bluuebunny/crossref_metadata_2025 huggingface dataset:
235.PAF induced changes in intracellular Ca2+ and membrane potential in the 2-cell mouse embryo
Authors: Li Y. , Day M. L. , O'Neill C.
Published: October 2010

Platelet-activating factor (PAF) is an autocrine survival factor for the preimplantation embryo. PAF induces a transient increase in intracellular Ca2+ ([Ca2+]i) in 2-cell embryos that is caused by the interdependent influx of external calcium and release of calcium from internal stores. A membrane current with L-type calcium channel properties is activated during PAF-induced calcium signalling. Since the L-type channel in many cell types is primarily voltage-gated we were interested to learn whether this was also the case in the 2-cell embryo. The present study investigated the relationship between the PAF-induced Ca2+ transient and changes in membrane potential (Em) in the 2-cell embryo.
The perforated whole-cell patch-clamp technique was used to detect changes in Em and standard calcium imaging techniques were used to measure changes in [Ca2+]i in 2-cell embryos from QS mice. Embryos were first loaded with Fluo-3 and then pretreated with PAF:acetylhydrolase to degrade the embryo derived PAF before patch clamping. Whole-cell perforated patch-clamping was performed by inclusion of 240mg/ml Nystatin in the pipette solution. Changes in Em and [Ca2+]i were recorded simultaneously after treatment of the embryo with PAF.
In 2-cell embryos PAF induced a change in Em, consisting of an initial small depolarisation of 2.4 � 0.2 mV (42 � 4 sec after addition of PAF) followed by one or more transient hyperpolarisations of -8 � 1 mV (100 � 9 sec after addition of PAF). Transient increases in [Ca2+]i paralleled the membrane hyperpolaristions and were initiated at 84 � 8 sec after addition of PAF. These responses to PAF were seen in 58% of 2-cell embryos (n = 52). It is not yet clear whether these changes in Em account for the activation of calcium influx through the L-type channel. The results show for the first time that the 2-cell embryo is an electrically active organism.
236.Differential expression of monocarboxylate cotransporter proteins in preimplantation embryos
Authors: Jansen S. , Pantaleon M. , Kaye P.
Published: October 2010

During preimplantation development mouse embryos demonstrate a switch in substrate preference. Pyruvate consumption, high during the first few cleavage stages, declines as the morula develops to a blastocyst, when glucose becomes the preferred substrate. Whilst pyruvate utilisation has been well characterised, changes in the function and expression of pyruvate transporters during this crucial period remain unclear. Pyruvate, lactate and other monocarboxylates are transported across mammalian cell membranes via a specific H+-monocarboxylate cotransporter (MCT). Fourteen members of this family have been identified of which MCT1, MCT2 and MCT4 are well characterised. Although mRNA expression profiles are known during early mouse development (1,2), the specific roles of each protein isoform are unknown. In order to understand these, the expression pattern for each isoform and their cellular localisation during preimplantation development have been determined. Mouse embryos were freshly collected from superovulated Quackenbush mice at 24, 48, 72 and 96 h post-hCG and expression of MCT1, MCT2 and MCT4 analysed by confocal laser scanning immunohistochemistry. Our results confirm that all three MCT proteins are expressed in preimplantation embryos. Immunoreactivity for MCT1 and MCT2 appears diffuse throughout the cytoplasm of cleavage stage embryos. As development proceeds, MCT1 localised to the basolateral membranes of morulae and blastocysts, whilst stronger MCT2 expression was found on the apical trophectoderm as well as the inner cell mass. MCT4 immunoreactivity on the other hand is apparent at cell-cell contact sites in cleavage stage embryos and morulae, but it is not apparent in the blastocyst. The demonstration of different expression patterns for MCT1, MCT2 and MCT4 in mouse embryos implies specific functional roles for each in the critical regulation of H+, pyruvate and lactate transport during preimplantation development.
(1) Harding EA, Day ML, Gibb CA, Johnson MH, Cook DI (1999) The activity of the H+-monocarboxylate cotransporter during pre-implantation development in the mouse. Eur. J. Physiol. 438, 397–404. (2) H�rubel F, El Mouatassim S, Gu�rin P, Frydman R, M�n�zo Y (2002) Genetic expression of monocarboxylate transporters during human and murine oocyte maturation and early embryonic development. Zygote 10, 175–181.
238.Determination of differentially displayed oxygen-sensitive genes in bovine blastocysts
Authors: Harvey A. J. , Kirstein M. , Navarrete-Santos A. , Kind K. L. , Fischer B. , Thompson J. G.
Published: October 2010

Oxygen-regulated gene expression in the bovine embryo contrasts markedly with that observed in the mouse. Under low (2%) oxygen moderate changes in gene expression are observed in the bovine blastocyst, compared with 3- to 4-fold increases in the mouse. We have determined that these moderate gene expression changes are most likely regulated by Hypoxia-Inducible Factor (HIF)-2 transcription factor activity in the bovine, in the absence of HIF1, although HIF2 target genes are largely unknown. The aim of this study was to screen, by differential display RT-PCR, for putative oxygen-regulated transcripts that might confer developmental competence in blastocysts cultured under varying oxygen atmospheres post compaction.
In vitro-produced bovine blastocysts were generated using standard protocols. Compact morulae were randomly allocated to treatments under either 2%, 7% or 20% oxygen for 72 h from Day 5. Blastocyst RNA was isolated using TriReagent and samples were reverse transcribed using Superscript II. cDNA was amplified using 10-mer primers in reactions containing 32Pα-labelled dCTP. Resulting bands were detected by autoradiography, excised, purified and ligated into pGEMT vectors for transformation and sequencing. Seven clones were identified as having high homology with known sequences in GenBank. Real-time PCR was undertaken to confirm oxygen-regulation using Sybr green master mix.
Myotrophin mRNA was significantly increased following 2% oxygen culture, compared with 20% cultured blastocysts (P�<�0.01), as was GLUT1 (P�<�0.01). The expression of anaphase-promoting complex showed a significant association with oxygen, being higher in 2% cultured blastocysts (P�<�0.05). Acetyl-coA-acetyltransferase I, chronic myelogenous leukemia tumor antigen (CML66), cyclin I, NADH dehydrogenase subunit 2 and ribonucleotide reductase M1, genes identified using differential display, were not altered by post compaction oxygen concentration.
This study has identified potentially HIF2-specific regulated genes, and supports the hypothesis that reduced oxygen concentrations post-compaction may influence bovine embryo development through oxygen-regulated changes in gene expression.
239.Sensitivity of bovine morulae and blastocysts to heat shock in vitro
Authors: Naik V. , Jonsson N. N. , Phillips N. J. , Kaye P.
Published: October 2010

Thermotolerance of blastocysts and morulae is greater than that of zygotes. However, lower rates of pregnancy have been recorded when morulae rather than blastocysts were transferred. The aim of this study was to determine if morulae displayed lower thermotolerance than blastocysts in vitro. Embryos were produced from oocytes collected from abattoir-sourced ovaries. On Day 7 post fertilisation, embryos were classified as morulae or blasotcysts and subjected to either an increase in temperature from 39�C to 41.5�C over 1 h followed by a decrease to 39�C over 2 h (HS) or maintained at 39�C (NHS), using a water jacketed CO2 incubator. The number of embryos progressing to expanded and hatched blastocysts was recorded after 48 h further culture at 39�C. After arcsine transformation, the proportions progressing were submitted to a general linear model using adjusted sum of squares for tests of difference. Factors were embryonic stage, treatment and sire and the interaction terms of stage and sire with treatment. The Kruskall-Wallis test was also applied to the untransformed, non-parameteric data set. Non-parameteric , univariant analysis indicated non-significant effects of treatment (NHS median proportion progressing =72%, HS = 64%, P = 0.12) and of sire (NHS = 78%, HS = 64%, P = 0.168), while the effect of stage was highly significant (morulae = 35%, blastocysts = 79%, P�=�0.0000). However, according to the general linear model, treatment and stage were significant factors (F�=�5.39 and 38.3, respectively, and P�=�0.032 and 0.000, respectively) and sire approached significance (F�=�4.09, P�=�0.058). Neither of the interaction terms was significant. It was concluded that embryos, which were morulae on Day 7, were less likely to progress to expanded or hatched blastocysts and that heat shock reduced developmental progression.
247.Molecular basis of oocyte - paracrine signalling that promotes mouse granulosa cell proliferation
Authors: Gilchrist R. B. , Ritter L. J. , Myllymaa S. , Kaivo-Oja N. , Amato F. , Ritvos O. , Mottershead D. G.
Published: October 2010

Oocytes regulate follicle growth and development by secreting paracrine growth factors that act on granulosa cells (GC). We have recently determined that growth differentiation factor-9 (GDF-9) accounts for ~50% of the total mitogenic activity of oocytes, the remaining portion is as yet uncharacterised. This study was conducted to identify the receptor/signalling system utilised by oocytes to promote GC proliferation. We used an established oocyte-secreted mitogen bioassay, where denuded oocytes are co-cultured with primed-mouse mural GC. In this system, oocytes, GDF-9, TGF-b1 and activin-A all promoted GC DNA synthesis in a dose-dependent manner, but bone-morphogenetic protein-6 (BMP-6) and BMP-7 did not. The type-II receptor for GDF-9 is BMPRII and using real-time RT-PCR, cumulus cells (CC) and mural GC were found to express equivalent levels of BMPRII mRNA. We tested the capacity of the receptor ectodomain (ECD) to neutralise oocyte-stimulated mural GC proliferation. The BMPRII ECD antagonised both oocyte and GDF-9 bioactivity in a dose-dependent manner, completely abolishing activity of both mitogens at 1 ug/mL. The BMPRII ECD did not antagonise TGF-β and partially antagonised activin-A bioactivity, demonstrating its specificity. The TGFβR-II ECD, activin R-II ECD and activin R-IIB ECD all failed to neutralise oocyte- or GDF-9-stimulated GC DNA synthesis, whereas they did antagonise the activity of their respective ligands. The BMPRII ECD also completely antagonised oocyte-stimulated CC DNA synthesis. Using this oocyte-factor bioassay with mural GC transfected with Smad luciferase reporter constructs, we found that oocytes, GDF-9 and TGF-β (but not BMP-6) activated the Smad2/3 pathway. Consistent with this, oocytes and GDF-9 led to phosphorylation of GC Smad2 molecules as detected by Western blot. Conversely the Smad1/5/8 pathway was activated by BMP-6, but not by GDF-9, TGF-β nor surprisingly by oocytes. This study provides evidence that BMPRII is a key receptor for transmitting the paracrine actions of oocytes in GC. However, oocyte-secreted factors do not activate the BMP intracellular signalling pathway but rather the TGF-β/activin intracellular pathway.
262.How the elephant got its epididymis
Authors: Jones R. C.
Published: October 2010

An idea has been promoted for more than 30 years, that the elephant does not have an epididymis (1,2), that each derivative of the mesonephric (Wolffian) ducts only develops into a ductus deferens. Whilst the location of some of the duct provides some support for this idea, it is considered that the interpretation is misleading in implying that the testes only have ejaculatory ducts that don't function like the epididymides of other mammals. Modern interpretation indicates that all mammals have epididymides, but some species exhibit adaptations of the epididymides that are determined by the structure and general physiology of the species, and the reproductive strategy of the males to achieve paternity. The modern interpretation of epididymal evolution is based on comparative cytological studies of duct differentiation, the functions of the ducts in regulating their milieu and effecting post-testicular sperm development and storage, and the biological significance of the epididymis in natural selection. It has been shown that most of the more than 50 m length of the elephant's ductuli epididymides do lie on or close to the testis. However, more importantly, it has been shown that mature elephants have epididymides that are structurally differentiated into an initial segment (a feature that is unique to the mammalian epididymis) and 'middle' segments where sperm mature, and a terminal segment that is adapted for sperm storage both in structure and in the regulation of the luminal milieu. Each ductus epididymidis is connected to the urethra by a short ductus deferens.
(1) Short, R.V., Mann, T, and Hay, M. (1967) J. Reprod. Fertil. 13, 517–535. (2) Short, R.V. (2003) Reprod. Fertil. Devel. 15 (Supplement), 76.
265.Matrix metalloproteinases in the mouse model of menstruation: effect of doxycycline inhibition
Authors: Shen J. , Zhang J. , Kaitu'u T. J. , Brasted M. , Salamonsen L. A.
Published: October 2010

Strong correlative evidence supports a role for matrix metalloproteinases (MMP) in the tissue breakdown at menstruation. Because menstruation occurs only in women and a few old-world primates, it has not been possible to examine the functional significance of potentially key mediators of this process. To this end, we developed a mouse model for menstruation (1), in which ovariectomised mice are subjected to a decidualising stimulus: injection of oil into the uterine lumen following appropriate hormone-priming. During the 24 h following withdrawal of progesterone (P), the decidualised tissue progressively breaks down, in a manner that morphologically resembles that of human endometrium at menstruation. The aims of the present study were to examine the pattern of MMP expression during the time from progesterone withdrawal until complete tissue breakdown, and to determine whether administration of doxycycline, (a known MMP modulator), 3 h prior to P withdrawal, affected the expression or activity of the MMPs or restrained the tissue destruction. MMP-3 was present at foci in the decidual zone: these were initially associated with the restructuring at decidualisation and subsequently with the tissue destruction. MMP-7 was detected both in epithelium and in leukocytes, predominantly neutrophils. These were first apparent in the basal zone during the earliest stages of tissue instability, and dramatically increased in numbers as breakdown progressed. MMP-9 was found only in leukocytes, predominantly neutrophils and some macrophages, with greatly increased numbers with time. Zymography revealed a dramatic increase in both latent and active MMP-9 as tissue breakdown proceeded. Doxycycline reduced immunoreactive MMP-3 but not MMP-7 or MMP-9 in the tissue, and also decreased gelatinase activity. However, no apparent effect on tissue breakdown was observed. Further studies with a more potent MMP inhibitor are required to fully establish the importance of MMPs in these processes.
(1) Brasted et al. (2003) Biol. Reprod. 69, 1273.
267.Leukocyte matrix metalloproteinase expression is regulated in response to selected chemokines: implications for break-through bleeding
Authors: Morison N. B. , Jones R. L. , Hannan N. J. , Salamonsen L. A.
Published: October 2010

Break-through bleeding is a problem common to many women using long-acting progestin-only contraceptives such as Implanon, and is the main reason many women discontinue using them. Our previous studies (1) identified a number of chemokines (chemoattractive cytokines) that are highly expressed during the menstrual cycle with the capacity to selectively attract the leukocyte subsets present in each phase. Leukocytes produce factors thought to be critical for endometrial breakdown and remodelling, including matrix metalloproteinases (MMPs). We hypothesised that progestins act on endometrial epithelial cells to activate chemokine expression. The chemokines attract leukocytes into the tissue and stimulate their production of MMPs, contributing to tissue breakdown and break-through bleeding. The present study investigated the role of appropriately selected chemokines on leukocyte MMP production. The eosinophilic cell line EOL-1 and the mast cell line HMC1 were treated with four chemokines (fractalkine, HCC-1, MCP-3 and IL-8) at a range of concentrations. MMP production and activation were analysed by gelatin and casein zymography of culture medium. No response was observed from the mast cells under any of the treatments. However, there was a significant increase in latent MMP-9 production by eosinophils in response to increasing concentrations of IL-8 and MCP-3. Results show a greater than 2-fold increase in the amount of latent MMP-9 in response to IL-8 or MCP-3 (300�ng/mL and 80�ng/mL respectively) compared to non-treated controls. These chemokines had no significant effect on levels of active MMP-9 or latent and active forms of MMP-2. HCC-1 and fractalkine had little effect on either MMP-9 or MMP-2 production by these cells. These data support the hypothesis that MMP production by leukocytes is regulated, at least in part, by selective chemokines expressed during menstruation. Future studies will expand the leukocyte subtypes tested and investigate the role of synthetic progestins in chemokine expression from endometrial epithelial cells.
(1) Jones R. L. et al. (2003) Proceedings 34th Annual Conference of Society for Reproductive Biology. Reprod. Fertil. Dev. 15 (suppl.), A85.
268.Identifying markers for stromal stem/progenitor cells in human endometrium
Authors: Schwab K. E. , Gargett C. E.
Published: October 2010

The endometrium is divided into upper functionalis, which rapidly grows then differentiates before being shed, and lower basalis, from which cyclical regeneration begins. A small proportion of endometrial stromal cells have been identified with clonogenic activity, a functional property of stem cells (1). We hypothesised that stromal stem/progenitor cells expressing known stem cell markers reside in the basalis. The aims of this study were to: (1) investigate the clonogenic activity of human endometrial stromal cell populations enriched and depleted for known stem cell markers, and (2) identify a marker that will differentiate basalis from functionalis stroma. Endometrial tissue acquired from 23 ovulating women undergoing hysterectomy was digested with collagenase to produce single cell suspensions. Leukocytes and epithelial cells were removed, and stromal cells analysed by flow cytometry, FACS sorted into enriched and depleted populations, and cultured for clonal analysis as described (1). Markers analysed included stem cell markers, STRO-1, CD133, CD45 and CD34, and an endometrial stromal cell marker, CD90 (2).
Immunohistochemical analysis of CD90 was performed on full thickness human endometrial tissue. CD45– endometrial stromal cell populations contained 2.13 � 0.65% (n = 13) STRO-1+, and 5.43 � 1.42% (n = 16) CD133+ cells. Stromal cell populations enriched (0.65 � 0.42%) and depleted (0.95 � 0.58%) for STRO-1 showed no significant difference (P = 0.19, n = 5) for clonogenic activity. Surprisingly, clonogenicity of CD133+ stromal cells (0.74 � 0.56%) was lower than CD133– (3.89 � 1.35%) cells (P = 0.03, n = 6). Immunohistochemical staining showed strong CD90 staining in the functionalis, with lighter staining in the basalis. These observations were confirmed by flow cytometric analysis which identified two distinct populations (n = 9), CD90low (19.55 � 4.35%) and CD90hi (74.71 � 5.20%). Clonogenic analysis of these two populations is underway. Interestingly, dual-colour flow cytometry showed the CD133+ cells to be CD90low (n = 7). Further analysis suggests that the CD90lowCD133+ population are CD45–CD34+, suggesting endothelial progenitor cells. This study identified CD90 as a marker that distinguishes basalis and functionalis stroma, and demonstrated that STRO-1 and CD133 are not functional markers for clonogenic endometrial stromal stem/progenitor cells.
(1) Chan RW, Schwab KE, Gargett CE (2004) Biol. Reprod. 70, in press. (2) Fernandez-Shaw S, Shorter SC, Naish CE, Barlow DH, Starkey PM (1992) Hum. Reprod. 7,156–161.
273.Insulin-like growth factor treatment of pregnant guinea pigs during early pregnancy promotes fetal growth
Authors: Sferruzzi-Perri A. N. , Owens J. A. , Robinson J. S. , Roberts C. T.
Published: October 2010

Insulin-like growth factor (IGF)-II is an important regulator of growth in many tissues and is abundantly expressed in the placenta during pregnancy. Gene ablation studies performed in mice have shown that IGF-II deficiency results in both impaired fetal and placental growth, whereas deficiency in IGF-I reduces fetal growth only. Conversely, maternal IGF supplementation in early pregnancy in the guinea pig increases placental and fetal size by mid pregnancy. This study aimed to determine whether these anabolic effects persist into late pregnancy after cessation of treatment. On Day 20 of pregnancy, mothers were anaesthetised and a mini osmotic pump was implanted subcutaneously, to deliver 1mg/kg/day IGF-I (n = 7), IGF-II (n = 9) or vehicle (n = 7) for 17 days. Guinea pigs were killed on Day 62 of pregnancy (term ~67 days). Fetal and placental weights, and maternal and fetal body composition, were measured. Total litter size was unaffected by IGF treatment; however, IGF-II increased the number of viable fetuses by 26% (P = 0.01). After adjusting for the number of viable pups per litter, maternal IGF treatment increased fetal growth by increasing abdominal circumference, crown-rump length and fetal weight (fetal weight: IGF-I 79+/–2.5 g; IGF-II 78+/–2.6 g; vs vehicle 68+/–2.5 g, P = 0.02). IGF treatment did not alter absolute or relative fetal organ weights. IGF-I reduced placental weight by 9% and IGF-II increased it by 9%, but not significantly. IGF-I increased the fetal weight�:�placental weight ratio (19+/–0.9 vs 15+/–0.9, respectively P = 0.043). IGF treatment did not affect maternal weight gain during pregnancy nor net carcass weight; however, IGF-I reduced maternal lung and adipose tissue weights. In conclusion, maternal IGF-II treatment during early pregnancy improved fetal growth into late gestation, possibly by modulating placental efficiency. As poor placental development is implicated in fetal growth restriction, increasing maternal IGF abundance in early to mid pregnancy may be a potential therapeutic approach to placental insufficiency.
276.Targets of the action of nuclear transport factors in spermatogenesis
Authors: Efthymiadis A. , Hogarth C. A. , Szczepny A. , Loveland K. L. , Jans D. A.
Published: October 2010

During spermatogenesis, precise and orderly switches in gene expression are required. The movement of transcription factors (TFs) and nuclear proteins into and out of the nucleus is highly regulated, thus determining the extent and timing of gene expression. Most nuclear transport events are mediated by members of the importin superfamily that specifically recognise their cargoes and facilitate the passage of receptor-substrate complexes through the nuclear pore complex (NPC) which is made up of nucleoporin proteins. Eight human importin α isoforms are known that function in heterodimeric form with importin β whilst there are 20 members of the importin β family, which mediate the nuclear import or export of a very diverse set of protein or RNA cargoes. Understanding of importin and TF/chromatin component interaction during spermatogenesis should identify potential developmental switches, critical steps in the spermatogenic process. We are interested in the expression of different importins during spermatogenesis, and their specific nuclear import/export substrates as candidates in developmental switches. Preliminary analysis has shown that the nuclear import factors importin β1 and 3 are not only expressed in germ cells, but also alter their cellular distribution during maturation. In a yeast two-hybrid screen, using truncated importin β3 as bait and a library made from adult mouse testis, we identified a transcriptional repressor gene involved in cell cycle regulation, and an enzyme of the purine nucleotide biosynthesis pathway as candidate binding partners. Further studies will focus on elucidating the biological significance of these interactions in spermatogenesis.
282.Differential expression of the relaxin receptor (LGR7) in the mammary apparatus of the lactating tammar wallaby (Macropus eugenii)
Authors: Gwyther J. , Gehring H. M. , Parry L. J.
Published: October 2010

Growth and development of the mammary apparatus (nipple and mammary gland) are important aspects of lactation. Macropodid marsupials can suckle young of two different ages simultaneously, a phenomenon known as asynchronous lactation. As a result, the type of milk produced and the structure of the two mammary glands supporting young of different ages vary considerably. A role for the peptide hormone relaxin in lactation has been demonstrated in relaxin receptor (LGR7)-deficient mice, which fail to deliver milk to their offspring due to impaired nipple development (1). This study investigated the distribution of LGR7 in the different mammary glands and nipples during asynchronous lactation in the tammar wallaby. The specific aim was to determine if the age of the pouch young influences LGR7 gene expression. Tissues were collected from the mammary apparatus sustaining the neonate and an older pouch young in the same mother, between Days 0 and 21 of lactation (n = 5/stage). A partial sequence (250-bp) of the tammar LGR7 was first obtained from a region close to the N-terminus of the soluble ectodomain, with 82% amino acid homology compared to the human LGR7 sequence. LGR7 gene expression was then measured by quantitative-PCR, using a TaqMan probe in the Opticon 2 thermal cycler (MJ Research, GeneWorks). Expression of LGR7 was upregulated in both the nipple and mammary gland supporting the neonate between 5 and 11 days after birth. There was no difference in LGR7 expression between these two tissues. However, LGR7 mRNA concentrations were significantly (P < 0.05: paired t-test) higher in the mammary apparatus supporting the neonate compared with that of the older young. These data suggest that a local stimulus, such as the continuous sucking by the neonate, may influence LGR7 expression in the mammary apparatus.
(1) Krajnc-Franken et al. (2004) Mol. Cell. Biol. 24, 687–696.
287.Differential expression of plasminogen activation cascade components in human preterm delivery with and without preterm premature rupture of the fetal membranes
Authors: Nowell C. J. , Murthi P. , Kalionis B. , Moses E. K. , Brennecke S. P.
Published: October 2010

Previous studies from our laboratory have shown that plasminogen activation cascade (PAC) components are involved in the rupture of fetal membranes at term. The aim of this study was to test the levels of 4 key components of the PAC in fetal membranes collected from preterm deliveries (PTD) with and without preterm premature rupture of the fetal membranes (PPROM). In contrast to previous studies of the PAC in fetal membranes which have focused on labour at term, this study has examined PTD with PPROM (n = 10) and PTD without PPROM (n = 10).
Using immunohistochemistry, immunoblotting and zymography, we examined the expression and activity levels of key components of the PAC; plasminogen, PAI-2, uPA and uPAR for expression and plasminogen, plasmin and uPA for activity. The data presented here show significant changes in the expression and activity of PAC components in PTD + and – PPROM samples. When compared to term labour control samples both PTD + and – PPROM show increased expression and activity of PAC components. When PTD + and – PPROM are compared to each other differences in expression and activity can be seen. uPA expression and activity increases in PTD – PPROM, relative to term delivery samples, and increases further in PTD + PPROM samples. Immunohistochemical analysis reveals the uPAR is expressed at very low levels in the amnion epithelium, basement membrane and mesenchyme/intermediate zone of PTD + PPROM but is highly expressed in PTD – PPROM.
These data demonstrate differential expression of the PAC in PTD + PPROM cases compared with PTD – PPROM cases, which suggests different aetiologies and/or mechanistic pathways for the two types of PTD. These differences may be of importance in defining optimal clinical management of the two situations.
288.Labour-associated changes in human fetal membranes and placental adrenomedullin mRNA expression
Authors: Alghafra A. K. , Gude N. M. , King R. G. , Brennecke S. P.
Published: October 2010

We have recently shown that total adrenomedullin (AdM) concentrations are elevated in choriodecidual and amniotic tissues, but not in placental, in response to human labour at term and pre-term (1). Therefore, the present study was designed to determine whether AdM mRNA expression was increased with labour in term and pre-term samples by using Northern Blot Analysis. Placentas were collected either at elective caesarean section (not-in-labour, NIL), after normal vaginal deliveries and at caesarean section during labour (in-labour, IL) from women with singleton pregnancies at term (>36 weeks gestation, 39.2 � 0.2) or pre-term (between 24 to 36 weeks gestation, 32 � 0.4), but with otherwise uneventful antenatal courses.
There were significant labour-induced increases in AdM mRNA expressions in amnion and choriodecidua in both groups, pre-term and term (see table in PDF file).
No difference was found in those who had vaginal deliveries with those who had caesarean sections after the commencement of labour at pre-term. Amongst term samples in labour all subjects were delivered vaginally uneventfully, except two cases where forceps were used in the third stage of labour.
AdM mRNA levels are increased in association with both term and pre-term labour, both in amnion and choriodecidua. We postulate that increased production of AdM by fetal membranes in association with labour may be involved in fetal and/or maternal adaptations to labour. For example, AdM may compensate for local-acting vasoconstrictor substances that are increased during labour, or it may act on the fetal lung in preparation for extra-uterine life.
(1) Al-Ghafra A, Gude NM, Brennecke SP, King RG (2003) Labour-associated changes in adrenomedullin content in human placenta and fetal membranes. Clin. Sci. 105, 419–423.
289. Comparison of lymphocyte subsets and function in the rat and mouse testis
Authors: Aridi D. , Pellicci D. , Hutchinson P. , Hedger M. P.
Published: November 2010

Testicular leukocytes are assumed to be involved in immunological surveillance against infection and tumours as well as regulation of local immune responses. They are implicated in mechanisms that make the testis a successful site for tissue transplantation in both rats and mice. Our previous studies using multi-colour fluorescence flow cytometric analysis to examine isolated testicular leukocytes in the rat testis have established the existence of a significant population of predominantly CD8+ T cells and a comparable number of lymphocytes expressing natural killer (NK) cell markers (NK and NKT cells). The functional activity of these testicular NK and NKT cells subsequently has been confirmed by a standard flow cytometric cytotoxicity assay using an NK-sensitive tumour cell line (YAC-1) and an NKT-sensitive tumour cell line (U937). Similar analyses of mouse testicular leukocytes have shown a slightly different pattern. The data indicate that mouse testicular lymphocytes comprise T cells, NK cells, and NKT cells, similar to the rat testis. However, while the apparent numerical densities of T cells in rat and mouse testes were similar, the numbers of NK and NKT cells were considerably lower in the mouse. Mouse testicular NKT cells were positive for staining with the tetramer CD1d/αGC, which is used to identify classical NKT cells, whereas rat NKT cells did not stain for this marker. Moreover, the CD8/CD4 T cell ratio in the mouse testis displayed a skewing towards the CD4+ subset. These data highlight the possibility that the immunological environment, and hence the course of immunological events, might be quite different in the testes of the two species. The reasons for these differences are not clear, however they should be taken into account when considering studies of testicular immune processes. Finally, comparative studies of immunological process in the testes of rats and mice may be very informative.
111. THE ROLE OF L-PROLINE IN PREIMPLANTATION MOUSE EMBRYO DEVELOPMENT IN VITRO
Authors: Ozsoy S. , Morris M. B. , Day M. L.
Published: November 2010

Amino acids are known to play important roles in preimplantation embryo development, including regulation of cell volume and metabolism. Inclusion of l-glutamine, glycine and betaine in embryo culture medium has been shown to improve development in vitro by acting as organic osmolytes, thereby regulating cell volume. The purpose of the present study was to examine the effect of l-proline on preimplantation mouse embryo development in vitro. One-cell stage embryos were cultured in modified HTF, at low density (1 embryo/100 μL) and high density (1 embryo/μL) in the presence and absence of amino acids. Development of the embryos was scored every 24 h until the blastocyst stage. At low density, l-proline significantly increased the proportion of embryos developing to the blastocyst stage. This effect was abolished by culture at high density, suggesting that l-proline was activating a pathway similar to that involved in autocrine signalling by trophic factors in the preimplantation embryo. The improvement in development observed in the presence of l-proline was not due to its action as an organic osmolyte since the osmolality of the medium was 270 mOsm. Furthermore, glycine and betaine, which are known to act as osmolytes in embryos, had no effect on blastocyst development. In embryonic stem cells L-proline is taken up by an amino acid transporter and is involved in the regulation of growth and differentiation (1). The present data suggest that l-proline may have a similar, important role in preimplantation development.
(1) JM Washington, J Rathjen, F Felquer, A Lonic, MD Bettess, N Hamra, L Semendric, BSN Tan, J-A Lake, RA Keough, MB Morris and PD Rathjen (2010) Am J Physiol Cell Physiol 298: C982–C992.
Can translocations be used to mitigate human - wildlife conflicts?
Authors: Massei Giovanna , Quy Roger J. , Gurney Joanne , Cowan Dave P.
Published: November 2010

Growing public concerns about lethal methods to mitigate human–wildlife conflicts place increasing constraints on wildlife management options. Translocations, perceived as humane and non-lethal solutions, are increasingly advocated to resolve these conflicts. The present study summarises the literature on translocations of wild mammals, with particular emphasis on ‘problem’ animals, reviews the impact of translocations on survival, behaviour, animal welfare and potential spread of diseases, and evaluates the feasibility and effectiveness of translocations to mitigate human–wildlife conflicts. Translocations may have a detrimental impact on survival rates and lead to extreme dispersal movements. In some species, stress-related capture results in substantial mortality. In other species, homing causes animals to leave the release area. In addition, some animals resume the nuisance behaviour at the release site. Individuals that survive a translocation may suffer from malnutrition, dehydration, decreased immunocompetence and predation. Supportive measures such as acclimatisation pens and provision of food and shelter can drastically reduce post-release dispersal movements and mortality, although the adoption of these measures increases the cost of translocation. Translocations have the potential to spread diseases to conspecifics, humans, domestic animals and livestock. Health surveillance, seldom implemented, is likely to add significantly to the cost of translocation. Very few studies have reported the costs of translocations or addressed which stakeholders are expected to pay for translocating problem animals. Alternative management options are rarely mentioned. Despite the perceived humaneness of translocations to mitigate human–wildlife conflicts, the fate of translocated animals has been rarely monitored. In addition, very few studies have mentioned whether and for how long the conflict was resolved. We suggest that determining whether the translocation leads to the resolution of the problem should be the main criterion to evaluate the success of the translocation of problem animals. We propose a list of criteria to assist decisions regarding the suitability, effectiveness and humaneness of translocations to manage problems posed by wild mammals.
Breaking the Bond between Stimulant Use and Risky Sex: A Qualitative Study
Authors: Lyons Thomas , Chandra Gopika , Goldstein Jerome , Ostrow David G.
Published: October 2010

Stimulant-using men who have sex with men (MSM) are at increased risk for human immunodeficiency virus (HIV) transmission, and are more likely to practice unprotected anal sex than MSM who do not use methamphetamine and/or crack cocaine. In this paper the authors report on interviews with stimulant-using men who have sex with men who have participated in Crystal Meth Anonymous and other 12-step groups, focusing on those who did not have unprotected anal intercourse during a 6-month follow-up period and their reasons for doing so. The authors find 4 common themes cited: a diminished sexual drive; exclusive sex with a primary partner; greater sense of responsibility/commitment to safer sex; and most commonly of the four, an overall healthier sex life. Participants’ use of terms such as “healthy,” “enjoyable,” and “fulfilling” to describe sex not on stimulants, and avoidance of these terms for sex on stimulants, suggests a distinct dimension of sexual experience.
Teaching Condom Use Skills: Practice is Superior to Observation
Authors: Calsyn Donald A. , Hatch-Maillette Mary A. , Doyle Suzanne R. , Cousins Sarah , Chen TeChieh , Godinez Melinda
Published: October 2010

Men exposed to a condom skills practice exercise were hypothesized to perform better on condom skills measures than those exposed only to a demonstration or to no intervention. As part of a larger National Institute on Drug Abuse (NIDA) Clinical Trials Network HIV Prevention protocol, men in substance abuse treatment were administered male and female condom use skills measures (MCUS, FCUS) at preintervention, 2 weeks, 3 months, and 6 months postintervention. The MCUS and FCUS scores were compared for 3 intervention exposure groups (demonstration only [DO, n = 149], demonstration plus practice [D+P; n = 112], attended no sessions [NS, n = 139]) across the 4 assessment time points using a mixed effects linear regression model. There is a statistically significant intervention group-by-time effect ( P < .0001) for both the MCUS and FCUS. Post hoc, pairwise linear trends across time indicated that for both the MCUS and the FCUS, the D+P group is significantly superior to the DO group and the NS group.
Inside the Physician's Black Bag: Critical Ingredients of Brief Alcohol Interventions
Authors: Grossberg Paul , Halperin Abigail , Mackenzie Sara , Gisslow Mary , Brown David , Fleming Michael
Published: October 2010

Brief primary care interventions structured around patient workbooks have been shown to be effective in modifying hazardous drinking behavior. However, the critical ingredients of such interventions are not well understood, possibly contributing to their underutilization. Seventeen campus-based clinicians trained in a brief, workbook-based alcohol intervention participated in a qualitative study to identify the most promising clinician-patient interaction components within this shared approach, utilizing a focus group with the clinicians and ranking of the 24 workbook ingredients. Based on the clinicians’ collective experience, consensus emerged around the perceived strength of 5 main components: ( 1 ) providing a summary of the patient's drinking level, ( 2 ) discussing drinking likes and dislikes, ( 3 ) discussing life goals, ( 4 ) encouraging a risk-reduction agreement, and ( 5 ) asking patients to track their drinking (on cards provided for this purpose). This is the first paper to examine primary care physician perspectives on potentially critical components of effective brief alcohol intervention.
United Nations Office on Drugs and Crime International Network of Drug Dependence Treatment and Rehabilitation Resource Centres: Treatnet
Authors: Tomás-Rosselló Juana , Rawson Richard A. , Zarza Maria J. , Bellows Anne , Busse Anja , Saenz Elizabeth , Freese Thomas , Shawkey Mansour , Carise Deni , Ali Robert , Ling Walter
Published: October 2010

Key to the dissemination of evidence-based addiction treatments is the exchange of experiences and mutual support among treatment practitioners, as well as the availability of accurate addiction training materials and effective trainers. To address the shortage of such resources, the United Nations Office on Drugs and Crime (UNODC) created Treatnet, a network of 20 drug dependence treatment resource centers around the world. Treatnet's primary goal is to promote the use of effective addiction treatment practices. Phase I of this project included ( 1 ) selecting and establishing a network of geographically distributed centers; ( 2 ) conducting a capacity-building program consisting of a training needs assessment, development of training packages, and the training of 2 trainers per center in 1 content area each; and ( 3 ) creating good-practice documents. Data on the training activities conducted by the trainers during their first 6 months in the field are presented. Plans for Phase II of the Treatnet project are also discussed.
Feasibility of Emergency Department Bilingual Computerized Alcohol Screening, Brief Intervention, and Referral to Treatment
Authors: Vaca Federico , Winn Diane , Anderson Craig , Kim Doug , Arcila Mauricio
Published: October 2010

The purpose of this study was to assess the feasibility of utilizing a computerized alcohol screening and intervention (CASI) kiosk in an emergency department (ED). An interactive English and Spanish audiographical computer program, developed for used on a mobile computer cart, was administered to 5103 patients. Patients who screened at risk (19%) also received a fully computer-guided brief negotiated interview (BNI) and a printed personal alcohol reduction plan. A higher percentage of younger patients, and males (31% versus 16% females), screened at risk or dependent. Patient surveys indicated CASI was easy to use and over 75% did not prefer a medical professional over the computer. The ED-based bilingual computerized alcohol screening, brief intervention, and referral to treatment required little time to administer, was acceptable to patients, identified at-risk and dependent drinkers, and was able to provide personalized feedback and brief intervention.
Barbexaclone Abuse in a Cannabis Ex-User
Authors: Darcın Asli Enez , Dılbaz Nesrin , Okay Ihsan Tuncer
Published: October 2010

Abuse of drugs including addictive ingredients is common among patients with initial addiction history. This article reports a patient who had experienced a panic attack due to cannabis intoxication and has began to abuse an antiepileptic drug barbexaclone after he had quitted cannabis.
The dirty work of higher education
Authors: Parry Gareth
Published: October 2010

Through his early studies of the character of adult schools and community colleges in California, Burton Clark launched a set of ideas, sociological and educational, that served as baseline concepts in the study and practice of American higher education. His book The Open Door College
ranks among the classic accounts of a type of organisation – the public two-year college – which came to prominence throughout the United States in the post-war period. Its consideration of the roles played by such institutions in the larger education structure and society has
been at the centre of international, theoretical and empirical debates for half a century. Foremost among its arguments was the cooling-out function, a conception that enjoyed wide circulation over many decades. Rather less attention has been paid to its analysis of organisational determinacy
and the special problems of institutions that straddle secondary and higher education. These are the mass enterprises that do much of the dirty work of higher education. Unlike in the Clark corpus, they do not always receive the seriousness of scholarly treatment accorded to other kinds of
educational organisation.
Burton Clark's The Higher Education System: Academic Organization in Cross-National Perspective
Authors: Brennan John
Published: October 2010

In The Higher Education System, Burton Clark provides a model for the organisational analysis of higher education institutions and systems. Central to the model are the concepts of knowledge, beliefs and authority. In particular, Clark examines how different interest groups both
inside and outside the university shape and subvert the management of change. Within the university, Clark notes the tensions between the 'enterprise' and the 'discipline' and at the system level between the state authority, the market and the academic oligarchy. In considering the applicability
of Clark's model to an understanding of today's higher education systems and institutions, one can note a weakening of boundaries both within higher education institutions and between them and other institutions of society. Arguably, there has been a lessening of the organisational distinctiveness
of universities and an invasion by the language and ideas of the business world. The broadening of the social functions of modern higher education systems may be one of the reasons why academic authority seems to be subject to greater external challenge. Nevertheless, much of Clark's analytic
model remains highly relevant to our understanding of higher education systems and institutions even if their empirical manifestations have changed over the intervening years. Clark's model shares much in common with a more recent analysis of the changing relationship between higher education
and society conducted as part of a recent project of the European Science Foundation, although changes in emphasis and in authority relationships are also revealed.
Perspectives on higher education after a quarter of a century
Authors: Williams Gareth
Published: October 2010

Higher education is a field of study. It does not have an integral cognate discipline that defines its extent and borders. Contributions to understanding higher education come from, among others, sociologists, historians, political theorists, economists and students of science policy
and cultural studies. These were the disciplines whose exponents were assembled by Burton Clark in UCLA to summarise for a wider audience the contributions their disciplines were making to scholarship and research into higher education in the early 1980s. This paper reviews these disciplinary
approaches and Clark's reactions to them in the light of later developments in higher education and its study. It concludes by claiming that the contributors showed remarkable prescience in identifying many of the main developments in higher education studies over the following quarter of
a century. It also asserts that although there has been a huge expansion in higher education studies since then, and they now have all the attributes of an academically respectable field of study throughout the world, they remain dependent on their foundation disciplines for their intellectual
rigour.