Methods for recovery of microorganisms and intact microbial polar lipids from oil-water mixtures: laboratory experiments and natural well-head fluids.

Most of the world's remaining petroleum resource has been altered by in-reservoir biodegradation which adversely impacts oil quality and production, ultimately making heavy oil. Analysis of the microorganisms in produced reservoir fluid samples is a route to characterization of subsurface biomes and a better understanding of the resident and living microorganisms in petroleum reservoirs. The major challenges of sample contamination with surface biota, low abundances of microorganisms in subsurface samples, and viscous emulsions produced from biodegraded heavy oil reservoirs are addressed here in a new analytical method for intact polar lipids (IPL) as taxonomic indicators in petroleum reservoirs. We have evaluated the extent to which microbial cells are removed from the free water phase during reservoir fluid phase separation by analysis of model reservoir fluids spiked with microbial cells and have used the resultant methodologies to analyze natural well-head fluids from the Western Canada Sedimentary Basin (WCSB). Analysis of intact polar membrane lipids of microorganisms using liquid chromatography-mass spectrometry (LC-MS) techniques revealed that more than half of the total number of microorganisms can be recovered from oil-water mixtures. A newly developed oil/water separator allowed for filtering of large volumes of water quickly while in the field, which reduced the chances of contamination and alterations to the composition of the subsurface microbial community after sample collection. This method makes the analysis of IPLs (or indirectly microorganisms) from well-head fluids collected in remote field settings possible and reliable. To the best of our knowledge this is the first time that IPLs have been detected in well-head oil-water mixtures.

Nitric oxide-dependent cilia regulatory enzyme localization in bovine bronchial epithelial cells.

Airway epithelial-derived nitric oxide (NO), through the activation of nucleotide cyclases and downstream kinases, stimulates ciliary beating, yet the precise locations of these enzymes are unknown. We hypothesized that these NO-activated enzymes are located within, or adjacent to, the ciliary axoneme. Immunohistochemistry of intact ciliated cells revealed that endothelial-type nitric oxide synthase (eNOS), the RII isoform of the cAMP-dependent protein kinase (PKA-RII), the type I isoform of the cGMP-dependent protein kinase (PKG-I), and guanylate cyclase beta (GC-beta) all colocalized with pericentrin to the basal body. In contrast, the PKA-RI isoform and the PKG-II isoform localized to ciliary axonemes. Western blot analysis of isolated demembranated ciliary preparations detected eNOS, GC-beta, and both isoforms of PKA and PKG. An A-kinase-anchoring protein was also detected. Our findings suggest that these enzymes are sequestered close to their points of action into a discrete ciliary metabolon, enabling targeted phosphorylation and efficient upregulation of ciliary beating.

Ebola: Urgent Need, Rapid Response.

STATEMENT: The recent Ebola outbreak brought to bear the true nature of our ever-shrinking global health care community. Geography can no longer be considered a barrier to the spread of a highly communicable disease. Health care providers must be prepared to care for these patients regardless of location or environment. The simulation community responded rapidly to the Ebola epidemic with creative and timely solutions for training and systems optimization. Successful programs focused on 2 primary aims. First, the programs equipped the health care workers with essential knowledge for caring for their patients and protecting themselves. Second, they provided feedback to the system itself, identifying gaps in protocol and solutions for logistic challenges these patients often present. By having simulation integrated into the patient safety culture of our health care systems, we set in place a powerful tool that can respond nimbly to a high-stakes threat as evidenced by the success of these Ebola programs.

Continuous hemoglobin monitoring during massive blood transfusion in a multivisceral pediatric transplant patient.

A 10 year old, 25 kg girl, who was formerly a conjoined twin at the heart, diaphragm, liver, duodenum, bile ducts, and intestine, was admitted for a repeat liver, small bowel, pancreas, and kidney multivisceral transplant after having intestinal and liver failure. Intraoperatively, the patient had excessive bleeding, coagulopathy, and acidosis. Pulse co-oximetry was used for continuous monitoring of hemoglobin (Hb) during the procedure. Although noninvasive Hb appeared to follow a trend that correlated with arterial Hb concentration, it did not show accurate agreement with measured values from intermittent blood gas analysis. It may not be reliable during cases with abnormal physiology, rapid blood loss, and massive transfusion.