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Unlike regular time evolution governed by the Schr\"odinger equation, standard quantum measurement appears to violate time-reversal symmetry. Measurement creates random disturbances (e.g., collapse) that prevents back-tracing the quantum state of the system. The effect of these disturbances is explicit in the results of subsequent measurements. In this way, the joint result of sequences of measurements depends on the order in time in which those measurements are performed. One might expect that if the disturbance could be eliminated this time-ordering dependence would vanish. Following a recent theoretical proposal [A. Bednorz et al 2013 New J. Phys. 15 023043], we experimentally investigate this dependence for a kind of measurement that creates an arbitrarily small disturbance, weak measurement. We perform various sequences of a set of polarization weak measurements on photons. We experimentally demonstrate that, although the weak measurements are minimally disturbing, their time-ordering affects the outcome of the measurement sequence for quantum systems., Comment: 11 pages, 3 figures

Context: Production of 3-carbon units (as lactate) by granulosa cells (GCs) is important in follicular and oocyte development and may be modulated by metformin. Objective: The aim of the study was to examine the action of metformin on GC lactate production and potential mediation via AMP-activated protein kinase (AMPK). Design: GCs were prepared from follicular aspirates. After exposure to metformin and other potential modulators of AMPK in culture, aspects of cellular function were examined. Setting: The study was conducted in a private fertility clinic/university academic center. Patients: Women undergoing routine in vitro fertilization participated in the study. Interventions: All agents were added in culture. Main Outcome Measures: Lactate output of GCs was measured. Cell extracts were prepared after culture, and phosphorylated forms of AMPK and acetyl CoA carboxylase (ACC) were assayed using Western analysis. Results: Metformin led to a rapid increase in lactate production by GCs [minimum effective dose, 250 μm; maximum dose studied, 1 mm (1.22-fold; P < 0.01)]. This dose range of metformin was similar to that required for stimulation of phospho-AMPK in GCs [minimum effective dose, 250 μm; maximum effect, 500 μm (2.01-fold; P < 0.001)]. Increasing phospho-ACC, as a representative downstream target regulated by AMPK, was apparent over a lower range (minimum effective dose, 31 μm; maximum effect, 250 μm; P < 0.001). A level of metformin (125 μm) insufficient for the stimulation of lactate output when used alone potentiated the effects of suboptimal doses of insulin on lactate production. Adiponectin (2.5 μg/ml) had a small but significant effect on lactate output. Conclusions: Metformin activates AMPK in GCs, stimulating lactate production and increasing phospho-ACC. Metformin also enhances the action of suboptimal insulin concentrations to stimulate lactate production.

The isoforms of sterol regulatory element-binding proteins (SREBP) (1a, 1c, and 2) are key transcriptional regulators of lipid biosynthesis. We examined their regulation by gonadotropin and insulin in human granulosa cells. After removal of leukocytes, granulosa cells were exposed to hormonal additions for 16 h starting on d 2 of culture. Progesterone, lactate, and IGF binding protein-1 were measured in culture medium and cellular mRNA measured by competitive RT-PCR. Addition of human chorionic gonadotropin (hCG) (100 ng/ml) stimulated progesterone production (7.0-fold, P < 0.001 vs. control), whereas lactate was increased by hCG (1.6-fold, P < 0.001) and insulin (1.4-fold, P < 0.001; 1000 ng/ml). Insulin decreased IGF binding protein-1 production by 85% (P < 0.001). There were no significant effects on the expression of SREBP-1a but significant increases in mRNA for SREBP-1c with insulin (6.3-fold), hCG (10.4-fold) and in combination (15.2-fold; P < 0.01 for all comparisons). No consistent effects on SREBP-2 were observed. The expression of mRNA for fatty acid synthase, a target gene for SREBP-1c, was increased by hCG (24-fold, P = 0.006) and insulin (19-fold, P = 0.024), which also increased the level of cellular, total fatty acid (1.34-fold; P = 0.03). Thus, hCG and insulin cause a switch toward expression of the SREBP-1c isoform with consequent effects on fatty acid synthesis. We suggest that high circulating insulin, associated with clinically defined insulin resistance, may up-regulate SREBP-1c expression in the ovary.

Human granulosa cells (GC), prepared from follicular aspirates using a non-enzymic method, were maintained in culture on chamber slides in a defined medium without additional attachment factors or extracellular matrix (ECM). In this system, GC clustered to a limited extent and attached only loosely to the substratum necessitating medium replacement through repeated partial changes to avoid cell loss. Using this new culture system, cell size and progesterone production per cell increased, consistent with continuing luteinization. These processes were associated with maintenance and deposition of endogenous ECM components. Thus, pericellular heparan sulphate proteoglycan (HSPG) was clearly visible by immunocytochemistry around the luteinized GC after culture. Also progressive accumulation of laminin (particularly alpha(2)-, beta(1)- and gamma(1)-subunits) during culture was shown by Western blotting of GC extracts. Small patches of collagen IV, shown to be already present between freshly prepared GC, were maintained in culture. A clear effect of gonadotrophin on the maintenance of progesterone production in culture was paralleled by an apparent increased pericellular deposition of HSPG. To conclude, luteinization and maintenance of the GC-derived layer of the corpus luteum is likely to involve deposition and conservation of pericellular ECM components.

Human ovarian granulosa cells were cultured on a basement membrane preparation (Matrigel) to investigate the role of extracellular matrix components in granulosa cell cluster formation. Time-lapse videomicroscopy of these cultures revealed a rapid aggregation of cells which was initiated during the first 2-4 h of culture so that by 8 h most of the granulosa cells were incorporated into clusters. Further amalgamation then occurred with the transfer of cells along 'bridges' between combining clusters. The clustering process, which was complete by about 24 h, was accompanied by reorganisation of matrix which was visualised by immunolabelling of laminin. Clustering cells appeared to gather matrix which became distributed around the clusters. Confocal microscopy showed matrix to be present over the surface of each cluster as well as around the base apparently anchoring the aggregate to the culture surface. Results suggest the potential for active rearrangement of matrix by granulosa-derived cells during corpus luteum development.

Corpus luteum formation is characterized by a period of extensive vascularization, as capillaries in the thecal layer of the collapsed follicle following ovulation invade the previously avascular granulosa layer. In order to study these processes in vitro we have developed an endothelial cell preparation from the specific microvasculature of the ovarian follicle. Follicular aspirates, obtained at oocyte collection for in-vitro fertilization (IVF), were filtered to obtain fragments of follicle wall. These were set in Matrigel and then cultured allowing the growth of capillary-like structures through the matrix. Upon emergence from the Matrigel the growing cells formed monolayers with the characteristic cobble-stone morphology of endothelial cells. Immunocytochemistry demonstrated the presence of a range of endothelial-specific markers including von Willebrand factor (vWF), Ulex europeus agglutinin (UEA)-1, CD31 and E-selectin, as well as VCAM-1, which is normally associated with stimulated endothelial cells. RT-PCR analysis showed the expression of two receptors for vascular endothelial growth factor (flt-1 and KDR), and the endothelial nitric oxide synthase, adding further evidence of their identity as human ovarian microvascular endothelial cells (HOMEC). Thus, the novel preparative procedure described now allows the generation of HOMEC cultures from readily available material resulting from IVF procedures.

The rapid introduction of laparoscopic cholecystectomy has been associated with an apparently increased incidence of bile duct injury which has provoked worldwide concern. The true incidence and mechanism of iatrogenic ductal injury during the development of this procedure remain unclear. To assess this, the introduction of laparoscopic cholecystectomy in the West of Scotland has been audited prospectively over a 5-year period. All cases of biliary ductal injury have been independently reviewed. Some 48 surgeons undertaking laparoscopic cholecystectomy in 19 hospitals submitted prospective data between September 1990 and September 1995. A total of 5913 laparoscopic cholecystectomies were attempted with 98·3 per cent completion of data collection. During this period 37 laparoscopic bile duct injuries occurred. The annual incidence peaked at 0·8 per cent and has fallen to 0·4 per cent in the final year of audit. Injuries occurred after a median personal experience of 51 (range 3–247) laparoscopic chole cystectomies in 22 surgeons. Major bile duct injuries occurred in 20 of 37 patients, giving an incidence of 0·3 per cent. Five mechanisms for laparoscopic ductal injury were identified, including tenting, confluence and diathermy injuries as well as the classical and variant classical types. Ductal injuries were discovered at operation in 18 patients with consequent repair giving a good clinical outcome in 17. Contributory factors (severe inflammation, aberrant anatomy and poor visualization) were present in only 13 of 37 cases. This audit suggests that, at least in the introductory period, laparoscopic cholecystectomy is associated with an overall bile duct injury rate higher than that reported previously after open cholecystectomy, although the incidence of major ductal injury is similar. The late downward trend in bile duct injury, however, suggests there may be a prolonged learning curve for this procedure. Improved understanding of the mechanism of injury may lead to yet further reductions in this complication.

Granulosa cells were isolated from follicular aspirates collected at ovum recovery for in vitro fertilization. Cells were cultured in a defined medium on artificial extracellular matrix (Matrigel) in the presence or absence of hCG as a model for corpus luteum function. Release of cells from this culture system is reduced by hCG and this effect may be mediated through an inhibition of extracellular matrix degradation. Using zymography and western blot analysis, we confirm the identity of matrix metalloproteinases-2 and -9 in culture media. Matrix metalloproteinase-9 was the predominant gelatinase in freshly prepared granulosa cells and in culture media, and also represented a major metalloproteinase component in homogenates of early and mid-luteal phase samples of corpora lutea. Quantitative analysis of matrix metalloproteinases in culture media, obtained throughout the 14 day culture period and expressed per microgram of DNA, showed that matrix metalloproteinase-2, undetectable on day 2, rose throughout the culture period and that this rise was significantly inhibited by hCG. In contrast, matrix metalloproteinase-9 was clearly detectable on day 2 and remained relatively constant throughout much of the culture (day 2 to day 12) in the presence of gonadotrophin. Significantly increased production of matrix metalloproteinase-9 (day 6 to day 12) was evident in the absence of hCG. Our results provide further evidence for the hypothesis that the rescue of the corpus luteum in early pregnancy involves the maintenance of cellular function through the stabilization of the extracellular matrix.

We have previously shown that human granulosa cells cultured on a thin layer of extracellular matrix (ECM) are lost from culture in the absence of gonadotrophin. We now examine the effect of plating ECM onto glass or plastic. Such a comparison revealed that loss of cells from control cultures was more rapid when ECM was on glass, whereas cultures maintained with human chorionic gonadotrophin (HCG) showed greater stability when ECM was on plastic. The dose range of HCG required for the effect on cell retention was similar to that required for stimulation of progesterone production. Electron microscopy of cells freshly released as clusters revealed that many cells appeared undamaged, and confocal microscopy of cells stained with propidium iodide showed an absence of fragmented nuclei. Taken together, this evidence suggests that apoptosis is not the cause of cell release. We conclude that cells are released from culture, not as a result of cell death, but via an active process suppressed by HCG.

Granulosa cells were prepared from follicular aspirates obtained at oocyte collection for in-vitro fertilization (IVF) and maintained in culture. Substantial loss of cells from the culture surface occurred in the absence of gonadotrophin when cells were maintained on a thin layer of extracellular matrix (ECM) using a defined, serum-free medium. This cell loss was clearly and significantly reduced in the presence of human chorionic gonadotrophin (HCG) by days 4-6 of culture, and occurred in conjunction with loss of ECM. Analysis of culture medium by zymography using gelatin as substrate demonstrated the presence of metalloproteinases (MMP), MMP-9 (gelatinase B) appearing as the predominant band. Measurement of overall gelatinase activity in culture media revealed a progressive fall in gelatinase expressed on a per cell basis in media from HCG-treated cultures and this was less marked in controls. This suppression of gelatinase activity was consistent with an observed increase in production of tissue inhibitor of metalloproteinase-1 (TIMP-1) by HCG-treated cells, which was significant by days 6-8 of culture. We speculate that stabilization of the ECM may be an important aspect of HCG action in the corpus luteum.

We describe a XUV spectrometer for the study of dense hot microplasmas at wavelengths between ≈50 and ≈300 Å. It uses a commercially fabricated grazing incidence flat-field reflection grating with 1200 grooves per millimeter. The spectral resolution was optimized by imaging the source on a narrow slit with the help of a curved grazing incidence mirror. The instrument was tested with a laser-produced plasma as a source. The limit of the resolving power due to imaging aberrations of the flat-field grating ranges from 1500 at 50 Å to 3600 at 200 Å and has been achieved with a 5-μm slit. We also measured and calculated the grating efficiencies for the first to fifth diffraction order as a function of wavelength.

1. A gamma scintigraphic study has been carried out on the precorneal residence and pharmacodynamic action of a radiolabelled New Ophthalmic Delivery System (NODS) containing pilocarpine nitrate in 12 healthy volunteers. 2. The NODS was radiolabelled with the soluble marker technetium-99m labelled diethylenetriaminepentaacetic acid, to mark the release characteristics of soluble drugs contained within the matrix. 3. The relationship between the precorneal residence time of the marker and the duration of drug effect on intraocular pressure and pupil diameter was monitored. Results obtained following administration of the NODS were compared with those obtained after administration of a 25 microliters drop of a 2% w/v pilocarpine nitrate solution. Each formulation was administered to one eye only, the other eye acting as a control. 4. Dissolution of the radiolabel from the NODS in vivo showed considerable intersubject variation with half-times of dissolution ranging from 46 s to 833 s (mean +/- s.d. -280 +/- 217 s), the mean (+/- s.d.) half-time of clearance of the radiolabel from the NODS and corneal region of interest was 406 +/- 214 s whereas the radiolabelled solution had a mean (+/- s.d.) ocular surface residence time of 2.9 +/- 1.5 s. 5. Pupil diameter and intraocular pressure were measured for 5 h post-administration of the NODS and the solution. After both treatments pupil diameter was significantly constricted in the test eye when compared with the control eye (P less than 0.001; Student's paired t-test). Pupil diameter was constricted by 52% after administration of the NODS and by 35% after administration of the solution.(ABSTRACT TRUNCATED AT 250 WORDS)

In primates, corpus luteum development involves both gonadotrophin stimulation and exposure to low density lipoprotein (LDL) delivered through vascularization of the granulosa cell-derived layer. These regulatory influences were modelled in vitro using granulosa cells obtained during in-vitro fertilization (IVF) cycles controlled with gonadotrophin releasing hormone (GnRH) analogue, human menopausal gonadotrophin (HMG) and human chorionic gonadotrophin (HCG). Granulosa cells were cultured in defined medium on extracellular matrix. Without gonadotrophin or LDL in the medium, progesterone production declined progressively. With LDL alone, there was a short-lived elevation of progesterone output which subsequently declined. Culture with HCG alone resulted in a relatively unchanged rate of steroid production over 5 days despite morphological development. This contrasted with a marked and sustained increase in progesterone output over the same time when granulosa cells were cultured with combined HCG/LDL. Cultures were challenged with combined HCG/LDL on day 5. Where initial incubation included HCG, the challenge resulted in a recovery of progesterone output to values comparable to those of granulosa cells exposed to continuous HCG/LDL. Initial incubation without gonadotrophin led to a reduced response. Results suggest that LDL delivery to granulosa cells of the early corpus luteum causes a short-lived period of progesterone production. Sustained luteinization of granulosa cells and maintenance of gonadotrophin responsiveness requires continued exposure to gonadotrophin in the luteal phase.

Human granulosa cells, from women undergoing ovum collection for in-vitro fertilization (IVF), will luteinize in vitro and provide a model for investigating the antigonadotrophic action of a prostaglandin F2 alpha (PGF2 alpha) analogue, cloprostenol, on granulosa-derived luteal cells. The granulosa cells were cultured in a defined medium and exposed to treatments during a preincubation period of 0 to 3 days and a final incubation with low density lipoprotein (LDL) from days 3 to 4. In the absence of human chorionic gonadotrophin (HCG), progesterone production was low, whereas exposure to HCG in the final incubation resulted in a 10-fold increase in progesterone concentrations. The inclusion of cloprostenol with HCG in the final incubation significantly (P less than 0.05) inhibited HCG-stimulated progesterone production. Exposure to HCG during the preincubation prevented the antigonadotrophic action of cloprostenol in the final incubation. The antigonadotrophic action of cloprostenol was retained when the granulosa cells were exposed to cloprostenol during the preincubation. Omission of LDL from the final incubation lowered the production of progesterone but the pattern of responses to HCG and cloprostenol were similar. Prevention of the antigonadotrophic action of cloprostenol after exposure to HCG may be a mechanism through which chorionic gonadotrophin can prevent regression of the corpus luteum in early pregnancy. Cloprostenol does not appear to inhibit LDL-stimulated steroidogenesis in human granulosa cells.

The size distribution of marmoset luteal cells was determined on Days 6, 14 and 20 after ovulation in non-pregnant cycles and in early pregnancy. Image analysis was used to estimate the cell diameter of dispersed cells prepared from the marmoset corpus luteum (CL). Steroidogenic cells showed a size distribution consistent with one population of cells. There was a significant increase in mean cell diameter (P less than 0.05) from Day 6 to Day 14 in pregnant and non-pregnant animals with no further increase on Day 20. Micrographs of marmoset luteal tissue showed cells of greater than 10 micron containing the organelles typical of steroid-producing cells, and smaller non-steroidogenic cells surrounding the steroid-producing cells. On the basis of microscopy, there were no areas within the CL where cell composition was noticeably different. In contrast, micrographs of human luteal tissue showed two types of steroidogenic cell; most cells were similar to those in the marmoset CL but a smaller population of smaller cells could be distinguished around the periphery and along vascular septa. It is likely that these smaller and larger types of steroidogenic cells are of theca and granulosa cell origin respectively, the two cell populations differing in the degree of electron density and amount of rough endoplasmic reticulum. A distinguishing feature between marmoset and human luteal cells was the shape of the mitochondrian which were considerably rounder in marmoset luteal cells. The origin of steroidogenic cells in the marmoset CL is unclear, although in marmosets and man the luteal cell types display morphological characteristics distinct from the large and small luteal cells described for CL of the domestic ungulates.

Background Computed tomography (CT) has become established in the assessment of paediatric blunt abdominal trauma. However, advances in diagnostic imaging necessitate reassessment of the role of available diagnostic modalities. Methods Experience at a paediatric teaching hospital over a 5-year period was reviewed, with direct comparison of CT against ultrasonographic imaging in 26 children presenting with acute blunt abdominal trauma. Results Intra-abdominal injury was diagnosed by CT in 23 of 24 patients compared with 21 on ultrasonography, although ultrasonography identified organ-specific injury in only 12 of 24 patients. CT was superior in the assessment of the multiply injured child, and identified spinal and pelvic injuries in three patients. CT augmented plain chest radiography in ten patients with associated thoracic injuries. Conclusion CT is the imaging modality of choice in children with severe abdominal trauma but ultrasonography is a reasonable technique to arouse diagnostic suspicion in less severe injuries or where CT is unavailable or delayed.

The Fallopian tube provides the environment for early embryo growth, a process which is influenced by insulin-like growth factors (IGFs) in the tubal fluid. Whether the bioavailability of tubal IGFs is modulated by locally produced IGF-binding protein (IGFBP-1) is not clear. An explant culture system from human Fallopian tube mucosa was, therefore, developed enabling the potential for IGFBP-1 production by this tissue to be examined directly. Initial characterization of the system established that the explants maintained responsiveness to steroids. Thus, oviduct-specific glycoprotein production, a major product of the oviduct in vivo, continued to be made via an estrogen-sensitive pathway in the culture. The presence of mRNA for IGFBP-1 was established within the explants by the use of quantitative RT-PCR and IGFBP-1 protein was measured by enzyme-linked immunosorbent assay. Although insulin and estradiol had no consistent effect on IGFBP-1, addition of progesterone had a significant inhibitory effect on IGFBP-1 production, both at the mRNA and protein levels. A dose-range of progesterone revealed an incremental inhibitory effect of progesterone on IGFBP-1 output (maximal effect, 25-50 nmol/l), consistent with physiological inhibition of this process during the luteal phase. We suggest that progesterone might, therefore, play a role in controlling the bioavailability of IGFs to the embryo during early development within the Fallopian tube.

Computed tomography (CT) has become established in the assessment of paediatric blunt abdominal trauma. However, advances in diagnostic imaging necessitate reassessment of the role of available diagnostic modalities.Experience at a paediatric teaching hospital over a 5-year period was reviewed, with direct comparison of CT against ultrasonographic imaging in 26 children presenting with acute blunt abdominal trauma.Intra-abdominal injury was diagnosed by CT in 23 of 24 patients compared with 21 on ultrasonography, although ultrasonography identified organ-specific injury in only 12 of 24 patients. CT was superior in the assessment of the multiply injured child, and identified spinal and pelvic injuries in three patients. CT augmented plain chest radiography in ten patients with associated thoracic injuries.CT is the imaging modality of choice in children with severe abdominal trauma but ultrasonography is a reasonable technique to arouse diagnostic suspicion in less severe injuries or where CT is unavailable or delayed.

Human granulosa cells were maintained in culture with extracellular matrix in the presence or absence of human chorionic gonadotrophin (HCG) using a defined culture medium. Such cultures are maintained by gonadotrophin in a manner suggesting that features of 'luteal rescue' may be occurring in vitro. Western analysis of culture medium demonstrated that the granulosa cells produced tissue inhibitor of metalloproteinases (TIMP)-1 but not TIMP-2. The presence of TIMP-1 in cultured cells was also detected immunocytochemically. Immunoassay of TIMP-1 output revealed that HCG exposure for 7 days caused a 2-fold increase in TIMP-1 production versus control reaching maximum at approximately 1 ng HCG/ml. The sensitivity of this response to HCG was similar to that observed for stimulation of progesterone production. Delayed addition of HCG, from day 4 of culture, elicited increases in TIMP-1 which were evident within 24 hours, and were not explained by changes in cell replication or survival. Removal of HCG from cultures previously luteinized with HCG for 6 days resulted in a fall in TIMP-1 production. Thus TIMP-1 production by luteinized granulosa cells in culture is gonadotrophin dependent. We speculate that prolonged cellular function associated with 'luteal rescue' may result from increased extracellular matrix stability mediated by up-regulation of TIMP-1 production.

The rapid introduction of laparoscopic cholecystectomy has been associated with an apparently increased incidence of bile duct injury which has provoked worldwide concern. The true incidence and mechanism of iatrogenic ductal injury during the development of this procedure remain unclear. To assess this, the introduction of laparoscopic cholecystectomy in the West of Scotland has been audited prospectively over a 5-year period. All cases of biliary ductal injury have been independently reviewed. Some 48 surgeons undertaking laparoscopic cholecystectomy in 19 hospitals submitted prospective data between September 1990 and September 1995. A total of 5913 laparoscopic cholecystectomies were attempted with 98.3 per cent completion of data collection. During this period 37 laparoscopic bile duct injuries occurred. The annual incidence peaked at 0.8 per cent and has fallen to 0.4 per cent in the final year of audit. Injuries occurred after a median personal experience of 51 (range 3-247) laparoscopic cholecystectomies in 22 surgeons. Major bile duct injuries occurred in 20 of 37 patients, giving an incidence of 0.3 per cent. Five mechanisms for laparoscopic ductal injury were identified, including tenting, confluence and diathermy injuries as well as the classical and variant classical types. Ductal injuries were discovered at operation in 18 patients with consequent repair giving a good clinical outcome in 17. Contributory factors (severe inflammation, aberrant anatomy and poor visualization) were present in only 13 of 37 cases. This audit suggests that, at least in the introductory period, laparoscopic cholecystectomy is associated with an overall bile duct injury rate higher than that reported previously after open cholecystectomy, although the incidence of major ductal injury is similar. The late downward trend in bile duct injury, however, suggests there may be a prolonged learning curve for this procedure. Improved understanding of the mechanism of injury may lead to yet further reductions in this complication.

Granulosa cells were prepared from follicular aspirates obtained at oocyte collection for in-vitro fertilization (IVF) and maintained in culture. Substantial loss of cells from the culture surface occurred in the absence of gonadotrophin when cells were maintained on a thin layer of extracellular matrix (ECM) using a defined, serum-free medium. This cell loss was clearly and significantly reduced in the presence of human chorionic gonadotrophin (HCG) by days 4-6 of culture, and occurred in conjunction with loss of ECM. Analysis of culture medium by zymography using gelatin as substrate demonstrated the presence of metalloproteinases (MMP), MMP-9 (gelatinase B) appearing as the predominant band. Measurement of overall gelatinase activity in culture media revealed a progressive fall in gelatinase expressed on a per cell basis in media from HCG-treated cultures and this was less marked in controls. This suppression of gelatinase activity was consistent with an observed increase in production of tissue inhibitor of metalloproteinase-1 (TIMP-1) by HCG-treated cells, which was significant by days 6-8 of culture. We speculate that stabilization of the ECM may be an important aspect of HCG action in the corpus luteum.

Ultrashort 1.25 TW Fourier-transform limited puises are generated using chirped pulse amplification in a Cr:LiSAF laser system incorporating a regenerative amplifier and two additional double-pass amplifiers. The aperture of the final amplifier (25 mm) can support pulse energies up to the Joule level. Incorporating an additional preamplifier into the system will lead to the generation of 10 TW pulses in the near future.

We report the first operation of a flashlamp-pumped LiSrAlF6:Cr3+ regenerative amplifier for chirped 100fs pulses leading to a single chirped pulse energy of 2.5mJ.

The biological activity of deglycosylated human chorionic gonadotrophin (hCG) prepared by treatment of the native hormone with anhydrous hydrogen fluoride was evaluated using suspensions of dispersed cells from biopsies of human corpus luteum obtained during the luteal phase of normal menstrual cycles. A reproducible pattern of response to hCG in terms of progesterone production by luteal cells was established for a range of luteal ages. Deglycosylation of hCG led to a diminished level of maximum response to the hormone. Co-incubation of luteal cells with a level of hCG just sufficient to elicit a maximum response and increasing concentrations of deglycosylated hCG led to a progressive inhibition of the hormonal response; at a concentration of 103 ng deglycosylated hCG/ml (a tenfold excess of deglycosylated hCG over the native hormone), hCG-induced progesterone production was reduced by about 50%. Deglycosylated hCG therefore acts as a partial antagonist for the action of hCG on human luteal cells. J. Endocr. (1984) 101, 327–332

In this talk, measurements of thermal transport in spherical geometry using time-resolved x-ray spectroscopy are presented. The time dependence of the mass ablation rate (m) is determined by following the progress of the ablation surface through thin layers of material embedded at various depths below the surface of the target. These measurements made with 6, 12 and 24 uv (351 nm) beams from OMEGA are compared to previous thermal transport data and are in qualitative agreement with detailed LILAC hydrodynamic code simulations which predict a sharp decrease in m after the peak of the laser pulse. Viewgraphs of the talk comprise the report.

Results are presented of measurements made of the fractional absorption of short (∼1 ns) intense (2×1014 W cm−2) 10.6‐μm laser radiation by plasmas produced from massive plane targets. Two complementary techniques were used: infrared light accounting and plasma calorimetry. The methods are shown to provide consistent results, demonstrating peak absorptions of up to 50%. The beam‐polarization and irradiance‐angle dependence of the absorption is consistent with a resonance absorption model. Plasma calorimetry also yields a detailed spatial distribution of the particle emission, while infrared calorimetry shows that the scattered laser light distribution is strongly dependent on the polarization of the incident laser beam.

Suspensions of luteal cells were prepared from samples of human corpora lutea obtained during the luteal phase of menstrual cycles. Addition of oxytocin (1 μmol/l) to the various cell preparations had no effect on either basal production of progesterone or on steroidogenic responses to a range of concentrations of gonadotrophin. J. Endocr. (1985) 104, 149–151

An investigation of sparks produced in a number of gases by means of a transversely excited atmospheric‐pressure (TEA) CO2 laser has revealed the existence of a clearly visible plasma filament which propagates at an almost constant velocity in the same direction as the laser beam.

Sequential two‐dimensional photographs of a laser‐produced plasma in air with effective individual exposure times of [inverted lazy s] 5 psec have been obtained by utilizing a focal‐plane optical Kerr effect shutter.

Progesterone production was assessed following short-term incubations of luteal cell suspensions prepared from tissue samples of human corpora lutea obtained at specific times throughout the luteal phase of the menstrual cycle. Luteal cells responded rapidly and sensitively to human chorionic gonadotrophin (HCG; concentration required for 50% maximum response, 0·1–1·0 i.u./ml) with a maximum level of response (five- to tenfold higher than basal production) similar to that elicited by human LH or N6,02-dibutyryl cyclic AMP. In the absence of gonadotrophin or in the presence of sub-maximal (but not maximal) concentrations of HCG, progesterone production by mid-luteal phase cells was stimulated by prostaglandin F2α (1 μmol/l), an effect not observed during the late-luteal phase. l-Adrenaline and l-isoprenaline failed to elicit significant increases in the level of progesterone production.

Cell suspensions were prepared from tissue samples of human corpora lutea obtained during the mid- and late-luteal phase of the menstrual cycle. Both oestradiol and progesterone production by dispersed cells were stimulated by similar concentrations of human chorionic gonadotrophin (hCG). As the degree of stimulation of production by hCG was greater for progesterone than for oestradiol (five- to tenfold compared with two- to threefold higher than basal production), the ratio of progesterone to oestradiol produced varied according to the level of trophic stimulation. A comparison of cell suspensions prepared from mid- and late-luteal phase corpora lutea, exposed to the same concentration of hCG (10 i.u./ml) in vitro, did not reveal a shift to oestradiol production in the late-luteal phase. Provision of additional testosterone during incubation raised the level of oestradiol production by dispersed luteal cells. At an optimum concentration of testosterone (1 μmol/l), oestradiol synthesis was not raised further in the presence of hCG or N6, O2-dibutyryl cyclic AMP, suggesting a lack of induction or activation of the aromatase system by gonadotrophin in short-term cultures. Basal and stimulated levels of progesterone production were not significantly impaired in the presence of testosterone.

Dispersed marmoset luteal cells were incubated for 2 h and progesterone production measured after exposure to hCG, cloprostenol, dibutyryl cAMP, PGF-2 alpha, PGF-2, adrenaline or melatonin. The cells were studied on Days 6, 14 and 20 after ovulation in conception and non-conception cycles. Luteal cells from Day 14 non-pregnant marmosets were compared with human luteal cells taken in the mid-luteal phase. All the treatments stimulated progesterone production including cloprostenol, which is luteolytic when administered to the marmoset in vivo, but the degree of response varied with the stage of the cycle or pregnancy and between marmoset and human luteal cells. In the marmoset, overall analysis of the effect of the treatments showed that, on Day 6 after ovulation, there was no significant effect of any of the treatments in cells from pregnant or non-pregnant animals. In contrast, luteal cells from non-pregnant animals on Day 14 showed a significant response to the treatments (F (8,41) = 2.79, P less than 0.0145) whereas cells from pregnant Day-14 animals were responsive; in cells from pregnant animals, the control production of progesterone was high and already equivalent to the levels stimulated by the treatments. By Day 20, cells from pregnant animals produced lower control concentrations of progesterone than did those on Day 14 and there was a significant overall effect of the treatments (F (8,33) = 3.78, P less than 0.003). These results show that the marmoset CL gains responsiveness to treatment between Days 6 and 14 after ovulation in the non-pregnant cycle. In pregnancy, on Day 14, 2 days after attachment of the embryo, the high control concentrations of progesterone and absence of response to treatment suggest that an embryo message may have affected the CL, providing an endogenous stimulus.

The energetics of this pumping scheme are shown. Short-pulse (50 to 100 ps) laser irradiation of an appropriate x-ray flashlamp medium generates broad-band emission in the range of 300 to 800 eV which preferentially photoionizes Ne to the /sup 2/S state of Ne II creating an inversion at approximately 27 eV. Although this approach does not depend on precise spectral overlap between the x-ray pump radiation and the medium to be pumped, it does require that the x-ray medium remain un-ionized prior to photoionization by the soft x-ray emission. Well-controlled focus conditions are required to ensure that the x-ray medium is not subjected to electron or x-ray preheat prior to irradiation by the soft x-ray source. The magnitude of the population inversion is predicted to be critically dependent upon rapid photoionization of the two states; therefore, ultra-short pulse irradiation of the laser flashlamps is required.

Summary. Suspensions of luteal cells were prepared by collagenase dispersion of guinea-pig corpora lutea obtained at specific times during the oestrous cycle. Luteal cells incubated with hCG produced increased amounts of progesterone. For Days 3\p=n-\13of the oestrous cycle, the concentrations of hCG required for 50% of the maximum response were within the range, 1 \m=x\10\m=-\3 to 7 \m=x\10\m=-\3 i.u./ml, showing no marked loss of sensitivity to hCG with increasing luteal age. PGF-2\g=a\(1 \g=m\mol/1),had no effect on basal production of progesterone but significantly inhibited hCG\x=req-\ stimulated progesterone production by luteal cells isolated on Days 7, 9, 10, 12 and 13 of the cycle. This concentration of PGF-2\g=a\had no significant effect on progesterone production by luteal cells prepared earlier in the cycle (Days 3 and 5). It is concluded that (a) the luteolytic action of PGF-2\g=a\in the guinea-pig is mediated, at least in part, by direct action on luteal cells, and (b) the cells from newly formed corpora luteal are resistant to the direct inhibitory action of PGF-2\g=a\

Human granulosa cells were prepared from follicular aspirates obtained during oocyte collection for in-vitro fertilization. Following several days in culture, cells were washed and then progesterone output was measured in 2-h incubations. After culture for 3 days, incubated cells responded well to human chorionic gonadotrophin (hCG) and prostaglandin (PG) E2 with similar levels of maximum response. Exposure of cultured cells to epidermal growth factor (EGF) for 2 days (days 3–5) led to substantial increases both in basal production and in responses to hCG and PGE2 during subsequent incubations. These effects of EGF were not accompanied by measurable increases in DNA levels in cultures over this time. Results may point to a possible paracrine role for EGF-like factors modulating the activity of cells forming the early corpus luteum. Journal of Endocrinology (1989) 121, 397–402

SUMMARY A cell suspension was prepared from normal rat adrenal tissue by collagenase digestion of decapsulated adrenal glands. Corticosterone produced by these cells was assayed both by competitive protein binding and by radioimmunoassay (range: 0–1 ng; sensitivity: 30 pg). Before incubation, the medium and cells equivalent to one gland contained 27·7 ± 3·2 ng corticosterone which increased to 36·3 ± 7·0 ng corticosterone after 1 h of incubation. Although a normal sigmoid log dose—response curve was obtained using adrenocorticotrophin (ACTH) (ED50:1·2 × 10−4 i.u./ml), it was observed that log (ACTH concentration) was directly proportional to log (corticosterone produced) between 1 × 10−7 and 1 × 10−3 i.u. ACTH/ml. A similar linear relationship was demonstrated for the effect of adenosine 3′,5′-monophosphate on steroidogenesis. Time-course studies revealed that after addition of ACTH there was a definite lag (3 min) before onset of increased steroidogenesis; within 5 min a constant corticosterone production rate was achieved. β1–24 Adrenocorticotrophin was shown to retain its ability to stimulate steroidogenesis when diazotized to beads of polyacrylamide too large to enter cells. Evidence is provided that the activity of the β1–24 ACTH—polyacrylamide beads is not due to biologically active peptides cleaved from the complex. It is concluded that ACTH can stimulate steroidogenesis without entering its target cells.

Much of the interest currently expressed in CO2 lasers of extremely high peak power arises because of the possibility of their incorporation into various schemes aimed at producing thermonuclear fusion. Proposals in which uniform irradiation by a specially shaped laser pulse of a solid or hollow spherical target induces a high compression of the thermonuclear material have lead to predicted required laser energies which should be realizable in the near future (l,2). In addition the possibility of using high power CO2 lasers to heat lower density, magnetically confined plasmas to thermonuclear conditions has also been considered (3). In both these approaches there is implicit the development of efficient, large aperture uniform CO2 laser beams capable of sustaining high power pulses of controllable shape and duration. Since the development of pulsed, transversely excited high pressure gas lasers(4,5), several different schemes have been devised for the excitation of large volumes of laser gas mixtures with high optical gain and energy extraction. At the present time the most promising approaches to this problem appear to be those of the so-called “double discharge” technique (6,7), the electron beam controlled discharge(8), and discharges incorporating U.V. photo-preionization(9). The present paper will discuss the principal features of the latter technique, and, in addition, various methods of producing ultrashort pulses with U.V. photo-preionized CO2 lasers will be described. Some initial results on the amplification of ultrashort pulses, obtained with an oscillator-plifier laser system, will also be described.

1. The powerful anti-inflammatory glucocorticoid triamcinolone acetonide, administered to rats at 20 and 2.5mg/kg, leads to a decrease in the incorporation in vivo of [3H]uridine and [32P]orthophosphate into hind-limb skeletal muscle. 2. At the higher dose, this decrease in the rate of incorporation of precursors into RNA precedes a decrease in the incorporating ability of muscle ribosomes, which commences about 4–5h after drug administration, but is unaccompanied by any changes in the concentration of tissue ATP or free amino acids. 3. The ribosomal dysfunction extends to polyribosomes, which can only be successfully isolated from the muscle of triamcinolone-treated animals after the addition of α-amylase to the tissue homogenate to remove glycogen. 4. The specific radioactivity of muscle protein labelled in vivo with 14C-labelled amino acids does not decrease progressively after triamcinolone administration. After 2h there is an apparent stimulation of incorporation which leads to an overall discrepancy between measurements of protein-synthetic activity made in vivo and in vitro. 5. There is a significant increase in muscle-glycogen concentration between 8 and 12h after the administration of triamcinolone acetonide (20mg/kg), although a significant decrease occurs after 4h. The fall in glycogen concentration may be due to a decrease in the rate of synthesis of protein essential for glucose uptake into the tissues. 6. As judged by (a) incorporation of 14C-labelled amino acids into protein, (b) [3H]uridine and [32P]-orthophosphate incorporation into RNA, (c) the rate of induction of tryptophan pyrrolase and (d) changes in the pool sizes of taurine and tryptophan, the responses in liver followed the same time-course as those in muscle after administration of the drug.

Decisions concerning the modality of choice for children with ESRD are not made in a random fashion. Intuition, technical expertise, past experiences and enthusiasm for new modalities color one's perception of the optimal treatment choice. We propose a framework where treatment alternatives are examined systematically. A feasibility equation for the treatment of choice has been developed which scores 4 variables: quality of life, medical factors, transplantability and management issues. Scores are then tabulated for treatment choice. The model was used to examine 19 children ages 6 wks.-16 yrs. over 10 yrs. Five pts. whom the authors had no input in decision-making were examined retrospectively, 11 were scored retrospectively where the authors decided the modality and 3 pts. were studied prospectively. Twenty four decisions were audited. Early pts. had only 3 possible dialysis alternatives, centre peritoneal, centre hemo. and home hemo., whereas the more recent pts. had the additional choices of home peritoneal cycler, continuous ambulatory peritoneal and self-care hemo. We believe that this model can be helpful at different points in time. Initially it encourages a review of all possible treatment choice. Later it may be used as an ongoing assessment tool pointing to other alternatives as more Information is gathered and thus minimize the need for decision-making at the time of a crisis. Finally, it serves as a tool to choose other modes of dialysis after the treatment of choice fails.

A passive aperture‐scalable device is described which provides high contrast enhancement of a laser pulse over baseline optical noise while maintaining efficient net transmission of the principal laser pulse. The technique, which is based upon the nonresonant phenomenon of self‐induced ellipse rotation, is capable of providing effective gain isolation for virtually any high‐power laser system. The constrast enhancement capabilities of the device are demonstrated experimentally using a single ultrashort laser pulse obtained from a cavity‐dumped passively mode‐locked Nd : glass laser. In addition, a discussion is given concerning the ability of the device to temporally compress or stretch the optical pulse.