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ObjectiveTo investigate the efficacy of automated control of inspired oxygen (FiO2) by Predictive Intelligent Control of Oxygenation (PRICO) on the Fabian ventilator in maintaining oxygen saturation (SpO2) in preterm infants on high flow nasal cannula (HFNC) support.DesignSingle-centre randomised two-period crossover study.SettingTertiary neonatal intensive care unit.Patients27 preterm infants (gestational age (GA) 0.25.InterventionA 24-hour period on automated FiO2-control with PRICO compared with a 24-hour period on routine manual control (RMC) to maintain a SpO2 level within target range of 88%–95% measured at 30 s intervals.Main outcome measuresPrimary outcome: time spent within target range (88%–95%). Secondary outcomes: time spent above and below target range, in severe hypoxia (SpO2 98%), mean SpO2 and FiO2 and manual FiO2 adjustments.Results15 patients received PRICO-RMC and 12 RMC-PRICO. The mean time within the target range increased with PRICO: 10.8% (95% CI 7.6 to 13.9). There was a decrease in time below target range: 7.6% (95% CI 4.2 to 11.0), above target range: 3.1% (95% CI 2.9 to 6.2) and in severe hypoxia: 0.9% (95% CI 1.5 to 0.2). We found no difference in time spent in severe hyperoxia. Mean FiO2 was higher during PRICO: 0.019 (95% CI 0.006 to 0.030). With PRICO there was a reduction of manual adjustments: 9/24 hours (95% CI 6 to 12).ConclusionIn preterm infants on HFNC support, automated FiO2-control by PRICO is superior to RMC in maintaining SpO2 within target range. Further validation studies with a higher sample frequency and different ventilation modes are needed.

The consistory was a crucial institution in early modern Reformed churches. This article examines the nominations and elections of elders and deacons in the Dutch Reformed consistories of Kampen and Wemeldinge, shedding light on who was being nominated and elected and how such processes functioned in these churches. In particular, research into the Kampen consistory records demonstrates the importance given to the office of elder despite little theological backing for such a hieararchy; this was true to a lesser extent in Wemeldinge. In addition, the Kampen civil authorities played a significant role in the life of the consistory, most notably through the service of burgomasters as elders. The presence of burgomasters on the consistory is not present in Wemeldinge, indicating a more separate relationship between the church and state. In both Kampen and Wemeldinge, the elections of elders and deacons were unique and responded to the challenges and priorities of the individual contexts and communities.

ObjectiveTo evaluate incidence of minimally invasive surfactant therapy (MIST) failure, identify risk factors and assess the impact of MIST failure on neonatal outcome.DesignRetrospective cohort study. MIST failure was defined as need for early mechanical ventilation (SettingTwo tertiary neonatal intensive care centres in the Netherlands.PatientsInfants born between 24 weeks’ and 31 weeks’ gestational age (GA) (n=185) with MIST for respiratory distress syndrome.InterventionsMIST procedure with poractant alfa (100–200 mg/kg).Main outcome measuresContinuous positive airway pressure (CPAP) failure after MIST in the first 72 hours of life.Results30% of the infants failed CPAP after MIST. In a multivariate logistic regression analysis, four risk factors were independently associated with failure: GA Furthermore, MIST failure was independently associated with an increased risk of severe intraventricular haemorrhage.ConclusionWe observed moderate MIST failure rates in concordance with the results of earlier studies. Absence of corticosteroids and lower surfactant dose are risk factors for MIST failure that may be modifiable in order to improve MIST success and patient outcome.

Preterm birth is associated with increased mortality and morbidity. Tocolytic drugs, such as indomethacin, are often used to postpone preterm delivery. Indomethacin has been proven to be more effective than other tocolytic agents in terms of delaying birth but is often prescribed with caution because of its potential association with adverse neonatal outcomes. We aim to study the effects of antenatal indomethacin on neonatal outcomes after controlling for potential confounders, as compared to nifedipine and/or atosiban.In this cohort study, we performed a retrospective analysis of maternal and neonatal data. Women were included if they received indomethacin, nifedipine or atosiban as a tocolytic drug for imminent preterm labor and gave birth at a gestational age (GA) between 23Four hundred seventy-four women, delivering 610 infants were investigated. The incidence of the following adverse neonatal outcomes were significantly higher after indomethacin use: neonatal death (The presumed association between antenatal indomethacin exposure and several adverse neonatal outcomes may be based upon indication bias. Taking important confounding factors, such as GA at birth and neonatal birth weight into account, antenatal indomethacin exposure does not result in a higher incidence of adverse neonatal outcomes. However, there may be a higher risk for spontaneous intestinal perforation.

Objective: The aim of this research was to assess the quality and inter- and intra-observer agreement of tracings obtained by three different techniques for uterine contraction monitoring: the external tocodynamometer (TOCO), the intrauterine pressure catheter (IUPC) and a recently introduced method based on electrohysterography (EHG). Study design: We included 150 uterine activity registrations from a previous prospective observational study (W3 study), conducted at Máxima Medical Centre in Veldhoven, the Netherlands. Term singleton pregnant women were simultaneously monitored with TOCO, IUPC and EHG during labor. Six clinicians, blinded to the source (TOCO, IUPC, or EHG) and subject, evaluated all tracings that were subsequently presented in random order. They annotated contractions and assigned each tracing a score for interpretability of 2 (good), 1 (moderate) or 0 (poor). To evaluate inter-observer agreement, we calculated kappa values for the qualitative assessment, and intraclass correlation coefficients (ICC) for the number of contractions annotated by clinicians. Four clinicians repeated this procedure to evaluate intra-observer agreement. Results: IUPC tracings received the highest quality rating, with a mean score of 1.95, followed by a mean score of 1.60 for EHG and 0.80 for TOCO (p < 0.05). Mean weighted kappa values were 0.63 for TOCO and 0.45 for EHG. The average number of contractions that was picked up by clinicians was 59.8 for the intrauterine pressure catheter, 49.8 for EHG and 26.4 for TOCO. The ICC of the intrauterine pressure catheter was significantly higher than the external methods, regarding both inter- and intra-observer agreement (0.98 and 0.99 respectively). Conclusion: IUPC recordings scored best regarding quality, inter- and intra-observer agreement. However, due to safety issues, in many countries this technique is not used anymore. The quality of TOCO was rated as poor and many contractions were missed as compared to the gold standard. From a clinical interpretational point of view, EHG is favorable to TOCO. EHG recordings were assigned higher quality scores, but with less agreement between clinicians. An explanation could be that EHG is a relatively new technique, while IUPC and the TOCO are being used for decades. Building experience with EHG (training) is therefore recommended.

Objective: To evaluate the influence of maternal obesity on the performance of external tocodynamometry and electrohysterography. Study design: In a 2-hour measurement during term labor, uterine contractions were simultaneously measured by electrohysterography, external tocodynamometry, and intra-uterine pressure catheter. The sensitivity was compared between groups based on obesity (non-obese/obese/morbidly obese) or uterine palpation (good/moderate/poor), and was correlated to maternal BMI and abdominal circumference. Result: We included 14 morbidly obese, 18 obese, and 20 non-obese women. In morbidly obese women, the median sensitivity was 87.2% (IQR 74–93) by electrohysterography and 45.0% (IQR 36–66) by external tocodynamometry (p < 0.001). The sensitivity of electrohysterography appeared to be non-influenced by obesity category (p = 0.279) and uterine palpation (p = 0.451), while the sensitivity of tocodynamometry decreased significantly (p = 0.005 and p < 0.001, respectively). Furthermore, the sensitivity of both external methods was negatively correlated with obesity parameters, being non-significant for electrohysterography (range p-values 0.057–0.088) and significant for external tocodynamometry (all p-values < 0.001). Conclusions: Electrohysterography performs significantly better than external tocodynamometry in case of maternal obesity.

Objective Current uterine monitoring techniques have major drawbacks that could be avoided when using electrohysterography for uterine monitoring. Recently, a new electrohysterography method has been developed, providing a real-time tocogram on standard cardiotocography monitors. The diagnostic characteristics of this novel method need to be determined and compared to conventional methods We hypothesised that electrohysterography can perform better than external tocodynamometry due to the adhesive properties of the contact electrodes (less motion sensitive), and the improved signal acquisition through subcutaneous tissue (less obesity sensitive). Study design In this prospective diagnostic accuracy study, uterine contractions of labouring women were simultaneously monitored by three different monitoring techniques: electrohysterography, external tocodynamometry, and intra-uterine pressure catheter as method of reference. We performed a two-hour measurement during first and/or second stage of term labour. The contractions of each method were automatically detected by a computer-based algorithm. As the applied method had not been described in literature before, an interim analysis was performed to minimise exposure to the invasive pressure catheter. The main outcome parameter was the sensitivity of electrohysterography in comparison to external tocodynamometry for uterine contraction detection, tested by the Wilcoxon signed rank test. Results Uterine contractions of 48 term labouring women were simultaneously monitored by electrohysterography, external tocodynamometry, and intra-uterine pressure catheter. The study was terminated after the interim analysis as the sensitivity of electrohysterography was significantly higher compared to external tocodynamometry: median 89.5% (interquartile range (IQR); 82–93) and 65.3% (IQR; 53–81) respectively, p

A side effect of radiotherapy in the head and neck area is a reduction of the capillary blood flow and with it, a reduction in local defenses. Depending on the duration and intensity of the radiation, hypoxia, hypocellularity and hypovascularity, may occur, resulting in an increased risk of infection. Hyposalivation, a commonly occurring phenomenon after radiotherapy, leads to a higher caries sensitivity. To keep oral health at an acceptable level as much as possible, teeth are checked by a dentist prior to radiotherapy. Non-essential teeth and teeth with pathology are extracted, in order to prevent future problems. Dental treatment in the area treated with radiation will nevertheless sometimes be necessary after radiotherapy. Because the risk of infection is high and may result in the loss of part of the jaw, antibiotic prophylaxis is started prior to invasive treatment. In general, amoxicillin 500 mg 3dd 1 is chosen for 14 days. After treatment, wound healing should be checked by the specialist.Radiotherapie in het hoofd-halsgebied heeft als neveneffect dat de doorbloeding en daarmee de afweer lokaal vermindert. Afhankelijk van de duur en de intensiteit van de bestraling kan onder andere hypoxie, hypocellular iteit en hypovasculariteit optreden, met een verhoogd infectie risico. Hyposalivatie, een veel voorkomend verschijnsel na radiotherapie, geeft een hogere gevoeligheid voor cariës. Om de mondgezondheid zoveel mogelijk op een aanvaardbaar peil te houden, wordt voorafgaand aan de radiotherapie het gebit gesaneerd. Niet vitale gebitselementen en gebitselementen met pathologie worden geëxtraheerd om toekomstige problemen te voorkomen. Toch zal na radiotherapie een tandheelkundige behandeling in het bestraalde gebied soms nodig zijn. Omdat het infectiegevaar zo groot is, en tot verlies van een deel van de kaak kan leiden, wordt voorafgaand aan invasieve behandelingen gestart met AB-profylaxe. In het algemeen wordt gekozen voor amoxicilline 500 mg 3dd gedurende 14 dagen. Na de behandeling dient controle op de wondgenezing door de specialist plaats te vinden.

Background: Data on long-term consequences of preterm birth on pain coping later in life are limited. Aim: To assess whether gestational age, birth weight and neonatal disease severity have effect on pain coping style in adolescents born preterm or with low birth weight.Study design: Observational, longitudinal study (Project On Preterm and SGA-infants, POPS-19).Subjects: We analyzed data of 537 adolescents at the age of 19 years, who were born at a gestational age < 32 weeks or with a birth weight < 1500 g.Outcome measures: Participants completed the pain coping questionnaire (PCQ) that assesses pain coping strategies in three higher-order factors: approach ("to deal with pain"), problem-focused avoidance ("to disengage from pain") and emotion-focused avoidance ("expression of pain"). Furthermore, their pain coping effectiveness, pain controllability and emotional reactions to pain were assessed. All participants completed an IQ testResults: Univariate analysis showed no significant correlation between length of stay, sepsis and necrotizing enterocolitis and any of the higher-order factors. Approach was only correlated with IQ Problem-focused avoidance was, in the multiple regression analysis (including gestational age, IVH and IQ), only correlated with IQ For emotion-focused avoidance (including birth weight, SGA, NH, respiratory support and IQ) three independent predictors remained: NH was positively correlated, while respiratory support and IQ were negatively correlated with emotion-focused avoidance.Conclusions: Early neonatal characteristics and neonatal disease severity have limited effect on pain coping style in adolescence. Higher IQ was associated with the use of adaptive coping strategies, while maladaptive strategies were used less. (C) 2016 Published by Elsevier Ireland Ltd.

Objective To evaluate preferences from patients and users on 3 uterine monitoring techniques, during labor. Study Design Women in term labor were simultaneously monitored with the intrauterine pressure catheter, the external tocodynamometer, and the electrohysterograph. Postpartum, these women filled out a questionnaire evaluating their preferences and important aspects. Nurses completed a questionnaire evaluating users' preferences. Results Of all 52 participating women, 80.8% preferred the electrohysterograph, 17.3% the intrauterine pressure catheter and 1.9% the external tocodynamometer. For these women, the electrohysterograph scored best regarding application and presence during labor (p Conclusion Patients prefer the electrohysterograph over the external tocodynamometer and the intrauterine pressure catheter, while healthcare providers report ambiguous results.

Chronic pain is common after groin hernia surgery, affecting approximately 10% of patients. The type of anesthesia has been shown to affect short term pain (within 30 days) where Local Anesthesia ( ...

Objective: To investigate whether incomplete umbilical cord blood gas (UCBG) analysis occurs more often than the incomplete reporting of the Apgar score, and risk factors associated with the incomplete values. Methods: A total of 8824 infants born alive after 26 weeks’ gestation between January 2009 and April 2013 were included. We extracted data on five-minute Apgar score, UCBG analysis, gestational age, mode of delivery, time of delivery and multiple pregnancy. Univariate and multivariable logistic regression analyses were performed. Results: Five-minute Apgar score was incomplete in 15 cases (0.2%) and UCBG analysis in 1960 cases (22.2%), p

Elevated atmospheric CO2 levels and increasing nitrogen deposition both stimulate plant production in terrestrial ecosystems. Moreover, nitrogen deposition could alleviate an increasing nitrogen limitation experienced by plants exposed to elevated CO2 concentrations. However, an increased rate of C flux through the soil compartment as a consequence of elevated CO2 concentrations has been suggested to limit C sequestration in terrestrial ecosystems, questioning the potential for terrestrial C uptake to mitigate increasing atmospheric CO2 concentrations. Our study used data from 77 published studies applying elevated CO2 and/or N fertilization treatment to monitor carbon storage potential in grasslands, and considered the influence of management practices involving biomass removal or irrigation on the elevated CO2 effects. Our results confirmed a positive effect of elevated CO2 levels and nitrogen fertilization on plant growth, but revealed that N availability is essential for the increased C influx under elevated CO2 to propagate into belowground C pools. However, moderate nutrient additions also promoted decomposition processes in elevated CO2, reducing the potential for increased soil C storage. An important role was attributed to the CO2 response of root biomass in soil carbon responses to elevated CO2, since there was a lower potential for increases in soil C content when root biomass increased. Future elevated CO2 concentrations and increasing N deposition might thus increase C storage in plant biomass, but the potential for increased soil C storage is limited.

Background Tranexamic acid reduces blood loss and transfusion requirements in cardiac surgery but may increase the risk of coronary graft thrombosis. We previously reported the 30-day results of a trial evaluating tranexamic acid for coronary artery surgery. Here we report the 1-year clinical outcomes. Methods Using a factorial design, we randomly assigned patients undergoing coronary artery surgery to receive aspirin or placebo and tranexamic acid or placebo. The results of the tranexamic acid comparison are reported here. The primary 1-year outcome was death or severe disability, the latter defined as living with a modified Katz activities of daily living score of less than 8. Secondary outcomes included a composite of myocardial infarction, stroke, and death from any cause through to 1 year after surgery. Results The rate of death or disability at 1 year was 3.8% in the tranexamic acid group and 4.4% in the placebo group (relative risk, 0.85; 95% confidence interval, 0.64-1.13; P = .27), and this did not significantly differ according to aspirin exposure at the time of surgery (interaction P = .073). The composite rate of myocardial infarction, stroke, and death up to 1 year after surgery was 14.3% in the tranexamic acid group and 16.4% in the placebo group (relative risk, 0.87; 95% CI, 0.76-1.00; P = .053). Conclusions In this trial of patients having coronary artery surgery, tranexamic acid did not affect death or severe disability through to 1 year after surgery. Further work should be done to explore possible beneficial effects on late cardiovascular events.

The LH surge induces a multitude of events that are essential for ovulation and corpus luteum formation. The transcriptional responses to the LH surge of preovulatory granulosa cells (GCs) are complex and still poorly understood. In this study, a genome-wide bovine oligo array was used to determine how the gene expression profile of GCs is modulated by the LH surge. GCs from three different stages were used to assess the short- and long-term effects of this hormone on follicle differentiation: 1) 2 h before induction of the LH surge, 2) 6 h and 3) 22 h after the LH surge. The results obtained were a list of differentially expressed transcripts for each GC group. To provide a comprehensive understanding of the processes at play, biological annotations were used to reveal the different functions of transcripts, confirming that the LH surge acts in a temporal manner. The pre-LH group is involved in typical tasks such as cell division, development, and proliferation, while the early response to the LH surge included features such as response to stimulus, vascularization, and lipid synthesis, which are indicative of cells preparing for ovulation. The late response of GCs revealed terms associated with protein localization and intracellular transport, corresponding to the future secretion task that will be required for the transformation of GCs into corpus luteum. Overall, results described in this study provide new insights into the different transcriptional steps that GCs go through during ovulation and before luteinization.

Cumulus cells (CCs) are essential for oocytes to reach full development competency and become fertilized. Many major functional properties of CCs are triggered by gonadotropins and governed by the oocyte. Consequently, cumulus may reflect oocyte quality and is often used for oocyte selection. The most visible function of CCs is their ability for rapid extracellular matrix expansion after the LH surge. Although unexplained, LH induces the final maturation and improves oocyte quality. To study the LH signaling and gene expression cascade patterns close to the germinal vesicle breakdown, bovine CCs collected at 2 h before and 6 h after the LH surge were hybridized to a custom-made microarray to better understand the LH genomic action and find differentially expressed genes associated with the LH-induced oocyte final maturation. Functional genomic analysis of the 141 overexpressed and 161 underexpressed clones was performed according to their molecular functions, gene networks, and cell compartments. Following real-time PCR validation of our gene lists, some interesting pathways associated with the LH genomic action on CCs and their possible roles in oocyte final maturation, ovulation, and fertilization are discussed. A list of early potential markers of oocyte competencyin vivoandin vitrois thereafter suggested. These early biomarkers are a preamble to understand the LH molecular pathways that trigger the final oocyte competence acquisition process in bovine.

Summary Transvaginal ultrasound-guided follicle aspiration was performed repeatedly in 5 Dutch Warmblood mares during a time span of 3 months (mid-June-mid-September 1992). During 24 aspiration sessions (4–7/mare), 200 follicles were aspirated and 34 oocytes retrieved, giving oocyte recovery rates of 12.3% and 24.4% when using single channel needles or double barrel needles respectively. There was no significant effect of cycle stage on oocyte recovery rates. Follicle aspiration performed without any hormonal pretreatment gave significantly higher oocyte recovery rates of 26.9%, compared with 8.2 and 15.3% for pretreatments with prostaglandin F2α and human chorionic gonadotrophin respectively. Measurements of daily plasma progesterone concentrations in the mares revealed that follicle aspiration induced luteinisation of the follicles within 1–6 days after aspiration, resulting in a progesterone pattern similar to that of a normal oestrous cycle with ovulation when follicles were aspirated during oestrus, or in prolonged dioestrus in the mares in which follicle aspiration was performed during the luteal phase. The procedure was well tolerated by the animals and no clinical side effects were noted. This technique provides a non-invasive, relatively simple method for the repeated puncture of multiple immature and preovulatory follicles in the mare and the possibility of obtaining up to 6 oocytes per mare in a single follicle aspiration session.

The present paper describes LH and FSH secretion, follicle development and ovulation in sows that were subjected to a limited nursing regimen. From Day 14 of lactation, 32 sows were separated from their piglets for 12 h every day (intermittent suckling; IS). Half the sows had boar contact during separation. Nine of 32 sows ovulated spontaneously within 14 days from initiation of IS. The frequency of LH pulses on the first day of IS tended to be higher in anovulatory sows (6.3 v. 4.2 pulses per 12 h; P < 0.10); other characteristics of LH secretion were similar to sows that ovulated. The characteristics of FSH secretion did not differ over the 8-h sampling period. Boar contact did not influence either LH and FSH secretion or the number of sows that ovulated. Up to 58% of anovulatory sows showed an increase in follicle size after initiation of IS and, 4 days after the initiation of IS, one-third still had follicles similar in size to those in ovulatory sows. However, the oestradiol concentration in anovulatory sows did not increase. We conclude that FSH and LH stimulation in anovulatory sows is not limiting for normal follicle development, but that ovarian follicles are not responsive to increased LH secretion.

Objective: To investigate the reliability of length measurements of the mandible by comparing orthopantomograms (OPTs) with lateral cephalograms. Design: Observational study. Setting: OPTs and lateral cephalograms were taken of 20 human dry skulls. Four orthodontists and four maxillofacial surgeons located landmarks on all radiographs using a computer program for cephalometric measurements. Intraobserver and interobserver variability in locating landmarks was assessed, as well as positioning of the skulls prior to radiography between the x-ray assistants. Magnification differences between the left and right side of the mandible on the OPT were determined for five skulls. Kappa statistics were used to calculate the intraclass correlation coefficient for intraobserver and interobserver differences. An F test was used to assess differences between methods and between type of observer. Results: No significant differences were found in the magnification factor of the left and right side of the mandible. Compared with a lateral cephalogram, the OPT had comparable reliability in measuring mandibular distances condylion-gonion, gonion-menton, and condylion-menton. No significant differences were observed between the x-ray assistants in taking the OPTs and lateral cephalograms or in repositioning the skulls. Significant differences were found between orthodontists and maxillofacial surgeons for landmark measurements. Conclusion: An OPT is as reliable as a lateral cephalogram for linear measurements of the mandible (condylion-gonion, gonion-menton, and condylion-menton).

Zusammenfassung:Gegenstand und Ziel: Zuchthündinnen der Rasse Deutscher Schäferhund weisen relativ häufig einen kurzen Sexualzyklus von 3–4 Monaten bei vorberichtlicher Suboder Infertilität auf. Ziel der vorliegenden Studie war, durch Analyse gelbkörperund graviditätsrelevanter Hormone bei solchen Tieren und normal zyklischen Hündinnen gegebenenfalls bestehende hormonelle Dysregulationen aufzudecken. Material und Methoden: 22 nicht tragende und 19 tragende Hündinnen der Rasse Deutscher Schäferhund wurden jeweils in Gruppen mit normalem (Dauer > 6 Monate, Kontrolle) und kurzem Läufigkeitsintervall (< 5 Monate) eingeteilt. Während der Lutealphase bzw. Gravidität wurden die Konzentrationen von Progesteron, Prolaktin und Relaxin bestimmt. Ergebnisse: Die nicht graviden Hündinnen mit verkürztem Zyklus wiesen signifikant geringere Progesteronkonzentrationen auf als die normozyklischen Tiere. Bei den tragenden Hündinnen mit verkürztem Zyklus war es aufgrund einer durchgeführten Progesteronsubstitution nicht möglich, die Gelbkörperaktivität zu charakterisieren. Bei diesen Tieren ergaben sich von Tag 20 bis 60 signifikant geringere Prolaktinund Relaxinkonzentrationen als bei den graviden Kontrolltieren. Zudem bestand eine erhöhte Inzidenz an Fruchtverlusten. Schlussfolgerung: Die Ergebnisse deuten auf eine herabgesetzte Gelbkörperfunktion bei den kurzzyklischen Hündinnen sowohl in der autonomen als auch in der prolaktinabhängigen Lutealphase hin, die wahrscheinlich auf einem hypophysären Mangel beruht. Klinische Relevanz: Die relativ hohe Inzidenz kurzer Zyklen innerhalb einer Rasse spricht für einen genetischen Hintergrund. Die tierärztliche Betreuung solcher Hündinnen sollte primär auf die sorgfältige Diagnostik der Gelbkörperfunktion zur Selektion von Tieren mit intakten Ovarien für den Zuchteinsatz gerichtet sein.

The basal and gonadotropin releasing hormone (GnRH)-induced plasma concentrations of follicle stimulating hormone (FSH) and luteinizing hormone (LH) were studied in four anestrous and four ovariectomized (OVX) bitches. Blood samples were obtained via jugular venipuncture 40min before and 0, 10, 20, 30, 60, 90, and 120min after the i.v. administration of synthetic GnRH in a dose of 10microg/kg body weight. The basal plasma FSH and LH concentrations were significantly higher in the OVX bitches than in the anestrous bitches. In the anestrous bitches, the plasma FSH concentration was significantly higher than the pretreatment level at 10, 20, and 30min, whereas the plasma LH concentration was significantly elevated at 10 and 20min. The maximal GnRH-induced plasma FSH concentration in the anestrous bitches did not surpass the lowest plasma FSH concentration in the OVX bitches, whereas the GnRH-induced plasma LH concentrations in the anestrous bitches overlapped with the basal plasma LH concentrations in the OVX bitches. In the OVX bitches, GnRH administration did not induce a significant change in the plasma FSH concentration, whereas the plasma LH concentration increased significantly at 10 and 20min. In conclusion, the results of the present study indicate that in anestrous bitches GnRH challenge results in increased plasma levels of both FSH and LH, whereas in the OVX bitches, in which the basal plasma FSH and LH concentrations are higher, only a rise in the plasma LH concentration is present after GnRH stimulation. The results also suggest that a test to measure plasma concentration of FSH in single samples appears to have potential in verification of neuter status in bitches.

This paper presents LH, estradiol, and cortisol in 12 sows that were separated from their piglets for 12 h a day, beginning around 2 w of lactation, until weaning (intermittent suckling, IS). To separate sows from their piglets, the sows either were moved to a different unit (total separation), or were only inhibited from suckling their piglets by a physical barrier (physical separation). Blood samples were frequently collected during 4.5 consecutive days. At the start of IS, four sows showed advanced follicle growth. In the eight remaining sows, total separation resulted in 4/4 sows ovulating, while physical separation resulted in 2/4 sows ovulating. Total and physical separation resulted in different LH secretion patterns. Total separation resulted in a lower amplitude of LH pulses than physical separation throughout the period of sampling (0.26 versus 0.53 ng/ml; P

The aim of this study was to investigate the effects of treatment with medroxyprogesterone acetate (MPA) on canine adenohypophyseal function. Five Beagle bitches were treated with MPA (10mg/kg, every 4 weeks) and their adenohypophyseal function was assessed in a combined adenohypophyseal function test. Four hypophysiotropic hormones (CRH, GHRH, GnRH, and TRH) were administered before and 2, 5, 8, and 11 months after the start of MPA treatment, and blood samples for determination of the plasma concentrations of ACTH, cortisol, GH, IGF-1, LH, FSH, prolactin, alpha-MSH, and TSH were collected at -15, 0, 5, 10, 20, 30, and 45 min after suprapituitary stimulation. MPA successfully prevented the occurrence of estrus, ovulation, and a subsequent luteal phase. MPA treatment did not affect basal and GnRH-induced plasma LH concentrations. The basal plasma FSH concentration was significantly higher at 2 months after the start of MPA treatment than before or at 5, 8, and 11 months after the start of treatment. The maximal FSH increment and the AUC for FSH after suprapituitary stimulation were significantly higher before treatment than at 5, 8, and 11 months of MPA treatment. Differences in mean basal plasma GH concentrations before and during treatment were not significant, but MPA treatment resulted in significantly elevated basal plasma IGF-1 concentrations at 8 and 11 months. MPA treatment did not affect basal and stimulated plasma ACTH concentrations, with the exception of a decreased AUC for ACTH at 11 months. In contrast, the maximal cortisol increment and the AUC for cortisol after suprapituitary stimulation were significantly lower during MPA treatment than prior to treatment. MPA treatment did not affect basal plasma concentrations of prolactin, TSH, and alpha-MSH, with the exception of slightly increased basal plasma TSH concentrations at 8 months of treatment. MPA treatment did not affect TRH-induced plasma concentrations of prolactin and TSH. In conclusion, the effects of chronic MPA treatment on adenohypophyseal function included increased FSH secretion, unaffected LH secretion, activation of the mammary GH-induced IGF-I secretion, slightly activated TSH secretion, suppression of the hypothalamic-pituitary-adrenocortical axis, and unaffected secretion of prolactin and alpha-MSH.

In this study we investigated the temporal relationship between ovulation, egg formation, oviposition and the changes in plasma concentrations of progesterone, luteinizing hormone and estradiol-17β during the egg laying cycle in farmed ostriches. In 10 egg-producing birds, transcutaneous ultrasound scanning was performed at 3 h intervals and blood sampling at hourly intervals during a period of at least 48 h (one egg laying cycle). In hens (n = 8) that ovulated during the observational period, the ovulated egg was first detected 2 h after oviposition; thus, ovulation occurred shortly after oviposition in all birds. During the period between two consecutive ovipositions, the developing egg remained for 9 h in the proximal part (infundibulum, magnum or isthmus) and for 39 h in the distal part of the oviduct (uterus). In ovulating hens, plasma progesterone concentrations showed a characteristic and consistent profile: from basal levels of around 0.1 ng/ml concentrations started to increase 12 h before oviposition, reached an average maximum of 3.5 ng/ml at 3 h before oviposition and returned to basal levels 3 h and 30 min after oviposition. Changes in plasma luteinizing hormone and estradiol-17β concentrations showed comparable patterns of elevation and decline relative to the timing of oviposition and ovulation. However, variation in their individual basal concentrations was generally larger and peak values were less conspicuous than those of progesterone. In non-ovulating hens (n = 2) neither progesterone, nor luteinizing hormone nor estradiol-17β showed elevations to peak concentrations before oviposition. These data demonstrate that during the egg laying cycle of ostriches, events such as ovulation, egg development and oviposition evolve according to a rather strict time schedule, and that progesterone, luteinizing hormone and estradiol-17β reach peak concentrations shortly before ovulation. Additionally, our findings also show that on-farm ultrasound scanning is a useful technique to discriminate between ovulating and non-ovulating hens.

The possibility that differences in hormonal regimes between the two oviducts in the cow around ovulation affects secretory activity of the oviduct epithelial cells and/or sperm-oviduct binding was studied. Oviducts were collected immediately after slaughter at 6 hr before to 5 hr after timed ovulation of 14 normally cyclic cows that had been inseminated (n = 6) or not (n = 8) and material obtained from the same cows was processed in three ways. First, in vivo, after artificial insemination of the cows, low numbers of sperm cells (approx. 15 per oviduct) were found within the entire oviducts as observed by scanning electron microscopy (SEM). Almost all sperm were located in the isthmus and then only on ciliated cells and showed without exception fully matured, intact morphology. Secretory activity of noninseminated oviduct epithelia was induced after ovulation which was most predominant in the pockets of the ipsi-lateral ampulla compared to the contra-lateral ampulla (P < 0.01). Second, ex vivo, explants dissected from oviducts of the noniseminated cows were incubated with sperm. In all cases, the sperm bound to the explants in a similar pattern as observed in vivo and this binding was strictly fucose-dependent. The main difference with in vivo experiments was the high numbers of sperm bound at any site of the oviduct ( approximately 3,000 cells per mm(2)) indicating the high sperm binding capacity of the oviduct epithelia. Ovulation induced a striking drop in sperm binding capacity in the oviducts and was most pronounced in the isthmus ( approximately 1,300 cells per mm(2); P < 0.001) and to a lesser extent in the ampulla ( approximately 2,000 cells per mm(2), P < 0.01). Third, in vitro, pieces of tissue dissected from oviducts of the noninseminated cows were cultured to mono-layers. Culturing epithelial cells resulted in loss of their normal morphological appearance. In all cases, the sperm binding capacity in monolayers was very low (

In the present study, the pulsatile serum profiles of prolactin, LH and testosterone were investigated in eight clinically healthy fertile male beagles of one to six years of age. Serum hormone concentrations were determined in blood samples collected at 15 min intervals over a period of 6 h before (control) and six days before the end of a four weeks treatment with the dopamine agonist cabergoline (5 microg kg(-1) bodyweight/day). In addition, the effect of cabergoline administration was investigated on thyrotropin-releasing hormone (TRH)-induced changes in the serum concentrations of these hormones. In all eight dogs, the serum prolactin concentrations (mean 3.0 +/- 0.3 ng ml(-1)) were on a relatively constant level not showing any pulsatility, while the secretion patterns of LH and testosterone were characterised by several hormone pulses. Cabergoline administration caused a minor but significant reduction of the mean prolactin concentration (2.9 +/- 0.2 ng ml(-1), p < 0.05) and did not affect the secretion of LH (mean 4.6 +/- 1.3 ng ml(-1) versus 4.4 +/- 1.7 ng ml(-1)) or testosterone (2.5 +/- 0.9 ng ml(-1) versus 2.4 +/- 1.2 ng ml(-1)). Under control conditions, a significant prolactin release was induced by intravenous TRH administration (before TRH: 3.8 +/- 0.9 ng ml(-1), 20 min after TRH: 9.1 +/- 5.9 ng ml(-1)) demonstrating the role of TRH as potent prolactin releasing factor. This prolactin increase was almost completely suppressed under cabergoline medication (before TRH: 3.0 +/- 0.2 ng ml(-1), 20 min after TRH: 3.3 +/- 0.5 ng ml(-1)). The concentrations of LH and testosterone were not affected by TRH administration. The results of these studies suggest that dopamine agonists mainly affect suprabasal secretion of prolactin in the dog.

Compared with other domestic animals, relatively little is known about the changes in, and temporal relations between, reproductive hormones around the time of ovulation in the domestic bitch. Therefore, plasma concentrations of luteinizing hormone (LH), follicle-stimulating hormone (FSH), estradiol-17beta, progesterone, prolactin (PRL), and alpha-melanocyte-stimulating hormone (alpha-MSH) were determined one to six times daily from the start of the follicular phase until 5 days after the estimated day of ovulation in six Beagle bitches. In all bitches, the pre-ovulatory LH surge was accompanied by a pre-ovulatory FSH surge. A pre-ovulatory PRL or alpha-MSH surge was not observed. The pre-ovulatory FSH and LH surges started concomitantly in four bitches, but in two bitches the FSH surge started 12 h earlier than the LH surge. The FSH surge (110+/-8 h) lasted significantly longer than the LH surge (36+/-5 h). In contrast with the pre-ovulatory FSH surge, the pre-ovulatory LH surge was bifurcated in four of six bitches. The mean plasma LH concentrations before (1.9+/-0.4 microg/L) and after (1.9+/-0.3 microg/L) the LH surge were similar, but the mean plasma FSH concentration before the FSH surge (1.6+/-0.3 U/L) was significantly lower than that after the FSH surge (3.1+/-0.2 U/L). In most bitches the highest plasma estradiol-17beta concentration coincided with or followed the start of the pre-ovulatory LH surge. In five of the six bitches the plasma progesterone concentration started to rise just before or concurrently with the start of the LH surge. In conclusion, the results of this study provide evidence for the differential regulation of the secretion of LH and FSH in the bitch. In addition, the interrelationship of the plasma profiles of estradiol-17beta and LH suggests a positive feedback effect of estradiol-17beta on LH surge release. The start of the pre-ovulatory LH surge is associated with an increase in the plasma progesterone concentration in this species.

Transcription is arrested in the bovine oocyte within the first few hours of in vitro maturation, thus the stored maternal mRNAs accumulated in the oocyte are essential to sustain development until the Maternal-Zygotic Transition. In vivo matured oocytes have superior blastocyst formation rates than in vitro matured oocytes, suggesting that the mRNA content of these oocytes is of higher quality. To determine which transcripts may be associated with developmental competence, a Suppressive Subtractive Hybridization was performed between oocytes collected by ovariectomy at 6 hr post-LH surge and oocytes from slaughterhouse collected after 6 hr of maturation, resulting in a library enriched in these functionally important mRNAs. The clones were spotted onto a cDNA microarray and transcripts potentially associated with developmental competence were hybridized onto these slides. Hybridizations were performed with transcripts up-regulated in oocytes cultured for 6 hr in the presence or absence of rFSH in vitro, and secondly with transcripts up regulated in early-cleaving embryos versus those at the one-cell stage at 36 hr postfertilization. From these hybridizations, 13 candidates were selected. Their functional association with embryonic competence was validated by measuring their relative transcript levels by quantitative real-time PCR in eight different conditions: oocytes cultured with or without rFSH, early--versus late-cleaving embryos, and oocytes from different follicle sizes (1-3, 3-5, 5-8, and >8 mm of diameter). The gene candidates CCNB2, PTTG1, H2A, CKS1, PSMB2, SKIIP, CDC5L, RGS16, and PRDX1 showed a significant quantitative association with competence compared to BMP15, GDF9, CCNB1, and STK6.

Calves derived from IVP embryos may suffer from the large offspring syndrome that has been related to effects of in vitro culture on the intrinsic quality of the embryo. Limited information is available on the role of the placenta in such cases. In this study, bovine pregnancy-associated glycoprotein (bPAG) was used as a marker to test whether placental function is influenced by the route of embryo production. Therefore, from day 7 until day 119 of ongoing gestations, resulting from transfer of MOET ( n = 53), IVP-co-culture ( n = 21) and IVP–SOF ( n = 38) embryos, bPAG levels were compared in peripheral plasma of recipients. Plasma progesterone levels were compared as well. From day 25 of gestation onwards, bPAG could be detected in all recipients and the levels were significantly influenced by the day of gestation. Although IVP calves were significantly heavier than the in vivo produced calves, this difference was not reflected in the bPAG profiles of the embryo production groups. Yet, the mean bPAG level of the three last sampling moments (days 105–119) tended to be positively related to the birth weight of the calves, irrespective of the embryo production technique. Progesterone concentrations were not influenced by route of embryo production, but were significantly affected by parity of the recipient and day of gestation.

The present study aimed to test the generally accepted view that a follicular wave starts with follicles newly recruited from the population smaller than 3 mm, which later compete for dominance. According to this view, subordinate follicles are expected to be too atretic to join the next follicular wave. Ten cows were ovariectomized shortly prior to the LH surge, thus around the start of the first follicular wave of the cycle. Per cow, on average, 14.4 follicles of >/=3 mm were dissected. Follicular health was determined on the basis of four parameters: 1) judgment of the degree of atresia by stereomicroscope, 2) incidence of apoptotic nuclei among the granulosa cells, 3) estradiol and progesterone concentrations, and 4) insulin-like growth factor-I (IGF-I) binding proteins (IGFBPs)-2, -4, and -5 concentrations in the follicular fluid. In addition to the preovulatory follicle, 3.1 other follicles, mainly sized 3-4.5 mm, were found to be healthy based on the proportion of apoptotic nuclei, and concentrations of estradiol/progesterone, and IGFBPs. The ability of these follicles to respond with growth on the preovulatory and periovulatory FSH surges was supported by a comparison to the follicular population of four cows 31-68 h after the LH surge. The present results point to an alteration of the view on the follicular wave. The larger follicles during the first days of the follicular wave are, in general, derived from follicles that also joined the previous wave. A portion of these growing follicles are estradiol active and compete for dominance. Other growing follicles lack estradiol production and are probably derived from rather atretic follicles. The first newly recruited follicles do not reach the size of 3 mm before 31 h after the preovulatory FSH surge. At that time, the larger follicles are already competing for dominance.

In the present study, ribosomal RNA (rRNA) gene activation, monitored through nucleolus development, was studied by autoradiography following 3H-uridine incubation, transmission electron microscopy, and immunofluorescence confocal laser scanning microscopy of key nucleolar proteins involved in rRNA transcription (topoisomerase I, upstream binding factor, and RNA polymerase I) and processing (fibrillarin, nucleolin, and nucleophosmin) in in vivo developed, in vitro produced, and parthenogenetic bovine embryos. In general, in vivo developed embryos displayed formation of fibrillo-granular nucleoli during the 4th post-fertilization cell cycle. During the previous stages of development, nucleolus precursor bodies (NPBs) were observed. However, on some occasions the initial steps of nucleolus formation were observed already at the 2- and 4-cell stage in cases where such embryos were collected from superovulated animals together with later embryonic stages presenting nucleolar development and autoradiographic labeling. The in vitro produced embryos displayed very synchronous formation of fibrillo-granular nucleoli and autoradiographic labeling during the 4th cell cycle. In vivo developed and in vitro produced embryos displayed allocation of nucleolar proteins to fibrillar and granular compartments of the developing nucleoli during the 4th cell cycle. The parthenogenetic embryos typically displayed formation of fibrillo- granular nucleoli during the 5th cell cycle and autoradiographic labeling was not observed until the morula stage. Moreover, the 1-, 2-, and 4-cell parthenogenetic embryos practically lacked NPBs. On the other hand, parthenogenetic embryos displayed allocation of nucleoar proteins to nuclear entities during the 4th cell cycle. In conclusion, both in vivo developed and in vitro produced bovine embryos displayed activation of transcription and nucleolar development during the 4th cell cycle. However, in vivo developed embryos flushed together with later developmental stages displayed premature activation of these processes. Parthenogenetic bovine embryos, on the other hand, displayed a delayed activation. Mol. Reprod. Dev. 65: 73–85, 2003. © 2003 Wiley-Liss, Inc.

To determine the effect of group composition on backtest (Bt) responses and to determine the predictive value of the Bt for the physiological stress response to weaning and mixing, 814 pigs were backtested at 3, 10 and 17 days of age. Twenty-nine percent of all pigs were cross-fostered at 3 days according to Bt responses and groups were formed of animals with high responses (HR) only, low responses (LR) only or mixed groups of animals with high, intermediate and low responses (MISC). Original litters (OR, no cross-fostering) were used as controls. Cortisol responses were measured in saliva after weaning at 4 weeks of age and after moving and mixing at 9 weeks of age. In HR groups, mean Bt responses decreased after cross-fostering while in LR groups, mean Bt scores increased. In both groups, Bt responses of individual animals before and after cross-fostering were not correlated. In MISC and OR groups, all Bt scores were correlated. Weaning and mixing caused a significant rise in cortisol in all animals while moving or weighing did not. No relations were found between Bt scores and cortisol levels. We conclude that Bt behaviour can change according to the social environment between 3 and 10 days. This could be intentional, to form a varied group, or it might be caused by a change in the hypothalamic–pituitary–adrenal (HPA) function due to social stress. At an older age, this ability is lost and common farm practises such as regrouping, weaning and mixing of piglets at ages >10 days might have a negative effect on the piglets.

With the introduction of multiple ovulation, embryo recovery and transfer techniques (MOET) plus embryo freeze-thaw methods in the early 1980s, the breeding industry has the tools in hand to increase the number of calves from donors of high genetic merit. In the early 1990s, the introduction of ovum pick-up followed by in vitro embryo production (OPU-IVP) opened up even greater possibilities. Using these technologies, we challenge biological mechanisms in reproduction. Where normally one oocyte per estrous cycle will develop to ovulation, now numerous other oocytes that otherwise would have degenerated are expected to develop into an embryo. Completion of oocyte growth and pre-maturation in vivo before final maturation both appear to be essential phases in order to obtain competence to develop into an embryo and finally a healthy offspring. In order to increase oocyte quality and quantity in embryo production technologies, current procedures focus primarily on improving the homogeneity of the population of oocytes with regard to growth and state of pre-maturation at the start of a treatment. In the case of MOET, dominant follicle removal (DFR) before superovulation treatment improves the number of viable embryos per session from 3.9 to 5.4 in cows but not in heifers and a prolonged period of follicle development obtained by preventing release of the endogenous LH surge increases the number of ova but not the number of viable embryos per session. In the case of OPU-IVP, the frequency of OPU clearly affects quantity and quality of the collected oocytes and FSH stimulation prior to OPU every 2 weeks resulted in 3.3 embryos per session. Analysis of 7,800 OPU sessions demonstrated that the oocyte yield is dependent on the team, in particular, the technician manipulating the ovaries. It is concluded that an increased understanding of the processes of oocyte growth, pre- and final maturation will help to improve the efficiency of embryo technologies. However, somewhere we will meet the limits dictated by nature.

Bovine embryos produced in vitro differ substantially from embryos produced in vivo in the mRNA expression patterns of genes important for development. Several factors in the in vitro production systems have profound effects on embryonic mRNA expression patterns. The effects of the type of maturation on the expression pattern of genes important for development in blastocysts produced in vitro have not yet been investigated. The aim of the present study was to investigate the effects of various maturational protocols on the relative abundance of a panel of six marker genes, indicative of compaction and cavitation, metabolism, stress susceptibility and RNA processing, in bovine blastocysts produced in vitro. Four groups of blastocysts were analysed by a sensitive semi-quantitative RT-PCR assay. Blastocysts were produced in vitro from oocytes of different origin from: (1) 3-8 mm follicles; (2) preovulatory follicles before the LH surge; and (3) preovulatory follicles 24 h after the LH surge. The first two groups were matured in vitro, whereas the third group had undergone maturation in vivo. A fourth group comprised blastocysts developed entirely in vivo. Expression of glucose transporter 1 was significantly (P < 0.05) higher, and expression of desmocollin 2 and plakophilin tended to be higher (P < 0.1) for in vivo (group 4) compared with in vitro blastocysts (group 1), whereas no differences were found for heat shock protein 70.1, E-cadherin and poly(A) polymerase. Expression of the six transcripts did not differ among blastocysts produced in vitro from oocytes of groups 1, 2 and 3. Results indicate that alterations in the relative abundance of these transcripts in blastocysts produced in vitro cannot primarily be attributed to the origin of the oocyte, but are likely to have been induced by post-maturation or fertilization culture conditions.

We tested the challenge hypothesis for the hormonal regulation of aggression in chicks of the black-headed gull, Larus ridibundus. Chicks of this species are highly aggressive toward conspecifics, but never to peers that hatched from the same clutch (modal clutch size is three). Therefore, in the first experiment small families were housed together in large groups (challenged condition) and compared to families kept isolated (nonchallenged condition). As expected, in the challenged condition during the initial stage of territory establishment basal levels of testosterone M were clearly higher than those in the nonchallenged condition. In the second experiment we tested the effect of a short social challenge on short-term T-fluctuations. The design was based on an earlier experiment, showing that after temporary T-treatment chicks become very sensitive to social challenges while having low basal T-levels. We now show that these social challenges induce brief elevations in plasma T-levels. These peaks are similar to those in previously untreated chicks but untreated chicks do not respond with aggression to a challenge. Therefore, we conclude that the initial exposure to elevated T-levels increases the sensitivity to brief changes in T induced by social challenges. In this way exposure to T, that may be detrimental for development, is minimized while birds remain able to defend territories. This is the first report showing that the challenge hypothesis as established for adult birds, is also applicable for aggressive behavior in young birds outside the sexual context. Furthermore we suggest that a phase of priming with T is necessary to obtain the high behavioral responsiveness to a challenge. (C) 2002 Elsevier Science (USA).

The consequences of peri-parturient Monensin administration were studied in two dairy herds in ≥ 2 parity HF cows (1) calving in moderate body condition (score, BCS: 3.25-3.75) and producing ≥8500 kg FCM in the previous lactation (Herd K), as well as (2) in those being overfed before calving (BCS: ≥3.75), but producing less milk (5-8000 kg FCM) (Herd S). In both herds on days 249-256 of gestation half of the cows received an intraruminal constant release capsule of Monensin (Rumensin® capsule, ELANCO) (Monensin-K and Monensin-S cows), the others remained untreated controls (Control-K and Control-S cows). Blood samples were taken on day 1-3 after calving and again 4 times with 7 days intervals to assay certain hormones and metabolites. On days 1-3 and again on days 28-35 also the ACTH induced Cortisol response and TRH induced T4/T3 responses were determined. The resumption of ovarian cyclicity was followed by milk progesterone profiles. In the first 10 weeks after calving the milk production of Monensin-K cows (n=13) did not differ from that of Control-K cows (n=14). The cows lost their body weight continuously in the first 10 weeks of lactation in Herd S, whereas only in the first 5 weeks in Herd K. Monensin only slightly influenced the degree of weight loss (on both farms), and the circulating leptin level (determined only in Herd S). Significantly lower insulin, IGF-1, T4, T3, glucose and cholesterol levels as well as less elevated βOH-butyrate and non-esterified fatty acid concentrations were seen in Control-S (n=13) than in Monensin-S (n=ll) cows, first of all in the first two weeks after calving. These ionophor-related differences were less obvious or disappeared in Herd-K. Monensin shortened the postpartum acyclic and anestrous periods as well as the calving to re-conception interval but this effect was significant only in the Herd S.