Prolongation anormale d’un bloc fémoral analgésique: cas Clinique

La prolongation anormale d'un bloc nerveux peut être définie comme un dépassement du délai habituel de récupération sensitive ou motrice. A travers un cas clinique d'une prolongation anormale d'un bloc analgésique et une revue de la littérature, les auteurs discutent les facteurs de risque et les moyens de prévention de cette complication.Key words: Bloc nerveux fémoral, dexamethasone, prolongation anormal

The use of laser-assisted hatching in bovine in vitro produced embryos to improve pregnancy rate

In vitro produced (IVP) embryos not hatching from the zona pellucida (ZP) after transfer is one possible contributing factor of a lower pregnancy rate when compared to in vivo embryos. This study evaluates using a microscope objective mounted laser to cut the ZP prior to transfer into the recipient to assist hatching. The preliminary data evaluated the effect of laser treatment on IVP embryos and subsequent blastomere survival. In six replicates, bovine oocytes were in vitro produced according to the standard laboratory procedures of TransOva Genetics, Sioux Center, IA. On days 5, 6, and 7 of in vitro culture, embryos were randomly divided into 3 groups: no treatment (Control; n=63), sham ZP cut (Sham; n=68), or ZP cut (Cut; n=70). Control embryos were immediately returned to the incubator. Sham embryos were exposed to all conditions as Cut except laser assisted hatching. The XyClone® system was used to treat the Cut group using pulse strength of 90% and pulse length of 600 μsec. Embryos were returned to culture until day 8 when embryonic development and the percentage of live cells were determined and analyzed with Chi square. The number of developing embryos and the percentage of live cells per embryo showed no significant difference. Mean live cells ranged from 89-96% regardless of day of treatment. The laser assisted hatching effect on IVP embryo viability was evaluated by randomly dividing commercially produced embryos obtained from TransOva Genetics into two groups on day of transfer, Control or Cut. The ZP of treated embryos were cut with the laser using 80% pulse strength and pulse length of 500 μsec on day 7, immediately prior to transfer into estrous synchronized recipients. Ultrasonagraphy determined pregnancy rates. Thirty day pregnancy rates were 49.2% and 54.1% for Control (n= 189) and Cut (n=148) embryos, respectively, and were not statistically different (p > 0.05). However, 60 day Control pregnancy rate was 45.7% (n= 166) and the Cut group rate was 57.7% (n= 123) revealing a statistical difference (p < 0.05). These results demonstrate that the XyClone® system assisted hatching can improve 60 day pregnancy rates for IVP embryos by approximately 11 %

Association of oocyte and early embryo morphology with age and the establishment and maintenance of pregnancy after ICSI in mares

2010 Spring.Covers not scanned.Includes bibliographical references.Increasing maternal age in humans, horses and lab animals has been associated with a decrease in fertility. Oocyte quality and morphology have been implicated as primary causes of reduced fertility in older mares. Selected oocyte morphological parameters have been correlated with pregnancy development in humans and horses. Objective measurements of morphology to assess oocyte quality would provide a critical evaluation and help identify zygotes with the highest developmental potential for transfer, to optimize recipient utilization and pregnancy rates. The hypotheses of the research were that oocyte and early embryo morphology differ with donor mare age and correspond with developmental potential. Objectives for the first study were to compare: 1) oocyte donor age with oocyte morphology and developmental competency after ICSI, and 2) oocyte morphology with developmental competency (cleavage, early pregnancy, late pregnancy and pregnancy loss) after ICSI. Objectives for the second study were to compare developmental potential of ICSI produced embryos with: 1) oocyte donor age, and 2) cleavage characteristics, and 3) rate of embryonic development. Oocytes were collected from donor mares in a clinical ICSI programs. The mares were divided into the following age groups and fertility categories: 1) 3-13 yr with Known fertility, 2) 2-13 yr with Unknown fertility, 14-19 yr, 20-23 yr and 24-27 yr. Approximately 24 h after induction of follicle maturation, and oocytes were collected and cultured approximately 18 h before being stripped of cumulus cells. Photographic images (200x) were captured before oocytes were injected with sperm. Images of oocytes were measured using digital calipers within a computer software program. Ooplasm volume was larger (p<0.05) for oocytes from mares 14-19 yr and 20-23 yr than mares 3-13 yr Known than for mares 24-27 yr. Perivitelline space volume was similar between mares 3-13 yr Unknown and mares 20-23 yr, but was smaller (p<0.05) between mares 3-13 Unknown and the other age groups. Oocyte diameter (OD) was smaller (p=0.05) between oocytes from donors 3-13 yr Known and donors 14 -19 yr, but similar among all other groups. Inner zona pellucida diameter (IZPD) differed (p=0.03) only between mares 14-19 yr and mares 3-13 yr Unknown, with oocytes from mares 14- 19 yr having the largest numerical IZPD and mares 3-13 yr Unknown having the smallest IZPD. Ooplasm diameter (OpD) was smaller (p≤0.02) for oocytes from mares 3-13 yr Known than from mares 14-19 or 20-23 yr. The diameter of the zona pellucida with the surrounding matrix (ZPTM) was greater (p<0.05) for mares 3-13 yr Unknown than for all other groups. The rate of embryo development (hours per cell) prior to oviductal embryo transfers was faster (P<0.05) for embryos that did versus did not produce an early pregnancy and tended (P≤0. l) to be faster for embryos that did versus did not produce a late pregnancy. Embryonic vesicles that had a more rapid increase in diameter were more often (p<0.05) maintained to the late pregnancy stage. Donor mare age exerted a large effect on the development and outcome of pregnancies. Oocyte morphology was not a reliable indicator of oocyte developmental potential, although speed of early embryonic development was associated with embryonic competency

THE EVOLUTION OF IN VITRO FERTILIZATION IN THE UNITED STATES: A CLOSER LOOK AT MEDIA COVERAGE

Since the inception of in vitro fertilization in the United States in 1981 the U.S. media have covered its advances, moral debates and oddities. The reproductive technology, which manually combines both egg and sperm to form an embryo outside of a woman's uterus, sparks the public's curiosity in many ways - ethically, legally, emotionally and scientifically. Infertility, the disease that IVF overpowers, affects one million women each year. But the media rarely write in-depth articles that give readers and viewers information about the evolution of IVF and the years of trials and error that doctors and patients endured to improve the technique. Nor has the press covered the cost of IVF. That information is important to potential patients and the medical field and the history of research and medical development. This series of three magazine articles fills the information gap and also looks at the media's response to that gap.Master of Art

Somatic cell interspecies nuclear transfer

The low efficiency of the nuclear transfer (NT) procedure requires large number of oocytes to produce embryos and live offspring. A series of experiments were conducted to evaluate the ability of the bovine cytoplast to reprogram nuclei from horses and llamas. In a preliminary study, equine oocytes from small (\u3c20mm diameter) follicles were either pretreated with roscovitine or placed in maturation (IVM only) prior to NT. Roscovitine pretreatment did not improve nuclear maturation rates (roscovitine pretreatment 57% vs. IVM only 66%) and no fusion was obtained from roscovitine-pretreated oocytes after NT. Another preliminary study was conducted with the objective to produce llama NT embryos and to compare their development in two in vitro culture conditions (G1.2® vs. CR1aa). No difference was found in the number of embryos cleaved after 2 d of culture. This resulted in the first scientific report of somatic cell NT, in vitro culture and transfer of NT embryos in the llama. In the next experiment, adult horse and llama fibroblasts were injected into enucleated cow oocytes. The results showed the cow cytoplasm is capable of partially reprogramming nuclei from other species and support mitotic divisions. However, this study also showed a consistent embryonic developmental arrest at the 8- to 16- cell stage when horse or llama donor cells were used as donor nuclei. When a more closely related species of donor cell (banteng) and recipient oocyte (domestic cattle) were used for NT, no embryonic developmental arrest was found. Embryos progressed to achieve high blastocyst rates (banteng male cell line 28% vs. banteng female cell line 15%). Two banteng interspecies NT pregnancies were established and subsequently lost from the banteng male cell line. In the final study, the effect of a mixed mitochondrial population (heteroplasmy) on early embryonic development was investigated. Ooplasmic transfer performed in combination with NT procedure indicated presence of foreign mitochondria clustered in a small portion of the cytoplasm in early stages of embryo development. When goat ooplasm was transferred into interspecies (cow oocyte-goat donor cell) NT embryos, fusion and cleave rates were reduced suggesting an increased level of heteroplasmy or nuclear-ooplasmic incompatibilities

Fixed-time insemination of porcine luteinizing hormone-treated superovulated beef cows and the resynchronization of beef cows for fixed-time embryo transfer

Two trials were conducted to compare the effectiveness of fixed-time artificial insemination (AI) to AI based upon visual detection of estrus following superstimulation of donor beef cows. In Trial 1, multiparous beef cows (n = 31) were randomly allotted to one of three treatments following superstimulation and removal of an intravaginal progesterone insert (CIDR). Cows in the Control group were inseminated at 12 and 24 h after onset of estrus. Cows in the Estradiol group were injected with estradiol-17β (1 mg, im) at 12 h and inseminated at 24 and 36 h after CIDR removal. Cows in the pLH36 group were injected with porcine LH (Lutropin, 12.5 mg, im) at 24 h and inseminated at 36 and 48 h after CIDR removal. Mean numbers of viable embryos were 7.8, 3.6 and 9.6 for Control, Estradiol and pLH36 treatment groups, respectively (P > 0.10). In Trial 2, multiparous beef cows (n = 22) were randomly allotted to one of three treatments following superstimulation and removal of a CIDR. Sixteen of the cows were superstimulated a second time approximately 50 days later and allotted to one of the two treatments that differed from the initial treatment group. Cows in the Control group were inseminated at 12 and 24 h after onset of estrus. Cows in the two pLH groups were injected with porcine LH (Lutropin,12.5 mg, im) at 24 h after CIDR removal and were inseminated with either one unit of semen at 36 and 48 h (pLH36) or with two units of semen at 48 h (pLH48) after CIDR removal. Mean numbers of viable embryos were 3.0, 6.4 and 3.8 for Control, pLH36 and pLH48 treatment groups, respectively (P > 0.10). These data indicate that administration of pLH can facilitate use of fixed-time AI in superovulated beef cows without sacrificing embryo production. The second study evaluated the efficacy of resynchronizing beef cow recipients using CIDR devices for only 7 or 14 d. Recipient cows received CIDRs either on the day of transfer (n = 88) or 7 d post-transfer (n = 230). All CIDRs were removed on d 21 and cows were observed for estrus between d 22 and 24. Cows that displayed estrus were ultrasounded on d 30, those cows not pregnant that possessed a CL had an embryo transferred that day. Cows were later examined for pregnancies approximately 23 to 30 d later. There were no differences in pregnancy rates between cows with 7 or 14 d CIDRs and therefore data were combined. Pregnancy rates at two different ranches indicate that beef cow recipients can be successfully resynchronized by insertion of a CIDR without compromising pregnancy rates of transferred embryos. At Center Ranch the pregnancy rate for the first transfer was 57% while the resynchronized group that received the second transfer had a pregnancy rate of 55%. At Mound Creek Ranch the first transfer of embryos produced 59% pregnancy rates while the second transfer had a pregnancy rate of 71%. No significant differences (P > 0.05) were observed between the pregnancy rates of the initial transfer and those of the resynchronized transfer using only CIDRs, indicating that resynchronization using CIDRs can be used without reducing pregnancy rates

Natural and Anthropogenic Effects on Life History Characteristics in the Side-blotched Lizard (Uta stansburiana)

Survival is a prerequisite for successful reproduction, and is thus intertwined with fitness. Some physiological systems can improve survival, like the immune system, but compete with other processes for resources. Because animals evolved with resource limitation, it is important to understand how these resource-allocation decisions are made. To meet this end, I performed four investigations addressing how life-history characteristics shift in side-blotched lizards (Uta stansburiana) using laboratory studies and multi-year field sampling. First, I measured metabolic rates in response to different immune challenges and different energy states in male lizards. I found that, surprisingly, cutaneous biopsies were associated with a downregulation of metabolic rate, lipopolysaccharide injection did not affect metabolic rate, and the fastest-healing individuals had the largest decrease in metabolic rate, regardless of feeding treatment. In my second experiment, female lizards were challenged with a cutaneous biopsy and follicle-stimulating hormone to force a trade-off between the reproductive and immune systems. I learned that follicle-stimulating hormone increased metabolic rates and cutaneous biopsies decreased them, and that a combination of both treatments reduced metabolic rate the most. In my third experiment, I sampled wild lizards from northern populations in eastern Oregon and southern populations in southern Utah. In the first year, longer-lived northern lizards had higher circulating corticosterone concentration, decreased reproductive investment, and increased microbiocidal ability, but the subsequent year did not hold to these trends. A subsequent common-garden experiment revealed that southern lizards were faster at healing cutaneous wounds, but lost much more body mass than the northern individuals, which healed more slowly but maintained body mass. Finally, I have conducted a 5-year field investigation addressing the life-history trade-offs associated with urbanization, which reveals interesting effects of precipitation on survival and reproductive effort at urban and rural sites

Molecular Mechanisms of microRNA-1205 in Aggressive Prostate Cancer

Prostate cancer (PCa) is one of the most commonly diagnosed cancers among men in the United States. High mortality rates of PCa are associated with metastatic castration-resistant prostate cancer (mCRPC) due to the maintenance of androgen receptor (AR) signaling despite androgen deprivation therapies (ADTs). Resistance to second generation ADTs leads to the progression of AR-independent treatment related-neuroendocrine PCa (t-NEPC), which is observed in nearly 1 in 5 men with mCRPC and is associated with very poor outcomes. The 8q24 chromosomal locus is a region of very high PCa susceptibility that carries genetic variants associated with PCa aggressiveness. Located at this region is the PVT1 non-protein coding gene, implicated in PCa, and that encodes microRNA-1205 (miR-1205). As PVT1-encoded miRNAs are largely understudied, we sought to understand the role of miR-1205 to further implicate the importance of the 8q24 chromosomal locus in PCa. We observed that miR-1205 is underexpressed in a cohort of histologically confirmed PCa tissues, when compared to normal tissues and is also underexpressed in CRPC cells, when compared to non-CRPC cells. Our preliminary data suggest that a synthetic analog of miR-1205 inhibited tumor volume in a xenograft mouse model of CRPC, suggesting that miR-1205 may be a tumor suppressor in PCa. To understand the molecular mechanisms of miR-1205, we identified and validated FRYL as a molecular target of miR-1205 and demonstrated that miR-1205 can regulate FRYL mRNA stability. To identify additional molecular targets of miR-1205, a RNA pulldown assay was performed and sequencing of miR-1205 enriched targets (Arraystar INC) were identified (in collaboration with Dr. Konstantinos Krampis) in various PCa models. We discovered that miR-1205’s molecular signature may be involved in neural-associated pathways, including dendrite regulation and cell differentiation. Interestingly, FRYL is also predicted to regulate dendritic branching and is significantly overexpressed in NEPC tissue when compared to benign prostate tissues. Therefore, we hypothesized that miR-1205 and FRYL could be involved in PCa neuroendocrine differentiation (NED), a phenomenon that is observed in t-NEPC. Differentiation of PCa luminal cells into AR-negative neuroendocrine cells occurs as a mechanism of defense towards ADT’s. We examined miR-1205 regulation of FRYL in NED in vitro by culturing LNCaP androgen sensitive PCa cells under androgen deprivation conditions. We observed FRYL mRNA overexpresssion and significant underexpression of miR-1205 in LNCaP-NED cells when compared to undifferentiated LNCaP cells. To further characterize this mechanism, a miR-1205 mimic was used to overexpress miR-1205 in LNCaP cells. While FRYL and NEPC markers (AurA and Neuron specific enolase 2) expression decreased when miR-1205 was overexpressed, FRYL knockdown did not decrease NED markers, suggesting that miR-1205 regulates NED-specific targets in a FRYL-independent manner. In conclusion, this project provides novel insights into the role of miR-1205 in biological mechanisms that could have clinical relevance in PCa