Structural basis for dual-mode inhibition of the ABC transporter MsbA.

The movement of core-lipopolysaccharide across the inner membrane of Gram-negative bacteria is catalysed by an essential ATP-binding cassette transporter, MsbA. Recent structures of MsbA and related transporters have provided insights into the molecular basis of active lipid transport; however, structural information about their pharmacological modulation remains limited. Here we report the 2.9 Å resolution structure of MsbA in complex with G907, a selective small-molecule antagonist with bactericidal activity, revealing an unprecedented mechanism of ABC transporter inhibition. G907 traps MsbA in an inward-facing, lipopolysaccharide-bound conformation by wedging into an architecturally conserved transmembrane pocket. A second allosteric mechanism of antagonism occurs through structural and functional uncoupling of the nucleotide-binding domains. This study establishes a framework for the selective modulation of ABC transporters and provides rational avenues for the design of new antibiotics and other therapeutics targeting this protein family.

Effect on functional outcome of robotic assisted rehabilitation versus conventional rehabilitation in patients with complete spinal cord injury: a prospective comparative study.

STUDY DESIGN: Prospective Comparative Study. OBJECTIVE: This study aims to compare the functional outcomes of Robotic-assisted rehabilitation by Lokomat system Vs. Conventional rehabilitation in participants with Dorsolumbar complete spinal cord injury (SCI). SETTING: University level teaching hospital in a hilly state of northern India. METHODS: 15 participants with Dorsolumbar SCI with ASIA A neurology were allocated to robotic rehabilitation and 15 participants to conventional rehabilitation after an operative procedure. Pre-and Post-rehabilitation parameters were noted in terms of ASIA Neurology, Motor and sensory function scores, WISCI II score (Walking Index in SCI score), LEMS (Lower Extremity Motor Score), SCI M III score (Spinal Cord Independence Measure III score), AO Spine PROST (AO Patient Reported Outcome Spine Trauma), McGill QOL score (Mc Gill Quality of Life score), VAS score (Visual Analogue Scale) for pain and Modified Ashworth scale for spasticity in lower limbs. RESULTS: On comparing robotic group with conventional group there was a statistically significant improvement in Robotic-assisted rehabilitation group in terms of Motor score (p = 0.034), WISCI II score (p = 0.0001), SCIM III score (p = 0.0001), AO PROST score (p = 0.0001), Mc GILL QOL score (p = 0.0001), Max velocity (p = 0.0001) and Step length (p = 0.0001). Whereas LEMS score (p = 0.052), ASIA neurology (p = 0.264 (ASIA A); 1.000 (ASIA B); 0.053 (ASIA C)), VAS score (p = 0.099), Sensory score (p = 0.422) and Modified Ashworth scale for spasticity (p = 0.136) were not statically significant when comparing between two groups. CONCLUSION: Robot-assisted rehabilitation is superior than conventional rehabilitation in people living with SCI with AIS A neurology. Differences in the patient group, type of a lesion its and severity, duration from onset to initiation of rehabilitation, devices employed, administration of the therapies and regulation of interventions are likely the cause of variations in the findings seen in the literature for robotic assisted training. LEVEL OF EVIDENCE: III.

Design, optimization, and evaluation of a wet electrostatic precipitator (ESP) for aerosol collection.

In this study, we developed, optimized, and evaluated in lab and field experiments a wet electrostatic precipitator (ESP) for the collection of ambient PM2.5 (particulate matter with aerodynamic diameter < 2.5 µm) into ultrapure water by applying an electrostatic charge to the particles. We operated the wet ESP at different flow rates and voltages to identify the optimal operating conditions. According to our experimental measurements, a flow rate of 125 lpm and an applied positive voltage of 11 kV resulted in a lower ozone generation of 133 ppb and a particle collection efficiency exceeding 80-90% in all size ranges. For the field tests, the wet ESP was compared with the versatile aerosol concentration enrichment system (VACES) connected to a BioSampler, a PTFE filter sampler, and an OC/EC analyzer (Sunset Laboratory Inc., USA) as a reference. The chemical analysis results indicated the wet ESP concentrations of metal and trace elements were in very good agreement with those measured by the VACES/BioSampler and PTFE filter sampler. Moreover, our results showed comparable total organic carbon (TOC) concentrations measured by the wet ESP, BioSampler, and OC/EC analyzer, while somewhat lower TOC concentrations were measured by the PTFE filter sampler, possibly due to the limitations of extracting water-insoluble organic carbon (WIOC) from a dry substrate in the latter sampler. The comparable TOC content in the wet ESP and BioSampler samples differs from previous findings that showed higher TOC content in BioSampler samples compared to those collected by dry ESP. The results of the Dithiothreitol (DTT) assay showed comparable DTT activity in the VACES/BioSampler and wet ESP PM samples while slightly lower in the PTFE filter samples. Overall, our results suggest that the wet ESP could be a promising alternative to other conventional sampling methods.

Utility of clinicoradiological, microbiological, histopathological, and molecular methods in the diagnosis of spinal tuberculosis.

PURPOSE: The diagnosis of STB is mainly based on clinicoradiological observations substantiated by bacterial culture, staining, Gene Xpert, and histopathology. The purpose of the study was to correlate these methods to evaluate the effectiveness in the diagnosis of STB. METHODS: A total of 178 clinicoradiologically suspected cases of STB were included in the study. The specimens for diagnostic workup were collected either during surgery or by CT-guided biopsy. All these specimens were tested for tuberculosis through ZN staining, solid culture, histopathology, and PCR. The sensitivity, specificity, PPV, and NPV of each test were calculated using histopathology as a gold standard. RESULTS: Out of the 178 cases, a total of 15 cases were excluded from this study. Among the remaining 163 cases, TB was diagnosed in 143 [87.73%] on histopathology, 130 [79.75%] on Gene Xpert, 40 [24.53%] on culture, and 23 [14.11%] on ZN stain. The sensitivity, specificity, PPV, and NPV of Gene Xpert were 86.71, 70, 95.38, and 42.42%, respectively. The sensitivity, specificity, PPV, and NPV of AFB culture were 27.97, 100, 100, and 16.26%, respectively. The sensitivity, specificity, PPV, and NPV of AFB stain were 16.08, 100, 100, and 14.29%, respectively. Gene Xpert showed a moderate agreement [Ƙc = 0.4432] with histopathology. CONCLUSION: No single diagnostic modality can ascertain the diagnosis, and it is desirable to have a combination of diagnostic batteries for better results. A combination of Gene Xpert and histopathology aids in early and reliable diagnosis of STB.

Effect of Biomass Burning, Diwali Fireworks, and Polluted Fog Events on the Oxidative Potential of Fine Ambient Particulate Matter in Delhi, India.

We investigated the influence of biomass burning (BURN), Diwali fireworks, and fog events on the ambient fine particulate matter (PM2.5) oxidative potential (OP) during the postmonsoon (PMON) and winter season in Delhi, India. The real-time hourly averaged OP (based on a dithiothreitol assay) and PM2.5 chemical composition were measured intermittently from October 2019 to January 2020. The peak extrinsic OP (OPv: normalized by the volume of air) was observed during the winter fog (WFOG) (5.23 ± 4.6 nmol·min-1·m-3), whereas the intrinsic OP (OPm; normalized by the PM2.5 mass) was the highest during the Diwali firework-influenced period (29.4 ± 18.48 pmol·min-1·µg-1). Source apportionment analysis using positive matrix factorization revealed that traffic + resuspended dust-related emissions (39%) and secondary sulfate + oxidized organic aerosols (38%) were driving the OPv during the PMON period, whereas BURN aerosols dominated (37%) the OPv during the WFOG period. Firework-related emissions became a significant contributor (∼32%) to the OPv during the Diwali period (4 day period from October 26 to 29), and its contribution peaked (72%) on the night of Diwali. Discerning the influence of seasonal and episodic sources on health-relevant properties of PM2.5, such as OP, could help better understand the causal relationships between PM2.5 and health effects in India.

A role for music in cataract surgery: a systematic review.

PURPOSE: There have been several studies that explore the anxiolytic effects of music during cataract surgery. This is the first systematic review of literature highlighting the effects of music on patients undergoing cataract surgery without pharmacological anxiolytics. METHODS: A systematic review of literature using PRISMA methodology identified randomized controlled trials involving patients undergoing cataract surgery with music as an intervention during the procedure with outcomes measured using at least one assessment of anxiety level, pain score, satisfaction, or vital sign measurements. Studies using pharmacological anxiolytics or additional pain-relieving medications besides local anesthetic were excluded. RESULTS: Three studies, involving 523 patients, were included with patients undergoing cataract surgery with music intervention. Music intervention during cataract surgery has demonstrated use in reducing perceived pain, reducing anxiety, and improving vitals with increased benefit of reduced heart rate and blood pressures with binaural beat embedded music. CONCLUSIONS: The clinical implications of this review identify non-traditional approaches to reducing anxiety intraoperatively for cataract surgeries, offering clinicians alternatives to minimizing benzodiazepine and medication use risks.

Complexation of Iron and Copper in Ambient Particulate Matter and Its Effect on the Oxidative Potential Measured in a Surrogate Lung Fluid.

We investigated the complexation state of atmospheric iron and copper and its impact on the oxidative potential (OP) of ambient PM2.5 (PM, particulate matter). A novel fractionation scheme was developed to segregate three different fractions of Fe and Cu present in ambient PM2.5: (i) complexed with hydrophobic organic compounds, (ii) complexed with hydrophilic organic compounds, and (iii) free or inorganic metal fraction. A solid phase extraction (C-18) column was used to separate these fractions. The fractionation scheme applied to the ambient PM2.5 samples collected from an urban site showed that up to 70-90% of water-soluble Fe and Cu were complexed with the organic compounds. The capability of the complexes of Fe(II) and Cu(II) with Suwanee river fulvic acid (SRFA), a proxy for the atmospheric organic compounds, to generate reactive oxygen species (ROS) (·OH and H2O2) in a surrogate lung fluid (SLF) was measured. The complex of Fe-SRFA showed a strong synergistic effect in the generation of ·OH in SLF, while that of Cu-SRFA showed an additive effect. Overall, our results indicate that organic complexation of the metals in ambient PM could significantly alter their OP and needs to be considered while assessing their health impacts.

Review of Acellular Assays of Ambient Particulate Matter Oxidative Potential: Methods and Relationships with Composition, Sources, and Health Effects.

Oxidative stress is a potential mechanism of action for particulate matter (PM) toxicity and can occur when the body's antioxidant capacity cannot counteract or detoxify harmful effects of reactive oxygen species (ROS) due to an excess presence of ROS. ROS are introduced to the body via inhalation of PM with these species present on and/or within the particles (particle-bound ROS) and/or through catalytic generation of ROS in vivo after inhaling redox-active PM species (oxidative potential, OP). The recent development of acellular OP measurement techniques has led to a surge in research across the globe. In this review, particle-bound ROS techniques are discussed briefly while OP measurements are the focus due to an increasing number of epidemiologic studies using OP measurements showing associations with adverse health effects in some studies. The most common OP measurement techniques, including the dithiothreitol assay, glutathione assay, and ascorbic acid assay, are discussed along with evidence for utility of OP measurements in epidemiologic studies and PM characteristics that drive different responses between assay types (such as species composition, emission source, and photochemistry). Overall, most OP assays respond to metals like copper than can be found in emission sources like vehicles. Some OP assays respond to organics, especially photochemically aged organics, from sources like biomass burning. Select OP measurements have significant associations with certain cardiorespiratory end points, such as asthma, congestive heart disease, and lung cancer. In fact, multiple studies have found that exposure to OP measured using the dithiothreitol and glutathione assays drives higher risk ratios for certain cardiorespiratory outcomes than PM mass, suggesting OP measurements may be integrating the health-relevant fraction of PM and will be useful tools for future health analyses. The compositional impacts, including species and emission sources, on OP could have serious implications for health-relevant PM exposure. Though more work is needed, OP assays show promise for health studies as they integrate the impacts of PM species and properties on catalytic redox reactions into one measurement, and current work highlights the importance of metals, organic carbon, vehicles, and biomass burning emissions to PM exposures that could impact health.

Disrupting Gram-Negative Bacterial Outer Membrane Biosynthesis through Inhibition of the Lipopolysaccharide Transporter MsbA.

There is a critical need for new antibacterial strategies to counter the growing problem of antibiotic resistance. In Gram-negative bacteria, the outer membrane (OM) provides a protective barrier against antibiotics and other environmental insults. The outer leaflet of the outer membrane is primarily composed of lipopolysaccharide (LPS). Outer membrane biogenesis presents many potentially compelling drug targets as this pathway is absent in higher eukaryotes. Most proteins involved in LPS biosynthesis and transport are essential; however, few compounds have been identified that inhibit these proteins. The inner membrane ABC transporter MsbA carries out the first essential step in the trafficking of LPS to the outer membrane. We conducted a biochemical screen for inhibitors of MsbA and identified a series of quinoline compounds that kill Escherichia coli through inhibition of its ATPase and transport activity, with no loss of activity against clinical multidrug-resistant strains. Identification of these selective inhibitors indicates that MsbA is a viable target for new antibiotics, and the compounds we identified serve as useful tools to further probe the LPS transport pathway in Gram-negative bacteria.

Exploration of Pyrrolobenzodiazepine (PBD)-Dimers Containing Disulfide-Based Prodrugs as Payloads for Antibody-Drug Conjugates.

A number of cytotoxic pyrrolobenzodiazepine (PBD) monomers containing various disulfide-based prodrugs were evaluated for their ability to undergo activation (disulfide cleavage) in vitro in the presence of either glutathione (GSH) or cysteine (Cys). A good correlation was observed between in vitro GSH stability and in vitro cytotoxicity toward tumor cell lines. The prodrug-containing compounds were typically more potent against cells with relatively high intracellular GSH levels (e.g., KPL-4 cells). Several antibody-drug conjugates (ADCs) were subsequently constructed from PBD dimers that incorporated selected disulfide-based prodrugs. Such HER2 conjugates exhibited potent antiproliferation activity against KPL-4 cells in vitro in an antigen-dependent manner. However, the disulfide prodrugs contained in the majority of such entities were surprisingly unstable toward whole blood from various species. One HER2-targeting conjugate that contained a thiophenol-derived disulfide prodrug was an exception to this stability trend. It exhibited potent activity in a KPL-4 in vivo efficacy model that was approximately three-fold weaker than that displayed by the corresponding parent ADC. The same prodrug-containing conjugate demonstrated a three-fold improvement in mouse tolerability properties in vivo relative to the parent ADC, which did not contain the prodrug.

Synergistic and Antagonistic Interactions among the Particulate Matter Components in Generating Reactive Oxygen Species Based on the Dithiothreitol Assay.

We assessed the interactions among the particulate matter (PM) components in generating the reactive oxygen species (ROS) based on a dithiothreitol (DTT) assay. We started with the standard solutions of known redox-active substances, i.e., quinones (9,10-phenanthraquinone, 1,2-naphthoquinone, 1,4-naphthoquinone, and 5-hydroxy-1,4-naphthoquinone) and metals [Fe (II), Mn (II), and Cu (II)]. Both DTT consumption and hydroxyl radical (·OH) generation were measured in the DTT assay. The interactions of Fe were additive with quinones in DTT consumption but strongly synergistic in ·OH generation. Cu showed antagonistic interactions with quinones in both DTT consumption and ·OH generation. Mn interacted synergistically with quinones in DTT oxidation but antagonistically in ·OH generation. The nature of the interactions of these metals (Fe, Mn, and Cu) with ambient humic-like substances (HULIS) resembled that with quinones, although the intensity of interactions were weaker in DTT consumption than ·OH generation. Finally, we demonstrated that the DTT consumption capability of ambient PM can be well explained by HULIS, three transition metals (Fe, Mn, and Cu), and their interactions, but ·OH generation involves a contribution (∼50%) from additional compounds (aliphatic species or metals other than Fe, Mn, and Cu) present in the hydrophilic PM fraction. The study highlights the need to account for the interactions between organic compounds and metals, while apportioning the relative contributions of chemical components in the PM oxidative potential.

Rethinking Dithiothreitol-Based Particulate Matter Oxidative Potential: Measuring Dithiothreitol Consumption versus Reactive Oxygen Species Generation.

We measured the rate of generation of reactive oxygen species (ROS) [hydroxyl radicals (•OH) and hydrogen peroxide (H2O2)] catalyzed by ambient particulate matter (PM) in the dithiothreitol (DTT) assay. To understand the mechanism of ROS generation, we tested several redox-active substances, such as 9,10-phenanthrenequinone (PQ), 5-hydroxy-1,4-naphthoquinone (5H-1,4NQ), 1,2-naphthoquinone (1,2-NQ), 1,4-naphthoquinone (1,4-NQ), copper(II), manganese(II), and iron (II and III). Both pure compounds and their mixtures show different patterns in DTT oxidation versus ROS generation. The quinones, known to oxidize DTT in the efficiency order of PQ > 5H-1,4NQ > 1,2-NQ > 1,4-NQ, show a different efficiency order (5H-1,4NQ > 1,2-NQ ≈ PQ > 1,4-NQ) in the ROS generation. Cu(II), a dominant metal in DTT oxidation, contributes almost negligibly to the ROS generation. Fe is mostly inactive in DTT oxidation, but shows synergistic effect in •OH formation in the presence of other quinones (mixture/sum > 1.5). Ten ambient PM samples collected from an urban site were analyzed, and although DTT oxidation was significantly correlated with H2O2 generation (Pearson's r = 0.91), no correlation was observed between DTT oxidation and •OH formation. Our results show that measuring both DTT consumption and ROS generation in the DTT assay is important to incorporate the synergistic contribution from different aerosol components and to provide a more inclusive picture of the ROS activity of ambient PM.

Highly Acidic Ambient Particles, Soluble Metals, and Oxidative Potential: A Link between Sulfate and Aerosol Toxicity.

Soluble transition metals in particulate matter (PM) can generate reactive oxygen species in vivo by redox cycling, leading to oxidative stress and adverse health effects. Most metals, such as those from roadway traffic, are emitted in an insoluble form, but must be soluble for redox cycling. Here we present the mechanism of metals dissolution by highly acidic sulfate aerosol and the effect on particle oxidative potential (OP) through analysis of size distributions. Size-segregated ambient PM were collected from a road-side and representative urban site in Atlanta, GA. Elemental and organic carbon, ions, total and water-soluble metals, and water-soluble OP were measured. Particle pH was determined with a thermodynamic model using measured ionic species. Sulfate was spatially uniform and found mainly in the fine mode, whereas total metals and mineral dust cations were highest at the road-side site and in the coarse mode, resulting in a fine mode pH < 2 and near neutral coarse mode. Soluble metals and OP peaked at the intersection of these modes demonstrating that sulfate plays a key role in producing highly acidic fine aerosols capable of dissolving primary transition metals that contribute to aerosol OP. Sulfate-driven metals dissolution may account for sulfate-health associations reported in past studies.

Ambient Size Distributions and Lung Deposition of Aerosol Dithiothreitol-Measured Oxidative Potential: Contrast between Soluble and Insoluble Particles.

Ambient particulate matter may upset redox homeostasis, leading to oxidative stress and adverse health effects. Size distributions of water-insoluble and water-soluble OPDTT (dithiothreitol assay, measure of oxidative potential per air volume) are reported for a roadside site and an urban site. The average water-insoluble fractions were 23% and 51%, and 37% and 39%, for fine and coarse modes at the roadside and urban sites, respectively, measured during different periods. Water-soluble OPDTT was unimodal, peaked near 1-2.5 µm due to contributions from fine-mode organic components plus coarse-mode transition metal ions. In contrast, water-insoluble OPDTT was bimodal, with both fine and coarse modes. The main chemical components that drive both fractions appear to be the same, except that for water-insoluble OPDTT the compounds were absorbed on surfaces of soot and non-tailpipe traffic dust. They were largely externally mixed and deposited in different regions in the respiratory system, transition metal ions predominately in the upper regions and organic species, such as quinones, deeper in the lung. Although OPDTT per mass (toxicity) was highest for ultrafine particles, estimated lung deposition was mainly from accumulation and coarse particles. Contrasts in the phases of these forms of OPDTT deposited in the respiratory system may have differing health impacts.

α-Aryl pyrrolidine sulfonamides as TRPA1 antagonists.

A series of α-aryl pyrrolidine sulfonamide TRPA1 antagonists were advanced from an HTS hit to compounds that were stable in liver microsomes with retention of TRPA1 potency. Metabolite identification studies and physicochemical properties were utilized as a strategy for compound design. These compounds serve as starting points for further compound optimization.

Simplified Bryostatin Analogues Protect Cells from Chikungunya Virus-Induced Cell Death.

Chikungunya virus (CHIKV) is a mosquito-borne alphavirus showing a recent resurgence and rapid spread worldwide. While vaccines are under development, there are currently no therapies to treat this disease, except for over-the-counter (OTC) analgesics, which alleviate the devastating arthritic and arthralgic symptoms. To identify novel inhibitors of the virus, analogues of the natural product bryostatin 1, a clinical lead for the treatment of cancer, Alzheimer's disease, and HIV eradication, were investigated for in vitro antiviral activity and were found to be among the most potent inhibitors of CHIKV replication reported to date. Bryostatin-based therapeutic efforts and even recent anti-CHIKV strategies have centered on modulation of protein kinase C (PKC). Intriguingly, while the C ring of bryostatin primarily drives interactions with PKC, A- and B-ring functionality in these analogues has a significant effect on the observed cell-protective activity. Significantly, bryostatin 1 itself, a potent pan-PKC modulator, is inactive in these assays. These new findings indicate that the observed anti-CHIKV activity is not solely mediated by PKC modulation, suggesting possible as yet unidentified targets for CHIKV therapeutic intervention. The high potency and low toxicity of these bryologs make them promising new leads for the development of a CHIKV treatment.

The cryptophycins as potent payloads for antibody drug conjugates.

The cryptophycins are a potent class of cytotoxic agents that were evaluated as antibody drug conjugate (ADC) payloads. Free cryptophycin analog 1 displayed cell activity an order of magnitude more potent than approved ADC payloads MMAE and DM1. This potency increase was also reflected in the activity of the cryptophycin ADCs, attached via a either cleavable or non-cleavable linker.

Organic aerosols associated with the generation of reactive oxygen species (ROS) by water-soluble PM2.5.

We compare the relative toxicity of various organic aerosol (OA) components identified by an aerosol mass spectrometer (AMS) based on their ability to generate reactive oxygen species (ROS). Ambient fine aerosols were collected from urban (three in Atlanta, GA and one in Birmingham, AL) and rural (Yorkville, GA and Centerville, AL) sites in the Southeastern United States. The ROS generating capability of the water-soluble fraction of the particles was measured by the dithiothreitol (DTT) assay. Water-soluble PM extracts were further separated into the hydrophobic and hydrophilic fractions using a C-18 column, and both fractions were analyzed for DTT activity and water-soluble metals. Organic aerosol composition was measured at selected sites using a high-resolution time-of-flight AMS. Positive matrix factorization of the AMS spectra resolved the organic aerosol into isoprene-derived OA (Isop_OA), hydrocarbon-like OA (HOA), less-oxidized oxygenated OA, (LO-OOA), more-oxidized OOA (MO-OOA), cooking OA (COA), and biomass burning OA (BBOA). The association of the DTT activity of water-soluble PM2.5 (WS_DTT) with these factors was investigated by linear regression techniques. BBOA and MO-OOA were most consistently linked with WS_DTT, with intrinsic water-soluble activities of 151 ± 20 and 36 ± 22 pmol/min/µg, respectively. Although less toxic, MO-OOA was most widespread, contributing to WS_DTT activity at all sites and during all seasons. WS_DTT activity was least associated with biogenic secondary organic aerosol. The OA components contributing to WS_DTT were humic-like substances (HULIS), which are abundantly emitted in biomass burning (BBOA) and include highly oxidized OA from multiple sources (MO-OOA). Overall, OA contributed approximately 60% to the WS_DTT activity, with the remaining probably from water-soluble metals, which were mostly associated with the hydrophilic WS_DTT fraction.

Reactive Oxygen Species Generation Linked to Sources of Atmospheric Particulate Matter and Cardiorespiratory Effects.

Exposure to atmospheric fine particulate matter (PM2.5) is associated with cardiorespiratory morbidity and mortality, but the mechanisms are not well understood. We assess the hypothesis that PM2.5 induces oxidative stress in the body via catalytic generation of reactive oxygen species (ROS). A dithiothreitol (DTT) assay was used to measure the ROS-generation potential of water-soluble PM2.5. Source apportionment on ambient (Atlanta, GA) PM2.5 was performed using the chemical mass balance method with ensemble-averaged source impact profiles. Linear regression analysis was used to relate PM2.5 emission sources to ROS-generation potential and to estimate historical levels of DTT activity for use in an epidemiologic analysis for the period of 1998-2009. Light-duty gasoline vehicles (LDGV) exhibited the highest intrinsic DTT activity, followed by biomass burning (BURN) and heavy-duty diesel vehicles (HDDV) (0.11 ± 0.02, 0.069 ± 0.02, and 0.052 ± 0.01 nmol min(-1) µg(-1)source, respectively). BURN contributed the largest fraction to total DTT activity over the study period, followed by LDGV and HDDV (45, 20, and 14%, respectively). DTT activity was more strongly associated with emergency department visits for asthma/wheezing and congestive heart failure than PM2.5. This work provides further epidemiologic evidence of a biologically plausible mechanism, that of oxidative stress, for associations of adverse health outcomes with PM2.5 mass and supports continued assessment of the utility of the DTT activity assay as a measure of ROS-generating potential of particles.

Design, synthesis, and evaluation of potent bryostatin analogs that modulate PKC translocation selectivity.

Modern methods for the identification of therapeutic leads include chemical or virtual screening of compound libraries. Nature's library represents a vast and diverse source of leads, often exhibiting exquisite biological activities. However, the advancement of natural product leads into the clinic is often impeded by their scarcity, complexity, and nonoptimal properties or efficacy as well as the challenges associated with their synthesis or modification. Function-oriented synthesis represents a strategy to address these issues through the design of simpler and therefore synthetically more accessible analogs that incorporate the activity-determining features of the natural product leads. This study illustrates the application of this strategy to the design and synthesis of functional analogs of the bryostatin marine natural products. It is specifically directed at exploring the activity-determining role of bryostatin A-ring functionality on PKC affinity and selectivity. The resultant functional analogs, which were prepared by a flexible, modular synthetic strategy, exhibit excellent affinity to PKC and differential isoform selectivity. These and related studies provide the basic information needed for the design of simplified and thus synthetically more accessible functional analogs that target PKC isoforms, major targets of therapeutic interest.