Microvessels of the blood-brain barrier (BBB) regulate transport into the brain. The highly specialized brain microvascular endothelial cells, a major component of the BBB, express tight junctions and efflux transporters which regulate paracellular and transcellular permeability. However, most existing models of BBB microvessels fail to exhibit physiological barrier function. Here, using (iPSC)-derived human brain microvascular endothelial cells (dhBMECs) within templated type I collagen channels we mimic the cylindrical geometry, cell-extracellular matrix interactions, and shear flow typical of human brain post-capillary venules. We characterize the structure and barrier function in comparison to non-brain-specific microvessels, and show that dhBMEC microvessels recapitulate physiologically low solute permeability and quiescent endothelial cell behavior. Transcellular permeability is increased two-fold using a clinically relevant dose of a p-glycoprotein inhibitor tariquidar, while paracellular permeability is increased using a bolus dose of hyperosmolar agent mannitol. Lastly, we show that our human BBB microvessels are responsive to inflammatory cytokines via upregulation of surface adhesion molecules and increased leukocyte adhesion, but no changes in permeability. Human iPSC-derived blood-brain barrier microvessels support quantitative analysis of barrier function and endothelial cell dynamics in quiescence and in response to biologically- and clinicallyrelevant perturbations.

Three-dimensional (3D) tissue-engineered models of the blood-brain barrier (BBB) recapitulate in vivo shear stress, cylindrical geometry, and cell-ECM interactions. Here we address four issues associated with BBB models: cell source, barrier function, cryopreservation, and matrix stiffness. We reproduce a directed differentiation of brain microvascular endothelial cells (dhBMECs) from two fluorescently labeled human induced pluripotent stem cell lines (hiPSCs) and demonstrate physiological permeability of Lucifer yellow over six days. Microvessels formed from cryopreserved dhBMECs show expression of BBB markers and maintain physiological barrier function comparable to non-cryopreserved cells. Microvessels displaying physiological barrier function are formed in collagen I hydrogels with stiffness matching that of human brain. The dilation response of microvessels was linear with increasing transmural pressure and was dependent on matrix stiffness. Together these results advance capabilities for tissue-engineered BBB models.

No abstract

Acute abdominal pain in the horse is a common emergency presenting to equine practices. The wide variety of etiologies makes prognosticating survival a challenge. A retrospective, multi-institutional clinical study was performed to determine clinical parameters associated with survival of horses with colic, and to use them to develop a colic survival scoring system. The scoring system was then validated using clinical data in the prospective portion of the study. Medical records from 67 horses presenting for acute abdominal pain were evaluated to develop the colic assessment score. Twenty eight variables were compared between survivors and non-survivors and entered into logistic regression models for survival. Of these, six variables were included in the colic assessment score. A total colic assessment score range was from 0 to 12, with the highest score representing the lowest probability of survival. The optimal cutoff value to predict survival was seven resulting in an 86% sensitivity and 64% specificity with a positive predictive value of 88% and a negative predictive value of 57%. Data from 95 horses presenting for abdominal pain to two equine hospitals was then collected prospectively to validate the colic assessment score. Horses from the prospective portion of the study that received a score >7 were classified as predicted to die and those with a score ≤7 were predicted to survive. The classification was compared to the actual outcome, of which the sensitivity, specificity, positive and negative predictive values of the colic assessment score were 84, 62, 88, and 52%, respectively.

One barrier to breast cancer diagnosis in low-resource settings is that devices for core needle biopsy (CNB) are either disposable and expensive, or reusable and susceptible to internal contamination. Through interviews with field workers and verification experiments, we identified that a common, commercially available, reusable CNB device allows contaminants to enter the driver chamber during firing, necessitating laborious cleaning of the entire device after every use. We introduce a novel CNB device attachment that eliminates this contamination mode and interfaces with existing commercial reusable drivers and low-cost disposable needles. This attachment repositions the driver–needle connection to the exterior of the driver, preventing retrograde flow of blood. Using an unmodified commercial CNB, we replicate chamber contamination by firing into a body fluid-mimicking glycerol solution. Prototypes were tested for their performance in eliminating this contamination. We tested the effectiveness of a cleaning procedure to reduce trace contamination by using a fluorescent dye and measuring the intensity of fluorescence after cleaning. The device's ability to reliably and consistently biopsy tissue with the novel attachment was evaluated using breast tissue models. In these tests, a reusable CNB with our attachment exhibited no measurable internal contamination, and maintained full biopsy functionality as measured by tissue sample weight and length. Minimizing internal device contamination would simplify the cleaning process for reusable biopsy devices. This would improve the accessibility of breast cancer biopsies in low- and middle-income countries (LMICs).

A 12-year pregnant Thoroughbred mare at ~ 340 days of pregnancy was presented for evaluation of lethargy, anorexia, and colicsigns. Reproductive examination was unremarkable; however, uroperitoneum was diagnosed via transabdominal ultrasonography,serum biochemistry, and peritoneal fluid analysis. Mare was medically stabilized for 24 hours. To promote fetal maturation, 80mg dexamethasone was given prior to Caesarean section and exploratory celiotomy. A ventral urinary bladder wall rupture andmultiple descending colon mesenteric rents were identified and repaired. Foal was delivered alive, treated for neonatal maladjustmentsyndrome, and discharged after 14 days. Seven days after surgery, severe complications of the mare led to its abrupt declinein health and sudden death.