Seasonal Variation of Melatonin Concentration and mRNA Expression of Melatonin-Related Genes in Developing Ovarian Follicles of Mares Kept under Natural Photoperiods in the Southern Hemisphere.

This study investigated the seasonal variations in mRNA expression of FSH (Fshr), LH (Lhr) receptors, melatonin (Mt1 and Mt2) receptors, melatonin-synthetizing enzymes (Asmt and Aanat) and melatonin concentration in developing follicles from mares raised in natural photoperiods. For one year, ultrasonographic follicular aspiration procedures were performed monthly, and small (<20 mm), medium (20 to 35 mm) and large (>35 mm) follicles were recovered from five mares. One day before monthly sample collections, an exploratory ultrasonography conducted to record the number and the size of all follicles larger than 15 mm. The total number of large follicles were higher during the spring/summer (8.2 ± 1.9) than during autumn/winter (3.0 ± 0.5). Compared to autumn/winter seasons, there was an increase of Fshr and Aanat mRNA expressions in small, medium and large follicles, an increase of Lhr and Asmt mRNA expressions in medium and large follicles and an increase of Mt1 and Mt2 mRNA expressions in small and large follicles during spring/summer. The melatonin levels in follicular fluid were also higher during the spring/summer seasons. The present data show that melatonin locally upregulates the mRNA expression of Mt1 and Mt2 receptors and melatonin-forming enzymes in mare developing follicles during reproductive seasons.

Changes in Oviductal Cells and Small Extracellular Vesicles miRNAs in Pregnant Cows.

Early embryonic development occurs in the oviduct, where an ideal microenvironment is provided by the epithelial cells and by the oviductal fluid produced by these cells. The oviductal fluid contains small extracellular vesicles (sEVs), which through their contents, including microRNAs (miRNAs), can ensure proper cell communication between the mother and the embryo. However, little is known about the modulation of miRNAs within oviductal epithelial cells (OECs) and sEVs from the oviductal fluid in pregnant cows. In this study, we evaluate the miRNAs profile in sEVs from the oviductal flushing (OF-sEVs) and OECs from pregnant cows compared to non-pregnant, at 120 h after ovulation induction. In OF-sEVs, eight miRNAs (bta-miR-126-5p, bta-miR-129, bta-miR-140, bta-miR-188, bta-miR-219, bta-miR-345-3p, bta-miR-4523, and bta-miR-760-3p) were up-regulated in pregnant and one miRNA (bta-miR-331-5p) was up-regulated in non-pregnant cows. In OECs, six miRNAs (bta-miR-133b, bta-miR-205, bta-miR-584, bta-miR-551a, bta-miR-1193, and bta-miR-1225-3p) were up-regulated in non-pregnant and none was up-regulated in pregnant cows. Our results suggest that embryonic maternal communication mediated by sEVs initiates in the oviduct, and the passage of gametes and the embryo presence modulate miRNAs contents of sEVs and OECs. Furthermore, we demonstrated the transcriptional levels modulation of selected genes in OECs in pregnant cows. Therefore, the embryonic-maternal crosstalk potentially begins during early embryonic development in the oviduct through the modulation of miRNAs in OECs and sEVs in pregnant cows.

Resiliency of equid H19 imprint to somatic cell reprogramming by oocyte nuclear transfer and genetically induced pluripotency†.

Cell reprogramming by somatic cell nuclear transfer and in induced pluripotent stem cells is associated with epigenetic modifications that are often incompatible with embryonic development and differentiation. For instance, aberrant DNA methylation patterns of the differentially methylated region and biallelic expression of H19-/IGF2-imprinted gene locus have been associated with abnormal growth of fetuses and placenta in several mammalian species. However, cloned horses are born with normal sizes and with no apparent placental anomalies, suggesting that H19/IGF2 imprinting may be epigenetically stable after reprogramming in this species. In light of this, we aimed at characterizing the equid H19 gene to determine whether imprinting is altered in somatic cell nuclear transfer (SCNT)-derived conceptuses and induced pluripotent stem cell (iPSC) lines using the mule hybrid model. A CpG-rich region containing five CTCF binding sites was identified upstream of the equine H19 gene and analyzed by bisulfite sequencing. Coupled with parent-specific and global H19 transcript analysis, we found that the imprinted H19 remains monoallelic and that on average the methylation levels of both parental differentially methylated regions in embryonic and extra-embryonic SCNT tissues and iPSC lines remained unaltered after reprogramming. Together, these results show that, compared to other species, equid somatic cells are more resilient to epigenetic alterations to the H19-imprinted locus during SCNT and iPSC reprogramming.

Acute exposure to hyperosmotic conditions reduces sperm activation by urine in the yellowtail tetra Astyanax altiparanae, a freshwater teleost fish / Exposição aguda a condições hipereosmoticas reduz ativação do sêmen por urina em lambari do-rabo-amarelo (Astyanax altiparanae), um teleósteo de água doce

In freshwater fish with external fertilization, sperm sampling can be contaminated with urine, which triggers motility and gives rise to decreased fertilization success. The maintenance of freshwater fish in hyperosmotic conditions may reduce urine production and improve sperm quality. Thus, the aim of this work was to verify if acute exposure to various NaCl concentrations improves sperm quality in the yellowtail tetra Astyanax altiparanae. Spermiation was induced using a single dose of carp pituitary gland (5 mg kg-1) and the males were maintained at various NaCl concentrations: NaCl 0.00% (control), NaCl 0.45% (hypoosmotic), NaCl 0.9% (isosmotic) and NaCl 1.0% (hyperosmotic) for 6 h at 26 °C. Sperm was collected and verified for activation by urine and motility traits. At 0.00%, 0.45%, and 0.90%, the sperm was motile just after sampling, indicating activation by urine. Surprisingly, at hyperosmotic conditions, no activation was observed. Other sperm and motility parameters did not show any statistical differences, including sperm viability (P = 0.7083), concentration (P = 0.9030), total motility (P = 0.6149), VCL (curvilinear velocity; P = 0.1216), VAP (average path velocity; P = 0.1231) and VSL (straight-line velocity; P = 0.1340). Our results indicate that acute maintenance at hyperosmotic conditions eliminates sperm activation by urine and maintains sperm quality. Such a new procedure is interesting for both basic and applied sciences, including reproductive practice in fish.(AU)

Em peixes de água doce com fertilização externa, a amostragem de espermatozoides pode ser contaminada pela urina, o que desencadeia motilidade e gera menor sucesso na fertilização. A manutenção de peixes de água doce em condições hiperosmóticas pode reduzir a produção de urina e melhorar a qualidade do esperma. Assim, o presente trabalho foi delineado para verificar se a exposição aguda a várias concentrações de NaCl melhora a qualidade do esperma no tetra-amarelo Astyanax altiparanae. A espermiação foi induzida usando uma dose única de hipófise da carpa (5 mg kg-1) e os machos foram mantidos em várias concentrações de NaCl: NaCl 0,00% (controle), NaCl 0,45% (hipoosmótico), NaCl 0,9% (isosmótico) e NaCl 1,0% (hiperosmótico) por seis horas a 26 °C. O esperma foi colhido e verificado quanto à ativação por urina e traços de motilidade. Em 0,00%, 0,45%, 0,90% os espermatozóides eram móveis logo após a amostragem, indicando ativação pela urina. Surpreendentemente, em condições hiperosmóticas, nenhuma ativação foi observada. Outros parâmetros espermáticos e de motilidade não mostraram diferenças estatísticas, incluindo viabilidade espermática (P = 0,7083), concentração (P = 0,9030), motilidade total (P = 0,6149), VCL (Velocidade Curvilinear; P = 0,1216), VMD (Velocidade Média de Deslocamento; P = 0,1230) e VLR (Velocidade em linha Reta; P = 0,1340). Nossos resultados indicam que a manutenção aguda em condições hiperosmóticas elimina a ativação do esperma pela urina e mantém a qualidade do esperma. Esse novo procedimento é interessante para as ciências básicas e aplicadas, incluindo a prática reprodutiva em peixes.(AU)

Use of color-Doppler ultrasonography for selection of recipients in timed-embryo transfer programs in beef cattle.

We aimed to study the association between CL characteristics assessed by color-Doppler ultrasonography (Doppler-US) at the time of embryo transfer (ET) and pregnancy rate (P/ET) in beef recipients. Estrous cycles of crossbred beef recipients were synchronized for timed-ET. On the day of ET (Day 7), CL area, proportion of luteal blood perfusion (BP), and the relationship between the largest dominant follicle (DF) and CL (ipsilateral or contralateral) were determined. Animals (n = 444) received an in vitro produced embryo from Nelore donors, placed in the uterine horn ipsilateral to the CL. Recipients were split retrospectively in three subgroups according to CL area [small (<3 cm2), medium (3-4 cm2), or large (>4 cm2)] and three subgroups according to luteal signals of BP [low (≤40%), medium (45-50%) or high (≥55%)]. Pregnancy was detected on Days 30-45 by transrectal ultrasonography and P/ET was analyzed considering the effects of cow's category (suckling or non-suckling), CL area, luteal BP and side of DF. P/ET increased along with BP category [low, 45.9%B, (62/135); medium, 54.1%AB (93/172); and high, 58.4%A (80/137)]. When luteal BP was evaluated as a continuous variable, a significant (P < 0.05) linear and positive effect was observed on P/ET. A greater (P < 0.05) CL area and serum progesterone concentrations were observed in the medium and high BP than in the low BP category. Although an effect of luteal size category was not significant on P/ET [small, 49% (76/155); medium, 59.7% (83/139); and large, 50.7% (75/148); P > 0.1], when CL area was evaluated as a continuous variable, a quadratic effect (P < 0.05) indicated a positive relationship between P/ET and CL area until luteal tissue reached 4.07 cm2, followed by a negative relationship. The location of the first-wave DF in relation to the CL did not affect P/ET (P > 0.1). In conclusion, Doppler-US is an innovative tool that has the potential to be used for selection of suitable embryo recipients based on luteal BP. Selection of recipients that have a greater chance of maintaining pregnancy will increase the success of timed-ET programs.

Ultrasonography-accessed luteal size endpoint that most closely associates with circulating progesterone during the estrous cycle and early pregnancy in beef cows.

The aim was to evaluate the associations between circulating P4 concentrations, corpus luteum (CL) size (diameter, area or volume) and blood perfusion (BP) in cows. In Experiment 1, Pearson's correlations (P < 0.05) with P4 concentrations were observed during CL development (D8) for total area (TA; r = 0.76), luteal area (ACL; r = 0.72), total and luteal diameter (TD and DCL respectively; r = 0.46). During mid-late diestrus, there was a positive correlation (P < 0.05) only at D15 with TA and ACL (r > 0.60), TD, total volume (TV) and luteal volume (VCL; r > 0.434). During luteal regression, the correlation was only observed at D18 for ACL (r = 0.478) and D20 with several variables. In Experiment 2, CL weight and ACL had the greatest correlation with P4 (r > 0.6). In Experiment 3, TA and ACL were the variables that were most closely correlated with serum P4 concentrations at D7 in recipient cows. Correlation coefficients were greater for luteal measurements when there were compact compared with cavitary CLs. In Experiment 4, there was no correlation (P > 0.05) between P4 and any of the variables measured on D4 and D7 in recipient cows detected in estrus. On D18 to D20, all CL characteristics were correlated (P < 0.05) with plasma P4, and luteal BP and BP area were more closely (P < 0.05) correlated than ACL. In conclusion, CL perimeter area measurements had the greatest association with luteal function during CL development; whereas for BP there was a greater correlation with P4 than luteal size during luteolysis.

Effect of hCG application at different moments of the estrous cycle on corpus luteum and uterine vascularization and serum progesterone concentration in mares.

Establishment of pregnancy after embryo transfer is the ultimate goal of an embryo transfer program and increasing pregnancy rates and reducing pregnancy loss are mandatory. The utilization of treatments to improve conception rates in recipient mares has been the focus of several research groups over the last years and the results are controversial. Some studies using human chorionic gonadotrophin (hCG) found promising results. Our hypothesis was that hCG administration would cause an additional stimulation on luteal function, uterine and luteal vascularization and progesterone concentration, and the mares would have increased uterine and cervix tone. Therefore, in the present study the effects of hCG administration to induce ovulation, on day 0 (day of ovulation) or day 5 postovulation were evaluated on corpus luteum characteristics, reproductive tract vascularization, and serum progesterone concentration from ovulation until day 15 postovulation. Groups were: G1: (control) - no hCG; G2: 2500 IU of hCG to induce ovulation when a follicle greater than 35mm and uterine edema were detected; G3: 2500 IU hCG on day 0; G4: 2500 IU hCG on day 5 postovulation. Twelve mares were randomly assigned to each group, during consecutive cycles, in a Latin Square experimental design, in a total of 48 cycles. Doppler ultrasound evaluations were performed daily from day 0 until day 15 postovulation, including mesometrial vascularity, endometrial vascularity and corpus luteum vascularity. Blood samples were collected for serum progesterone concentration. Data was analyzed using the Proc Glimmix SAS Procedure for nonparametric variables and Proc Mixed for parametric parameters. There was no treatment effect for all variables studied (P > 0.05). Characteristics were only affected by day (P < 0.05). It can be concluded that hCG administration at the time points suggested in the current study did not alter the characteristics evaluated.

Grooves surrounding the micropyle decrease the inseminating dose in fish.

In fish with external fertilization, sperm must reach the oocyte through the micropyle to enter the cytoplasm. Fertilization success is then influenced by characteristics of oocytes or sperm. In this study, we evaluated oocyte morphology and sperm motility parameters and their effects on the inseminating dose in a teleost fish Astyanax altiparanae. Interestingly, we found one of the lowest yet described inseminating doses in teleosts (2390 spermatozoa oocyte-1 ml-1). Such a fertilization efficacy may be explained by the long duration of sperm motility (>75 s), the small oocyte diameter (695.119 µm), large micropyle diameter (7.57 µm), and the presence of grooves on the oocyte surface that guides spermatozoon to the fertilization area. Additionally, we have described for the first time a structure that combines grooves on the chorion surface and a ridge in the micropylar area.

Fatty Acid Binding Protein 3 And Transzonal Projections Are Involved In Lipid Accumulation During In Vitro Maturation Of Bovine Oocytes.

Oocytes that undergo in vitro maturation (IVM) are metabolically abnormal and accumulate excess lipid content. However, the mechanism of lipid accumulation and the role of cumulus cells in this process are unclear. Recently, it was shown that fatty acid binding proteins (FABPs) performed intra- and extracellular fatty acid transport. We postulated that FABP3 might be responsible for fatty acid transport from cumulus cells to the oocytes via transzonal projections (TZPs) during IVM. Transcript and protein levels of FABP3 were analyzed in both in vivo- and in vitro-matured cumulus-oocyte-complexes and were increased in IVM samples. Further analysis showed increased lipid content in oocytes and cumulus cells in IVM samples compared to in vivo-derived. We therefore speculated that altered traffic of fatty acids via FABP3 during IVM was the mechanism leading to the excess of lipids accumulated within IVM oocytes. Furthermore, we demonstrated an increase in FABP3 levels and lipid content during the first 9 h of IVM, further strengthening the possibility of fatty acid transport via FABP3 and TZPs. Additionally, disruptions of TZPs during IVM decreased lipid accumulation in oocytes. Our results shed light on a possible mechanism involving FABP3 and TZPs that causes excess lipid accumulation in oocytes during IVM.

Follicular dynamics, ovarian vascularity and luteal development in mares with early or late postpartum ovulation.

Follicular development and deviation processes during the postpartum period are not completely known in horses. Thus, we aimed to study the characteristics of follicular dynamics and ovarian vascular perfusion during the postpartum period in mares that demonstrated estrous behavior and had early (<10 days) or late (≥10 days) postpartum ovulation. Ten mares were scanned daily by transrectal ultrasonography from the first day postpartum (d1) to the sixteenth day after the first postpartum ovulation (D0 = ovulation). The animals were split in the early (n = 3) and late (n = 7) ovulation groups (average interval between parturition and ovulation: 8.0 ± 0.0 and 14.7 ± 1.2 days, respectively). For the follicular growth, no difference (P > 0.1) was detected between the groups when the data were normalized for the days preceding the first postpartum ovulation (from D-7 to D-1). However, when the data were normalized to days postpartum, the dominant follicle was larger (P < 0.05) in the early-ovulated group in all days during this period (d1 to d7). The number of follicles >25 mm diameter was greater (P < 0.05) in the early-ovulated group during the first 3 days postpartum, and the late-ovulated mares showed greater number of follicles with 20-25 mm during d4 to d7. For blood flow characteristics, no difference (P > 0.1) was detected between groups in vascular perfusion of the dominant follicle or in the ovarian pedicle ipsilateral to the largest follicle. Similarly, no difference (P > 0.1) was detected in progesterone concentrations, corpus luteum (CL) area and vascular perfusion of the CL. Pregnancy rate did not differ (P > 0.1) between the early (3/3; 100%) and late (5/7; 71.4%) groups. Therefore, the characteristics of the follicle growth on the preceding days of ovulation were similar between the early- and late-ovulated mares and were consistent with the follicular dynamics expected in non-pregnant and non-lactating mares. However, when the data were analyzed for the days relative to parturition, greater follicle development was present in mares that ovulate earlier during the postpartum period (<10 days). The results suggest that important events may occur before parturition, resulting in early follicle development, mainly in those mares that show estrus and ovulate within 10 days postpartum.

Fertility and uterine hemodynamic in cows after artificial insemination with semen assessed by fluorescent probes.

Fluorescent probes (propidium iodide, Hoechst 33342, fluorescein isothiocyanate-conjugated Pisum sativum agglutinin, and 5,5',6,6'-tetrachloro-1,1',3,3'-tetraethylbenzimidazolcarbocyanine iodide) were used to simultaneously evaluate the integrity of plasma and acrosomal membranes as well as mitochondrial membrane function in cryopreserved bovine semen and to verify its influence on fertility and postinsemination uterine vascularization. One hundred eighty-two Nellore cows were distributed for artificial insemination (AI) using semen batches separated according to the cell percentage presenting intact plasma membrane, intact acrosome, and high mitochondrial function (IPIAH): group G (44.5% IPIAH, n = 68), group M (23.0% IPIAH, n = 56), and group R (8.5% IPIAH, n = 58). The uterine hemodynamic was evaluated by Doppler sonogram in three periods: 30 hours before AI, 4 and 24 hours after AI were considered the resistance index and the uterine vascularization score. The pregnancy rate of group G (64.7%) was greater (P > 0.05) compared with group R (36.2%), but both did not differ from group M (50.0%). There was no effect (P > 0.05) of semen quality on uterine vascularization. Greater vascularization was noticed 4 hours after AI than 30 hours before and 24 hours after AI. Semen evaluation using fluorescent probes contributes to predicting fertilizing potential of semen. The use of semen with less percentage of IPIAH sperm does not alter uterine hemodynamic in cows.

O uso de indicadores da fase pré-analítica como ferramenta da gestão laboratorial / The use of indicators in the pre-analytical phase as a laboratory management tool

Introduction: Efficient laboratory services are the basis of modern health systems. Scientific innovations have contributed to substantial improvements in the laboratory environment, but errors still persist. These errors are classified as pre-analytical, analytical and post-analytical, according to the time of occurrence. Objective: To evaluate the frequency of pre-analytical errors in the clinical laboratory service of a military hospital. Methods: A total of 329,582 tests were performed in the clinical laboratory of Hospital Naval Marcílio Dias (HNMD) from August to October 2012, and pre-analytical errors were documented. Results: The most frequent cause of the observed pre-analytical errors was hemolysis (27.54%), followed by samples not received (25.43%) and insufficient sample volume (18.49%). The samples from the Integrated Home Care Service (SIAD) showed the highest frequency of errors (3.38%), followed by those from the inpatient (0.76%) and the outpatient departments (0.21%). Conclusion: Our study demonstrates the importance of managing laboratory pre-analytical quality in order to ensure service excellence...

Introdução: Serviços laboratoriais efetivos são a base dos sistemas de saúde modernos. Inovações científicas têm contribuído para melhorias substanciais no meio laboratorial, mas os erros ainda persistem. Esses erros são classificados como pré-analíticos, analíticos e pós-analíticos, dependendo do momento de sua ocorrência. Objetivo: Avaliar a frequência de erros pré-analíticos ocorridos no serviço de análises clínicas de um hospital militar. Métodos: Um total de 329.582 exames foram realizados no laboratório de análises clínicas do Hospital Naval Marcílio Dias no período de agosto a outubro de 2012, e os erros pré-analíticos documentados. Resultados: Os erros pré-analíticos mais observados foram decorrentes da hemólise (27,54%), seguidos de material não recebido (25,43%) e amostra insuficiente (18,49%). As amostras oriundas do Serviço Integrado de Atendimento Domiciliar (SIAD) foram as que apresentaram a maior frequência de erros (3,38%), seguidas pelo setor de pacientes internos (0,76%) e ambulatoriais (0,21%), respectivamente. Conclusão: Nosso estudo demonstra a importância da gestão da fase pré-analítica na garantia da qualidade laboratorial, de maneira a assegurar um serviço de excelência...

Characteristics of the equine embryo and fetus from days 15 to 107 of pregnancy.

In spite of numerous, substantial advances in equine reproduction, many stages of embryonic and fetal morphological development are poorly understood, with no apparent single source of comprehensive information. Hence, the objective of the present study was to provide a complete macroscopic and microscopic description of the equine embryo/fetus at various gestational ages. Thirty-four embryos/fetuses were aged based on their crown rump length (CRL), and submitted to macroscopic description, biometry, light and scanning microscopy, as well as the alizarin technique. All observed developmental changes were chronologically ordered and described. As examples of the main observed features, an accentuated cervical curvature was observed upon macroscopic examination in all specimens. In the nervous system, the encephalic fourth ventricle and the encephalic vesicles forebrain, midbrain, and hindbrain, were visualized from Day 19 (ovulation = Day 0). The thoracic and pelvic limbs were also visualized; their extremities gave rise to the hoof during development from Day 27. Development of other structures such as pigmented optical vesicle, liver, tail, cardiac area, lungs, and dermal vascularization started on Days 25, 25, 19, 19, 34, and 35, respectively. Light and scanning microscopy facilitated detailed examinations of several organs, e.g., heart, kidneys, lungs, and intestine, whereas the alizarin technique enabled visualization of ossification. Observations in this study contributed to the knowledge regarding equine embryogenesis, and included much detailed data from many specimens collected over a long developmental interval.

Analgesia controlada pelo paciente com morfina ou fentanil no pós-operatório de reconstruçäo de ligamentos do joelho: estudo comparativo / Patient controlled analgesia with morphine or fentanyl in knee ligament reconstruction postoperative period: a comparative study

Justificativa e objetivos - a morfina ou o fentanil têm sido utilizados para analgesia pós-operatória, por via peridural, de forma contínua com bomba de infusäo, permitindo também que a analgesia seja controlada pelo paciente (ACP). O objetivo deste estudo foi avaliar a qualidade da analgesia, o consumo de opióides e os efeitos colaterais do emprego de infusäo contínua e de ACP com morfina ou fentanil, no pós-operatório de pacientes submetidos a reconstruçäo de ligamento do joelho, sob anestesia peridural com bupivacaína associada ao fentanil. Método - participaram do estudo 42 pacientes com idade entre 16 e 50 anos, estado físico ASA I e II, divididos aleatoriamente em dois grupos de 21: Grupo M (Morfina) e Grupo F (Fentanil). Todos os pacientes foram submetidos a anestesia peridural com bupivacaína a 0,5 por cento (100 mg) associada a fentanil (100 µg). Ao final da cirurgia os pacientes receberam morfina (Grupo M) ou fentanil (Grupo F) em regime de ACP, por via peridural, através de bomba de infusäo. No Grupo M foi utilizada soluçäo fisiológica (93 ml) contendo morfina 2 mg (2ml) e bupivacaína 0,5 por cento (5 ml). No Grupo F foi utilizada soluçäo fisiológica (85 ml) contendo fentanil 500 µg (10 ml), bupivacaína (5 ml a 5 por cento) sem adrenalina. Para os dois grupos foi programado inicialmente infusäo contínua de 4 ml.h elevado a menos um, com doses de 2 ml em bolus liberado pelo paciente num intervalo de 30 minutos. Foram comparados os seguintes parâmetros: dor; número de bolus acionados; consumo de opióides; bloqueio motor; sedaçäo e efeitos colaterais (prurido, náuseas, vômitos e retençäo urinária). Resultados - Näo houve necessidade de suplementaçäo anestésica no per-operatório. A maioria dos pacientes (85,7 por cento) obteve excelente grau de analgesia pós-operatória (EAV 0a 2), sem diferença significativa entre os grupos. A média de bolus acionados näo diferiu nos dois grupos. O consumo médio de morfina foi de 1,663 ñ 0,384 mg sendo o de fentanil de 0,435 ñ 0,083 mg. Näo houve bloqueio motor após a instituiçäo da ACP. A incidência de sedaçäo e de outros efeitos colaterais foi também significativamente maior no grupo da morfina