Nonsurgical collection of embryos in Shiba goats

A method of nonsurgical embryo collection in the Shiba goat, a native Japanese miniature goat breeding nonseasonally, was developed. The apparatus used for flushing the uterus was made on the model of the two-way catheter for cows. Embryo collection was performed on days 5 to 7 in 37 females superovulated with PMSG and hCG and resulted in successful recovery of 69 embryos in 19 females (51.4%). The average number of embryos collected from each successful female was 3.6. The recovery rate of embryos calculated on the basis of the number of embryos recovered and corpora lutea observed by culdoscopy in 15 successful females was 89.5%. This nonsurgical method seem to be efficient enough for collecting morulae and blastocysts in Shiba goats.     

Sequential uterine horn versus simultaneous total uterine flush to recover bovine embryos nonsurgically

Beef cows were superovulated with follicle stimulating hormone (FSH) to compare two nonsurgical methods of embryo recovery from the uterus. In the first method each uterine horn was independently flushed with physiological saline solution (PSS) through a Foley catheter passed through the cervix and into the uterine horn. In the second method both uterine horns were simultaneously flushed with PSS by passing the catheter into the uterine body. In both methods, the numbers of ovulations were determined after embryo collection by counting the corpora lutea (CL) on both ovaries of each cow through a flank incision. Independent flushing (n = 19) averaged 6.4 embryos and 16.1 CL per cow for a recovery rate of 40%. Simultaneous flushings (n = 22) averaged 5.4 embryos and 17.7 CL per cow for a recovery rate of 31%. This difference between the recovery rates of the two flushing methods was not significant (P>0.05).     

Pulmonary hypertension induced with an intra-arterial balloon: an alternate mechanism

The Laks catheter is a triple-lumen balloon catheter used to distend the canine main pulmonary artery while recording right ventricular pressure and the arterial pressure distal to the balloon. A rise in arterial pressure reported to occur during distension has been attributed to vasoconstriction rather than passive obstruction by the balloon. We tested this in six anesthetized dogs by inflating the Laks catheter-balloon while recording pressure distal to the balloon from the Laks catheter as well as from additional catheters in right and left pulmonary arteries placed retrogradely through lobar branches following thoracotomy. We found that balloon inflation increased pressures in the arterial port of the Laks catheter and in the left pulmonary artery catheter but reduced it in the right pulmonary artery. Tightening a snare around the right pulmonary artery had the same effects on pressures. Similar results were obtained while cardiac output was controlled by left ventricular bypass perfusion in four dogs. We conclude that the Laks catheter-balloon obstructs flow to the right lung and that the arterial pressure rise recorded in it during balloon inflation cannot be distinguished from that caused by occlusion of the right pulmonary artery.     

Fertility of donor mares following nonsurgical collection of embryos.

Embryos were collected nonsurgically on Day 7 or 8 after ovulation from 7 Quarter horse mares using a modified 30-ml Foley catheter to flush the uterine horn ipsilateral to the recent ovulation with 500 ml TCM-199 containing Hepes buffer. After collection, the uteri were infused with nitrofurazone to reduce the chances of infection due to the procedure. Eleven collections from 7 mares resulted in recovery of 9 embryos and nonsurgical transfer of 4 of these resulted in the birth of one foal. After collections, 8 oestrous cycles averages 22.75 days and 2 extended oestrous cycles were 43 and 59 days long respectively. Of 6 mares mated after one or two embryo collections, 5 conceived to a single service and the sixth during the third oestrus in which she was covered.     

Antioxidant metabolism in Xenopus laevis embryos is affected by stratospheric balloon flight

To test the effects of low levels of radiation from space on living organisms, we flew Xenopus laevis embryos at different stages of development on a stratospheric balloon (BI.R.BA mission). After recovery, different parameters were analyzed to assess the effects of flight, with particular regard to oxidative stress damage. Because of failed temperature control during flight, the flight shielded embryos (FC) could not be used for biochemical or morphological comparisons. In contrast, the incubation conditions (i.e. temperature, containers, volumes) for the flight embryos (F) were parallel to those for the ground controls. Mortality data show that younger embryos (16 h) flown on the balloon (F) are more sensitive to radiation exposure than older ones (40 h and 6 days). Exposure during flight lowered the antioxidant potential in all embryos, particularly older ones. These preliminary data demonstrate that flight on a stratospheric balloon might affect antioxidant metabolism, though it is not yet possible to correlate these results with low radiation exposure during flight.     

A Method for Collection of Bile in the Conscious Sheep

A method to collect bile directly from the hepatic duct is described for use in the sheep. The technique is a combination of the intestinal re-entrant cannulae and a catheter from the duodenal lumen to the hepatic duct. The cystic duct is ligated near its junction with the common bile duct. The catheter is fixed in the proximal visible end of the hepatic duct. One plastic cannula is fixed to the duodenum opposite to the opening of the common bile duct and the other is fixed in the same way about 15 cm posterior to the first one. The two plastic cannulae fixed together with a plastic tube serve as an extra-abdominal anastomosis. During the collection periods the bile duct catheter is passed through an opening in the wall of the connection tube into a collection bag that is fixed to the plastic cannulae. Between the collection periods the catheter ends in the lumen of the anastomosis.     

Nonsurgical collection of blastocysts from dairy goats

In two trials, eight attempts were made to collect fertilized ova from dairy goats by nonsurgical methods. In both trials the cervix of each doe was dilated by inserting a Laminaria japonica tent device into the cervical canal prior to flushing. In Trial 1, an attempt was made to collect embryos from four nonsuperovulated does by flushing phosphate-buffered saline (PBS) through a rigid pipette. Little fluid and no embryos were recovered from the does. All four donors were in estrus two days after the procedure. In the second trial, FSH-superovulated does were collected on day 5 following estrus. The donors were anesthetized, and a modified Foley catheter was passed through the cervical canal. In three does, a 24 ga. two-way Foley was stiffened with a size 10 (French) polypropylene catheter which penetrated the Foley, extending 7 cm beyond the tip. Recovery of flushing medium with this device was minimal, and laparotomy of one doe revealed a punctured uterus. Replacement of this device with a different catheter, through which a polypropylene catheter (size 5 Fr.) penetrated only 1 to 2 cm, resulted in satisfactory return of infused PBS, and recovery of two blastocysts and one degenerated ovum from this doe. Use of the same device on a second doe without laparotomy resulted in collection of seven blastocysts and three degenerated ova. Of three observed donors that received Laminaria tents (including one which was not flushed) two were in estrus three days after the procedure, while unused synchronized recipients showed normal cycle lengths. Surgical transfer of two blastocysts from each donor to each of two synchronized recipients resulted in the birth of twin kids from one recipient doe. The study demonstrates the feasibility of embryo collection from dairy goats by nonsurgical means.     

Static and dynamic operating characteristics of a pericardial balloon.

Previously, we developed a balloon transducer to measure the constraint of the pericardium (i.e., pericardial pressure) on the surface of the heart. It was validated physiologically in that it was shown to measure a pressure equal to the difference between the left ventricular end-diastolic pressure measured before and after pericardiectomy at the same left ventricular volume. To define its static operating characteristics, we loaded the balloon nonuniformly with weights that covered fractions of the balloon surface and found that the balloon accurately recorded the average stress if the stress was applied over at least 23% of its surface. To test its performance when curved, we placed it in large and small cylinders (minimum diameter 31 mm) and found that the balloon accurately recorded the stress. To define its dynamic operating characteristics, we applied sinusoidal stresses and found that its frequency response was limited only by that of the connecting catheter. When better dynamic response is required, we introduce a micromanometer-tipped catheter to obtain a unity-gain frequency response that is flat to 200 Hz.     

Management of obstructed balloon catheters.

Failure of a balloon catheter to deflate is not uncommon and prevents its removal. Methods of overcoming the problem include traction, bursting the balloon by overinflation, dissolving it with solvents, puncturing it percutaneously with a needle, or puncturing it with a wire stylet passed through the catheter. All except the last technique have major disadvantages and are of questionable safety. Transcatheter puncture of the balloon was used in 16 patients to remove obstructed balloon catheters without any technical difficulty, distress to the patient, or complication. The procedure is safe, simple, and does not require an anaesthetic. If necessary it could be performed safely by nursing or paramedical staff without the patient having to be admitted to hospital. It is the method of choice for the management of this problem.     

Collection of tubal stage bovine embryos by means of endoscopy. A technique report

Here we describe the development and optimization of endoscopy-mediated transvaginal access for collecting ova and embryos from the bovine oviduct. The novel technique was developed in three experimental setups: In Experiment 1 embryos were collected unilaterally from nonstimulated heifers. We flushed the oviducts of superovulated heifers unilaterally (Experiment 2) and bilaterally (Experiment 3). In Experiment 1 the oviducts of 18 heifers were successfully cannulated, which resulted in the collection of twelve 1-cell to 8-cell embryos and one empty zona. Unilateral flushing of 13 animals (Experiment 2) resulted in 84 ova with 6.3 +/- 3.2 observed ovulation sites. Bilateral flushing of 25 animals (Experiment 3) resulted in 293 ova plus 10 empty zonae from 11.8 +/- 5.4 ovulation sites. Given our experience from these studies we optimized the technical equipment by improving the flushing metal catheter (Experiment 4). The novel catheter hermetically sealed the lumen of the ampulla at the moment, the medium was flushed through the oviduct. This resulted in a visible flow of medium via oviducts toward the embryo filter connected to an embryo flushing catheter that was fixed in the uterine horns. Our endoscopy-guided method is minimally invasive and facilitates the flushing of tubal stage embryos.     

Patency of the cervical canal in cross-bred female Zebu x Brown Swiss selected for non-surgical recovery or transfer of embryos

A test to evaluate the degree of patency of the cervical canal was carried out on 260 Zebu x Brown Swiss cows and heifers that were assigned for non-surgical recovery or transfer of embryos. A total of 204 females (78.5%) were found to have a patent cervix (P <0.05), compared with 56 (21.5%) which had a blockage or obstruction when an attempt was made to pass a catheter through the cervical canal or the "cervical knot" of the purebred and cross-bred Zebu cows. A smaller percentage (12.89%) of non-patent cervices (P <0.05) were detected in multiparas, compared with 25 and 27.5% in primiparas and nullipara, respectively. The test for cervical patency is carried out on Bos indicus cattle to screen the animals before non-surgical embryo transfer manipulations and to attain better results in the application of non-surgical recovery and transfer of embryos.     

Recent developments in pig embryo transfer.

Porcine embryo transfer has been performed for approximately 50 years, and surgical methods have proven to be reliable for collection and transfer of embryos. However, surgical collection and transfer have the disadvantage of being less useful on the farm. Recently, new procedures for both collection and transfer of embryos have been developed to improve usefulness. The surgical procedure has been refined to a minimally invasive procedure, using endoscopy for collection and transfer of embryos. A nonsurgical procedure for embryo collection has also been devised, but is limited to use in sows with surgically shunted (shortened) uterine horns. Nonsurgical embryo transfer procedures have been developed recently and have proven to be successful. The nonsurgical procedures are preferable to surgical procedures from an animal welfare point of view and because these procedures can be performed on farms without the need for special facilities.     

Effect of pulmonary blood flow on leukocyte uptake and release by dog lung

The effect of pulmonary blood flow on leukocyte uptake and release by the lung was examined in 10 anesthetized spontaneously breathing dogs. Pulmonary arterial and pulmonary venous blood was sampled with catheters placed into the right ventricle and aorta, respectively. Pulmonary blood flow was lowered by inflating a balloon catheter located in the inferior vena cava. In five experiments simultaneous blood samples were drawn from the right ventricle and aorta at 10-s intervals during a control period, a 2- to 3-min period of low flow, and a recovery period. In five additional experiments, less frequent samples were taken over periods of 15-60 min. Total leukocyte concentrations and differential counts were determined for each blood sample. The study shows that large numbers of leukocytes become sequestered within the lung when pulmonary blood flow is low and that an equivalent number of cells are released from the lung after deflation of the balloon catheter. Both the polymorphonuclear leukocytes and the lymphocytes were taken up by the lung when pulmonary blood flow was reduced. We conclude that pulmonary blood flow has a marked effect on the uptake and release of leukocytes by the dog lung.     

In vitro simulations of embryo transfer in a laboratory model of the uterus

Embryo transfer (ET) is the final manual intervention during which the newly formed embryo is placed within the uterus by a transcervical catheter. The loading of the syringe-catheter complex with the transferred volume consists of the transfer media (which contains the embryos) separated by air spaces on both sides. The dynamics involved in injecting the syringe-catheter complex is not well understood nor has it been investigated to date. We developed an in vitro experimental setup for simulations of ET into a rigid transparent uterine model. The catheter was loaded in sequences of liquid and air as it is in the clinical setting. The transferred liquid was colored with a dye and its dispersion within the uterine cavity was recorded by a video camera. The results demonstrated, for the first time, the importance of having a gas phase in the catheter load. The resulting air bubbles within the uterus were carried upward towards the fundus by buoyant forces, thereby dragging behind them the transferred liquid which contained the embryos. This could be expected to substantially increase the probability for the embryos to be present near the fundal wall at the time window for implantation. There was also evidence of a dependency of the rate of injection upon the catheter load into the uterus: a low speed generated several air bubbles which led to more of the transferred liquid being carried towards the fundal end, thus possibly enhancing the potential for implantation.     

Numerical study of the uniformity of balloon-expandable stent deployment

Stents are small tubelike structures, implanted in coronary and peripheral arteries to reopen narrowed vessel sections. This endovascular intervention remains suboptimal, as the success rate is limited by restenosis. This renarrowing of a stented vessel is related to the arterial injury caused by stent-artery and balloon-artery interactions, and a local subsequent inflammatory process. Therefore, efforts to optimize the stent deployment remain very meaningful. Several authors have studied with finite element modeling the mechanical behavior of balloon-expandable stents, but none of the proposed models incorporates the folding pattern of the balloon. We developed a numerical model in which the CYPHER stent is combined with a realistic trifolded balloon. In this paper, the impact of several parameters such as balloon length, folding pattern, and relative position of the stent with respect to the balloon catheter on the free stent expansion has been investigated. Quantitative validation of the modeling strategy shows excellent agreement with data provided by the manufacturer and, therefore, the model serves as a solid basis for further investigations. The parametric analyses showed that both the balloon length and the folding pattern have a considerable influence on the uniformity and symmetry of the transient stent expansion. Consequently, this approach can be used to select the most appropriate balloon length and folding pattern for a particular stent design in order to optimize the stent deployment. Furthermore, it was demonstrated that small positioning inaccuracies may change the expansion behavior of a stent. Therefore, the placement of the stent on the balloon catheter should be accurately carried out, again in order to decrease the endothelial damage.     

Calculation of dose distribution near an innovative concentric balloon catheter for endovascular brachytherapy

Purpose: Using a radioactive solution-filled catheter for intravascular irradiation has the potential problem of chemical and radiological toxicity in the case of a balloon rupture. In order to reduce this risk, an innovative concentric balloon catheter was developed.Methods and Materials: The concentric balloon was made by inner and outer balloons filled with saline and radioactive solution, respectively. The optimal inner radius was determined by comparing the dose rate reduction vs. the volume reduction for various inner and outer radii for 188Re, 32P, and 90Y solutions.Results: For a balloon with an outer radius of 1.5 mm, there was no advantage of a concentric balloon. For balloons with outer radii of 3.0 and 5 mm, the optimal inner radius was 1.5 and 3 mm, respectively.Conclusions: With the newly designed concentric balloon, the risk of toxicity can be reduced while keeping the dose rate high enough so that the treatment times within tolerable limits are still maintained.     

An Aortic Tampon for Emergency Control of Ruptured Abdominal Aneurysm

Ruptured abdominal aneurysm has now become a common surgical emergency, frequently amenable to successful resection and cure. The final result is often marred, however, by the effects of renal, coronary or cerebral ischemia resulting from dangerous hypotension during transportation of the patient to a vascular centre.An aortic catheter has been developed which is passed by way of a brachial artery cut-down so that it rests in the abdominal aorta. The balloon at its tip is then filled with sodium diatrizoate (Hypaque) so that it completely obstructs the aortic lumen just above the level of the aneurysm. Accurate positioning of the balloon to carefully preserve renal blood flow is facilitated by fluoroscopic control. The use of this procedure in three patients has been very satisfactory, with a dramatic return of consciousness and of normal blood pressure, without the need for further blood replacement. Subsequent surgery with dissection of the aneurysm was aided by the presence of the palpable inflated balloon.     

A treasure house of comparative embryology

The Embryo Collection of the Hubrecht Laboratory is a treasure house of comparative embryology. It is the largest and most important collection of its kind in the world, and consists of thousands of vertebrate embryos stored in alcohol, or prepared as histological sections. Many elusive species are included in the collection, some represented by complete developmental series. The accompanying archives offer a remarkable insight into the methods used to collect embryos form wild animals, as well as the motives behind the founders of the collection. Carefully maintained, documented and catalogued, the collection is available for study by all interested scientists. We argue that this collection is one of the greatest biodiversity resources in existence.     

Wedge pressure in large vs. small pulmonary arteries to detect pulmonary venoconstriction

Pulmonary arterial wedge pressure measures the pressure where blood flow resumes on the venous side. By occlusion of a large artery, the point where blood flow resumes will be in or near the left atrium. However, by occlusion of a small artery, it is possible to shift the point where flow resumes to a more proximal site in the veins and thus measure a pressure within the small veins. Increased pulmonary venous pressure, as a result of partial obstruction in the large veins, may not be detected by wedging a Swan-Ganz catheter in a large artery but may be detected by wedging in a small artery. We demonstrated this phenomenon in open-chest dogs by mechanically obstructing the left lower lobar vein or by infusing histamine to cause a generalized pulmonary venoconstriction. The wedge pressure measured by a 7-F Swan-Ganz catheter, with its balloon inflated in the main left lower lobar artery, nearly equaled left atrial pressure. On the other hand, the wedge pressure measured with a 7-F, 5-F, or a PE-50 catheter advanced into a small artery (without a balloon) was considerably higher than left atrial pressure. These results suggest that high resistance in the pulmonary veins can be demonstrated with the Swan-Ganz catheter by comparing the pressures obtained with the catheter wedged in a small and large artery.     

Minimally invasive transabdominal collection of preimplantation embryos from the common marmoset monkey (Callithrix jacchus)

A novel, minimally invasive, transabdominal embryo collection method (transabdominal method) was developed as an alternative to a standard abdominal incision for embryo collection in the common marmoset. The abdominal incision method was used for 304 flushes using 36 female animals, whereas the transabdominal method was used for 488 flushes using 48 females; successful embryo collection rates were 48.0% and 48.4% (P > 0.05), respectively. These techniques were successfully duplicated at another institute (German Primate Center, DPZ). At that institution, successful embryo collection rates were 88.9% and 77.8% for the abdominal incision and transabdominal methods, respectively (P > 0.05), whereas the average numbers of preimplantation embryos obtained per flush were (mean ± SD) 1.91 ± 0.35 and 1.71 ± 0.14 (P > 0.05). The transabdominal method reduced animal stress, did not require incisional wound healing, and enabled successive embryo recoveries to be done much sooner. More embryos in early developmental stages (zygotes/morulae) were recovered using the transabdominal method (76.1%) than the abdominal incision method (52.6%, P < 0.01). In contrast, recovery of arrested or abnormal embryos was not significantly different between these two methods (9.8% and 8.3%). To verify developmental ability of embryos recovered by the transabdominal method, transfer of 28 normal embryos to 14 surrogate mothers yielded a nidation rate of 57%. Five females sustained term pregnancies and eight neonates were born. This novel transabdominal method will facilitate progress in marmoset developmental biology and embryology.