Relationships between serum electrolyte concentrations and ileus: A joint clinical and mathematical modeling study.

AIM: Prolonged postoperative ileus (PPOI) occurs in around 15% of patients after major abdominal surgery, posing a significant clinical and economic burden. Significant fluid and electrolyte changes may occur peri-operatively, potentially contributing to PPOI; however, this association has not been clearly elucidated. A joint clinical-theoretical study was undertaken to evaluate peri-operative electrolyte concentration trends, their association with ileus, and predicted impact on bioelectrical slow waves in interstitial cells of Cajal (ICC) and smooth muscle cells (SMC). METHODS: Data were prospectively collected from 327 patients undergoing elective colorectal surgery. Analyses were performed to determine associations between peri-operative electrolyte concentrations and prolonged ileus. Biophysically based ICC and SMC mathematical models were adapted to evaluate the theoretical impacts of extracellular electrolyte concentrations on cellular function. RESULTS: Postoperative day (POD) 1 calcium and POD 3 chloride, sodium were lower in the PPOI group (p < 0.05), and POD3 potassium was higher in the PPOI group (p < 0.05). Deficits beyond the reference range in PPOI patients were most notable for sodium (Day 3: 29.5% ileus vs. 18.5% no ileus, p = 0.04). Models demonstrated an 8.6% reduction in slow-wave frequency following the measured reduction in extracellular NaCl on POD5, with associated changes in cellular slow-wave morphology and amplitude. CONCLUSION: Low serum sodium and chloride concentrations are associated with PPOI. Electrolyte abnormalities are unlikely to be a primary mechanism of ileus, but their pronounced effects on cellular electrophysiology predicted by modeling suggest these abnormalities may adversely impact motility recovery. Resolution and correction of electrolyte abnormalities in ileus may be clinically relevant.

A Pipeline for the Registration of Calcium Transient Data to Structural Networks of the Interstitial Cells of Cajal.

Interstitial cells of Cajal (ICC) generate electrical pacemaker activity in the gastrointestinal (GI) tract known as slow waves, which regulate GI motility. ICC express both the Kit receptor tyrosine kinase protein and a Ca2+-activated Cl--channel, encoded by the anoctamin1 (Ano1) protein, which is an essential contributor to the Ca2+ cycling of ICC and slow wave pacemaking. Recent dye-loading imaging studies have demonstrated Ca2+ transients in ICC in isolated tissue preparations. The main aim of this study was to develop a method that allows Ca2+ transients to be registered to structural ICC network data. Confocal image stacks of ICC labeled for Kit or Ano1 and Ca2+ recording data were processed using a thresholding protocol. The Ca2+ transients were then registered to the ICC structural network. First, a general idea of the placement was found by mapping the field-of-view of the Ca2+ transient data to the distorted tissue that contained the ICC network image. The errors in the registration were then corrected for by warping the internal Ca2+ transient field according to the structural network. In data sets from tissues with induced, targeted knockdown of Ano1 expression in a subset of ICC, agreement between the Ca2+ transient data and structural network was 68 ± 10%. This level of agreement allowed selective extraction of Ca2+ data from Ano1-positive (Ano1+) and Ano1-negative (Ano1-) ICC. In the future, this technique will allow investigation into the functional properties of ICC in relation to the level of knockdown of specific ICC associated proteins.

A Method for Multi-day Tracking of Gastrointestinal Smooth Muscle Contractile Patterns in Organotypic Culture.

Gastrointestinal (GI) motility is a key component of digestive health, and it is a complex both rhythmic and arrhythmic process governed by many physiological factors. The ability to accurately track the movements of the GI tissues in vitro over multiple days would provide valuable insights into GI tract physiology. A correlational analysis tracking algorithm was developed and applied to intestinal smooth muscle tissues that were maintained in organotypic culture. Physiologically relevant metrics, such as frequency, amplitude and motility index were independently assayed to quantify smooth muscle contractions. The results were validated by manually detected frequency determined using a standard software package. The algorithm was capable of tracking the changes in contractions of the same tissues over six days. This proof-of-concept study demonstrates the feasibility of long-term tracking of GI motility in organotypic cultures over multiple days. The approach allows study of the effects on GI smooth muscle contractility of direct controlled targeting of the cells and molecules in the GI tunica muscularis.

A mathematical model of the effects of anoctamin-1 loss on intestinal slow wave entrainment.

The interstitial cells of Cajal (ICC) generate electrophysiological events called slow waves that regulate the motility of the gastrointestinal (GI) tract. Recent studies have demonstrated that the Ca2+-activated Cl- -channel, encoded by the anoctamin1 (Ano1) protein, has a major role in regulating intestinal slow waves and motility. The main aim of this study was to develop a multi-scale mathematical model capable of simulating both normal slow wave entrainment and the effects of Ano1 knockout (KO) on the normal activity. A biophysically-based cell model was adapted to simulate the effects of Ano1 KO at the cellular level. A 10mm one-dimensional (1D) model was then developed to simulate entrained intestinal slow wave propagation. Cellular KO at levels of 100% and 20% were applied to a varying-sized middle region of the 1D model. The main finding was that the level of loss of entrainment increased as both cellular and spatial Ano1 KO levels increased, mostly manifesting as ectopic activation. In the future, this model will be extended and used in combination with Ca2+ -imaging data to quantitatively investigate the effects of Ano1 loss in ICC.