Application of a Modified 3D-PCSI-MS Ion Source to On-Site, Trace Evidence Processing via Integrated Vacuum Collection.

Trace evidence, including hair, fibers, soil/dust, and gunshot residue (GSR), can be recovered from a crime scene to help identify or associate a suspect with illegal activities via physical, chemical, and biological testing. Vacuum collection is one technique that is employed in recovering such trace evidence but is often done so in a targeted manner, leaving other complementary, chemical-specific information unexamined. Here, we describe a modified 3D-printed cone spray ionization (3D-PCSI) source with integrated vacuum collection for on-site, forensic evidence screening, allowing the processing of targeted physical traces and nontargeted chemical species alike. The reported form factor allows sample collection, onboard extraction, filtration, and spray-based ionization in a singular vessel with minimal handling of evidence by the operator. Utilizing authentic forensic evidence types and portable MS instrumentation, this new method was characterized through systematic studies that replicate CSI applications. Reliability in the form of false positive/negative response rates was determined from a modest, user-blinded data set, and other attributes, such as collection efficacy and detection limit, were examined.

Flipping the switch: dynamic modulation of membrane transporter activity in bacteria.

The controlled entry and expulsion of small molecules across the bacterial cytoplasmic membrane is essential for efficient cell growth and cellular homeostasis. While much is known about the transcriptional regulation of genes encoding transporters, less is understood about how transporter activity is modulated once the protein is functional in the membrane, a potentially more rapid and dynamic level of control. In this review, we bring together literature from the bacterial transport community exemplifying the extensive and diverse mechanisms that have evolved to rapidly modulate transporter function, predominantly by switching activity off. This includes small molecule feedback, inhibition by interaction with small peptides, regulation through binding larger signal transduction proteins and, finally, the emerging area of controlled proteolysis. Many of these examples have been discovered in the context of metal transport, which has to finely balance active accumulation of elements that are essential for growth but can also quickly become toxic if intracellular homeostasis is not tightly controlled. Consistent with this, these transporters appear to be regulated at multiple levels. Finally, we find common regulatory themes, most often through the fusion of additional regulatory domains to transporters, which suggest the potential for even more widespread regulation of transporter activity in biology.

Rapid screening of high-priority N-nitrosamines in pharmaceutical, forensic, and environmental samples with paper spray ionization and filter cone spray ionization-mass spectrometry.

RATIONALE: The burgeoning concern of N-nitrosamine (NAM) contamination found in various pharmaceutical compositions has increased the demand for rapid and reliable screening methods to better assess the breadth of the problem. These carcinogenic compounds are also found in food, water, and soil, and they have been used in poison-related homicides. METHODS: A combination of complementary, ambient ionization methods, paper spray ionization (PSI) and filter cone spray ionization (FCSI)-mass spectrometry (MS), was characterized towards trace-level residue screening of select NAMs (e.g., N-nitrosodimethylamine, N-nitrosodiethylamine, N-nitrosodibutylamine) directly from complex and problematic matrices of interest, including prescription and over-the-counter tablets, drinking water, soil, and consumable goods. Spectral data for analyte confirmation and detection limit studies were collected using a Thermo LCQ Fleet ion trap mass spectrometer. RESULTS: PSI-MS and FCSI-MS readily produced mass spectral data marked by their simplicity (e.g., predominantly protonated molecular ions observed) and congruence with traditional electrospray ionization mass spectra in under 2 min. per sample. Both methods proved robust to the complex matrices tested, yielding ion signatures for target NAMs, as well as active pharmaceutical ingredients for analyzed tablets, flavorants inherent to food products, etc. Low part-per-million detection limits were observed but were shown dependent on sample composition. CONCLUSIONS: PSI-MS and FCSI-MS were successful in detecting trace-level NAMS in complex liquid- and solid-phase matrices with little to no prior preparation. This work suggests that these methodologies can provide a means for assessing problematic pharmaceutical adulterants/degradants for expedited quality control, as well as enhancing environmental stewardship efforts and forensic investigations.

Production of diverse brGDGTs by Acidobacterium Solibacter usitatus in response to temperature, pH, and O2 provides a culturing perspective on brGDGT proxies and biosynthesis.

Branched glycerol dialkyl glycerol tetraethers (brGDGTs) are bacterial membrane lipids that are frequently employed as paleoenvironmental proxies because of the strong empirical correlations between their relative abundances and environmental temperature and pH. Despite the ubiquity of brGDGTs in modern and paleoenvironments, the source organisms of these enigmatic compounds have remained elusive, requiring paleoenvironmental applications to rely solely on observed environmental correlations. Previous laboratory and environmental studies have suggested that the globally abundant bacterial phylum of the Acidobacteria may be an important brGDGT producer in nature. Here, we report on experiments with a cultured Acidobacterium, Solibacter usitatus, that makes a large portion of its cellular membrane (24 ± 9% across all experiments) out of a structurally diverse set of tetraethers including the common brGDGTs Ia, IIa, IIIa, Ib, and IIb. Solibacter usitatus was grown across a range of conditions including temperatures from 15 to 30°C, pH from 5.0 to 6.5, and O2 from 1% to 21%, and demonstrated pronounced shifts in the degree of brGDGT methylation across these growth conditions. The temperature response in culture was in close agreement with trends observed in published environmental datasets, supporting a physiological basis for the empirical relationship between brGDGT methylation number and temperature. However, brGDGT methylation at lower temperatures (15 and 20°C) was modulated by culture pH with higher pH systematically increasing the degree of methylation. In contrast, pH had little effect on brGDGT cyclization, supporting the hypothesis that changes in bacterial community composition may underlie the link between cyclization number and pH observed in environmental samples. Oxygen concentration likewise affected brGDGT methylation highlighting the potential for this environmental parameter to impact paleotemperature reconstruction. Low O2 culture conditions further resulted in the production of uncommon brGDGT isomers that could be indicators of O2 limitation. Finally, the production of brGTGTs (trialkyl tetraethers) in addition to the previously discovered iso-C15-based mono- and diethers in S. usitatus suggests a potential biosynthetic pathway for brGDGTs that uses homologs of the archaeal tetraether synthase (Tes) enzyme for tetraether synthesis from diethers.

Comparative analysis of off-road vehicle crashes in children: motorcycles versus quad bikes.

OBJECTIVE: To characterise and compare off-road motorcycle and quad bike crashes in children in New South Wales (NSW), Australia. METHODS: A retrospective, cross-sectional study was performed of children aged 0-16 years, admitted to hospitals in NSW, from 2001 to 2018 following an injury sustained in an off-road motorcycle or quad bike crash, using linked hospital admissions, mortality and census data.Motorcycle and quad bike injuries were compared regarding: demographics; incidence; body region injured and type of injury; injury severity based on the survival risk ratio; length of stay and mortality. RESULTS: There were 6624 crashes resulting in hospitalisation; 5156 involving motorcycles (77.8%) and 1468 involving quad bikes (22.2%). There were 10 fatalities (6 from motorcycles and 4 from quad bikes). The rates of injury declined over the study period for motorcycles, but not for quad bikes.Motorcycle riders were more likely than quad bike riders to have lower limb injuries (OR 1.49, p<0.001) but less likely to have head/neck (OR 0.616, p<0.001), abdominal (OR 0.778, p=0.007) and thoracic (OR 0.745, p=0.003) injuries. Quad bike crashes resulted in higher injury severity (mean International Classification Injury Severity Score 0.975 vs 0.977, p=0.03) and longer hospital stay (mean 2.42 days vs 2.09 days, p=0.01). CONCLUSIONS: There are significant differences between quad bike and motorcycle crashes in injury type and affected body region. While quad bike injuries in children were more severe, there were almost four times more hospitalisations from motorcycles overall. The overall larger burden of motorcycle crashes suggests a greater focus of injury prevention countermeasures for two-wheeled riders is needed.

Structural basis of ion - substrate coupling in the Na+-dependent dicarboxylate transporter VcINDY.

The Na+-dependent dicarboxylate transporter from Vibrio cholerae (VcINDY) is a prototype for the divalent anion sodium symporter (DASS) family. While the utilization of an electrochemical Na+ gradient to power substrate transport is well established for VcINDY, the structural basis of this coupling between sodium and substrate binding is not currently understood. Here, using a combination of cryo-EM structure determination, succinate binding and site-directed cysteine alkylation assays, we demonstrate that the VcINDY protein couples sodium- and substrate-binding via a previously unseen cooperative mechanism by conformational selection. In the absence of sodium, substrate binding is abolished, with the succinate binding regions exhibiting increased flexibility, including HPinb, TM10b and the substrate clamshell motifs. Upon sodium binding, these regions become structurally ordered and create a proper binding site for the substrate. Taken together, these results provide strong evidence that VcINDY's conformational selection mechanism is a result of the sodium-dependent formation of the substrate binding site.

The Impact of Concomitant Ipsilateral Knee Pain on Functional Outcomes in Total Ankle Arthroplasty.

In end-stage ankle arthritis, little is known about the impact of concomitant knee pathology, including the impact of ipsilateral knee pain on total ankle arthroplasty (TAA) outcomes. The aim of this study was to determine the prevalence of ipsilateral preoperative knee pain in patients undergoing TAA and analyze its impact on patient-reported functional outcome measures (PROMs). A retrospective review was performed on the Vancouver End Stage Ankle Arthritis Database at a single institution. In total, 114 patients were studied, with patient demographics collected preoperatively, including the presence or absence of knee pain. Postoperative follow-up was performed at 5 years, primarily analyzing disease-specific PROMs, including the Ankle Osteoarthritis Score (AOS) and Ankle Arthritis Score (AAS). Multivariate mixed-effects linear regression models compared the scores between the groups. In total, 31 patients (27.2%) presented with concomitant ipsilateral knee pain. Despite more females in the knee pain group (64.5% vs 36.1%) there were no other significant differences at baseline between the knee pain and no knee pain groups in terms of demographics or baseline primary disease specific PROMs. At 5 years, the patients with knee pain had significantly worse AAS (37.9 ± 23.8 vs 21.2 ± 16.3, P = .004) and AOS total scores (38.1 ± 24.1 vs 21.9 ± 15.5, P = .005) compared with the no-knee pain group. Both groups improved significantly from baseline across all outcome measures; however, the magnitude of improvement was less in the knee pain group. Our study demonstrated that over one-quarter of patients with end-stage ankle arthritis undergoing TAA present with ipsilateral concomitant knee pain. If present, it is associated with worse functional outcomes at the 5-year mark. Further studies are needed to evaluate if knee pain influences complications, implant failure rates, and survival.Levels of Evidence: Level III.

Topological analysis of a bacterial DedA protein associated with alkaline tolerance and antimicrobial resistance.

Maintaining membrane integrity is of paramount importance to the survival of bacteria as the membrane is the site of multiple crucial cellular processes including energy generation, nutrient uptake and antimicrobial efflux. The DedA family of integral membrane proteins are widespread in bacteria and are associated with maintaining the integrity of the membrane. In addition, DedA proteins have been linked to resistance to multiple classes of antimicrobials in various microorganisms. Therefore, the DedA family are attractive targets for the development of new antibiotics. Despite DedA family members playing a key physiological role in many bacteria, their structure, function and physiological role remain unclear. To help illuminate the structure of the bacterial DedA proteins, we performed substituted cysteine accessibility method (SCAM) analysis on the most comprehensively characterized bacterial DedA protein, YqjA from Escherichia coli. By probing the accessibility of 15 cysteine residues across the length of YqjA using thiol reactive reagents, we mapped the topology of the protein. Using these data, we experimentally validated a structural model of YqjA generated using evolutionary covariance, which consists of an α-helical bundle with two re-entrant hairpin loops reminiscent of several secondary active transporters. In addition, our cysteine accessibility data suggest that YqjA forms an oligomer wherein the protomers are arranged in a parallel fashion. This experimentally verified model of YqjA lays the foundation for future work in understanding the function and mechanism of this interesting and important family.

Thermostability-based binding assays reveal complex interplay of cation, substrate and lipid binding in the bacterial DASS transporter, VcINDY.

The divalent anion sodium symporter (DASS) family of transporters (SLC13 family in humans) are key regulators of metabolic homeostasis, disruption of which results in protection from diabetes and obesity, and inhibition of liver cancer cell proliferation. Thus, DASS transporter inhibitors are attractive targets in the treatment of chronic, age-related metabolic diseases. The characterisation of several DASS transporters has revealed variation in the substrate selectivity and flexibility in the coupling ion used to power transport. Here, using the model DASS co-transporter, VcINDY from Vibrio cholerae, we have examined the interplay of the three major interactions that occur during transport: the coupling ion, the substrate, and the lipid environment. Using a series of high-throughput thermostability-based interaction assays, we have shown that substrate binding is Na+-dependent; a requirement that is orchestrated through a combination of electrostatic attraction and Na+-induced priming of the binding site architecture. We have identified novel DASS ligands and revealed that ligand binding is dominated by the requirement of two carboxylate groups in the ligand that are precisely distanced to satisfy carboxylate interaction regions of the substrate-binding site. We have also identified a complex relationship between substrate and lipid interactions, which suggests a dynamic, regulatory role for lipids in VcINDY's transport cycle.

Paediatric off-road vehicle injury in rural and regional Australia.

OBJECTIVE: Off-road riding of quad bikes and motorcycles is common among children across rural and remote Australia, but is a significant source of injury and hospitalisation. An in-depth analysis of paediatric off-road vehicle crashes was undertaken to inform injury prevention countermeasures by characterising injury patterns and sources of injury. DESIGN: This is a prospective in-depth case series. PARTICIPANTS: Participants are children aged 16 and under who have been hospitalised due to injury sustained from the use of an off-road motorcycle or quad bike in New South Wales, Australia. INTERVENTIONS: Crash investigation techniques (medical data, structured interview, vehicle and crash site inspection) were used to ascertain details of the crash event, protective gear, injury information and contributory factors. RESULTS: Thirty children were recruited, 27 boys and 3 girls, ranging in age from 4 to 16 years, having crashed on off-road motorcycles (n = 27) or quads (n = 3). Most (73.3%) were participating in unstructured social riding. A total of 67 separate injuries were observed, with overall Injury Severity Scores between 1 and 35. There were high rates of wearing helmets and motorcycle-specific garments. The most commonly injured areas were the upper and lower extremities. The most common sources of injury were from impacting the ground, obstacles/other riders or the vehicle. CONCLUSION: This study demonstrates the patterns of riding and injury in rural paediatric off-road vehicle riders, occurring despite high rates of helmet/protective gear use. This underscores the need for investigation into the injury mitigation and fit properties of protective gear and the inherent risks for physically and developmentally maturing children.

Rapid, in situ detection of chemical warfare agent simulants and hydrolysis products in bulk soils by low-cost 3D-printed cone spray ionization mass spectrometry.

Chemical warfare agents (CWAs) are toxic chemicals that have been used as disabling or lethal weapons in war, terrorist attacks, and assasinations. The Chemical Weapons Convention (CWC) has prohibited the use, development, production, and stockpiling of CWAs since its initiation in 1997, however, the threat of deployment still looms. Detection of trace CWAs post-deployment or post-remediation, in bulk matrices such as soil, often requires lengthy sample preparation steps or extensive chromatographic separation times. 3D-printed cone spray ionization (3D-PCSI), an ambient ionization mass spectrometric (MS) technique, provides a rapid, simple, and low-cost method for trace CWA analysis in soil matrices for both in-laboratory and in-field detection. Described here is the utilization of conductive 3D-printed cones to perform both rapid sampling and ionization for CWA simulants and hydrolysis products in eight solid matrices. The analysis of trace quantities of CWA simulants and hydrolysis products by 3D-PCSI-MS coupled to both a commercial benchtop system and a field-portable MS system is detailed. Empirical limits of detection (LOD) for CWA simulants on the benchtop MS ranged from 100 ppt to 750 ppb and were highly dependant on solid matrix composition, with the portable system yielding similar spectral data from alike matrices, albeit with lower sensitivity.

The Impact of Concomitant LOW Back Pain (LBP) on Functional Outcomes in Total Ankle Arthroplasty (TAA).

Although numerous studies have demonstrated that concomitant low back pain (LBP) is associated with worse functional outcomes in patients undergoing total hip and knee arthroplasty, no study has analyzed its impact on patients undergoing total ankle arthroplasty (TAA). The aim of this study was to determine the prevalence of LBP in people undergoing TAA and analyze its impact on patient reported functional outcome measures (PROMs). A retrospective review was performed on data from the Vancouver End Stage Ankle Arthritis Database. In total, 87 patients undergoing TAA were studied, with patient demographics collected preoperatively, including the absence or presence of LBP. Postoperative follow-up was performed at 5 years, primarily analyzing disease-specific PROMs including the Ankle Osteoarthritis Score and Ankle Arthritis Score. The Short Form-36 was used as a secondary outcome measure to assess global function. Multivariable linear mixed-effects regression models were conducted to compare the PROM between patients with LBP with those without LBP. In total, 30 patients (35%) presented with concomitant LBP. There were no significant differences at baseline between the LBP group and no LBP group in terms of demographics or baseline primary disease-specific PROMs. At 5 years, the patients with LBP had significantly worse Ankle Arthritis Score (32 ± 23 vs 22 ± 17, p = .03), Ankle Osteoarthritis Score Total (34 ± 23 vs 22 ± 16, p = .01), and Short Form-36 physical (PCS) components summaries (33 ± 12 vs 44 ± 9, p = .001) compared to the no-LBP group. Both groups improved significantly from baseline across all outcome measures. Our study demonstrated that the prevalence of concomitant LBP in end stage ankle arthritis undergoing TAA is similar to that described in arthritic knees and hips. If present, it can be associated with worse functional outcomes in the intermediate term. However it is not a contraindication to surgery, with patients still experiencing significant improvements from baseline. Further studies are needed to evaluate if LBP influences complications, implant failure rates and survival.

FIELDABLE MASS SPECTROMETRY FOR FORENSIC SCIENCE, HOMELAND SECURITY, AND DEFENSE APPLICATIONS.

Mass spectrometry is commonly used in forensic chemistry laboratories for sensitive, definitive analysis. There have been significant efforts to bring mass spectrometry analysis on-site through the development of ruggedized, fieldable instruments. Testing samples in the field is of particular interest in forensic science, homeland security, and defense applications. In forensic chemistry, testing seized drugs in the field can significantly improve efficiencies in processing of related criminal cases. The screening of passengers and luggage at transportation hubs is a critical need for homeland security for which mass spectrometry is well suited to provide definitive answers with low false positive rates. Mass spectrometry can yield reliable data for military personnel testing sites for potential chemical weapons release. To meet the needs of the forensic and security communities fieldable mass spectrometers based on membrane inlet systems and hybrid gas chromatography systems have been developed and commercialized. More recently developed ambient ionization mass spectrometry methods can eliminate the time, equipment, and expertise associated with sample preparation, and so are especially appealing for on-site analysis. We describe the development of fieldable mass spectrometry systems, with emphasis on commercially available systems that have been deployed for on-site analysis of seized drugs, chemical warfare agents, explosives, and other analytes of interest to the forensic and security communities. © 2020 John Wiley & Sons Ltd. Mass Spec Rev.

Solvent accessibility changes in a Na+-dependent C4-dicarboxylate transporter suggest differential substrate effects in a multistep mechanism.

The divalent anion sodium symporter (DASS) family (SLC13) plays critical roles in metabolic homeostasis, influencing many processes, including fatty acid synthesis, insulin resistance, and adiposity. DASS transporters catalyze the Na+-driven concentrative uptake of Krebs cycle intermediates and sulfate into cells; disrupting their function can protect against age-related metabolic diseases and can extend lifespan. An inward-facing crystal structure and an outward-facing model of a bacterial DASS family member, VcINDY from Vibrio cholerae, predict an elevator-like transport mechanism involving a large rigid body movement of the substrate-binding site. How substrate binding influences the conformational state of VcINDY is currently unknown. Here, we probe the interaction between substrate binding and protein conformation by monitoring substrate-induced solvent accessibility changes of broadly distributed positions in VcINDY using a site-specific alkylation strategy. Our findings reveal that accessibility to all positions tested is modulated by the presence of substrates, with the majority becoming less accessible in the presence of saturating concentrations of both Na+ and succinate. We also observe separable effects of Na+ and succinate binding at several positions suggesting distinct effects of the two substrates. Furthermore, accessibility changes to a solely succinate-sensitive position suggests that substrate binding is a low-affinity, ordered process. Mapping these accessibility changes onto the structures of VcINDY suggests that Na+ binding drives the transporter into an as-yet-unidentified conformational state, involving rearrangement of the substrate-binding site-associated re-entrant hairpin loops. These findings provide insight into the mechanism of VcINDY, which is currently the only structurally characterized representative of the entire DASS family.

The current role of mass spectrometry in forensics and future prospects.

Mass spectrometry (MS) techniques are highly prevalent in crime laboratories, particularly those coupled to chromatographic separations like gas chromatography (GC) and liquid chromatography (LC). These methods are considered "gold standard" analytical techniques for forensic analysis and have been extensively validated for producing prosecutorial evidentiary data. However, factors such as growing evidence backlogs and problematic evidence types (e.g., novel psychoactive substance (NPS) classes) have exposed limitations of these stalwart techniques. This critical review serves to delineate the current role of MS methods across the broad sub-disciplines of forensic science, providing insight on how governmental steering committees guide their implementation. Novel, developing techniques that seek to broaden applicability and enhance performance will also be highlighted, from unique modifications to traditional hyphenated MS methods to the newer "ambient" MS techniques that show promise for forensic analysis, but need further validation before incorporation into routine forensic workflows. This review also expounds on how recent improvements to MS instrumental design, scan modes, and data processing could cause a paradigm shift in how the future forensic practitioner collects and processes target evidence.

Tafazzin deficiency impairs CoA-dependent oxidative metabolism in cardiac mitochondria.

Barth syndrome is a mitochondrial myopathy resulting from mutations in the tafazzin (TAZ) gene encoding a phospholipid transacylase required for cardiolipin remodeling. Cardiolipin is a phospholipid of the inner mitochondrial membrane essential for the function of numerous mitochondrial proteins and processes. However, it is unclear how tafazzin deficiency impacts cardiac mitochondrial metabolism. To address this question while avoiding confounding effects of cardiomyopathy on mitochondrial phenotype, we utilized Taz-shRNA knockdown (TazKD ) mice, which exhibit defective cardiolipin remodeling and respiratory supercomplex instability characteristic of human Barth syndrome but normal cardiac function into adulthood. Consistent with previous reports from other models, mitochondrial H2O2 emission and oxidative damage were greater in TazKD than in wild-type (WT) hearts, but there were no differences in oxidative phosphorylation coupling efficiency or membrane potential. Fatty acid and pyruvate oxidation capacities were 40-60% lower in TazKD mitochondria, but an up-regulation of glutamate oxidation supported respiration rates approximating those with pyruvate and palmitoylcarnitine in WT. Deficiencies in mitochondrial CoA and shifts in the cardiac acyl-CoA profile paralleled changes in fatty acid oxidation enzymes and acyl-CoA thioesterases, suggesting limitations of CoA availability or "trapping" in TazKD mitochondrial metabolism. Incubation of TazKD mitochondria with exogenous CoA partially rescued pyruvate and palmitoylcarnitine oxidation capacities, implicating dysregulation of CoA-dependent intermediary metabolism rather than respiratory chain defects in the bioenergetic impacts of tafazzin deficiency. These findings support links among cardiolipin abnormalities, respiratory supercomplex instability, and mitochondrial oxidant production and shed new light on the distinct metabolic consequences of tafazzin deficiency in the mammalian heart.

Integrating SERS and PSI-MS with Dual Purpose Plasmonic Paper Substrates for On-Site Illicit Drug Confirmation.

Forensic laboratory backlogs are replete with suspected drug samples. Shifting analysis toward the point of seizure would save significant time and public funds. Moreover, a two-tiered identification strategy for controlled substance testing that relies on two independent, discerning methods could entirely circumvent the need for forensic laboratory testing. To this end, we coupled Raman spectroscopy and paper spray ionization mass spectrometry (PSI-MS) on a single instrumental platform. Both methods are capable of ambient analysis with fieldable instruments, yet Raman is often limited to bulk analysis. Critical to this work is the development of a gold nanoparticle (AuNP)-embedded paper swab to extend the capability of Raman spectroscopy to trace evidence via surface-enhanced Raman scattering (SERS). Plasmonic papers are characterized with respect to SERS signals and compatibility with PSI-MS analysis. Proof-of-principle is established with the identification of five representative drugs, and detection limits on the scale of 1-100 ng are achieved for both PSI-MS and SERS. The integrated SERS-PSI-MS system achieved 99.8% accurate chemical identification in a blind study consisting of 500 samples. Additionally, we demonstrate facile discrimination of several JWH-018 isomers via SERS even when MS and MS2 spectra are indistinguishable. Successful coupling of SERS and PSI-MS to enable on-site chemical analysis by two independent methods can potentially lead to a desirable paradigm shift in the handling of drug evidence.

Filter Cone Spray Ionization Coupled to a Portable MS System: Application to On-Site Forensic Evidence and Environmental Sample Analysis.

The complexity of field-borne sample matrices and the instrumental constraints of portable mass spectrometers (MS) often necessitate that preparative steps are added prior to ambient MS methods when operated on-site, but the corresponding decrease in throughput and experimental simplicity can make field operation impractical. To this end, we report a modified ambient MS method, filter cone spray ionization (FCSI), specifically designed for simple, yet robust, processing of bulk forensic evidence and environmental samples using a fieldable MS system. This paper-crafted source utilizes low-cost laboratory consumables to produce a conical structure that serves as a disposable, spray-based ionization source. Integrated extraction and filtration capabilities mitigate sample heterogeneity and carryover concerns and expedite sample processing, as characterized through the analysis of a variety of authentic forensic evidence types (e.g., abused pharma tablets, counterfeit/adulterated tablets, crystal-based drugs, synthetic marijuana, toxicological specimens) and contaminated soil samples. The data presented herein suggests that the FCSI-MS design could prove robust to the rigors of field-borne, bulk sample screening, overcoming the inefficiencies of other ambient MS methods for these sample classes. Novel applications of FCSI-MS are also examined, such as the coupling to trace evidence vacuum filtration media.

Ligand Exchange/Scrambling Study of Gold(I)-Phosphine Complexes in the Solid Phase by DESI-MS Analysis.

Only a few analytical techniques are available for the characterization of mechanochemical synthetic reaction products. We demonstrate here that DESI-MS is a powerful technique for this purpose, combining the selectivity of MS-based assays with the simplicity and in situ analysis capability of ambient ionization methods. In this work, we report that auranofin, a gold-based drug, and its precursor triethylphosphine gold(I) chloride undergo a complex array of ligand exchange/scrambling reactions with thiol-containing amino acids in the solid state. The products were readily characterized by DESI-MS analysis from the solid-phase reaction, clearly exhibiting ligand exchange and scrambling, with independent confirmation by solid state 13C-NMR. The thioglucose and triethylphosphine moieties exchanged with cysteine and its derivatives, whereas the glutathione replaced 2,3,4,6-tetra-o-acetyl-ß-1-D-glucopyranose only. It was concluded that ligand exchange and scrambling reactions can be carried out in the solid state, and some of the unique products reported in this study can be conveniently prepared through mechanochemical synthesis in good yields (> 98%), as demonstrated by synthesis of (L-cysteinato-S)-triethylphosphine gold(I) from triethylphosphine gold(I) chloride and L-cysteine.

Author Correction: The gut microbiota in infants of obese mothers increases inflammation and susceptibility to NAFLD.

An amendment to this paper has been published and can be accessed via a link at the top of the paper.