Factors affecting natural antimicrobial expression in the human female reproductive tract

The incidence of sexually transmitted infections has increased due to changes in lifestyle and contraceptive practices. The female reproductive tract is therefore increasingly being exposed to potential infection, thus the mechanisms involved in its defence are important. The first line of defence is provided via the mucosal epithelial surface that lines the reproductive tract and the innate immune system. The innate immune response is fundamental to the recognition of infectious agents and the implementation of further defence mechanisms. Natural antimicrobial peptides have been described as being central to the function of the innate immune response. These peptides have been found to be expressed across a wide range of mucosal surfaces, including the reproductive tract. It has been shown that these molecules have cyclical variation providing protective coverage across the menstrual cycle. Their expression appears to be governed by a variety of different effector mechanisms which involve cytokines, sex steroids and cell-cell interactions.The aims ofthis thesis are (1) to characterise the expression and regulation of a range of natural antimicrobials within the human endometrium using representative cell lines. (2) To investigate the role(s) played by the interaction between epithelial and stromal cells in the expression of these molecules. (3) To assess the effects of different inflammatory mediators on the production of antimicrobials and the elucidation of the underlying mechanisms. (4) To identify the interactions between antimicrobials and with other innate immune effector molecules. (5) To elucidate the role of sex steroids in the regulation of natural antimicrobials. (6) To examine the expression patterns of these molecules over time in response to infection. (7) To describe the expression of antimicrobials in human Fallopian tubes with and without an ectopic gestation. (8) To examine the expression of natural antimicrobials in the uterine decidua ofectopic and failed intra-uterine pregnancy.Mimics of infection such as lipopolysaccharide (LPS - representative of G -ve infection), lipoteichoic acid (LTA - representative of G +ve infection) and cytokines such as 1L-1ß and TNFα; were demonstrated to alter the mRNA expression of natural antimicrobial molecules in the endometrial cell line Hec-IA (human endometrial carcinoma). The expression of natural antimicrobials was further identified to be both phasic and temporal over time. Changes in expression levels were also observed in primary endometrial stromal cell lines. The interaction between the epithelial and stromal cells concurrently with the addition of mediators of inflammation also yielded a change in mRNA expression. The presence of stromal derived media and mimics of infection caused an increase and earlier expression of molecules such as elafin in the Flec-IA cell line. Flowever, with the addition of progesterone treated stromal derived media, a downregulation of elafin was observed. Further investigation suggested a role for progesterone either directly upon the epithelial cells, which proved to have high levels of genomic progesterone receptors (PR-A and PR-B) or via stromal mediated factor(s). Two such factors were identified and investigated further, TGFß-1 and MMP-7 (matrilysin). Hec-IA cells treated directly with progesterone demonstrated increased levels of elafin mRNA, however, the presence of stromal cells inhibited this effect. The treatment of Hec-IA cells with TGFß-1 concurrently with inflammatory stimuli decreased the expression of elafin. However, the addition of MMP-7 increased the expression of elafin mRNA.In the presence of progesterone endometrial stromal cells up-regulate the level of TGFß-1 and this has been shown to decrease the epithelial expression of both MMP7 and elafin.Antimicrobials are differentially expressed within the Fallopian tube collected during different stages of the ovarian cycle and this appeared to be governed by the circulating levels of the exogenous sex steroids. Many of these molecules were also observed to be increased in response to an ectopic pregnancy. The uterine decidua from an ectopic gestation was also demonstrated to show differential expression to that observed in the decidua of both surgically managed miscarriage and from termination.This work furthers our understanding in the expression and regulation of antimicrobials in the upper reproductive tract, including the role of paracrine mediated factors. The altered expression in the presence of an ectopic gestation both in the Fallopian tube and uterine decidua indicates that further research may offer preventive, diagnostic and treatment opportunities in adverse pregnancy outcomes

Networking acupuncture in Vietnam

This thesis proposes that medical anthropologists change the way we think about acupuncture in Vietnam. Acupuncture should not be conceived as a discrete medicophilosophical system as has been acupuncture’s textual identity in academic writings to date. Acupuncture is rather a performative network, in the sense used by Bruno Latour, constituted through energetic relationships between science, people, textbooks, classrooms, pedagogic practices, clinical technologies and much more. These come into interaction and their collaborations produce acupuncture in unexpected ways. This conclusion was generated through 15 months of ethnographic fieldwork with acupuncturists in Ho Chi Minh City and catchments from 2007-08. Fieldwork involved observing acupuncturists engage patients, participating in acupuncture classes and volunteering on acupuncture charity teaching and treating missions. A snowballing method was used to generate connections with a mobile and diverse group of medical specialists. First, it will be shown that in Vietnam, science and tradition were united in the creation of a New Medicine that must be considered on its own terms rather than as a grafting of two different types of medical system. The New Medicine modelled pedagogic and legitimacy-making practices which circulated in the city. Second, local formation of acupuncture objects and shaping of clinical treatment flatten out previously taken for granted hierarchies when describing clinical medical knowledge. The technology of vision was integral to the construction of such knowledge and when interrupted caused acupuncture to grind to a halt. Finally, person networks, after Mark Granovetter, were active in the city generating professional success and legality for practitioners but these will also be analysed using a Latourian approach. Recent ethnographic investigations of science and technology are used to help portray, more faithfully, the interactive dynamic of acupuncture experienced during fieldwork. Such writings extend the scope of what can be investigated as participating in the creation of medical realities in southern Vietnam. I argue that medical knowledge is a reality constructed through continual practices. Knowledge is not a commodity or eternally static entity, knowledge is what we do

Promoting young people's health through drama and dance

Peer reviewedPublisher PD

Laboratory models for studying ectopic pregnancy

PURPOSE OF REVIEW: Understanding of the aetiology of tubal ectopic pregnancy (tEP) remains incomplete. We aim to summarize the latest advances in laboratory models of tEP that we believe will, ultimately, contribute to improving the diagnosis and management of the condition. RECENT FINDINGS: Progress in proteome pre-fractionation and multidimensional protein identification technology has proved particularly effective in identifying novel biomarkers of tEP. These, and related global proteomic and genomic approaches, have as yet to be fully exploited in this context but do have substantial potential to inform future hypothesis driven studies. The majority of data generated since 2009 to explain the aetiology of tEP continues to derive from descriptive human ex-vivo studies. In-vitro models of Fallopian tube ciliary and smooth muscle function have improved to a limited degree, on the back of continuing advances in imaging and data acquisition. We believe that the recent development of a primary human Fallopian tube epithelium culture system represents the most significant recent advance in laboratory models for studying ectopic pregnancy. There remain no good animal models of tEP. SUMMARY: The establishment of a viable animal model of tEP remains the key obstacle to a complete understanding the aetiology of the condition

Modeling human liver biology using stem cell-derived hepatocytes

Stem cell-derived hepatocytes represent promising models to study human liver biology and disease. This concise review discusses the recent progresses in the field, with a focus on human liver disease, drug metabolism and virus infection

Generation of Functional Human Hepatic Endoderm from Human Induced Pluripotent Stem Cells

With the advent of induced pluripotent stem cell (iPSC) technology, it is now feasible to generate iPSCs with a defined genotype or disease state. When coupled with direct differentiation of defined lineage, such as hepatic endoderm (HE). iPSC would revolutionise the way we study human liver biology and generate efficient “off the shelf” models of human liver disease. Here we show the `proof of concept' that iPSC lines representing both male and female sexes and two ethnic origins can be differentiated to HE at efficiencies of between 70–90%, using a method mimicking a physiological condition. iPSC-derived HE exhibited hepatic morphology, and expressed the hepatic markers, Albumin and E-Cadherin as assessed by immuno-histochemistry. They also expressed alpha fetal protein (AFP), HNF4a, and a metabolic marker, Cyp7A1, demonstrating a definitive endodermal lineage differentiation. Furthermore, iPSC-derived hepatocytes produced and secreted the plasma proteins, fibrinogen, fibronectin, transthyretin (TTR) and AFP, an essential feature for functional HE. Additionally iPSC-derived HE supported both CYP1A2 and 3A4 metabolism, which is essential for drug and toxicology testing. CONCLUSION: This work is first to demonstrate the efficient generation of hepatic endodermal lineage from human iPSC that exhibits key attributes of hepatocytes, and the potential application of iPSC-derived HE in studying human liver biology. In particular, iPSC from individuals representing highly polymorphic variants in metabolic genes and different ethnic groups will provide pharmaceutical development and toxicology studies a unique opportunity to revolutionise predictive drug toxicology assays and allow the creation of in vitro hepatic disease models

Liver cell therapy: is this the end of the beginning?

The prevalence of liver diseases is increasing globally. Orthotopic liver transplantation is widely used to treat liver disease upon organ failure. The complexity of this procedure and finite numbers of healthy organ donors have prompted research into alternative therapeutic options to treat liver disease. This includes the transplantation of liver cells to promote regeneration. While successful, the routine supply of good quality human liver cells is limited. Therefore, renewable and scalable sources of these cells are sought. Liver progenitor and pluripotent stem cells offer potential cell sources that could be used clinically. This review discusses recent approaches in liver cell transplantation and requirements to improve the process, with the ultimate goal being efficient organ regeneration. We also discuss the potential off-target effects of cell-based therapies, and the advantages and drawbacks of current pre-clinical animal models used to study organ senescence, repopulation and regeneration

Development of a low cost electron gun

The purpose of this project is to design and optimize a rudimentary electron gun at a scale and cost that would be easily achievable in a home lab. For the electron gun to function properly, a high vacuum chamber was constructed from surplus components. A number of parts were custom machined including vacuum parts, power electronics, vacuum gauges and the cooling system. The gun consists of a thermionic cathode held at a high negative potential relative to a grounded anode. The system was characterized using a phosphor screen, a floating collector plate, and current-voltage measurements

Building a home from foam—túngara frog foam nest architecture and three-phase construction process

Frogs that build foam nests floating on water face the problems of over-dispersion of the secretions used and eggs being dangerously exposed at the foam : air interface. Nest construction behaviour of túngara frogs, Engystomops pustulosus, has features that may circumvent these problems. Pairs build nests in periodic bursts of foam production and egg deposition, three discrete phases being discernible. The first is characterized by a bubble raft without egg deposition and an approximately linear increase in duration of mixing events with time. This phase may reduce initial over-dispersion of foam precursor materials until a critical concentration is achieved. The main building phase is marked by mixing events and start-to-start intervals being nearly constant in duration. During the final phase, mixing events do not change in duration but intervals between them increase in an exponential-like fashion. Pairs joining a colonial nesting abbreviate their initial phase, presumably by exploiting a pioneer pair's bubble raft, thereby reducing energy and material expenditure, and time exposed to predators. Finally, eggs are deposited only in the centre of nests with a continuously produced, approximately 1 cm deep egg-free cortex that protectively encloses hatched larvae in stranded nests