Analysis of Selected Volatile Organic Compounds in Split and Nonsplit Swiss Cheese Samples Using Selected‐Ion Flow Tube Mass Spectrometry (SIFT‐MS)

Splits/cracks are recurring product defects that negatively affect the Swiss cheese industry. Investigations to understand the biophysicochemical aspects of these defects, and thus determine preventive measures against their occurrence, are underway. In this study, selected‐ion, flow tube mass spectrometry was employed to determine the volatile organic compound (VOC) profiles present in the headspace of split compared with nonsplit cheeses. Two sampling methodologies were employed: split compared with nonsplit cheese vat pair blocks; and comparison of blind, eye, and split segments within cheese blocks. The variability in VOC profiles was examined to evaluate the potential biochemical pathway chemistry differences within and between cheese samples. VOC profile inhomogeneity was most evident in cheeses between factories. Evaluation of biochemical pathways leading to the formation of key VOCs differentiating the split from the blind and eye segments within factories indicated release of additional carbon dioxide by‐product. These results suggest a factory‐dependent cause of split formation that could develop from varied fermentation pathways in the blind, eye, and split areas within a cheese block. The variability of VOC profiles within and between factories exhibit varied biochemical fermentation pathways that could conceivably be traced back in the making process to identify parameters responsible for split defect.     

Studies on apple and blueberry fruit constituents: do the polyphenols reach the colon after ingestion?

The aim of our studies was to determine the amount of polyphenols reaching the colon after oral intake of apple juice and blueberries. After a polyphenol-free diet healthy ileostomy volunteers consumed a polyphenol-rich cloudy apple juice while others consumed anthocyanin-rich blueberries. Ileostomy effluent was collected and polyphenols were identified using HPLC-DAD as well as HPLC-ESI-MS/MS; quantification was performed with HPLC-DAD. Most of the orally administered apple polyphenols were absorbed from or metabolized in the small intestine. Between 0 and 33% of the oral dose was recovered in the ileostomy bags with a maximum of excretion after 2 h. A higher amount of the blueberry anthocyanins under study (up to 85%, depending on the sugar moiety) were determined in the ileostomy bags and therefore would reach the colon under physiological circumstances. Such structure-related availability has to be considered when polyphenols are used in model systems to study potential preventive effects in colorectal diseases.     

Identification and differentiation of haze substances using Raman microspectroscopy

The identification of turbidity or haze is an important part of brewery analytics. Haze can be caused by various issues throughout the brewing process and identifying the composition can pinpoint the origin. Haze analytics are commonly based on enzymatic or microscopic methods, which can be inaccurate or laborious. Raman microspectroscopy (RMS) presents a promising alternative for detecting haze particles. It is fast, easy to use and requires little sample preparation. Here, the applicability of RMS at 532 nm and 785 nm to identify potentially haze-forming particles has been evaluated. At 532 nm, measurements were taken with standard microscope slides. Due to the high fluorescent background of normal glass at an excitation wavelength of 785 nm, fused quartz microscope slides were used at this wavelength. Starch, arabinoxylan, cellulose, yeast β-glucan, barley β-glucan, gliadin, ferulic acid, proline, glutamine, calcium oxalate and PVPP were identified at 532 nm. The same substances when analysed at 785 nm resulted in problems with weak carbohydrate spectra of the β-glucans and arabinoxylan. All the other substances could be analysed at 785 nm. Catechin, which produced fluorescence noise at 532 nm could be identified at 785 nm. Although there is an issue with the intense fluorescence noise of some beer components, Raman microspectroscopy has great potential in haze analysis and potentially in wider brewery analyses. © 2020 The Authors. Journal of the Institute of Brewing published by John Wiley & Sons Ltd on behalf of The Institute of Brewing & Distilling     

Cycloadditions of Trans-Cyclooctenes and Nitrones as Tools for Bioorthogonal Labelling

Trans-cyclooctenes (TCOs) represent interesting and highly reactive dipolarophiles for organic transformations including bioorthogonal chemistry. Herein we show that TCOs react rapidly with nitrones and that these reactions are bioorthogonal. Kinetic analysis of acyclic and cyclic nitrones with strained-trans-cyclooctene (s-TCO) shows fast reactivity and demonstrates the utility of this cycloaddition reaction for bioorthogonal labelling. Labelling of the bacterial peptidoglycan layer with unnatural d -amino acids tagged with nitrones and s-TCO-Alexa488 is demonstrated. These new findings expand the bioorthogonal toolbox, and allow TCO reagents to be used in bioorthogonal applications beyond tetrazine ligations for the first time and open up new avenues for bioorthogonal ligations with diverse nitrone reactants.     

Virtual Pharmacophore Screening Identifies Small-Molecule Inhibitors of the Rev1-CT/RIR Protein–Protein Interaction

Translesion synthesis (TLS) has emerged as a mechanism through which several forms of cancer develop acquired resistance to first-line genotoxic chemotherapies by allowing replication to continue in the presence of damaged DNA. Small molecules that inhibit TLS hold promise as a novel class of anticancer agents that can serve to enhance the efficacy of these front-line therapies. We previously used a structure-based rational design approach to identify the phenazopyridine scaffold as an inhibitor of TLS that functions by disrupting the protein–protein interaction (PPI) between the C-terminal domain of the TLS DNA polymerase Rev1 (Rev1-CT) and the Rev1 interacting regions (RIR) of other TLS DNA polymerases. To continue the identification of small molecules that disrupt the Rev1-CT/RIR PPI, we generated a pharmacophore model based on the phenazopyridine scaffold and used it in a structure-based virtual screen. In vitro analysis of promising hits identified several new chemotypes with the ability to disrupt this key TLS PPI. In addition, several of these compounds were found to enhance the efficacy of cisplatin in cultured cells, highlighting their anti-TLS potential.     

The quantum magnetism of individual manganese-12-acetate molecular magnets anchored at surfaces

The high intrinsic spin and long spin relaxation time of manganese-12-acetate (Mn(12)) makes it an archetypical single molecular magnet. While these characteristics have been measured on bulk samples, questions remain whether the magnetic properties replicate themselves in surface supported isolated molecules, a prerequisite for any application. Here we demonstrate that electrospray ion beam deposition facilitates grafting of intact Mn(12) molecules on metal as well as ultrathin insulating surfaces enabling submolecular resolution imaging by scanning tunneling microscopy. Using scanning tunneling spectroscopy we detect spin excitations from the magnetic ground state of the molecule at an ultrathin boron nitride decoupling layer. Our results are supported by density functional theory based calculations and establish that individual Mn(12) molecules retain their intrinsic spin on a well chosen solid support.     

Hemocompatibility of polymers for use in vascular endoluminal implants

Implanted polymers for cardiovascular applications may function as structural supports, barriers, or provide a means for local drug delivery. Several thermoplastic biodegradable drug delivery polymers are potential candidates for blood-contacting implant applications. For intravascular applications specifically, a criterion for material selection is the intrinsic hemocompatibility of the baseline polymer. As an initial screening approach for selection of polymers for in vivo use, thin films of polyesters: poly(ɛ-caprolactone) (PCL), poly(lactic acid) (PLA), poly(lactic-co-glycolic acid) (PLGA); polyanhydrides: poly(fatty acid dimer-co-sebacic acid) (PFAD:SA) and poly(biscarboxyphenoxypropane-co-sebacic acid) (PCPP:SA); and poly(ethylene glycol) (PEG)-ylated polyesters: PLA:PEG, PCL:PEG and PCL:PLA:PEG polymers were spin-cast on glass cover slips and placed in an in vitro flow system exposing them at a controlled shear to overflowing human whole blood. Platelet adherence, aggregate formation, and thrombus formation, as well as leukocyte adherence were assessed following 5 min of flow. At 5 min of flow the rank order of materials, in terms of least to most thrombogenic was: PCL < PFAD:SA < PCPP:SA < PLGA < PLA. All PEGylated materials, in general, had less thrombus formation than baseline unmodified materials.     

Ambulance transport rates after motor vehicle collision for older vs. younger adults: A population-based study

Older adults are at greater risk than younger adults for life-threatening injury after motor vehicle collision (MVC). Among those with life-threatening injury, older adults are also at greater risk of not being transported by emergency medical services (EMS) to an emergency department. Despite the greater risk of serious injury and non-transportation among older adults, little is known about the relationship between patient age and EMS transportation rates for individuals experiencing MVC. We describe transport rates across the age-span for adults seen by EMS after experiencing MVC using data reported to the North Carolina Department of Motor Vehicles between 2008 and 2011. Of all adults aged 18 years and older experiencing MVC and seen by EMS (n = 484,310), 36.3% (n = 175,768) were transported to an emergency department. Rates of transport for individuals seen by EMS after MVC increased only a small amount with increasing patient age. After adjusting for potential confounders of the relationship between patient age and the decision to transport (patient gender, patient race, air bag deployment, patient trapped or ejected, and injury severity), transport rates were: age 18–64 = 36.0% (95% confidence interval [CI], 35.9–36.2%); age 65–74 = 36.6% (95% CI, 36.0–37.1%); age 75–84 = 37.3% (95% CI, 36.5–38.1%), and age 85–94 = 38.2% (95% CI, 36.7–39.8%). In North Carolina between 2008 and 2011, the transportation rate was only slightly higher for older adults than for younger adults, and most older adults experiencing MVC and seen by EMS were not transported to the emergency department. These findings have implications for efforts to improve the sensitivity of criteria used by EMS to determine the need for transport for older adults experiencing MVC.     

Evaluation of the repeatability and reproducibility of a suite of qPCR-based microbial source tracking methods

Many PCR-based methods for microbial source tracking (MST) have been developed and validated within individual research laboratories. Inter-laboratory validation of these methods, however, has been minimal, and the effects of protocol standardization regimes have not been thoroughly evaluated. Knowledge of factors influencing PCR in different laboratories is vital to future technology transfer for use of MST methods as a tool for water quality management. In this study, a blinded set of 64 filters (containing 32 duplicate samples generated from 12 composite fecal sources) were analyzed by three to five core laboratories with a suite of PCR-based methods utilizing standardized reagents and protocols. Repeatability (intra-laboratory variability) and reproducibility (inter-laboratory variability) of observed results were assessed. When standardized methodologies were used, intra- and inter-laboratory %CVs were generally low (median %CV 0.1–3.3% and 1.9–7.1%, respectively) and comparable to those observed in similar inter-laboratory validation studies performed on other methods of quantifying fecal indicator bacteria (FIB) in environmental samples. ANOVA of %CV values found three human-associated methods (BsteriF1, BacHum, and HF183Taqman) to be similarly reproducible (p > 0.05) and significantly more reproducible (p < 0.05) than HumM2. This was attributed to the increased variability associated with low target concentrations detected by HumM2 (approximately 1–2 log₁₀copies/filter lower) compared to other human-associated methods. Cow-associated methods (BacCow and CowM2) were similarly reproducible (p > 0.05). When using standardized protocols, variance component analysis indicated sample type (fecal source and concentration) to be the major contributor to total variability with that from replicate filters and inter-laboratory analysis to be within the same order of magnitude but larger than inherent intra-laboratory variability. However, when reagents and protocols were not standardized, inter-laboratory %CV generally increased with a corresponding decline in reproducibility. Overall, these findings verify the repeatability and reproducibility of these MST methods and highlight the need for standardization of protocols and consumables prior to implementation of larger scale MST studies involving multiple laboratories.     

Natural Wax for Transient Electronics

Emerging classes of bioresorbable electronic materials serve as the basis for active biomedical implants that are capable of providing sensing, monitoring, stimulating, and other forms of function over an operating period matched to biological processes such as wound healing. These platforms are of interest because subsequent dissolution, enzymatic degradation, and/or bioresorption can eliminate the need for surgical extraction. This report introduces natural wax materials as long‐lived, hydrophobic encapsulation layers for such systems, where biodegradation eventually occurs by chain scission. Studies of wax stability as an encapsulation material demonstrate the ability to retain operation of underlying biodegradable electronic systems for durations between a few days to a few weeks during complete immersion in aqueous solutions in ex‐vivo physiological conditions. Electrically conductive composites result from the addition of tungsten micro/nanoparticles, as a conductive, printable paste with similar lifetimes. Demonstrations of these materials in partially biodegradable wireless light‐emitting diodes and near‐field communication circuits illustrate their use in functional bioresorbable electronic systems. Investigations in animal models reveal no signs of toxicity or other adverse biological responses.