Biomedical subjects

B D Murphy

Publications and source records attributed to B D Murphy.

At least 19 recordsLinked to original sources

PubMed2006

Identification of penumbra and infarct in acute ischemic stroke using computed tomography perfusion-derived blood flow and blood volume measurements.

BACKGROUND AND PURPOSE: We investigated whether computed tomography (CT) perfusion-derived cerebral blood flow (CBF) and cerebral blood volume (CBV) could be used to differentiate between penumbra and infarcted gray matter in a limited, exploratory sample of acute stroke patients. METHODS: Thirty patients underwent a noncontrast CT (NCCT), CT angiography (CTA), and CT perfusion (CTP) scan within 7 hours of stroke onset, NCCT and CTA at 24 hours, and NCCT at 5 to 7 days. Twenty-five patients met the criteria for inclusion and were subsequently divided into 2 groups: those with recanalization at 24 hours (n=16) and those without (n=9). Penumbra was operationally defined as tissue with an admission CBF <25 mL x 100 g(-1) x min(-1) that was not infarcted on the 5- to 7-day NCCT. Logistic regression was applied to differentiate between infarct and penumbra data points. RESULTS: For recanalized patients, CBF was significantly lower (P<0.05) for infarct (13.3+/-3.75 mL x 100 g(-1) x min(-1)) than penumbra (25.0+/-3.82 mL x 100 g(-1) x min(-1)). CBV in the penumbra (2.15+/-0.43 mL x 100 g(-1)) was significantly higher than contralateral (1.78+/-0.30 mL x 100 g(-1)) and infarcted tissue (1.12+/-0.37 mL x 100 g(-1)). Logistic regression using an interaction term (CBFxCBV) resulted in sensitivity, specificity, and accuracy of 97.0%, 97.2%, and 97.1%, respectively. The interaction term resulted in a significantly better (P<0.05) fit than CBF or CBV alone, suggesting that the CBV threshold for infarction varies with CBF. For patients without recanalization, CBF and CBV for infarcted regions were 15.1+/-5.67 mL x 100 g(-1) x min(-1) and 1.17+/-0.41 mL x 100 g(-1), respectively. CONCLUSIONS: We have shown in a limited sample of patients that CBF and CBV obtained from CTP can be sensitive and specific for infarction and should be investigated further in a prospective trial to assess their utility for differentiating between infarct and penumbra.

Acute Disease↗

PubMed2005

Expression of adiponectin and its receptors in swine.

Adiponectin is an adipocyte-derived hormone that plays an important role in lipid metabolism and glucose homeostasis. Objectives of this study were 1) to determine the presence and distribution of adiponectin and its receptors 1 and 2 (adipoR1 and adipoR2) in porcine tissues; 2) to characterize pig adiponectin, adipoR1, and adipoR2 mRNA levels in various fat depots from three different breeds of pigs; and 3) to study, in stromal-vascular cell culture, the effects of leptin and tumor necrosis factor-alpha (TNFalpha) on pig adiponectin, adipoR1, and adipoR2 gene expression. To this end, fat Chinese Upton Meishan (UM, n = 10), lean Ham Line (HL, n = 10), and Large White (LW, n = 10) gilts were used. We report the isolation of partial cDNA sequences of pig adipoR1 and adipoR2. Porcine-deduced AA sequences share 97 to 100% homology with human and murine sequences. Pig adipoR1 mRNA is abundant in skeletal muscle, visceral fat, and s.c. fat tissues, whereas adipoR2 mRNA is predominantly expressed in liver, heart, skeletal muscle, and visceral and s.c. fat tissues. Pig adiponectin mRNA levels in s.c. and visceral fat tissues were not associated with plasma insulin and glucose in fasting animals. Subcutaneous (r = -0.44, P < 0.05), visceral (r = -0.43, P < 0.05), and total body fat (r = -0.42, P < 0.05) weights were negatively correlated with adiponectin mRNA levels measured in visceral, but not s.c., fat. Pig adipoR1 and adipoR2 mRNA levels, in visceral fat, were less expressed in fat UM gilts than in the lean HL gilts (P < 0.05). Inverse associations were found between s.c. (r = -0.57, P < 0.01), visceral (r = -0.46, P < 0.05), and total body fat (r = -0.56, P < 0.01) weights and adipoR2 mRNA levels in visceral fat only. We were unable to find such associations for adipoR1 mRNA levels in the overall gilt population. The current study demonstrated that TNFalpha downregulates adiponectin and adipoR2, but not adi-poR1, mRNA levels in stromal-vascular cell culture. Moreover, leptin significantly decreased adiponectin mRNA levels, whereas there was no effect on adiponectin receptors. We conclude that adiponectin and adi-poR2 mRNA levels, but not adipoR1, are modulated in pig visceral fat tissues. Furthermore, our results indicate that TNFalpha interferes with adiponectin function by downregulation of adipoR2 but not of adipoR1 mRNA levels in pigs.

Adiponectin↗

PubMed2001

Characterization of swine leptin (LEP) polymorphisms and their association with production traits.

Four polymorphisms in the swine leptin (LEP) gene were characterized and evaluated for association with economically important production traits in Yorkshire, Landrace and Duroc pigs. Our results show that these polymorphisms are generally of low frequency or are absent in pig populations. Two polymorphisms (A2845T and T3469C) may be associated (P < 0.0078) with feed intake and growth rate traits in Landrace pigs.

Alleles↗

PubMed2001

Heat shock interferes with steroidogenesis by reducing transcription of the steroidogenic acute regulatory protein gene.

A key regulatory point in fine tuning of steroidogenesis is the synthesis of steroidogenic acute regulatory protein, which transfers cholesterol into mitochondria. Heat shock and toxic insults reduce steroidogenic acute regulatory protein, severely compromising steroid synthesis. As the molecular mechanisms for this reduction remain elusive, we tested the hypothesis that heat shock directly interferes with transcription of the steroidogenic acute regulatory protein gene. We show that, in mouse MA-10 Leydig tumor cells, heat shock caused drastic declines in (Bu)(2)cAMP-induced progesterone accumulation and steroidogenic acute regulatory protein transcript abundance. A proximal steroidogenic acute regulatory protein promoter fragment (-85 to +39) is sufficient to direct both cAMP inducibility and heat shock inhibition. Nuclear extracts from MA-10 cells displayed binding to this proximal promoter fragment as a low mobility complex in gel shift experiments. This complex disappeared in nuclear extracts taken at 5 and 10 min after initiation of heat shock and reappeared in extracts taken at 2 and 8 h. Similar low- mobility complexes formed on oligonucleotides representing the overlapping subfragments of the minimal steroidogenic acute regulatory protein promoter fragment sensitive to the heat shock effect. Extracts from heat-shocked MA-10 cells displayed reduced complex formation to each of the subfragments. We conclude that heat shock reduces progesterone synthesis, steroidogenic acute regulatory protein mRNA abundance, and steroidogenic acute regulatory protein promoter activity and disrupts binding of nuclear proteins to the proximal region of the steroidogenic acute regulatory protein promoter. Together these observations provide strong evidence for a mechanism of transcriptional inhibition in the down-regulation of steroidogenic acute regulatory protein expression by heat shock.

Animals↗

PubMed2001

Formation and early development of the corpus luteum in pigs.

Numerous corpora lutea form from the multiple follicles that ovulate during the oestrous cycle of pigs. Vascular elements invade the follicle from the theca compartment, first centripetally, and subsequently by lateral branching of centripetal veins and arteries. The vessels are the vehicle for dispersion of steroidogenic theca cells throughout the corpus luteum. Mitosis occurs in both the theca and granulosa layers before ovulation, and in luteal cells well into the luteal phase. Luteal cell proliferation undergoes gradual restriction as the corpus luteum matures, but the mechanisms of exit from the cell cycle are unknown. The extracellular ligands that direct luteinization and maintain the corpus luteum include LH, prolactin, insulin and insulin-like growth factors (IGFs). These ligands induce qualitative and quantitative changes in steroid output, with progesterone as the principal product. These changes upregulate the cholesterol synthetic pathways to increase substrate availability. The intracellular regulation of luteinization is complex. A model is presented in which LH stimulates arachidonic and lineoleic acid metabolism to produce ligands for the nuclear proteins of the peripheral peroxisome activator receptor family. These ligands have downstream effects on cell differentiation and exit from the cell cycle. Luteal function is maintained by interactions among ligands, cholesterol regulatory proteins and constitutively expressed and regulated transcription factors.

Animals↗

PubMed2000

Porcine leptin receptor: molecular structure and expression in the ovary.

The porcine leptin receptor complementary DNA was cloned and sequenced and the leptin receptor gene expression evaluated in the porcine ovary. An open reading frame of 3498 nt cDNA was amplified from pig liver mRNA by RT-PCR. Sequence homology with the extracellular, transmembrane, and cytoplasmic domains of human, mouse, rat, sheep, and cow leptin receptors varied between 45% and 90%. Leptin receptor mRNA was present in porcine kidney, liver, spleen, lung, brain, testis, uterus, ovary, corpus luteum (CL), theca, and granulosa cells. The abundance of leptin receptor transcripts and protein varied during luteinization of granulosa cells in vitro and in the CL during the pig luteal phase. In the postovulatory CL, both mRNA and protein were low but detectable, maximal expression was observed in the midcycle CL, and lowest abundance occurred in regressed CL. Leptin receptor mRNA was present in granulosa cells at isolation and increased in abundance as the cells luteinized over 96 hr in culture. Leptin receptor protein was detectable after 12 hr of in vitro luteinization. We conclude that leptin receptor is expressed in granulosa and luteal cells, and varies during pig ovarian cell differentiation.

Amino Acid Sequence↗

PubMed2000

Interferon-tau stimulates granulocyte-macrophage colony-stimulating factor gene expression in bovine lymphocytes and endometrial stromal cells.

Interferon-tau (IFN-tau), the antiluteolytic signal produced by the trophoblast prior to implantation in ruminants, exhibits immunomodulatory properties. It stimulates the production of prostaglandin (PG) E(2) in bovine endometrial cells via the induction of cyclooxygenase-2 (COX-2). We previously demonstrated that preconditioning lymphocytes with PGE(2) increases the expression of granulocyte-macrophage colony-stimulating factor (GM-CSF), a cytokine that promotes conceptus growth and survival. Our goal in the present study was to evaluate the impact of IFN-tau on the expression of GM-CSF in bovine peripheral blood lymphocytes (PBL) and endometrial epithelial and stromal cells. Changes in PGE(2) production and mRNA levels of COX-2 were also studied in PBL in response to IFN-tau. Gene expression was estimated by semiquantitative reverse transcription-polymerase chain reaction and Northern analysis. The expression of GM-CSF in PBL was stimulated by treatment with IFN-tau. Furthermore, GM-CSF mRNA levels were increased after preconditioning PBL for 3 days with IFN-tau, followed by a 12-h restimulation without IFN-tau. Inhibition rather than stimulation of PGE(2) production and COX-2 expression in PBL during treatment with IFN-tau suggests a direct effect on GM-CSF expression. Moreover, GM-CSF expression was stimulated in uterine stromal cells in response to IFN-tau. This study provides the first evidence for stimulation of GM-CSF expression by IFN-tau in both leukocytes and endometrial stromal cells. In view of the role of GM-CSF on fetal growth and survival, these results support the hypothesis that the conceptus mediates accommodation mechanisms in the uterus during early pregnancy by modulating the expression of beneficial cytokines at the fetomaternal interface.

Animals↗

PubMed2000

Models of luteinization.

Luteinization is essential to the success of early gestation. It is the process by which elements of the ovarian follicle, usually including both theca interna and granulosa cells, are provoked by the ovulatory stimulus to develop into the corpus luteum. Although there are significant species differences in luteinization, some elements pervade, including the morphological and functional differentiation to produce and secrete progesterone. There is evidence that luteinization results in granulosa cell exit from the cell cycle. The mechanisms that appear to control luteinization include intracellular signalling pathways, cell adhesion factors, intracellular cholesterol and oxysterols, and perhaps progesterone itself as a paracrine or intracrine regulator. Cell models of luteinization, along with some of the conflicting observations on the luteinization process, are discussed in this review.

Animals↗

PubMed1999

Physics of rotating gamma systems for stereotactic radiosurgery.

PURPOSE: The purpose of this study was to evaluate the characteristics of a new rotating gamma system for stereotactic radiosurgery by comparison with a well accepted system. METHODS AND MATERIALS: A novel gamma unit for stereotactic radiosurgery has been developed and distributed to 15 hospitals in China. The unit contains 30 cobalt-60 gamma radiation sources with initial activity of 200 Ci (7.4 x 10(12) Bq) each. The sources are positioned along 30 arcs, and rotate continuously as a group in an axis orthogonal to the patient's body. Measurements have been made on a representative unit installed in the Auhai Radiosurgery Center at the Beijing Navy General Hospital in the People's Republic of China. Ionization chambers calibrated by an American accredited dosimetry calibration laboratory were used for these measurements, as well as radiochromic film and thermoluminescent dosimeters. The unit tested utilizes collimators of nominal diameters of 4, 8, 14, and 18 mm. Radiochromic film samples from a Leksell Model U Gamma Knife were evaluated by the same laboratory and are presented for comparison. The treatment planning system was not evaluated. RESULTS: Radiation-absorbed dose rates and profiles measured for this unit are comparable to those previously measured with the same techniques for the Leksell Model U Gamma Knife units in San Diego and Atlanta. CONCLUSION: This unit is capable of producing well collimated beams of high energy photons, suitable for stereotactic radiosurgery. It has similar physical characteristics to those previously reported for the Leksell Model U Gamma Knife unit.

Cobalt Radioisotopes↗

PubMed1999

Homologous and heterologous ligands downregulate follicle-stimulating hormone receptor mRNA in porcine granulosa cells.

We investigated homologous and heterologous downregulation of FSH receptor mRNA in porcine granulosa cells from ovaries of immature pigs. Cultures were treated with 0, 40, or 200 ng/ml porcine FSH or medium and terminated at 24 hr intervals for Northern analysis of FSH receptor and cytochrome P450 side chain cleavage (P450scc) mRNA, and for radioimmunoassay of progesterone. Cells luteinized over 96 hr, and control cultures displayed increases in P450scc (8-10 fold) and FSH receptor (2 fold) mRNA and progesterone (100 fold). FSH reduced FSH receptor mRNA by 50-90%, increased P450scc mRNA 8 fold within 48 hr, and elevated progesterone logarithmically over 96 hr. Luteinized cells, (after 96 hr) received FSH or LH (1-200 ng/ml) or prostaglandin E2 (0.01-1.0 mg/ml) for 6 hr resulting in increased P450scc mRNA (2-8 fold), and progesterone (2-5 fold), and reduced FSH receptor mRNA. FSH (200 ng/ml) or the cAMP analog, dbcAMP (1 mM) for 0-24 hr reduced FSH receptor mRNA to 15% of control from 4-24 hr and elevated P450scc mRNA at 4 and 6 hr, respectively, to maxima at 12-24 hr. Forskolin (1-10 mM) increased P450scc mRNA (2-3 fold) and downregulated FSH receptor mRNA, effects reversed by the inhibitor of cAMP, rpcAMPs. Both epidermal growth factor, and the activator of the protein kinase C pathway, phorbol 12-myristate, 13-acetate (PMA) at 10 nM reduced FSH receptor mRNA. We conclude that downregulation of FSH receptor mRNA in luteinized granulosa cells is mediated by both homologous and heterologous ligands which employ cAMP, and that growth factors that activate the PKC pathway reduce FSH receptor and P450scc mRNA abundance.

Animals↗

PubMed1999

Down-regulation of oxytocin-induced cyclooxygenase-2 and prostaglandin F synthase expression by interferon-tau in bovine endometrial cells.

Oxytocin (OT) is responsible for the episodic release of luteolytic prostaglandin (PG) F2alpha from the uterus in ruminants. The attenuation of OT-stimulated uterine PGF2alpha secretion by interferon-tau (IFN-tau) is essential for prevention of luteolysis during pregnancy in cows. To better understand the mechanisms involved, the effect of recombinant bovine IFN-tau (rbIFN-tau) on OT-induced PG production and cyclooxygenase-2 (COX-2) and PGF synthase (PGFS) expression in cultured endometrial epithelial cells was investigated. Cells were obtained from cows at Days 1-3 of the estrous cycle and cultured to confluence in RPMI medium supplemented with 5% steroid-free fetal calf serum. The cells were then incubated in the presence or absence of either 100 ng/ml OT or OT+100 ng/ml rbIFN-tau for 3, 6, 12, and 24 h. OT significantly increased PGF2alpha and PGE2 secretion at all time points (p < 0.01), while rbIFN-tau inhibited the OT-induced PG production and reduced OT receptor binding in a time-dependent manner. OT increased the steady-state level of COX-2 mRNA, measured by Northern blot, which was maximal at 3 h (9-fold increase) and then decreased with time (p < 0.01). OT also caused an increase in COX-2 protein, which peaked at 12 h (11-fold increase), as measured by Western blot. Addition of rbIFN-tau suppressed the induction of COX-2 mRNA (89%, p < 0.01) and COX-2 protein (50%, p < 0.01) by OT. OT also increased PGFS mRNA, and this stimulation was attenuated by rbIFN-tau (p < 0.01). To ensure that the decrease in COX-2 was not solely due to down-regulation of the OT receptor, cells were stimulated with a phorbol ester (phorbol 12-myristate 13-acetate; PMA) in the presence and absence of rbIFN-tau. The results showed that rbIFN-tau also decreased PMA-stimulated PG production and COX-2 protein. It can be concluded that rbIFN-tau inhibition of OT-stimulated PG production is due to down-regulation of OT receptor, COX-2, and PGFS.

Animals↗

PubMed1999

Growth factor modulation of steroidogenic acute regulatory protein and luteinization in the pig ovary.

In vivo and in vitro luteinization were investigated in the porcine ovary, with emphasis on expression of steroidogenic acute regulatory protein (StAR). StAR mRNA and protein as well as cytochrome P450 side-chain cleavage mRNA (P450scc) increased during the luteal phase in the corpus luteum (CL) and were absent in regressed CL. Cytochrome P450 aromatase mRNA (P450arom) was not detectable at any time in CL. In vitro luteinization of granulosa cells occurred over 96 h in culture, during which P450arom mRNA was present at 1 h after cell isolation but not detectable at 6 h; and P450scc and StAR mRNAs were first detectable at 6 h and 48 h, respectively. Incubation of cultures with insulin-like growth factor I (IGF-I, 10 ng/ml), dibutyryl cAMP (cAMP, 300 microM), or their combination, induced measurable StAR mRNA at 24 h (p < 0.05), increased progesterone accumulation at 48 h, and elevated both StAR and P450scc expression through 96 h. Incubation of luteinized granulosa cells with epidermal growth factor (EGF, 10 nM) changed their phenotype from epithelioid to fibroblastic, eliminated steady-state StAR expression, and interfered with cAMP induction of StAR mRNA and progesterone accumulation. EGF had little apparent effect on P450scc mRNA abundance. It is concluded that StAR expression characterizes luteinization, and early luteinization is induced by cAMP and IGF-I in vitro. Further, EGF induces a morphological and functional phenotype that appears similar to an earlier stage of granulosa cell function.

Animals↗

PubMed1998

Cloning of leukemia inhibitory factor (LIF) and its expression in the uterus during embryonic diapause and implantation in the mink (Mustela vison).

Leukemia inhibitory factor (LIF) is essential for embryo implantation in mice. Whether LIF plays a role in termination of embryonic diapause and initiation of implantation in carnivores, especially in species with obligate delayed implantation such as the mink, is not known. The objectives of this study were to clone the LIF coding sequence in the mink and determine its mRNA abundance in the uterus through embryonic diapause, implantation, and early postimplantation. We show that the mink LIF cDNA contains 609 nt encoding a deduced protein of 203 amino acids. The homologies are 80.6, 90, 88.2, 87.6, and 86.8% in coding sequence and 79.2, 90.1, 91, 90.1 and 85.4% in amino acid sequence with mouse, human, pig, cow, and sheep respectively. Glycosylation sites and disulfide bonds present in other species are generally conserved in the mink LIF sequence. Quantitation by polymerase chain reaction amplification indicates that LIF mRNA is expressed in mink uterus just prior to implantation and during the first two days after implantation, but not during diapause or later after implantation pregnancy. The abundance of LIF mRNA was significantly higher in the uterus at the embryo expansion stage (P < 0.05) than at days 1-2 of postimplantation. By immunohistochemical localization it was shown that LIF is expressed in the uterine epithelial glands at time of embryonic expansion and in early postimplantation. The coincidence of LIF expression with implantation in this species suggests that LIF is involved in the implantation process, and may be a maternal signal which terminates obligate embryonic diapause.

Amino Acid Sequence↗

PubMed1998

Prostaglandin E2 regulates both interleukin-2 and granulocyte-macrophage colony-stimulating factor gene expression in bovine lymphocytes.

Prostaglandin E2 (PGE2) is known to inhibit interleukin-2 (IL-2) production by human peripheral blood lymphocytes (PBL) and to increase granulocyte-macrophage colony-stimulating factor (GM-CSF). In many species with hemochorial placentation, down-regulation of IL-2 appears necessary to impede early embryonic demise, whereas up-regulation of GM-CSF increases embryonic growth and survival. It is not known whether the same mechanisms are involved in a species with a less invasive placenta. PGE2 is synthesized during early bovine gestation by the endometrium and by the embryo, and it may therefore be involved in regulating IL-2 and GM-CSF in this species. Our goal was to evaluate the impact of PGE2 on cellular proliferation and on IL-2 and GM-CSF gene expression in bovine PBL. Incorporation of [3H]thymidine was used to study DNA synthesis. Gene expression was estimated by semiquantitative polymerase chain reaction using bovine-specific primers and by Northern analysis using amplified bovine cDNAs as probes. The DNA synthesis and IL-2 mRNA levels of bovine PBL stimulated by concanavalin A (ConA) were greatly reduced by PGE2 in direct-treatment studies. Under the same conditions, GM-CSF gene expression was also inhibited. However, pretreatment of PBL for 72 h with ConA and PGE2, followed, after washing, by an incubation with ConA alone for 12 h resulted in reduced DNA synthesis, stable expression of IL-2, and a dramatic increase of GM-CSF mRNA levels. This is the first evidence in the bovine model that direct treatment with PGE2 down-regulates IL-2 and GM-CSF mRNA levels and that preconditioning with PGE2 stimulates GM-CSF gene expression. We propose that PGE2, either from embryonic or from endometrial compartments, induces bovine PBL to undergo functional changes, affecting cellular proliferation and cytokine production in order to accommodate the developing conceptus.

Animals↗

PubMed1998

Differentiation of the corpus luteum of the mink (Mustela vison): mitogenic and steroidogenic potential of luteal cells from embryonic diapause and postimplantation gestation.

The mink corpus luteum (CL) involutes after ovulation and remains dormant, synthesizing low amounts of progesterone until reactivated to terminate embryonic diapause. We examined the mitotic and steroid synthetic capacity of luteal cells from the diapause and postimplantation phases of mink gestation. Cells from diapause divided in vitro, reaching confluence in 7-8 days. Three phenotypes were distinguishable: a fusiform cell in whorls, a hypertrophied epithelioid cell, and a small epithelioid cell. The first and second cell types divided in vitro after confluence, evidenced by localization of proliferating cell nuclear antigen (PCNA) in their nuclei. The small epithelioid cells were present in cell nests and showed no PCNA activity. Cells derived from reactivated CL did not reach confluence and had no PCNA activity. Progesterone accumulation was enhanced in luteal cells from diapause by LH, FSH, and dibutyryl (Bu2)cAMP, and by LH and (Bu2)cAMP in cells from reactivated CL. In luteal cells from the diapause phase of gestation, LH and (Bu2)cAMP induced increases in mRNA coding for steroidogenic acute regulatory protein, while cytochrome P450 side-chain cleavage enzyme mRNA was increased by prolactin, LH and (Bu2)cAMP. Cellular concentrations of 3beta-hydroxysteroid dehydrogenase-delta5-4-isomerase mRNA were increased by prolactin and (Bu2)cAMP. Thus, luteinization in the mink CL does not engender exit from the cell cycle, as both fusiform and hypertrophied cells from diapause divide in vitro. Reactivation appears to represent terminal differentiation. LH is capable of stimulating steroidogenesis in vitro in luteal cells from diapause, and prolactin and LH appear to have both specific and overlapping stimulatory effects on the CL of this species.

Animals↗

PubMed1998

Luteotropic hormone receptors in the ovary of the mink (Mustela vison) during delayed implantation and early-postimplantation gestation.

The reproductive cycle of the mink displays rigid seasonality and obligate embryonic diapause. After ovulation, the corpus luteum (CL) involutes, and it secretes basal progesterone until activated prior to implantation. To study changes in the relative abundance of luteal prolactin and LH receptor mRNA through gestation, ovaries and serum were collected from pregnant female mink at 2-day intervals (n = 3 per date) through embryonic diapause and CL activation (March 19-31) and at 5-day intervals through implantation and early-postimplantation gestation (March 31-April 15). To determine the effect of endogenous prolactin, mink received Alzet osmotic minipumps releasing 2 mg/day 2-bromo-alpha-ergocryptine (bromocriptine) or saline on March 19. Ovaries and serum were taken from 3 animals every 2 days until March 31. Prolactin receptor mRNA in ovaries was low during CL activation but increased 3-fold through embryo implantation. Its abundance correlated with prolactin binding to ovarian membranes and with circulating prolactin. Bromocriptine suppressed endogenous prolactin levels and prevented the increase in prolactin receptor mRNA. There was a transient peak in LH receptor mRNA in the ovaries at March 19-23, which declined to basal levels by March 25 and remained constant through midgestation. Bromocriptine prevented the preimplantation peak in LH receptor mRNA and reduced its abundance below pretreatment levels. The results suggest that prolactin up-regulates its receptor and maintains the LH receptor in the mink CL. The pattern of LH receptor mRNA argues for a role for LH in CL reactivation and termination of embryonic diapause in mink.

Animals↗

PubMed1998

Bedside placement of peripherally inserted central catheters: a cost-effectiveness analysis.

PURPOSE: To compare two strategies for placement of peripherally inserted central catheters (PICCs). In strategy A, all PICC placements were initially attempted at the patient's bedside by trained intravenous (i.v.) nurses, with failures referred to the interventional radiology (IR) service. In strategy B, all PICCs were placed by interventional radiologists under fluoroscopic and/or venographic guidance. MATERIALS AND METHODS: Decision analysis and cost-effectiveness models were constructed with commercially available software. Data used in the model were obtained from a review of the authors' experience with bedside placement of PICCs by an i.v. team and data obtained from the literature. RESULTS: The cost-effectiveness of strategy A relative to strategy B depends on (a) the ability of the i.v. team to access a vein at the patient's bedside, (b) the cost of fluoroscopy or the IR suite, and (c) the intended use of the PICC. CONCLUSION: If the cost of fluoroscopy or the IR suite exceeds $100, strategy A is more cost-effective for most intended PICC uses. If the cost is less than $75, strategy B is more cost-effective for all intended PICC uses. If the cost is between $75 and $100, the most cost-effective strategy depends on the intended use of the PICC and the need to have the tip placed at the junction of the superior vena cava and right atrium.

Catheterization, Central Venous↗

PubMed1998

Cloning, developmental expression, and immunohistochemistry of cyclooxygenase 2 in the endometrium during embryo implantation and gestation in the mink (Mustela vison).

Cyclooxygenase (COX) is the first rate-limiting enzyme in the biosynthesis of PGs. There are two isoforms, COX-1, a constitutive enzyme and COX-2, the induced form, products of two different genes. In this study, we report COX-2 complementary DNA cloning, uterine expression, and immunohistochemical localization in the mink uterus during postimplantation gestation. The open reading frame of mink COX-2 contains 1812 nucleotides encoding 604 amino acids. The homologies are 86%, 83%, 83%, 83%, 85%, and 84% in nucleotides and 86%, 87%, 87%, 85%, 86%, and 88% in amino acids with human, mouse, rat, guinea pig, sheep, and rabbit, respectively. All domains associated with biological activity are conserved in the mink. Northern analysis revealed a transcript of 4.2 kb for COX-2 in mink uterus and adrenal. Semiquantitative RT-PCR showed that COX-2 messenger RNA is not present during diapause. The abundance of COX-2 messenger RNA reached its maxima (P < 0.05) on days 3-5 of postimplantation, gradually decreased through day 9, and was not present thereafter. By immunohistochemistry, COX-2 was present in uterine epithelium, stroma, and necks of endometrial glands at sites of implantation. COX-2 expression appears to be induced in the endometrium by the embryo and may play a role in implantation and placentation in the mink.

Amino Acid Sequence↗