Clinical and financial impacts of abnormal liver biochemistry after liver transplantation.

OBJECTIVE: After liver transplant (LT), many investigations are needed to evaluate abnormal liver function test (LFT), which has poor specificity for graft function and complication. A single center retrospective audit of all adult single organ LT from 1/1/2015 to 31/12/2017 was performed. Demographic, clinical and investigation data from the LT database and electronic medical records and cost data from the hospital's Business Intelligence Unit were analyzed. Patients were classified into uncomplicated or complicated LFT by 2 independent investigators and the number, type, and cost of investigations in the first 30 post-operative days were analyzed. Investigations prior to liver biopsy was sub-analyzed. RESULTS: There was 170 LT with 87 cases of uncomplicated LFT (51.2%) and 83 cases of complicated LFT (48.8%). Most patients with complicated LFT had additional investigations (97.6%), most commonly cholangiogram (55.4%) and liver biopsy (LBx) (50.6%). The additional investigations cost was $1863.3 (95% CI 1289.0-2437.6). Although most LBx (73.8%) showed evidence of rejection, LBx was often not the initial investigation of choice. Current LFT based post-transplant monitoring is inefficient. It remains difficult to determine which patient will benefit from an early invasive procedure like LBx, using LFT alone without further imaging investigations.

Floseal-induced small bowel obstruction.

Floseal is a haemostatic agent designed to augment the body's natural clotting cascade response. We describe the first case in over a decade, and the first case with intra-operative images of early post-operative small bowel obstruction (SBO) associated with Floseal use in general surgery. A previously well man in his 30s underwent laparoscopic appendicectomy for clinical acute appendicitis. Floseal was applied to the right lateral abdominal wall for haemostasis. He developed a mechanical SBO, with diagnostic laparoscopy confirming a transition point between the caecum and terminal ileum, adherent to the area of the previously applied Floseal. He underwent adhesiolysis and uneventful recovery. We propose Floseal may exacerbate early post-operative inflammation and provide a nidus for early adhesion formation. We recommend removing excess Floseal not incorporated in the haemostatic clot and to consider Floseal as a differential in early post-operative SBO.

Low Levels of Hepatocyte-Specific Methylation in Cell-Free DNA Are a Strong Negative Predictor for Acute T Cell-Mediated Rejection Requiring Treatment Following Liver Transplantation.

Graft-derived cell-free DNA (gdcfDNA) quantification is a promising, minimally invasive tool for detecting acute T cell-mediated rejection (ATCMR) following liver transplantation (LT). We investigated the utility of measuring hepatocyte-specific methylation in cfDNA (HS-cfDNA) to quantify gdcfDNA, examining its accuracy in detecting ATCMR in a prospective, cross-sectional study. Blood was collected from LT recipients immediately prior to graft biopsy for suspected rejection. HS-cfDNA was quantified using droplet-digital polymerase chain reaction. Prebiopsy liver function tests (LFTs) and HS-cfDNA levels were correlated with biopsy results and the primary outcome of treated biopsy-proven acute rejection (tBPAR). A total of 51 patients were recruited; 37 had evidence of rejection on biopsy and 20 required treatment. As much as 11 patients needed inpatient treatment for rejection. HS-cfDNA significantly outperformed LFTs in identifying patients with tBPAR, particularly those needing inpatient treatment (area under the curve, 73.0%; 95% confidence interval, 55.4%-90.6%; P = 0.01). At a threshold of <33.5% of the total cfDNA fraction, HS-cfDNA had a specificity of 97%, correctly excluding tBPAR in 30/31 patients. Quantifying graft-specific methylation in cfDNA has a major advantage over previous gdcfDNA techniques: it does not require genotyping/sequencing, lending it greater feasibility for translation into transplantation care. Low levels of HS-cfDNA were a strong negative predictor for tBPAR (negative predictive value, 86%) and may have a future role in triaging patients prior to invasive graft biopsies.

Haskap Berry Phenolic Subclasses Differentially Impact Cellular Stress Sensing in Primary and Immortalized Dermal Fibroblasts.

It is generally accepted that dietary phenolics from fruits are of significant importance to human health. Unfortunately, there is minimal published data on how differences in phenolic structure(s) impact biological pathways at cellular and molecular levels. We observed that haskap berry extracts isolated with ethanol:formic acid:water or phenolic subclass fractions separated using different concentrations of ethanol (40% and 100%) impacted cell growth in a positive manner. All fractions and extracts significantly increased population doubling times. All extracts and fractions reduced intracellular free radicals; however, there were differences in these effects, indicating different abilities to scavenge free radicals. The extracts and fractions also exhibited differing impacts on transcripts encoding the antioxidant enzymes (CAT, SOD1, GPX1, GSS and HMOX1) and the phosphorylation state of nuclear factor-κB (NF-κB). We further observed that extracts and fractions containing different phenolic structures had divergent impacts on the mammalian target of rapamycin (mTOR) and sirtuin 1 (SIRT1). siRNA-mediated knockdown of SIRT1 transcripts demonstrated that this enzyme is key to eliciting haskap phenolic(s) impact on cells. We postulate that phenolic synergism is of significant importance when evaluating their dietary impact.

Phenolic composition and antioxidant activities of saskatoon berry fruit and pomace.

Total phenolic chromatographic indices (TPCI) of three commercially grown saskatoon berry varieties and a pomace from commercial juice production were determined. Northline was shown to have the highest TPCI of 504.2 mg/100 g FW. These results agreed with total phenolic content results for these varieties. The TPCI of the commercial pomace was 404.2 mg/100 g pomace indicating that a significant concentration of phenolics were present in this co-product, showing the commercial relevance of this material. A phenolic rich extract (PRE; 500 ppm) of the Northline variety was compared to BHT (0.02% w:w) and Rosamox (0.2% w:w) for delaying the oxidation of borage oil via rancimat analysis. Induction times were 1.46 h (borage oil), 1.44 h (Rosamox), 2.18 h (BHT), and 2.42 h (PRE), which was a ∼65% delay in the oxidation of borage oil. These results clearly support the value of this material as an antioxidant ingredient in foods, pharmaceuticals, nutriceuticals and cosmetics.

Structural identification of compounds for use in the detection of juice-to-juice debasing between apple and pear juices.

The ability to detect the undeclared addition of a juice of lesser economic value to one of higher value (juice-to-juice debasing) is a particular concern between apple and pear juices due to similarities in their major carbohydrate/polyol profiles. Fingerprint compounds for the detection of this type of adulteration were identified in both commercial apple and pear juices by HPLC-PDA, were isolated chromatographically, and structurally identified by LC-MS/MS. The apple juice fingerprint was identified as 4-O-p-coumarylquinic acid and two pear compounds as isorhamnetin-3-O-rutinoside and abscisic acid. Additionally, the HPLC-PDA profile of pear juices in combination with pear fingerprint compounds including arbutin could be used to identify samples originating from China versus those from other geographical locations.

Analysis of a series of chlorogenic acid isomers using differential ion mobility and tandem mass spectrometry.

Chlorogenic acids are among the most abundant phenolics found in the human diet. Of these, the mono-caffeoylquinic acids are the predominant phenolics found in fruits, such as apples and pears, and products derived from them. In this research, a comprehensive study of the electrospray ionization (ESI) tandem mass spectrometric (MS/MS) dissociation behavior of the three most common mono-caffeoylquinic acids, namely 5-O-caffeoylquinic acid (5-CQA), 3-O-caffeoylquinic acid (3-CQA) and 4-O-caffeoylquinic acid (4-CQA), were determined using both positive and negative ionization. All proposed structures of the observed product ions were confirmed with second-generation MS(3) experiments. Similarities and differences between the dissociation pathways in the positive and negative ion modes are discussed, confirming the proposed structures and the established MS/MS fingerprints. MS/MS dissociation was primarily driven via the cleavage of the ester bond linking the quinic acid moiety to the caffeic acid moiety within tested molecules. Despite being structural isomers with the same m/z values and dissociation behaviors, the MS/MS data in the negative ion mode was able to differentiate the three isomers based on ion intensity for the major product ions, observed at m/z 191, 179 and 173. This differentiation was consistent among various MS instruments. In addition, ESI coupled with high-field asymmetric waveform ion mobility spectrometry-mass spectrometry (ESI-FAIMS-MS) was employed for the separation of these compounds for the first time. By combining MS/MS data and differential ion mobility, a method for the separation and identification of mono-caffeoylquinic in apple/pear juice samples was developed with a run time of less than 1 min. It is envisaged that this methodology could be used to identify pure juices based on their chlorogenic acid profile (i.e., metabolomics), and could also be used to detect juice-to-juice adulteration (e.g., apple juice addition to pear juice).

Oligosaccharide formation during commercial pear juice processing.

The effect of enzyme treatment and processing on the oligosaccharide profile of commercial pear juice samples was examined by high performance anion exchange chromatography with pulsed amperometric detection and capillary gas chromatography with flame ionization detection. Industrial samples representing the major stages of processing produced with various commercial enzyme preparations were studied. Through the use of commercially available standards and laboratory scale enzymatic hydrolysis of pectin, starch and xyloglucan; galacturonic acid oligomers, glucose oligomers (e.g., maltose and cellotriose) and isoprimeverose were identified as being formed during pear juice production. It was found that the majority of polysaccharide hydrolysis and oligosaccharide formation occurred during enzymatic treatment at the pear mashing stage and that the remaining processing steps had minimal impact on the carbohydrate-based chromatographic profile of pear juice. Also, all commercial enzyme preparations and conditions (time and temperature) studied produced similar carbohydrate-based chromatographic profiles.

The physicochemical properties of legume protein isolates and their ability to stabilize oil-in-water emulsions with and without genipin.

The physicochemical and emulsifying properties of legume protein isolates prepared from chickpea (CPI), faba bean (FPI), lentil (LPI) and soy (SPI) were investigated in the presence and absence of genipin. Solubility was highest for CPI (~94 %), followed by LPI (~90 %), FPI (~85 %) and SPI (~50 %). Surface characteristics revealed similar zeta potentials (~ - 47 mV) for CPI, LPI and FPI, but lower for SPI (~ - 44 mV). Contrastingly, surface hydrophobicity was greatest for CPI (~137 arbitrary units, AU), followed by SPI/LPI (~70 AU) and FPI (~24 AU). A significant (from 16.73 to ~8.42 mN/m) reduction in interfacial tension was observed in canola oil-water mixtures in the presence of non-crosslinked legume protein isolates. The extent of legume protein isolate-genipin crosslinking was found to be similar for all isolates. Overall, creaming stability increased in the presence of genipin, with maximum stability observed for SPI (65 %), followed by FPI (61 %), LPI (56 %) and finally CPI (50 %).

Authenticity analysis of pear juice employing chromatographic fingerprinting.

Pear juice is predominately composed of carbohydrates/polyols (>95% of the total soluble solids), making it susceptible to adulteration by the addition of less expensive commercial sweeteners. In this research, the major carbohydrate and polyol (fructose, glucose, sucrose, and sorbitol) content of 32 pure pear juices representing five world producing regions and three years of production was determined. Additionally, methods employing oligosaccharide profiling to detect the debasing of these samples with four commercial sweeteners (HFCS 55 and 90, TIS, and HIS) were developed using capillary gas chromatography with flame ionization detection (CGC-FID) and high-performance liquid chromatography with pulsed amperometric detection (HPAE-PAD). Detection limits for the four commercial sweeteners ranged from 0.5 to 5.0% (v/v). In addition, the developed CGC-FID method could be used to (a) detect the addition of pear to apple juice via arbutin detection and (b) determine if a pear juice was produced using enzymatic liquefaction via the presence of O-ß-d-glucopyranosyl-(1→4)-d-glucopyranose (cellobiose), all within a single chromatographic analysis.

Total pancreatectomy for the treatment of pancreatic neoplasms.

BACKGROUND: Total pancreatectomy (TP) is an operation that has long been associated with high morbidity and mortality, and rarely advocated for treatment of pancreatic tumours. Because of the improvements in diabetes management, there has been renewed interest in TP for treatment of pancreatic neoplasm, with a need to reassess outcomes. METHODS: Fifteen patients (9%) underwent TP in a single Australian high-volume tertiary institution between August 2005 and January 2012. Pancreaticoduodenectomy (PD) was performed in 150 patients during the same time period. Indications and peri-operative outcomes of patients were compared. RESULTS: TP was performed for treatment of malignancy in 13 (87%) cases, for tumours involving the pancreatic neck. Complete tumour clearance achieved. Portal vein resections were more frequently required than in the PD group (5 (33%) versus 9 (6%); P = 0.004), as were blood transfusions (11 (73%) versus 28 (19%); P < 0.001), and median operative times were longer (10.5 versus 7.0 h; P < 0.001). Overall, complications were significantly greater in the TP group compared with the PD group (13 (87%) versus 86 (57%); P = 0.029), but the difference was mainly as a result of high grade I complications, in particular symptomatic hypoglycaemia (4 (27%) versus 0 (0%): P < 0.001) that could be easily managed. The overall lengths of stay and readmission rates were similar between groups. CONCLUSION: TP should be considered in selective cases for treatment of pancreatic neoplasm if it allows complete clearance. The procedure does not appear to be associated with significant increases in serious complications compared with PD.

Encapsulation of flaxseed oil using a benchtop spray dryer for legume protein-maltodextrin microcapsule preparation.

Flaxseed oil was microencapsulated employing a wall material matrix of either chickpea (CPI) or lentil protein isolate (LPI) and maltodextrin using a benchtop spray dryer. Effects of emulsion formulation (oil, protein and maltodextrin levels) and protein source (CPI vs LPI) on the physicochemical characteristics, oxidative stability, and release properties of the resulting capsules were investigated. Microcapsule formulations containing higher oil levels (20% oil, 20% protein, 60% maltodextrin) were found to have higher surface oil and lower encapsulation efficiencies. Overall, LPI-maltodextrin capsules gave higher flaxseed oil encapsulation efficiencies (∼88.0%) relative to CPI-maltodextrin matrices (∼86.3%). However, both designs were found to provide encapsulated flaxseed oil protection against oxidation over a 25 d room temperature storage study relative to free oil. Overall, ∼37.6% of encapsulated flaxseed oil was released after 2 h under simulated gastric fluid, followed by the release of an additional ∼46.6% over a 3 h period under simulated intestinal fluid conditions.

Microcapsule production employing chickpea or lentil protein isolates and maltodextrin: physicochemical properties and oxidative protection of encapsulated flaxseed oil.

Flaxseed oil was microencapsulated, employing a wall material matrix of either chickpea (CPI) or lentil protein isolate (LPI) and maltodextrin, followed by freeze-drying. Effects of oil concentration (5.3-21.0%), protein source (CPI vs. LPI) and maltodextrin type (DE 9 and 18) and concentration (25.0-40.7%), on both the physicochemical characteristics and microstructure of the microcapsules, were investigated. It was found that an increase in emulsion oil concentration resulted in a concomitant increase in oil droplet diameter and microcapsule surface oil content, and a decrease in oil encapsulation efficiency. Optimum flaxseed oil encapsulation efficiency (∼83.5%), minimum surface oil content (∼2.8%) and acceptable mean droplet diameter (3.0 µm) were afforded with 35.5% maltodextrin-DE 9 and 10.5% oil. Microcapsules, formed by employing these experimental conditions, showed a protective effect against oxidation versus free oil over a storage period of 25 d at room temperature.

Gelation in protein extracts from cold acclimated and non-acclimated winter rye (Secale cereale L. cv Musketeer).

A protein gel is a three-dimensional network consisting of molecular interactions between biopolymers that entrap a significant volume of a continuous liquid phase (water). Molecular interactions in gels occur at junction zones within and between protein molecules through electrostatic forces, hydrogen bonding, hydrophobic associations (van der Waals attractions) and covalent bonding. Gels have the physicochemical properties of both solids and liquids, and are extremely important in the production and stability of a variety of foods, bioproducts and pharmaceuticals. In this study, gelation was induced in phenol extracted protein fractions from non-acclimated (NA) and cold-acclimated (CA) winter rye (Secale cereale L. cv Musketeer) leaf tissue after repeated freeze-thaw treatments. Gel formation only occurred at high pH (pH 12.0) and a minimum of 3-4 freeze-thaw cycles were required. The gel was thermally stable and only a specific combination of chemical treatments could disrupt the gel network. SDS-PAGE analysis identified ribulose-1,5-bisphosphate carboxylase oxygenase (Rubisco) as the major protein component in the gel, although Rubisco itself did not appear to be a factor in gelation. Raman spectroscopy suggested changes in protein secondary structure during freeze-thaw cycles. Overall, the NA and CA gels were similar in composition and structure, with the exception that the CA gel appeared to be amyloidic in nature based on thioflavin T (ThT) fluorescence. Protein gelation, particularly in the apoplast, may confer protection against freeze-induced dehydration and potentially have a commercial application to improve frozen food quality.

A fast track recovery program significantly reduces hospital length of stay following uncomplicated pancreaticoduodenectomy.

CONTEXT: Factors affecting length of hospital stay after uncomplicated pancreaticoduodenectomy have not been reported. We hypothesized that patients undergoing uncomplicated pancreaticoduodenectomy treated by fast track recovery program would have a shorter length of hospital stay compared to those managed by a standard program. METHODS: Patients without surgical or medical complications following pancreaticoduodenectomy managed by fast track or standard protocols, between 2005 and 2011, were identified and prognostic predictors for length of hospital stay determined. RESULTS: Forty-one patients treated by pancreaticoduodenectomy had no medical or surgical complications during this period. Of these patients, 20 underwent fast track recovery program compared to 21 who underwent standard care. Patients in the standard group were more likely to have a feeding jejunostomy tube (P<0.001), pylorus preserving procedure (P=0.001) and a nasogastric tube in place longer than 24 hours postoperatively (P<0.001). The median postoperative length of stay was shorter in the fast track recovery program group (8 days, range: 7-16 days) versus 14 days, range: 8-29 days; P<0.001). There were three readmissions in the fast track recovery program related to abdominal pain and none in the standard group. The overall length of stay, accounting for readmissions, still remained significantly shorter in the fast track recovery program group (median 9 days, range: 7-17 days versus median14 days, range: 8-29 days ; P<0.001). There were no significant differences in discharge destination between groups. On multivariate analysis, the only factor independently associated with postoperative discharge by day 8 was fast track recovery program (OR: 37.1, 95% CI: 4.08-338; P<0.001). CONCLUSION: Fast track recovery program achieved significantly shorter length of stay following uncomplicated pancreaticoduodenectomy.

Development of lyophilized gemini surfactant-based gene delivery systems: influence of lyophilization on the structure, activity and stability of the lipoplexes.

PURPOSE: Cationic gemini surfactants have been studied as non-viral vectors for gene therapy. Clinical applications of cationic lipid/DNA lipoplexes are restricted by their instability in aqueous formulations. In this work, we investigated the influence of lyophilization on the essential physiochemical properties and in vitro transfection of gemini surfactant-lipoplexes. Additionally, we evaluated the feasibility of lyophilization as a technique for preparing lipoplexes with long term stability. METHODS: A gemini surfactant [12-7NH-12] and plasmid DNA encoding for interferon-γ were used to prepare gemini surfactant/pDNA [P/G] lipoplexes. Helper lipid DOPE [L] was incorporated in all formulation producing a [P/G/L] system. Sucrose and trehalose were utilized as stabilizing agents. To evaluate the ability of lyophilization to improve the stability of gemini surfactant-based lipoplexes, four lyophilized formulations were stored at 25˚C for three months. The formulations were analyzed at different time-points for physiochemical properties and in vitro transfection. RESULTS: The results showed that both sucrose and trehalose provided anticipated stabilizing effect. The transfection efficiency of the lipoplexes increased 2-3 fold compared to fresh formulations upon lyophilization. This effect can be attributed to the improvement of DNA compaction and changes in the lipoplex morphology due to the lyophilization/rehydration cycles. The physiochemical properties of the lyophilized formulations were maintained throughout the stability study. All lyophilized formulations showed a significant loss of gene transfection activity after three months of storage. Nevertheless, no significant losses of transfection efficiency were observed for three formulations after two months storage at 25 ˚C. CONCLUSION: Lyophilization significantly improved the physical stability of gemini surfactant-based lipoplexes compared to liquid formulations. As well, lyophilization improved the transfection efficiency of the lipoplexes. The loss of transfection activity upon storage is most probably due to the conformational changes in the supramolecular structure of the lipoplexes as a function of time and temperature rather than to DNA degradation.

Major carbohydrate, polyol, and oligosaccharide profiles of agave syrup. Application of this data to authenticity analysis.

Nineteen pure agave syrups representing the three major production regions and four processing facilities in Mexico were analyzed for their major carbohydrate, polyol, and oligosaccharide profiles, as well as their physicochemical properties (pH, °Brix, total acidity, percent total titratable acidity, and color). Additionally, the detection of intentional debasing of agave syrup with four commercial nutritive sweeteners (HFCS 55 and 90, DE 42 and sucrose) was afforded by oligosaccharide profiling employing both high performance anion exchange liquid chromatography with pulsed amperometric detection (HPAE-PAD) and capillary gas chromatography with flame ionization detection (CGC-FID). Results showed that the major carbohydrate and polyol in agave syrups were fructose and inositol with mean concentrations of 84.29% and 0.38%, respectively. Oligosaccharide profiling was extremely successful for adulteration detection with detection limits ranging from 0.5 to 2.0% for the aforementioned debasing agents. Also, all four of these possible adulterants could be detected within a single chromatographic analysis.

Lentil and chickpea protein-stabilized emulsions: optimization of emulsion formulation.

Chickpea and lentil protein-stabilized emulsions were optimized with regard to pH (3.0-8.0), protein concentration (1.1-4.1% w/w), and oil content (20-40%) for their ability to form and stabilize oil-in-water emulsions using response surface methodology. Specifically, creaming stability, droplet size, and droplet charge were assessed. Optimum conditions for minimal creaming (no serum separation after 24 h), small droplet size (<2 µm), and high net droplet charge (absolute value of ZP > 40 mV) were identified as 4.1% protein, 40% oil, and pH 3.0 or 8.0, regardless of the plant protein used for emulsion preparation.

Rapid detection of ciprofloxacin effects on Fusarium graminearum and F. avenaceum cells in modulating environmental pH using a reactive, non-toxic food-dye indicator.

The objective of the study was to assess the effect of ciprofloxacin antibiotic on the physiological or phenotypic characteristics of food-borne toxigenic Fusarium graminearum and F. avenaceum molds under in vitro conditions. In the presence of ciprofloxacin, Fusarium mycelia growth and morphology were altered based on the antibiotic concentration range used. Results showed that ciprofloxacin in concentrations ≥40µg/mL induced chlamydospore formation in Fusaria and as such, this antibiotic should be considered as an important abiotic stress factor and growth inhibitor. A novel method was investigated to correlate chlamydospore formation with the colour changes observed in FD&C Green Number 3, a common water soluble food dye. The antibiotic-treated F. graminearum and F. avenaceum isolates produced chamydospores, which in turn altered environmental pH with concomitant changes in the colour and intensity of the dye. The colour changes observed as a function of environmental pH were supported by instrumental methods (pH meter and spectroscopy), and a commercial pH indicator (thymol blue) results. In conclusion, we propose that FD&C Green Number 3 can be used as an accurate indicator for the rapid assessment of Fusarium molds when grown on ciprofloxacin antibiotic-containing substrate. Special emphasis should be given to an indirect risk assessment of antibiotic effects on toxic molds.