Angiogenesis and vascular function in the ovary.

Ovarian function is dependent on the establishment and continual remodelling of a complex vascular system. This enables the follicle and/or corpus luteum (CL) to receive the required supply of nutrients, oxygen and hormonal support as well as facilitating the release of steroids. Moreover, the inhibition of angiogenesis results in the attenuation of follicular growth, disruption of ovulation and drastic effects on the development and function of the CL. It appears that the production and action of vascular endothelial growth factor A (VEGFA) is necessary at all these stages of development. However, the expression of fibroblast growth factor 2 (FGF2) in the cow is more dynamic than that of VEGFA with a dramatic upregulation during the follicular-luteal transition. This upregulation is then likely to initiate intense angiogenesis in the presence of high VEGFA levels. Recently, we have developed a novel ovarian physiological angiogenesis culture system in which highly organised and intricate endothelial cell networks are formed. This system will enable us to elucidate the complex inter-play between FGF2 and VEGFA as well as other angiogenic factors in the regulation of luteal angiogenesis. Furthermore, recent evidence indicates that pericytes might play an active role in driving angiogenesis and highlights the importance of pericyte-endothelial interactions in this process. Finally, the targeted promotion of angiogenesis may lead to the development of novel strategies to alleviate luteal inadequacy and infertility.

Corpus luteum-endometrium-embryo interactions in the dairy cow: underlying mechanisms and clinical relevance.

Conception rates of dairy cows are currently declining at an estimated 1% every year. Approximately, 35% of embryos fail to prevent luteolysis during the first three weeks of gestation. Interactions between the corpus luteum, endometrium and embryo are critical to the successful establishment of pregnancy and inadequacies will result in the mortality of the embryo. For example, as little as a one day delay in the post-ovulatory rise of progesterone has serious consequences for embryo development and survival. Recently, we found that LH support, degree of vascularization and luteal cell steroidogenic capacity were not the major factors responsible for this luteal inadequacy, but are nevertheless essential for luteal development and function. Progesterone acting on its receptor in the endometrium stimulates the production of endometrial secretions on which the free-living embryo is dependent. However, their exact composition and effects of inadequate progesterone remains to be determined. The embryo is recognized through its secretion of interferon tau (IFNT), which suppresses luteolytic pulses of prostaglandin F(2 alpha). In the cow, it is most likely that IFNT inhibits oxytocin receptor up-regulation directly and does not require the prior inhibition of oestrogen receptor alpha (ESR1). Unravelling the precise luteal-endometrium and embryo interactions is essential for us to understand pregnancy establishment and development of strategies to reverse the declining fertility of dairy cows.

Factors associated with persistent risk of depression in older people following discharge from an acute cardiac unit.

BACKGROUND: This longitudinal study aims to describe the prevalence and characteristics associated with persistent risk of depression in a group of older, hospitalized patients. METHODS: We examined patients at two time-points: baseline and one month post-discharge from hospital. Patients in this study comprised those who had been admitted to the cardiology unit, with no cognitive impairment, aged 60 years and over, and those who were followed up at both time points (N = 155). Questionnaires administered included risk of depression (Geriatric Depression Scale-15; GDS-15), cognitive impairment (Mini-mental State Examination), social support (7-Item Subjective Social Support Index), co-morbidity (Charlson's Comorbidity Index), sociodemographic variables, physical functioning (Modified Barthel's Index) and clinical variables. RESULTS: The prevalence of risk of depression (GDS-15 score > or = 5) among older inpatients at baseline was 34%. At one month post-discharge this had fallen to 17% and this group was identified as those at persistent risk of depression. Factors associated with a risk of persistent depression were: hospitalization within the last six months; length of stay of four days or more; discharge diagnosis of angina; and impaired Subjective Social Support Score. CONCLUSION: Depression occurs commonly among older hospitalized patients and may resolve spontaneously. The identification of factors associated with persistent risk of depression can be helpful when looking at which patients may benefit most from screening and treatment for depression after discharge.

A novel physiological culture system that mimics luteal angiogenesis.

Luteal inadequacy is a major cause of poor embryo development and infertility. Angiogenesis, the formation of new blood vessels, is an essential process underpinning corpus luteum (CL) development and progesterone production. Thus, understanding the factors that regulate angiogenesis during this critical time is essential for the development of novel strategies to alleviate luteal inadequacy and infertility. This study demonstrates the development of a physiologically relevant primary culture system that mimics luteal angiogenesis. This system incorporates all luteal cell types (e.g. endothelial, steroidogenic cells, fibroblasts and pericytes). Using this approach, endothelial cells, identified by the specific marker von Willebrand factor (VWF), start to form clusters on day 2, which then proliferate and develop thread-like structures. After 9 days in culture, these tubule-like structures lengthen, thicken and form highly organized intricate networks resembling a capillary bed. Development of the vasculature was promoted by coating wells with fibronectin, as determined by image analysis (P<0.001). Progesterone production increased with time and was stimulated by LH re-enforcing the physiological relevance of the model in mimicking in vivo luteal function. LH also increased the area stained positively for VWF by twofold (P<0.05). Development of this endothelial cell network was stimulated by fibroblast growth factor 2 and vascular endothelial growth factor A, which increased total area of VWF positive staining on day 9, both independently (three- to fourfold; P<0.01) and in combination (tenfold; P<0.001). In conclusion, the successful development of endothelial cell networks in vitro provides a new opportunity to elucidate the physiological control of the angiogenic process in the developing CL.

Fibroblast growth factor 2 is more dynamic than vascular endothelial growth factor A during the follicle-luteal transition in the cow.

Luteal inadequacy is a major cause of infertility in a number of species. During the early luteal phase, progesterone production requires the rapid growth of the corpus luteum (CL), which is in turn dependent on angiogenesis. In the present study, we examined the temporal changes in vascular endothelial growth factor A (VEGFA), fibroblast growth factor 2 (FGF2) and secreted protein, acidic, cysteine-rich (osteonectin) (SPARC) during the follicular-luteal transition and CL development in the cow. Luteal VEGFA concentrations increased as the CL developed but were lower in the regressing CL. Conversely, luteal FGF2 concentrations were highest immediately postovulation in the collapsed follicle and declined as the CL developed. Furthermore, three FGF2 isoforms were present in the collapsed follicle, but only one isoform was detected in older CL. Interestingly, FGF2 concentrations increased in the regressing CL. Western blot analysis for SPARC showed the presence of two isoforms, which were constitutively expressed throughout CL development. Further studies investigated the regulation of FGF2 by LH, which showed that FGF2 concentrations in preovulatory follicular fluid were higher in those animals that had experienced an LH surge. Moreover, LH stimulated FGF2 production in dispersed luteal cells. Conversely, the LH surge had no effect on follicular fluid VEGFA concentrations. In conclusion, FGF2 was more dynamic than VEGFA and SPARC during the follicular-luteal transition, which suggests that FGF2 plays a key role in the initiation of angiogenesis at this time. Furthermore, it is likely that this is stimulated by the LH surge. The results also suggest that VEGFA and SPARC have a more constitutive, but essential, role in the development of the CL vasculature.

Assessment of maximal aerobic power and critical power in a single 90-s isokinetic all-out cycling test.

The purpose of this study was to establish the validity of a 90-s all-out test for the estimation of maximal oxygen uptake (V.O (2max)) and submaximal aerobic ability as represented by critical power. We hypothesized that the fall in power output by the end of the 90-s all-out test (end power) would represent the exhaustion of anaerobic work capability, and as such, would correspond with the critical power. Sixteen active individuals (mean +/- SD: 30 +/- 6 years; 69.6 +/- 9.9 kg) carried out a series of tests: (i) an incremental ramp test to determine V.O (2max), (ii) three fixed-work rate trials to exhaustion to determine critical power, and (iii) two 90-s all-out tests to measure end power and peak V.O (2). End power (292 +/- 65 W) was related to (r=0.89) but was significantly higher (p<0.01) than critical power (264 +/- 50 W). The mean +/- 95 % limits of agreement (29 +/- 65 W) were too low to use these variables interchangeably. The peak V.O (2) in the 90-s trial was significantly lower than the V.O (2max) (3435 +/- 682 ml x min (-1) vs. 3929 +/- 784 ml x min (-1); p<0.01); mean +/- 95 % limits of agreement was equal to 495 +/- 440 mL x min (-1). The 90-s all-out test cannot, therefore, assess both V.O (2max) and critical power in adult performers. The duration of all-out exercise required to allow V.O (2) to attain its maximum is longer than 90 s.

Endocrine and cellular characteristics of corpora lutea from cows with a delayed post-ovulatory progesterone rise.

The timing of the post-ovulatory progesterone rise is critical to the embryonic development and survival. The aim of this study was to determine the underlying causes of delayed post-ovulatory progesterone rises. Two groups of non-lactating dairy cows with early (n = 11) or late (n = 9) post-ovulatory progesterone rises were created by inducing luteolysis in the presence of either a large (> 10 mm) or small (< 10 mm) follicle, respectively. LH pulses were measured on days 4 (all cows) and 7 (n = 7, early; n = 5, late) (day 1= ovulation). The cows were slaughtered on day 5 (n = 4 each group) or 8 (n = 7, early; n = 5, late). Immunohistochemical analysis for endothelial cells (von Willebrand Factor, VWF), steroidogenic cells (3beta-HSD) and proliferation marker (Ki67) were performed. The basal progesterone production and LH responsiveness (0.001-100 ng/ml) of dispersed luteal cells was investigated. The luteal concentrations of FGF-2 and VEGF were measured by ELISA and RIA, respectively. There were no differences in LH pulse characteristics, area of VWF staining, proliferation index, steroidogenic cell characteristics, basal or LH-stimulated progesterone production by luteal cells between cows with an early or late progesterone rise (P > 0.10). However, the area of VWF staining increased from days 5 to 8, while the proliferation index decreased (P < 0.05). Furthermore, the luteal cells were more responsive to LH on day 8 (P < 0.01). Luteal concentrations of FGF-2 were higher on day 5 (P = 0.05), while VEGF was greater on day 8 (P < 0.01). In conclusion, we have clearly shown that LH support, degree of vascularization or luteal cell steroidogenic capacity were not the major factors responsible for inadequate secretion of progesterone by the developing bovine CL.

Validity of the two-parameter model in estimating the anaerobic work capacity.

The curvature of the power-time (P-t) relationship (W') has been suggested to be constant when exercising above critical power (CP) and to represent the anaerobic work capacity (AWC). The aim of this study was to compare W' to (1) the total amount of work performed above CP (W (90s)') and (2) the AWC, both determined from a 90s all-out fixed cadence test. Fourteen participants (age 30.5 +/- 6.5 years; body mass 67.8 +/- 10.3 kg), following an incremental VO(2max) ramp protocol, performed three constant load exhaustion tests set at 103 +/- 3, 97 +/- 3 and 90 +/- 2% P-VO(2max) to calculate W' from the P-t relationship. Two 90s all-out efforts were also undertaken to determine W (90s)' (power output-time integral above CP) and AWC (power output-time integral above the power output expected from the measured VO(2)). W' (13.6 +/- 1.3 kJ) and W (90s)' (13.9 +/- 1.1 kJ; P = 0.96) were not significantly different but were lower than AWC (15.9 +/- 1.2 kJ) by 24% (P = 0.03) and 17%, respectively (P = 0.04). All these variables were correlated (P < 0.001) but great extents of disagreement were reported (0.2 +/- 6.4 kJ between W' and W (90s)', 2.3 +/- 7.2 kJ between W' and AWC, and 2.1 +/- 4.3 kJ between W (90s)' and AWC). The underestimation of AWC from both W' and W (90s)' can be explained by the aerobic inertia not taking into consideration when determining the two latter variables. The low extents of agreement between W', W (90s)' and AWC mean the terms should not be used interchangeably.

The induction of a delayed post-ovulatory progesterone rise in dairy cows: a novel model.

A delayed rise in post-ovulatory progesterone is associated with poor embryo development in the cow, although the underlying cause of this aberrant luteal function is poorly understood. The objective of this study was to develop a novel model, in which a delayed progesterone rise could be induced by manipulating the dynamics of the follicular phase. Luteolysis was induced in 20 dairy cows in the presence of either a larger follicle > 10 mm (LF, n = 11) or a smaller follicle < 10 mm (SF, n = 9) and transrectal ultrasonography was performed to determine follicle and CL growth and timing of ovulation. Plasma progesterone and oestradiol were analysed 3x daily. Cows were slaughtered on either day 4 (n = 4 per group) or day 7 (SF, n = 5; LF, n = 7) after ovulation. The pre-ovulatory follicle was larger in the LF group than the SF group at luteolysis (13.5 +/- 0.4 mm versus 6.7 +/- 0.7 mm, P < 0.001) and ovulation (16.7 +/- 0.3 mm versus 13.6 +/- 0.6 mm, P < 0.001). The LF group experienced a shorter follicular phase and ovulated 36 h earlier than the SF group (P < 0.001). At luteolysis, plasma oestradiol concentrations were greater in the LF group (P < 0.001), although peak concentrations were not different (P > 0.05). Moreover, higher progesterone concentrations were observed in the LF group during the early luteal phase (P < 0.05). Luteal weights were positively correlated with plasma progesterone concentrations on day 5 (P < 0.05) but not day 8. In conclusion, a model has been developed which has shown that the dynamics of follicle development during the pre-ovulatory period is an important determinant of subsequent CL development and function.

The effects of undernutrition, in utero, on reproductive function in adult male and female sheep.

The aim of this study was to determine the effects of maternal undernutrition during pregnancy on adult reproductive function in male and female offspring. Groups of ewes were fed rations providing either 100% (High, H) or 50% (Low, L) of estimated metabolisable energy (ME) requirements for pregnancy, from mating until day 95 of gestation, and thereafter were conventionally managed. At 20 months of age, LH and FSH profiles, and LH responses to exogenous GnRH were measured in male and female offspring and, in males, testicular responses to exogenous LH (as measured by testosterone concentrations) were also measured. Undernutrition had no effect on the mean birth weights of lambs of either sex, or on testicular size in male animals at either 6 weeks or 20 months of age. L males exhibited significantly higher FSH concentrations than H males (P < 0.05) but there were no differences with treatment in FSH profiles in females, basal LH profiles or gonadotrophin responses to GnRH in offspring of either sex, and no difference in basal testosterone concentrations or in the testosterone response to exogenous LH administration in males. Semen quality at 20 months of age was unaffected by pre-natal undernutrition but ovulation rate was significantly reduced in L compared to H female offspring (P < 0.05). It is concluded that pre-natal undernutrition had no effect on male reproductive development and adult function, but reduced ovulation rate in female progeny. This effect was not associated with a change in gonadotrophin profiles or pituitary responsiveness.

Bilateral hypoglossal palsies: a late complication of curative radiotherapy.

A 57 year old man developed bilateral hypoglossal nerve palsies 6 years after radiotherapy for carcinoma of the uvula. Follow-up over 2 years has demonstrated no evidence of tumour recurrence and no sign of neurological improvement. Reactive fibrosis and vascular insufficiency secondary to radiation and may have lead to hypoglossal nerve compression and infarction.