active as an ethanolamine-specific kinase, i.e. it had no choline kinase activity. The K values for ethanolamine and ATP were found to be 18.4 +/- 0.9 and 219 29 mu M respectively. TbC/EK2 (T brucei choline/ethanolamine kinase 2), on the other hand, was found to be able to phosphorylate both ethanolamine and choline, even though choline was the preferred substrate, with a K-m 80 times lower than that of ethanolamine. The K. values for choline, ethanolamine and ATP were 31.4 +/- 2.6 mu M, 2.56 +/- 0.31 mu M and 20.6 +/- 1.96 mu M respectively. Further substrate specificity analysis revealed that both TbEK1 and TbC/EK2 were able to tolerate various modifications at the amino group, with the exception of a quaternary amine for TbEK1 (choline) and a primary amine for TbC/EK2 (ethanolamine). Both enzymes recognized analogues with substituents oil C-2, but substitutions oil C-1 and elongations of the carbon chain were not well tolerated.Publisher PDFPeer reviewe

Using microbes to recover rare earths with low environmental impact?

Using Microbes to recover Rare Earths with low environmental impact Barbara Palumbo Roe, Simon Gregory, Antoni Milodowski, Julia West, Joanna Wragg British Geological Survey, Nicker Hill, Nottingham NG12 5GG, UK Steve Banwart, Maria Romero González, Wei Huang, Emma Wharfe Kroto Research Institute, University of Sheffield, Sheffield S3 7HQ, UK John Harding, Colin Freeman, Shaun Hall Department of Materials Science and Engineering, University of Sheffield, Sheffield S1 3JD, UK Microbes play an important role in the fate and transport of rare earth elements (REE) in relation to the REE exploitation life cycle. A step change in understanding is needed for key mobilisation, concentration and fractionation processes such as bioleaching, biosorption and biomineralisation and how they can 1) be harnessed to recover REE in situ from low grade ores or secondary deposits, and 2) be quantified for reactive transport in environmental risk assessment and management of mining operations. Heap/in-situ leaching methods are relatively low impact mining technologies, requiring less energy (for comminution) and in the case of in-situ leaching have a minimal footprint. Furthermore, biologically-assisted leaching and separation processes represent a more sustainable alternative to chemical processes. We discuss the microbial potential to accelerate dissolution of REEs from source minerals, and how the natural selectivity of mineral and microbial surfaces as ligands for adsorption and biomineralisation of REE dissolved species could be exploited in the recovery of REEs from fluids

Morinda citrifolia

Introduction. Noni (Morinda citrifolia) has been used for many years as an anti-inflammatory agent. We tested the efficacy of Noni in women with dysmenorrhea. Method. We did a prospective randomized double-blind placebo-controlled trial in 100 university students of 18 years and older over three menstrual cycles. Patients were invited to participate and randomly assigned to receive 400 mg Noni capsules or placebo. They were assessed for baseline demographic variables such as age, parity, and BMI. They were also assessed before and after treatment, for pain, menstrual blood loss, and laboratory variables: ESR, hemoglobin, and packed cell volume. Results. Of the 1027 women screened, 100 eligible women were randomized. Of the women completing the study, 42 women were randomized to Noni and 38 to placebo. There were no significant differences in any of the variables at randomization. There were also no significant differences in mean bleeding score or pain score at randomization. Both bleeding and pain scores gradually improved in both groups as the women were observed over three menstrual cycles; however, the improvement was not significantly different in the Noni group when compared to the controls. Conclusion. Noni did not show a reduction in menstrual pain or bleeding when compared to placebo

Multiple metachronous malignancies, one patient with three primary malignancies: a case report

We present a 61 year old Para 4 woman who presented with stage II Infiltrating lobular carcinoma of the breast after modified radical mastectomy. She was treated with Tamoxifen for seven years. She was diagnosed with multiple myeloma during year seven post mastectomy because of wrist pain. She was treated with melphalan, prednisone and allopurinol which she tolerated well and the pain in the wrist improved. Tamoxifen was also stopped. Ten months later she presented with vaginal bleeding and was diagnosed with a poorly differentiated endometrial adenocarcinoma at hysteroscopic suction curettage and had an abdominal hysterectomy. Two years later the patient succumbed to metastatic endometrial cancer