Uterine fluid proteome changes during diapause and resumption of embryo development in roe deer (Capreolus capreolus).

The uterine microenvironment during pre-implantation presents a pro-survival milieu and is essential for embryo elongation in ruminants. The European roe deer (Careolus capreolus) pre-implantation embryo development is characterised by a 4-month period of reduced development, embryonic diapause, after which the embryo rapidly elongates and implants. We investigated the uterine fluid proteome by label-free liquid chromatography tandem mass spectrometry at four defined stages covering the phase of reduced developmental pace (early diapause, mid-diapause and late diapause) and embryo elongation. We hypothesised that embryo development during diapause is halted by the lack of signals that support progression past the blastocyst stage. Three clusters of differentially abundant proteins were identified by a self-organising tree algorithm: (1) gradual reduction over development; (2) stable abundance during diapause, followed by a sharp rise at elongation; and (3) gradual increase over development. Proteins in the different clusters were subjected to gene ontology analysis. 'Cellular detoxification' in cluster 1 was represented by alcohol dehydrogenase, glutathione S-transferase and peroxiredoxin-2. ATP-citrate synthase, nucleolin, lamin A/C, and purine phosphorylase as cell proliferation regulators were found in cluster 2 and 'cortical cytoskeleton', 'regulation of substrate adhesion-dependent cell spreading' and 'melanosome' were present in cluster 3. Cell cycle promoters were higher abundant at elongation than during diapause, and polyamines presence indicates their role in diapause regulation. This study provides a comprehensive overview of proteins in the roe deer uterine fluid during diapause and forms a basis for studies aiming at understanding the impact of the lack of cell cycle promoters during diapause.

Fatty acid profile of blood plasma and oviduct and uterine fluid during early and late luteal phase in the horse.

During early pregnancy, the secretome of both oviduct and uterus serves as exchange medium for signaling factors between embryo and mother and provides the embryo with nutrients. The preimplantation embryo can utilize the fatty acids (FA) therein via direct incorporation into cell membrane lipid bilayers and for energy production via ß-oxidation. The FA concentration and composition of the oviduct (OF) and uterine fluid (UF) might be regulated by ovarian hormones to meet the changing needs of the growing embryo. In our study, we analyzed the FA profile of blood plasma (BP) and reproductive fluid samples obtained post mortem from slaughtered mares by gas chromatography mass spectrometry. Cycle stage was determined by visual evaluation of the ovary and measurement of plasma progesterone. No major effect of cycle stage on the FA profile was observed. However, the composition of FA was different between BP and both OF and UF. While linoleic, stearic, oleic and palmitic acid were the four most prevalent FA in both BP and reproductive fluids, the latter contained higher concentrations of arachidonic, eicosapentaenoic and dihomo-γ-linolenic acid. The finding suggests selective endometrial transport mechanisms from plasma into the reproductive fluids or increased endometrial synthesis of selected FA.

Influence of tibial hybrid fixation on graft tension and stability in ACL double-bundle reconstruction.

PURPOSE: Initial graft tension in anterior cruciate ligament (ACL) reconstruction affects stability and tension loss at follow-up. This study investigated the influence of hybrid tibial fixation in 3-tunnel double-bundle ACL reconstruction on initial graft tension and tension change and stability under anterior and combined rotatory loads. METHODS: Eleven fresh-frozen cadaveric knees were reconstructed with an ACL double bundle using a 3-tunnel technique. Grafts were tightened to 80 N in 60° (AM bundle) and 15° (PL bundle) of flexion. Anterior tibial translation under 134 N of anterior shear load and translation under combined rotatory and valgus loads (10 Nm valgus stress, 4 Nm internal tibial torque) were determined at 0°, 30°, 60°, and 90° flexion. In addition, graft tension under continuous passive motion was determined. Intact, ACL-resected and ACL-reconstructed joints with either tibial extracortical graft fixation or extracortical plus supplemental aperture graft fixation (hybrid fixation) were tested. RESULTS: Hybrid fixation did not increase graft tension in either bundle during fixation or in motion without additional load. AM-bundle tension increased (p < 0.05) at 0° under combined rotatory and valgus loads and at 30° and 60° under both loading conditions without decreasing the anterior tibial translation. PL-bundle tension increased (p < 0.05) only at 90° under combined rotatory and valgus loads. CONCLUSIONS: Tibial hybrid fixation in 3-tunnel double-bundle ACL reconstruction increases time-zero AM- and PL-bundle tensions under loading conditions, generating greater construct stiffness. This could lead to a longer preservation of ACL-graft stability in clinical follow-up before bony incorporation.

[Magnetic resonance imaging and computed tomography : What is important in orthopedics and traumatology]. / Magnetresonanz- und Computertomographie : Indikationsstellung in Orthopädie und Unfallchirurgie.

BACKGROUND: Magnetic resonance imaging (MRI) and computed tomography (CT) are established complementary tools for cross-sectional imaging in addition to standard x­rays in orthopedics and traumatology. OBJECTIVE: Illustration of possible applications of MRI and CT in acute and chronic joint diseases. MATERIAL AND METHODS: Summary of the relevant literature with focus on the validity of MRI and CT in depicting joint trauma and pathologies. In addition, description of pitfalls in evaluation of the images. RESULTS: The main focus of CT is the detailed visualization of fractures and deformities; however, MRI is the primary imaging technique for depiction of soft tissue pathologies, such as ligament tears and chondral lesions. Both imaging techniques are valuable in orthopedics and traumatology and complement each other in the development of treatment algorithms. CONCLUSION: Both MRI and CT are essential and complementing cross-sectional imaging techniques in the diagnostic procedures for joint pathologies in orthopedics and traumatology.

Embryonic resorption in context to intragestational corpus luteum regression: a longitudinal ultrasonographic study in the European brown hare (Lepus europaeus PALLAS, 1778).

Embryonic resorption is frequently observed in polytocous mammals. Often it occurs as partial litter resorption affecting only single conceptuses of a whole litter. The aim of the study was to describe the incidence and morphology of embryonic resorption in the European brown hare (Lepus europaeus). In 154 pregnancies viable conceptuses, conceptuses undergoing resorption and CL of pregnancy were ultrasonographically monitored during the entire gestation period. Resorptions were classified into (1) "pre-implantation resorptions," (2) "peri-implantation resorptions," and (3) "post-implantation resorptions." The incidence of resorption in the pre-implantation period was 9%, in the peri-implantation period 9%, and in the post-implantation period 24%. Post-implantation resorptions were found up to late pregnancy stages when fetal development was already in progress. The highest daily incidence of resorption was on Day 8 of the 42-day pregnancy. In 91% of the cases, the regression of one CL was observed, while an embryo was undergoing resorption at the same time. The number of resorptions did not significantly differ from the number of CL in regression during gestation, suggesting an interesting one-resorption-to-one-regression relationship. The ultrasonographic appearance of the luteal regression during pregnancy was similar to the morphology characteristic for postpartal luteolysis.

In vivo tissue sampling of embryonic resorption sites using ultrasound guided biopsy.

In the polytocous European brown hare (Lepus europaeus) more than 23% of all successful implantations undergo embryonic resorption. The objective of the study was to establish a minimally invasive ultrasound guided biopsy technique to collect embryonic resorption tissue in vivo. The sampled material was genetically analysed to determine paternity and the sex of the embryo. Female hares were either mated or artificially inseminated and pregnancy was confirmed by ultrasound on day six post ovulation. Subsequent embryonic development was ultrasonographically monitored on a regular basis to detect embryos undergoing resorption. Cell material of the resorption site was collected under ultrasonographic control via transabdominal biopsy of the placenta or aspiration of resorption fluid. To avoid breathing movements during the biopsy, the animals were intubated and a short apnoea was evoked by assisted ventilation. The presence of embryonic cells in the biopsy material was confirmed by microsatellite analysis in 11 of the fluid samples (n = 28) and six of the placental samples (n = 8). The lower success rate in the fluid samples was attributed to the abundance of maternal cells which was confirmed by the analysis of fluid sample smears. Male sex of the embryos undergoing resorption was detected by SRY analysis for ten of the fluid samples and for one of the placental samples. The two biopsy techniques did not have any negative impact on the prenatal development of the healthy siblings nor did it influence the future breeding performance of the females that were biopsied.

Pregnancy monitoring in dogs and cats using 3D and 4D ultrasonography.

Three-dimensional (3D)/four-dimensional (4D) volume ultrasound is an established method in human medicine that offers various options for analysing and presenting ultrasound volume data. However, the successful application of the different 3D/4D imaging modalities in pregnant dogs and cats has not yet been reported in the literature. The main reasons for this are: (1) the high costs of 3D/4D ultrasound systems, (2) operation difficulties due to high breathing frequency in non-sedated animals and (3) the missing specific knowledge in veterinary medicine concerning how to perform high-quality volume scans. Automatically acquired ultrasound volume data sets were generated with two different ultrasound systems: the portable Voluson i and the stationary Voluson Expert 730. Different 3D/4D imaging modalities were tested in regard of their practicability in pregnancy monitoring in dogs and cats. Nine different volume imaging modalities were applied using the saved files. For the presentation of the static 3D volume data sets, we used the multiplanar, niche, surface, transparency, glass body, inversion, volume calculation and tomographic ultrasound imaging modes. For the dynamic 4D data, the surface and glass body modes were applied. By changing the human standard settings to the requirements of small animal anatomy, it was found that 3D/4D ultrasound has great potential for the characterization of pregnancy in queens and bitches. The 3D/4D technology offered advanced information about pregnancy status and birth prediction and improved the diagnostic confidence. By using standardized examination protocols, 3D/4D ultrasound will allow a reduction in examination time by generating even more relevant information. These benefits, combined with possible future cost reduction of commercial ultrasound systems, might lead to frequent utilization in routine pregnancy diagnostic and birth management in small animal practice.

Comparison of sonographically measured bowel wall vascularity, histology, and disease activity in Crohn's disease.

The purposes of this study was to provide a retrospective comparison of semiquantitatively measured bowel wall vascularity by power Doppler sonography, endoscopic-histopathological biopsy findings, and disease activity in patients with confirmed Crohn's disease. Thirty-two out of 1,332 patients with histologically confirmed Crohn's disease (18 female, 14 male; mean age 38.8 years) met the inclusion criteria: ileocolonoscopy with biopsy and power Doppler sonographic determination of bowel wall vascularity with assessment of disease activity within a period of 5 days. Sonographic determination of bowel wall vascularity was based on a semiquantitative score. Endoscopic bowel wall biopsy specimens were assessed using a self-developed inflammation score and the disease activity was calculated using Crohn's disease activity index (CDAI). A significant association (p < 0.05) was shown for results of histology and bowel wall vascularity in the terminal ileum (kappa = 0.66; sensitivity 95%; specificity 69%). There was no observed association between CDAI and histology, although there was an association between CDAI and bowel wall vascularity (sensitivity 82%). Increased bowel wall vascularity in the terminal ileum measured by power Doppler ultrasound reflects inflammatory activity in histologically examined bowel wall. Power Doppler ultrasound may be able to monitor activity changes of the bowel wall determined by pharmaceutical treatment.

Early embryo development in the elephant assessed by serial ultrasound examinations.

The elephant has an extraordinary long pregnancy, lasting 21 months. However, knowledge on embryo development is limited. To date, only single morphological observations of elephant embryo development associated with placentation are available, all lacking correlation to gestational age. The present study describes morphological characteristics of early embryo development in the elephant with exact biometric staging. Six pregnancies in five Asian and one African elephants with known conception dates were followed by 2D and 3D ultrasound, covering the embryonic period from ovulation to day 116 post-ovulation. The embryonic vesicle was earliest observed was on day 50 p.o. The proper embryo was not detected until day 62 p.o. Embryonic heartbeat was first observed on day 71 p.o. The allantois, which became visible as a single sacculation on day 71 p.o. was subdivided in four compartments on day 76 p.o. By day 95 p.o., head, rump, front and hind legs were clearly distinguished. Between days 95 and 103 p.o. the choriovitelline placenta was replaced by the chorioallantoic placenta. A physiological midgut herniation was transiently present between days 95 and 116 p.o. On the basis of the late appearance of the embryonic vesicle, delayed implantation in the elephant is discussed. The study provides a coherent description of elephant embryonic development, formation of the extraembryonic organs and their role in placenta formation, all of which are of interest for both comparative evolutionary studies and the improvement of assisted reproduction techniques.

Cerebrospinal fluid response to structured treatment interruption after virological failure.

OBJECTIVE: Structured antiretroviral treatment interruption (STI) has been advocated as a therapeutic strategy for HIV-1 infection. We report initial observations of cerebrospinal fluid (CSF) HIV-1 infection in five patients undergoing serial lumbar punctures (LPs) during STI undertaken following virological failure. DESIGN AND METHODS: In this prospective observational study we quantified HIV-1 RNA concentrations and assessed both phenotypic drug susceptibility profiles and genotypic antiviral drug resistance mutations in CSF and plasma during the period of treatment interruption. CSF white blood cells were also counted, and patients' neurological status monitored. RESULTS: In four of the patients, CSF HIV-1 concentration increased more rapidly than that of the plasma, with consequent reduction in the ratio between plasma and CSF viral loads (pVL : cVL). Three individuals developed robust, though asymptomatic CSF lymphocytic pleocytosis. In all patients the predominant HIV-1 quasispecies shifted simultaneously in CSF and plasma from a drug-resistant to a more drug-susceptible phenotype with identical and simultaneous changes in genotypes associated with drug resistance. CONCLUSIONS: STI may be accompanied by previously unrecognized changes in tissue viral exposures and lymphocyte traffic. Hence, despite 'virological failure' as evidenced by persistent plasma viremia, ongoing antiretroviral treatment prior to its interruption appeared to suppress CSF HIV-1 infection (indeed more effectively than that of plasma) and restrain lymphocyte traffic into the CSF. Simultaneous change of resistance mutations in CSF and plasma was likely due to re-emergence and overgrowth of pre-existing strains with ready exchange of virus between these two compartments, either facilitated by or provoking a local CSF lymphocytosis.

Venous Function in Patients with Venous Disease and Healthy Controls Before and After a Bathing Procedure and Subsequent Cold Stimulus

The object of this study was to determine whether bathing in thermal water acutely affects venous function in patients with venous insufficiency. Measurements were taken of venous function in patients with leg varicosities and healthy subjects before and after a thermal bath and exercise on two days, with subsequent application of a cold stimulus on 1 day. A total of 28 patients with varicose veins in one or both legs (n = 45 legs) and 30 healthy controls (n = 60 legs). On 2 consecutive days all subjects underwent a standardized bathing and exercise procedure in water at 34 degreesC for 20 minutes. On one day, determined randomly for each subject, a cold stimulus was applied to both lower legs after bathing. Measurements were done before subjects entered the bath and after the completion of bathing (including cold stimulus on one day). Without the cold stimulus, venous function of patients and controls did not change. After application of the cold stimulus the venous capacity of both patients and controls was significantly reduced compared with the basal levels. There was no change in maximum venous outflow in the controls, but that of the patient group decreased slightly compared with basal levels. Contrary to widespread belief, bathing in water at temperatures above 28 degreesC does not adversely affect venous function in patients with varicose veins. When applied in moderation (34 degreesC for 20 minutes) this physical therapy should not be regarded as contraindicated in this and related conditions, especially given its known hemodynamic benefits. Further studies are required to determine the long-term effects of repeated immersion.

Physical therapy improves venous hemodynamics in cases of primary varicosity: results of a controlled study.

Physical factors are known to influence hemodynamics in the veins of the lower extremities. In a controlled, randomized study the authors investigated the effects of combined physical therapy on varicose veins. Over a twenty-four-week period a treatment group consisting of 12 persons exercised under the supervision of a therapist twice a week for sixty minutes. This included muscle and joint activation by means of externally applied compression and cold-temperature stimuli (ie, thermosteresis). They also exercised once a day without supervision for fifteen minutes. During the same period a control group of 12 persons underwent the same measurements but no treatment. In the treatment group venous capacity decreased by an average of 16% from 4.9 +/- 0.3 (sd) mL/100 mL tissue to 4.1 +/- 0.4 (P < 0.005, U-test) while in the control group it remained practically unchanged at 4.8 +/- 0.4 vs 5.0 +/- 0.3. Venous refilling time in the lower extremities also increased in the treatment group, half refilling time rising from 7.8 +/- 1.0 to 11.3 +/- 0.9 seconds (P < 0.001) and total refilling time from 17.0 +/- 1.4 to 25.7 +/- 2.1 seconds (P < 0.001); these parameters remained virtually unchanged in the control group, with half refilling time dropping slightly from 7.7 +/- 1.1 to 7.1 +/- 1.3 seconds and total refilling time from 18.3 +/- 1.7 to 16.3 +/- 1.9 seconds. Patient self-rating scores obtained by use of a standardized questionnaire administered at baseline and at the end of week 24 improved in the treatment group only. The combined physical therapy was thus shown to be of long-term therapeutic value, improving venous function and reducing patients' symptoms. These findings indicate that for the further development of this combined treatment regimen it would be useful to identify the individual factors contributing to its efficacy and evaluate them separately.

Efficacy, safety, and cost of intravenous sedation versus general anesthesia in children undergoing endoscopic procedures.

We prospectively evaluated 226 patients under 18 years of age who underwent 296 procedures, and intravenous sedation and general anesthesia were compared in regard to efficacy, safety, and cost. Children 6 to 9 years of age required the highest doses of midazolam (0.14 +/- 0.04 mg/kg) and meperidine (2.5 +/- 0.8 mg/kg). A Relative Adequacy Scale, constructed to assess each patient's arousal and cooperation during intravenous sedation, revealed a 95% completion rate. Heart rate monitored before, during, and after the procedure was similar in both groups during the procedure, but a lower preprocedure heart rate was noted in older patients having intravenous sedation, suggesting less patient anxiety. Average charges, excluding endoscopist's and pathology fees, were $768.52 in the intravenous sedation group versus $1,965.42 in the general anesthesia group. Endoscopic procedures can be performed safely, effectively, and at a lower cost to the patient under intravenous sedation in a properly equipped and staffed pediatric endoscopy suite.

[CO2-induced acral blood flow and the oxygen partial pressure in arterial occlusive disease]. / CO2-induzierte Zunahme der akralen Durchblutung und des Sauerstoffpartialdruckes bei arterieller Verschlusskrankheit.

The effect of CO2-containing water on foot skin blood flow (laser-Doppler flux) and transcutaneous oxygen partial pressure (tcpO2) was measured in 16 patients (15 men and 1 woman; mean age 58 [49-71] years) with peripheral arterial occlusive disease (stage II of Fontaine: bilateral occlusions of the superficial femoral arteries). After 30 minutes lying down and 10 minutes sitting up, one lower leg was immersed in fresh water, the other in CO2-containing water (1400 mg CO2/kg H2O), both at 34 degrees C for 20 minutes. On the next day the measurements were repeated changing sides. Skin blood flow of the dorsum of the foot and rhythmic flow oscillations (flux motion) increased threefold in the CO2 water-bath, after a latency period averaging 2 minutes, within 3 minutes from 1.1 to 3.9 relative units (P less than 0.005). Average tcpO2 increased during the bath in CO2 water by 9 mm Hg (P less than 0.005). There was no change in fresh water. The increase in cutaneous blood flow and oxygen partial pressure, which persisted throughout the period in CO2 water, is interpreted as an increase in microcirculation and can explain part of the effectiveness of this largely empirical treatment.

[Increase in skin blood circulation and transcutaneous oxygen partial pressure of the top of the foot in lower leg immersion in water containing carbon dioxide in patients with arterial occlusive disease. Results of a controlled study compared with fresh water]. / Zunahme von Fussrücken-Hautdurchblutung und -transkutanem Sauerstoffpartialdruck während Unterschenkel-Immersion in kohlendioxidhaltigem Wasser bei Patienten mit arterieller Verschlusskrankheit: Ergebnisse einer gegen Süsswasser kontrollierten Studie.

Semi-quantitative Doppler laser flowmetry and measurement of transcutaneous oxygen partial pressure (TCPO2 in mmHg) are reliable, non-invasive methods of continuous measurement suitable for underwater use. We measured the effect of aqueous CO2 (succinate + sodium bicarbonate = Kao Bub; 1400 mg CO2 per kg water) compared with fresh water (both at 34 degrees C, depth of leg immersion 35 cm, immersion time 20 min) on circulation and TCPO2 in the feet of 15 patients with bilateral stage-II occlusion of the Aa. fem. superf. intraindividually in a randomised, crossover trial. Measurements were made at the same time of day on two consecutive days. No change in either cutaneous microcirculation or TCPO2 was observed during immersion in fresh water. During immersion in a CO2 bath both culaneous blood flow (as expressed by Doppler laser flux) and the amplitude of the Doppler laser vasomotion flux increased more than threefold (p less than 0.001, Wilcoxon), while sitting TCPO2 increased by over 10% (from 63 to 71 mmHg; p less than 0.001). The observed changes in oxygen dissociation and cutaneous microcirculation may help to provide an explanation for the well-known therapeutic effect of CO2 baths in all stages of occlusive arterial disease.