The vagina is one organ of the body which has not been studied exhaustively. Moreover, most of the studies found in the contemporary literature have been performed on women affected by a variety of genital diseases. In the present study the vaginal epithelium was examined with a histological method, morphometry, whereby cyclical changes related to hormonal variation during the menstrual cycle were demonstrated. Determination of the quantity of estrogen receptors in the vaginal epithelium on two occasions during the menstrual cycle revealed a significantly greater number in the follicular than in the luteal phase. The results of these studies indicate the presence of a menstrual variation in the vaginal epithelium comparable to that in the endometrium. Phenoxymethylpenicillin (pcV) was used as a marker substance to study the dynamics of the transport mechanisms into the vagina. PcV was found to accumulate in the vaginal fluid and high concentrations persisted for a long period of time. In hysterectomized women, the appearance of pcV in the vaginal fluid followed the same pattern. Consequently, the substance is transported through the vaginal wall and need not enter with the secretions from the internal genitalia. The greatest concentration of pcV was in the distal portion of the vagina, possibly due to the specific internal circulation of fluid within the vagina. Bacterial vaginosis as an example of an ‘ecological disease’ has been studied with regard to the formation of endotoxin, a constituent of the cell wall of Gram- negative bacteria. Large amounts of endotoxin were found and the clinical implication of this finding has been pointed out. Furthermore, the influence of pcV on the vaginal microbial flora of healthy women has been investigated. A change from a situation with predominance of lactobacilli to the appearance of Gram-negative rods was observed. In one of the women the lactobacilli disappeared completely and were replaced by E. coliand high levels of endotoxin in the vaginal fluid were found. This study demonstrates the complexity of the ecological balance of the vaginal microbial flora and illustrates the difficulty of defining a ‘normal’ vaginal condition. Is there any unquestionable state of ‘normality’ even in a healthy woman free from symptoms of genital disease? / <p>S. 1-22: sammanfattning, s. 25-64: 6 uppsatser</p> / digitalisering@umu

In the human uterus prostaglandins (PG) PGE2, PGD2, PGI2, PGF2α and Thromboxane A2 (TXA2), also termed prostanoids, are synthesised and deactivated to 15-keto PGE2, J2 metabolites, 6-keto-PGF1α, 15-keto PGF2α and TXB2 respectively. However, not all metabolites have been analysed simultaneously within the same tissue. The primary objective of this thesis was to determine full uterine prostanoid profiles in human non-pregnancy, pregnancy and parturition, to better understand these processes and find suitable tocolytic targets. In addition, ten cytokines in human cervico-vaginal fluid (CVF) were measured according to interval to labour to test their suitability as labour onset predictors, with a view to developing a test to determine women at risk of preterm labour. Prostanoid analysis was carried out in endometrium (n=9) and myometrium (n=15- 16) donated by non-pregnant women and lower segment myometrium obtained from pregnant women (before (n=14) and after labour onset (n=7)) by liquid chromatography coupled with electrospray ionisation mass spectrometry (LC/ESIMS/ MS). Cytokines produced by CVF collected from pregnant donors (20-41 weeks gestation, n=2-10) were investigated using Enzyme-Linked Immunosorbent Assay (ELISA) or Luminex®. Human endometrium produced greater concentrations of TXB2, PGE2 and PGF2α than myometrium in vitro (p<0.05). Fifteen prostanoids were detected in human myometrium. Production of 6-keto-PGF1α, PGE1 and PGF1α increased whilst 15- keto PGE2 and PGJ2 decreased at term pregnancy (37-41 weeks gestation) versus non-pregnancy (p<0.05). Myometrium from parturient donors synthesised TXB2 and PGE2 more abundantly than the non-labouring equivalent. Cytokine concentration was greatest in CVF sampled the week before labour, in particular Interleukin-6 (IL-6), Macrophage Inflammatory Protein-1α (MIP-1α) and Monocyte Chemotactic Protein-1 (MCP-1) (p<0.05). Endometrial TXB2, PGE2 and PGF2α could aid in proliferation of glandular epithelium prior to ovulation. Prostacyclin may facilitate prolongation of pregnancy to term and thromboxane could contribute to uterine stimulation during labour. Cervical dilation may be influenced by PGE2 in lower segment myometrium. MCP- 1, MIP-1α and IL-6 could mark a short interval to labour onset.

Vaginal fluid is most often found at crime scenes where a sexual assault has taken place or on clothing or other items collected from sexual assault victims or perpetrators. Because the victim is generally known in these cases, detection of vaginal fluid is not a matter of individual identification, as it might be for semen identification. Instead, linkages can be made between victim and suspect if the sexual assault was carried out digitally or with a foreign object (e.g., bottle, pool cue, cigarette, handle of a hammer or other tool, etc.). If such an object is only analyzed for DNA and the victim is identified, the suspect may claim that the victim’s DNA is present because she handled and/or is the owner of the object and not because it was used to sexually assault her; identification of vaginal fluid residue would alleviate such uncertainty. Most of the research conducted thus far regarding methods for the identification of vaginal fluid involves mRNA biomarkers and identification of various bacterial strains.1-3 However, these approaches require extensive sample preparation and laboratory analysis and have not fully explored the genomic differences among all body fluid RNAs. No existing methods of vaginal fluid identification incorporate both high specificity and rapid analysis.4 Therefore, a new rapid detection method is required. Surface-enhanced Raman spectroscopy (SERS) is an emerging technique with high sensitivity for the forensic analysis of various body fluids. This technique has the potential to improve current vaginal fluid identification techniques due to its ease-of-use, rapid analysis time, portability, and non-destructive nature. For this experiment, all vaginal fluid samples were collected from anonymous donors by saturation of a cotton swab via vaginal insertion. Samples were analyzed on gold nanoparticle chips.4 This nanostructured metal substrate is essential for the large signal-enhancement effect of SERS and also quenches any background fluorescence that sometimes interferes with normal Raman spectroscopy measurements.5 Vaginal fluid SERS signal variation of a single sample over a six-month period was evaluated under both ambient and frozen storage conditions. Vaginal fluid samples were also taken from 10 individuals over the course of a single menstrual cycle. Four samples collected at one-week intervals were obtained from each individual and analyzed using SERS. The SERS vaginal fluid signals showed very little variation as a function of time and storage conditions, indicating that the spectral pattern of vaginal fluid is not likely to change over time. The samples analyzed over the span of one menstrual cycle showed slight intra-donor differences, however, the overall spectral patterns remained consistent and reproducible. When cycle spectra were compared between individuals, very little donor-to-donor variation was observed indicating the potential for a universal vaginal fluid signature spectrum. A cross-validated, partial least squares – discriminant analysis (PLS-DA) model was built to classify all body fluids, where vaginal fluid was identified with 95.0% sensitivity and 96.6% specificity, which indicates that the spectral pattern of vaginal fluid was successfully distinguished from semen and blood. Thus, SERS has a high potential for application in the field of forensic science for vaginal fluid analysis.

In the human uterus prostaglandins (PG) PGE2, PGD2, PGI2, PGF2¿ and Thromboxane A2 (TXA2), also termed prostanoids, are synthesised and deactivated to 15-keto PGE2, J2 metabolites, 6-keto-PGF1¿, 15-keto PGF2¿ and TXB2 respectively. However, not all metabolites have been analysed simultaneously within the same tissue. The primary objective of this thesis was to determine full uterine prostanoid profiles in human non-pregnancy, pregnancy and parturition, to better understand these processes and find suitable tocolytic targets. In addition, ten cytokines in human cervico-vaginal fluid (CVF) were measured according to interval to labour to test their suitability as labour onset predictors, with a view to developing a test to determine women at risk of preterm labour. Prostanoid analysis was carried out in endometrium (n=9) and myometrium (n=15- 16) donated by non-pregnant women and lower segment myometrium obtained from pregnant women (before (n=14) and after labour onset (n=7)) by liquid chromatography coupled with electrospray ionisation mass spectrometry (LC/ESIMS/ MS). Cytokines produced by CVF collected from pregnant donors (20-41 weeks gestation, n=2-10) were investigated using Enzyme-Linked Immunosorbent Assay (ELISA) or Luminex®. Human endometrium produced greater concentrations of TXB2, PGE2 and PGF2¿ than myometrium in vitro (p<0.05). Fifteen prostanoids were detected in human myometrium. Production of 6-keto-PGF1¿, PGE1 and PGF1¿ increased whilst 15- keto PGE2 and PGJ2 decreased at term pregnancy (37-41 weeks gestation) versus non-pregnancy (p<0.05). Myometrium from parturient donors synthesised TXB2 and PGE2 more abundantly than the non-labouring equivalent. Cytokine concentration was greatest in CVF sampled the week before labour, in particular Interleukin-6 (IL-6), Macrophage Inflammatory Protein-1¿ (MIP-1¿) and Monocyte Chemotactic Protein-1 (MCP-1) (p<0.05). Endometrial TXB2, PGE2 and PGF2¿ could aid in proliferation of glandular epithelium prior to ovulation. Prostacyclin may facilitate prolongation of pregnancy to term and thromboxane could contribute to uterine stimulation during labour. Cervical dilation may be influenced by PGE2 in lower segment myometrium. MCP- 1, MIP-1¿ and IL-6 could mark a short interval to labour onset. / Allergan Inc.