M.A. Warren

Interleukin-11 (IL-11) in human endometrium: expression throughout the menstrual cycle and the effects of cytokines on endometrial IL-11 production in vitro

Interleukin-11 is a member of the IL-6 family of cytokines. Its presence in mouse decidua has been shown and experiments in genetically modified mice have suggested the importance of its receptor in stromal cell decidualization. In this study we used immunocytochemistry to determine expression of IL-11 in human endometrium. The effects of TNFalpha, IL-1alpha and TGFbeta on IL-11 production by epithelial and stromal cells was also investigated. Immunocytochemical staining in sections cut from 19 endometrial biopsies obtained throughout the menstrual cycle showed that IL-11 was expressed in both human endometrial epithelial and stromal cells, with epithelial staining being more intense than that seen in the stromal cells, at all times except the late secretory phase when the intensity was similar. Basal IL-11 production by cultured epithelial cells was greater than basal production by stromal cells. IL-1alpha, TNFalpha and TGFbeta (0.1-10 ng/ml) all caused a concentration-dependent increase in IL-11 production by both epithelial and stromal cells, but stimulated: basal values were greater for stromal than epithelial cells for all three cytokines. This work shows, for the first time, the presence of IL-11 within the human endometrium and that its production is controlled by other cytokines, which are postulated to play a role in implantation. Thus IL-11 may also play an important role in human endometrial function and embryo implantation.

A prospective, randomised, cross-over study comparing the effects of clomiphene citrate and cyclofenil on endometrial morphology in the luteal phase of normal, fertile women

Objective To examine the effect of two anti‐oestrogens, clomiphene citrate and cyclofenil, on endometrial morphology in the luteal phase.

Physiology: The production of placental protein 14 by human uterine tubal epithelial cells in culture

Cells prepared from the mucosal layer obtained from the fimbrial, proximal ampullary and distal ampullary regions of the human uterine (Fallopian) tube have been grown in monolayer culture. Immunocytochemistry with anti-cytokeratin, anti-vimentin or anti-CD 45 antibodies indicated that the overwhelming number of cells were epithelial in nature and were free of fibroblasts and leukocytes. Basal and steroid-stimulated placental protein 14 (PP14) production was investigated in tissue obtained from nine patients undergoing hysterectomy, by addition of oestradiol and/or progesterone to confluent cultures. Basal PP14 production varied considerably between experiments, probably due to differences between individuals from whom the tissue had been obtained. However, there was no difference in basal PP14 production by cells prepared from the fimbrial, proximal ampullary and distal ampullary parts of the tube obtained from the same patient. When total PP14 production by cells obtained from an individual uterine tube was pooled both progesterone and oestradiol significantly (P < 0.05) stimulated the production of PP14 but the effect of progesterone either alone or in the presence of oestradiol was numerically greater than that of oestradiol alone. Considering PP14 production by cells prepared from the different regions of the tube showed that cells from the fimbrial region were more responsive to steroid stimulation than cells prepared from either the proximal or the distal ampullary regions. All combinations of hormonal supplementation stimulated PP14 production by cells from the fimbrial region on all days measured (P < 0.05 - P < 0.001).(ABSTRACT TRUNCATED AT 250 WORDS)

Endometrial responses in artificial cycles: a prospective study comparing four different oestrogen dosages.

Objective To examine the endometrial response to four different regimens of oestrogen.

A study of endometrial morphology in women who failed to conceive in a donor insemination programme

Objective To examine the morphology of endometrium in women who failed to conceive after nine or more cycles of donor insemination treatment.

Endometrial responses in artificial cycles : a prospective, randomized study comparing three different progesterone dosages

Objective To examine the endometrial response to three different regimens of progesterone.

Undernutrition during early adult life significantly affects neuronal connectivity in rat visual cortex

Male black and white Hooded Lister rats were undernourished for 29 days during early adult life. Undernourished rats had 30% more synapses per neurone in the visual cortex than matched controls. It is suggested that undernutrition may cause a delay in the normal decline of this ratio.

Response of human spermatozoa to an internal calcium ATPase inhibitor, 2,5‐di(tert‐butyl) hydroquinone

This study has investigated the effect of elevating intracellular calcium levels, using an internal calcium ATPase inhibitor, 2,5-di(tert-butyl) hydroquinone (TBQ), on human sperm function. Isolated sperm samples from five fertile donors were incubated in a capacitating media for up to 6 hr. After 0, 3, and 6 hr incubation, sperm were exposed to a range of TBQ concentrations; 100 microM, 10 microM, and 1 microM, for a fixed incubation period of 5 min. Controls were run for each experiment where sperm were incubated for 5 min in the absence of TBQ. Sperm capacitation and the acrosome reaction were monitored prior to and after exposure to TBQ, using the Chlortetracycline assay. In addition, sperm motility was assessed at each time point and after sperm had been exposed to TBQ. The treatment of sperm with TBQ caused a significant increase in the number of capacitated sperm with an optimum response being achieved in the presence of 100 microM TBQ. However, sperm motility was found not to be effected by the addition of TBQ. The results from the present study suggest that elevating intracellular calcium levels in human sperm by short exposure to a high concentration of TBQ can rapidly accelerate the capacitation process. Furthermore, the observation that TBQ did not elicit a change in sperm motility suggests that TBQ may be highly specific in its mode of action by acting within the head region of human sperm.

Elevating intracellular calcium levels in human sperm using an internal calcium ATPase inhibitor, 2,5-di(tert-butyl) hydroquinone (TBQ), initiates capacitation and the acrosome reaction but only in the presence of extracellular calcium

The aim of this study was to investigate the effect of an internal calcium ATPase inhibitor, TBQ, on human sperm capacitation and the acrosome reaction during incubation in a calcium-depleted media. Sperm were isolated into and incubated for up to 6 hr in media depleted of Ca2+ and two Ca(2+)-containing media controls. At set time intervals, sperm in each media group were treated with 100 microM TBQ for 5 min. Afterwards, sperm were induced to acrosome react using the divalent cation ionophore, A23187, as a measure of sperm fertilizing potential. It was established, using the Chlortetracycline assay, that incubation of sperm in a Ca(2+)-depleted media inhibited or delayed sperm capacitation resulting in fewer spontaneous or A23187-induced acrosome reacted sperm. However, incubation of sperm in a Ca(2+)-depleted media did not appear to inhibit sperm motility. The treatment of sperm with TBQ during their incubation in Ca(2+)-depleted media was found to have very little effect resulting in low numbers of capacitated and acrosome reacted sperm. The results from this study suggest that human sperm have an obligatory requirement for extracellular calcium during capacitation and the acrosome reaction, but may require either very little extracellular Ca2+ to maintain motility or possess internal Ca2+ stores sufficient for their requirements. In addition, TBQ did not increase the number of capacitated and acrosome reacted sperm during incubation in a Ca(2+)-depleted media suggesting that the TBQ-effect of accelerating sperm capacitation is dependent on presence of extracellular Ca2+.

Morphology of the Human Endometrium in the Peri-Implantation Period

Although the histology and ultrastructure of the human endometrium are well documented, it is clear that morphometry can reveal subtle changes in cellular biology, especially when combined with suitable sampling and dating of the specimen. These changes reflect functional events, such as protein secretion, which may be studied using the endometrial flushing technique. Although it is likely that further insights into the basic cell physiology of the reproductive tract will result from the use of appropriate in vitro models, the role of endometrial biopsy for clinical monitoring remains essential.