Regulatory pathways controlling the endovascular invasive trophoblast cell lineage

Hemochorial placentation is characterized by trophoblast-directed uterine spiral artery remodeling. The rat and human both possess hemochorial placentation and exhibit remarkable similarities regarding the depth of trophoblast invasion and the extent of uterine vascular modification. In vitro and in vivo research methodologies have been established using the rat as an animal model to investigate the extravillous/invasive trophoblast lineage. With these research approaches, two signaling pathways controlling the differentiation and invasion of the trophoblast cell lineage have been identified: i) hypoxia/hypoxia inducible factor and ii) phosphatidylinositol 3-kinase/AKT/Fos like antigen 1. Dissection of these pathways has facilitated identification of fundamental regulators of the invasive trophoblast cell lineage.     

Regulation of bovine pyruvate carboxylase mRNA and promoter expression by thermal stress

Pyruvate carboxylase (PC) catalyzes the rate-limiting step in gluconeogenesis from lactate and is a determinant of tricarboxylic acid cycle carbon flux. Bovine PC 5' untranslated region (UTR) mRNA variants are the products of a single PC gene containing 3 promoter regions (P3, P2, and P1, 5′ to 3′) that are responsive to physiological and nutritional stressors. The objective of this study was to determine the direct effects of thermal stress on PC mRNA and gene expression in bovine hepatocyte monolayer cultures, rat hepatoma (H4IIE) cells, and Madin-Darby bovine kidney epithelial (MDBK) cells. Hepatocytes were isolated from 3 Holstein bull calves and used to prepare monolayer cultures. Rat hepatoma cells and MDBK cells were obtained from American Type Culture Collection, Manassas, VA. Beginning 24 h after initial seeding, cells were subjected to either 37°C (control) or 42°C (thermal stress) for 24 h. Treatments were applied in triplicate in a minimum of 3 independent cell preparations. For bovine primary hepatocytes, endogenous expression of bovine PC mRNA increased (P < 0.1) with 24 h of thermal stress (1.31 vs. 2.79 ± 0.49, arbitrary units, control vs. thermal stress, respectively), but there was no change (P ≥ 0.1) in cytosolic phosphoenolpyruvate carboxykinase (PEPCK-C) mRNA expression. Similarly, exposure of MDBK cells to thermal stress increased (P < 0.1) expression of bovine PC mRNA without altering (P ≥ 0.1) PEPCK-C mRNA expression. Conversely, there was no effect (P ≥ 0.1) of thermal stress on endogenous rat PC (0.47 vs. 0.30 ± 0.08, control vs. thermal stress) or PEPCK-C (1.61 vs. 1.20 ± 0.48, arbitrary units, control vs. thermal stress, respectively) mRNA expressions in H4IIE cells. To further investigate the regulation of PC, H4IIE cells were transiently transfected with bovine promoter-luciferase constructs containing either P1, P2, or P3, and exposed to thermal stress for 23 h. Activity of P1 was suppressed (P < 0.1) 5-fold, activity of P2 was unchanged (P ≥ 0.1), and activity of P3 was increased (P < 0.1) by 5.4-fold. These data indicate that response of bovine PC gene to thermal stress is through promoter regulation and suggest that there are unique characteristics of bovine PC promoters that may contribute to the physiological response to thermal stress.     

LDSO: a program to simulate pedigrees and molecular information under various evolutionary forces

Simulations are a major tool to evaluate new statistical methods and optimize experimental designs in the genomic era. However, this can only be achieved when the simulations are close enough to reality, as well as diverse enough to be realistic. For mapping studies, it is thus critical to re-create as much as possible the forces generating linkage (mutation, random drift, changes in population sizes, selection and pedigree structure) and the mechanisms producing trait genetic architecture (additivity, dominance, epistasis). We present here a computer program (ldso) simulating these phenomena. Optional outputs provide statistics on the linkage disequilibrium (LD) structure and the identity by descent between chromosomal segments, facilitating further data analyses. Furthermore, ldso enables the simulation of genomic data in known pedigrees, which sticks as precisely as possible to recent population history and structures of the long-range LD, allowing optimization of fine-mapping strategies.     

3D-X-ray microcomputer tomography and optical coherence tomography as methods for the localization of the blastoderm in the newly laid unincubated chicken egg

The routine culling of the male offspring of hybrid layer type chickens is met with increasing public disapproval for both ethical and legal reasons. Until now practice-oriented methods for reliable sex diagnosis prior to hatch could not be developed. Molecular genetical analysis of blastodermic cells can be used for sex determination in unincubated eggs; however, knowledge of the precise localization of the germinal disc is crucial for the extraction of a carefully directed cell biopsy. In principle, 3D-X-ray micro computed tomography (3D-CT) has been proven a suitable method to localize the germinal disk in the unincubated egg without damaging the egg shell. No negative effects on embryogenesis and hatching rate of irradiated hatching eggs were established. The pictorial representation of the germinal disk using optical coherence tomography (OCT) failed in the unopened egg. The egg shell formed an impenetrable barrier for the currently available measuring method which utilized near infrared (NIR) wavelength regions. After opening the egg shell, the germinal disk could be visualized without any difficulties. In conclusion, technical possibilities for localization of the germinal disk in the unincubated egg already exist, but regarding technical parameters, the procedures have to be adapted to the specific purpose.     

Comparison of conservative management and juvenile pubic symphysiodesis in the early treatment of canine hip dysplasia.

OBJECTIVES: To evaluate the efficacy of juvenile pubic symphysiodesis (JPS) in a clinical setting for the early treatment of canine hip dysplasia (CHD), and to identify its indications and contraindications. METHODS: The final degree of CHD using the FCI (Fédération Cynologique Internationale) CHD classification in 5 Grades (A, B, C, D, E) was assessed at skeletal maturity in two homogeneous groups of dogs assessed at the age of 14 to 22 weeks and selected according to their susceptibility to CHD; one group was treated with JPS and one group was conservatively managed. Two hundred seventeen puppies completed the study; 81 were treated with JPS (group 1) and 76 were conservatively managed (group 2). A third group of 60 puppies with normal hips was followed as a negative control group. RESULTS: In group 1, 43.2% of the puppies had regression or a lack of progression of the disease in the final evaluation (Grade A & B), 25.9% had mild CHD (Grade C) and 30.9% had moderate and severe CHD (Grade D & E). In group 2, 23.6% of the puppies did not show any development of the disease (Grade A & B), 21.1% had mild CHD (Grade C) and 55.3% developed moderate to severe CHD (Grade D & E). Further investigation was done by comparing the severity of early signs of susceptibility to CHD with the final FCI Grades at adulthood in both groups. CLINICAL SIGNIFICANCE: The JPS procedure increased the odds of arresting or limiting the progression of CHD in mild to moderate grades of CHD, while it was less effective or ineffective in more severe forms.     

Use of extensible internal device in the femur of young dogs.

An extensible internal device (EID) was developed to preserve growth plate during the treatment of fracture complications or segmental bone loss from tumour resection in children. Since this type of extensible, transphyseal, internal fixation device has only been used in a few paediatric cases; the aim of this study was to evaluate an in vivo canine study, a surgical application of this device, and its interference with longitudinal growth of the non-fractured distal femur. Ten clinically healthy two- to three-month-old poodles weighing 1.5-2.3 kg were used. Following a medial approach to the right distal femur, one extremity of the EID, similar to a T-plate, was fixed in the femoral condyle with two cortical screws placed below the growth plate. The other extremity, consisting of an adaptable brim with two screw holes and a plate guide, was fixed in the third distal of the femoral diaphysis with two cortical screws. The EID was removed 180 days after application. All of the dogs demonstrated full weight-bearing after surgery. The values of thigh and stifle circumferences, and stifle joint motion range did not show any difference between operated and control hindlimbs. The plate slid in the device according to longitudinal bone growth, in all but one dog. In this dog, a 10.5% shortening of the femoral shaft was observed due to a lack of EID sliding. The other dogs had the same longitudinal lengths in both femurs. The EID permits longitudinal bone growth without blocking the distal femur growth plate if appropriately placed.     

Outbreak of cryptosporidiosis among veterinary students

In January 2007, six veterinary students became infected with Cryptosporidium species, and records indicated that another student had been diagnosed in November 2006. It was established that the seven students had worked with cattle from the same farm. Microbiological tests indicated that they were infected with Cryptosporidium parvum. Subtyping by sequence analysis indicated a common source of infection for five of the students, but there was insufficient material to type the other two samples. Investigations indicated that the outbreak was caused by a lapse in hygiene, particularly handwashing, on a farm with enzootic C parvum in calves.     

A multicentric retrospective study of serum/plasma urea and creatinine concentrations in dogs using univariate and multivariate decision rules to evaluate diagnostic efficiency

BACKGROUND: Urea and creatinine are the most frequently used indirect markers in plasma and serum of glomerular filtration rate in dogs. Both have been shown to lack sensitivity but their diagnostic efficiency for the diagnosis of kidney disease has been minimally investigated. OBJECTIVE: The purpose of this retrospective study was to investigate the influence of possible factors of variation on both analytes and to determine whether specific decision rules should be drawn up for subpopulations of dogs. METHODS: The results of urea and creatinine measurements, breed, sex, age, and health status (healthy, renal disease, or nonrenal disease) of 3822 dogs were collected from the archives of 5 veterinary clinics. Data were analyzed with univariate and multivariate decision rules with and without adjustment. RESULTS: There were significant effects and interactions of almost all of the sources of variation. Slight improvements in diagnostic efficiency were obtained by adjusting the decision rules to these sources of variations. Univariate decision rules gave approximately the same diagnostic efficiency for urea and creatinine concentrations, with sensitivity and specificity in the range of 70% and 90%, respectively, using the upper limit of the reference interval as the threshold value. Multivariate decision rules provided only minor improvements in diagnostic efficiency. CONCLUSION: Simultaneous measurement of both urea and creatinine is of limited diagnostic value over the analysis of a single variable. Creatinine is the preferred analyte as it is affected by fewer extrarenal factors of variation.