This project begins with an overview of the female reproductive tract microenvironment. It outlines the microenvironment of the vaginal, cervical, and endometrial epithelium and the interactions with immune cells and hormone cycles. The review also outlines the models currently used to study the female reproductive tract. The second chapter of the thesis is a study of the effects of pathogenic and commensal bacteria P. micra, F. magna, and F. nucleatum on cervical epithelial cells. This study analyzes cytotoxic effects after 24 hour infection of these bacteria. This was assessed through crystal violet staining, conventional pcr of cDNA synthesized from extracted cervical RNA, and LDH analysis. There is also an attempted biofilm assay. It was concluded that bacteria P. micra, F. magna and F. nucleatum have cytotoxic potential. This was not expected as F. magna is largely understood to be a commensal bacteria in the vaginal microbiome.
X-ray free-electron lasers provide novel opportunities to conduct single particle analysis on nanoscale particles. Coherent diffractive imaging experiments were performed at the Linac Coherent Light Source (LCLS), SLAC National Laboratory, exposing single inorganic core-shell nanoparticles to femtosecond hard-X-ray pulses. Each facetted nanoparticle consisted of a crystalline gold core and a differently shaped palladium shell. Scattered intensities were observed up to about 7 nm resolution. Analysis of the scattering patterns revealed the size distribution of the samples, which is consistent with that obtained from direct real-space imaging by electron microscopy. Scattering patterns resulting from single particles were selected and compiled into a dataset which can be valuable for algorithm developments in single particle scattering research.
Viral protein U (Vpu) is a type-III integral membrane protein encoded by Human Immunodeficiency Virus-1 (HIV- 1). It is expressed in infected host cells and plays several roles in viral progeny escape from infected cells, including down-regulation of CD4 receptors. But key structure/function questions remain regarding the mechanisms by which the Vpu protein contributes to HIV-1 pathogenesis. Here we describe expression of Vpu in bacteria, its purification and characterization. We report the successful expression of PelB-Vpu in Escherichia coli using the leader peptide pectate lyase B (PelB) from Erwinia carotovora. The protein was detergent extractable and could be isolated in a very pure form. We demonstrate that the PelB signal peptide successfully targets Vpu to the cell membranes and inserts it as a type I membrane protein. PelB-Vpu was biophysically characterized by circular dichroism and dynamic light scattering experiments and was shown to be an excellent candidate for elucidating structural models.
Methods: Three databases were used to search for papers published after 2010 until October 2019 discussing obesity, overweight and TCM. No forms of Chinese medicine were excluded from the search. Studies were excluded if they did not meet the date criteria or if they overlapped with papers found in other databases.
Results: Six of the selected papers covered acupuncture (electro, balance or catgut embedding acupuncture methods) either jointly with other treatments or alone, one exclusively on moxibustion and three studies on three different types of TCM herbal medicine. Each study showed a statistically significant effect on body mass index (BMI) value decrease or total weight loss (TWL). The six acupuncture papers all showed statistical significance at the 95% CI against control groups (sham acupuncture or no acupuncture) and against before-treatment BMI values or TWL values.
Conclusion: Of the treatments reviewed, almost all acupuncture studies were shown to be consistently effective in treating overweight or obese individuals within this selection of studies, as well as in another meta-analysis. This may be due to acupuncture’s ties to a neuroendocrine mechanism. Future studies should further explore the neuroendocrine connection between acupuncture and weight loss. Herbal medication was also shown to have a significant effect in reducing weight in each study; however, two studies used mice or rats as subjects, therefore understanding the effects on human subjects is limited.