Approximate 2-fold Increase Research Articles

Influence of Static Magnetic Field on HeLa and Huo2 Cells in the Presence of Aloe vera Extract.

This research aimed to assess the impact of static magnetic field (SMF) on apoptosis rate and cell cycle progression in the presence of Aloe vera Crude Extract (ACE) in normal (Huo2) and cancer cells (HeLa). The specimens were split into one untreated group (control) and two experimental groups, including treatment with ACE (Alo) and compound treatment with SMF and ACE (Alo+SMF). MTT assay determined the IC50 value, and flow cytometry was employed to evaluate cell cycle distribution and apoptosis rates. Statistical analysis was carried out through a two-way ANOVA followed by Tukey's post hoc test. Our results showed that combination treatment with SMF (10 mT) and ACE (Alo+SMF) significantly inhibited the cell proliferation. This increased the cell number in G2/M stage and early apoptosis in cancer cells compared to ACE treated cells after 24 and 48h but reduced the number of Huo2 cells in G2/M phase and early apoptosis after 24h. The effect of AEC on HeLa cells was intensified with increasing the SMF exposure time, such that the early apoptosis rate in Alo+SMF group had an approximate 4-fold increase compared to Alo group. This research proposes that the combination treatment accelerates the apoptosis induction of HeLa cell. During the interphase, there were significant differences between the cancer and healthy cells concerning the cell cycle. Moreover, exposure time may play an important role in the impact SMF on both healthy and cancer cells in the presence of AEC.

Effect of Hepatic Impairment on the Pharmacokinetics of Itacitinib.

Itacitinib is a potent, selective JAK-1 inhibitor currently in development for the treatment of chronic graft-vs-host-disease in combination with corticosteroids. Itacitinib is primarily eliminated via cytochrome P450 3A metabolism with minimal renal elimination. The purpose of this open-label study was to investigate the effect of hepatic impairment, as determined by Child-Pugh grade, on itacitinib pharmacokinetics. All participants received a single 300-mg dose of itacitinib orally in the fasted state. Blood samples were collected serially through 96 hours after dosing; 4 hours after dosing, an additional sample was collected for protein binding determination. Participants with moderate hepatic impairment (N = 8) had an approximate 2.5-fold increase in total exposure (area under the plasma concentration-time curve from time 0 to infinity [AUC0-∞ ]) and an approximate 2-fold increase in maximal exposure (Cmax ) compared to those with normal hepatic function (N = 8) (geometric mean ratio, 2.51 [90% confidence interval (CI), 1.54-4.08] for AUC0-∞ and 1.95 [90%CI, 1.14-3.35] for Cmax ). Participants with severe hepatic impairment (N = 6) had an approximate 4-fold increase in total exposure (AUC0-∞ ) and an approximate 3.5-fold increase in maximal exposure compared to participants with normal hepatic function (geometric mean ratio, 4.08 [90%CI, 2.41-6.89] for AUC0-∞ and 3.48 [90%CI, 1.94-6.23] for Cmax ). Protein binding was similar between participants with moderate or severe hepatic impairment and participants with normal hepatic function, with average unbound fractions (percent free) of 25.7%, 31.5%, and 25.6%, respectively. There were no serious or fatal treatment-related adverse events. The results of this study combined with exposure, efficacy, and safety data from the pivotal study in the relevant patient population will inform final dosing recommendations.

Association Between Immunosuppression and Outcomes in Oral Cavity Squamous Cell Carcinoma.

To assess the effect of immunosuppression on recurrence and mortality outcomes in oral cavity squamous cell carcinoma (SCC) after initial surgical treatment. Retrospective cohort study. A single academic tertiary referral center. Patients with oral cavity SCC treated with initial surgery were included. Immunosuppressed versus nonimmunosuppressed groups were compared. Primary end points were 5-year overall recurrence and all-cause mortality. Secondary end points were recurrence subtypes (local, regional, and distant) and disease-specific mortality. Of 803 patients with oral cavity SCC, 71 (9%) were immunosuppressed from therapeutic drug use (n = 48) or systemic disease (n = 23). The immunosuppressed group consisted of patients with a history of transplant (21%), autoimmune or pulmonary disorder (45%), hematologic malignancy or myeloproliferative disorder (30%), and HIV infection (3%). After adjusting for baseline variables of age, sex, comorbidities, pathologic tumor characteristics, and adjuvant treatment, all recurrence and mortality outcomes were worse in the immunosuppressed group. The multivariate-adjusted hazard ratio for overall recurrence was 2.16 (95% CI, 1.50-3.12; P < .01), and all-cause mortality was 1.79 (95% CI, 1.15-2.78; P < .01) in Cox regression analysis. The 2 groups were then matched in a 1:5 ratio according to the same baseline variables. All end points apart from disease-specific mortality were significantly worse in the immunosuppressed group after matching. This study demonstrates that immunosuppression is associated with poor outcomes in oral cavity SCC, with an approximate 2-fold increase in rates of recurrence and mortality. Future studies are needed to assess the risks and benefits of adjusting therapeutic immunosuppression in this population.

Rapid bone repair with the recruitment of CD206+M2-like macrophages using non-viral scaffold-mediated miR-133a inhibition of host cells

microRNAs offer vast therapeutic potential for multiple disciplines. From a bone perspective, inhibition of miR-133a may offer potential to enhance Runx2 activity and increase bone repair. This study aims to assess the therapeutic capability of antagomiR-133a delivery from collagen-nanohydroxyapatite (coll-nHA) scaffolds following cell-free implantation in rat calvarial defects (7 mm diameter). This is, to the best of our knowledge, the first report of successful in vivo antagomiR uptake in host cells of fully immunocompetent animals without distribution to other off-target tissues. Our results demonstrate the localized release of antagomiR-133a to the implant site at 1 week post-implantation with increased calcium deposits already evident in the antagomiR-133a loaded scaffolds at this early timepoint. This was followed by an approximate 2-fold increase in bone volume versus antagomiR-free scaffolds and a significant 10-fold increase over the empty defect controls, after just 4 weeks. An increase in host CD206+ cells suggests an accelerated pro-remodeling response by M2-like macrophages accompanying bone repair with this treatment. Overall, this non-viral scaffold-mediated antagomiR-133a delivery platform demonstrates capability to accelerate bone repair in vivo – without the addition of exogenous cells – and underlines the role of M2 macrophage-like cells in directing accelerated bone repair. Expanding the repertoire of this platform to deliver alternative miRNAs offers exciting possibilities for a variety of therapeutic indications. Statement of SignificancemicroRNAs, small non-coding RNA molecules involved in gene regulation, may have potential as a new class of bone healing therapeutics as they can enhance the regenerative capacity of bone-forming cells. We developed a collagen-nanohydroxyapatite-microRNA scaffold system to investigate whether miR133a inhibition can enhance osteogenesis in rat MSCs and ultimately accelerate endogenous bone repair by host cells in vivo without pre-seeding cells prior to implantation. Overall, this off-the-shelf, non-viral scaffold-mediated antagomiR-133a delivery platform demonstrates capability to accelerate bone repair in vivo – without the requirement of exogenous cells – and highlights the role of CD206+M2 macrophage-like cells in guiding accelerated bone repair. Translating the repertoire of this platform to deliver alternative miRNAs offers exciting possibilities for a vast myriad of therapeutic indications.

Effects of suspended sediment on POCIS sampling rates

Effects of chemical uptake onto polar organic chemical integrative samplers (POCIS) exposed to total suspended solid (TSS) sediment concentrations of 0 and 3600 ppm were investigated for 12 pesticides at constant concentration, temperature, and flow velocity. The effects of sediment exposure on POCIS uptake were negligible for compounds with polyethersulfone-water partition coefficients greater than three (i.e., log KPESW > 3). However, significant effects were observed for 3 of 12 compounds tested, and the maximum effect was an approximate 4-fold increase in sampling rate for the sediment experiment relative to the control. Effects of sediment on the pesticide distribution between polyethersulfone (PES) membranes and Oasis HLB sorbent were also investigated. The fraction of pesticide accumulated on PES membranes was relatively low for most compounds and ranged from 0 to 33%. In contrast, four compounds with higher affinity for PES accumulated preferentially on the membranes (fraction ranging from 64 to 96%), suggesting that a sampling rate derived from the additive contribution of membrane extraction and the more typical extraction of analytes from HLB sorbent would improve the sensitivity of sampling rate estimations for these compounds. However, for these same compounds, the combined sampling rate, Rs (HLB + PES), was considerably more susceptible to a sediment effect than the traditional sampling rate determination, relying solely on extraction from HLB sorbent.

Identification of a Novel T‐lymphocyte Inflammatory Protein in Psychological Stress‐ Induced Hypertension

Post‐traumatic stress disorder (PTSD) is a psychiatric condition characterized by hyperarousal, anxiety, anhedonia, and depression. While the behavioral manifestations are often the focus of this disease, PTSD also alters physiological processes leading to elevated sympathetic tone and norepinephrine (NE) outflow, pronounced inflammation, and a &gt;50% increased risk in developing co‐morbid cardiovascular diseases like hypertension. Our laboratory previously reported that exposure to NE can modulate levels of T‐lymphocyte pro‐inflammatory cytokines, and understanding that hypertension is regulated in part due to inflammation, we posited a connection between the autonomic nervous, immune, and cardiovascular systems in PTSD patients. We hypothesized that psychological stress‐induced sympathoexcitation leads to increased pro‐inflammatory cytokine production from T‐lymphocytes, which impacts the development of hypertension. Utilizing a mouse‐model of PTSD‐like psychological trauma, lymphatic sympathoexcitation was confirmed by a 2.5 fold increase in NE and 4.1 fold increase in tyrosine hydroxylase content in spleens from stressed animals (p&lt;0.001). This increased sympathetic tone correlated with elevated pro‐inflammatory cytokines IL‐17A, IL‐6, and IL‐2 (+4.4, +3.1, and +3.0 fold, respectively, p&lt;0.01) both in circulation and also specifically within splenic T‐lymphocytes. Examination of hemodynamics in the stressed mice elucidated a stable increase in resting mean arterial pressure (+19.5 mmHg, p&lt;0.05) over the stress‐induction period with an even greater (+39.8 mmHg, p&lt;0.01) rise after stress context re‐exposure compared to control animals. To investigate the causal contribution of T‐lymphocytes in driving this hypertensive response, recombination activating gene 2 (Rag2) knock‐out animals, devoid of mature lymphocytes, were utilized. Rag2 knock‐out mice showed no changes in pro‐inflammatory cytokines, a 75% decrease in splenic TH levels (p&lt;0.01), and a blunted blood pressure response upon context re‐exposure (+8.8 mmHg, p&lt;0.01). Single cell RNAseq analysis of T‐lymphocytes from stressed wild‐type animals demonstrated a significant increase (p=8.6×10−19) in calprotectin (S100a8 and S100a9) mRNA levels compared to control. Calprotectin is both an intracellular and extracellular protein involved in many inflammatory processes, but has been thought to be granulocyte‐specific making its detection in T‐lymphocytes highly unexpected. We confirmed 40 and 30‐fold increases in S100a8 and S100a9 mRNA, respectively, in purified T‐lymphocytes from stressed animals by qRT‐PCR (p&lt;0.01). Additionally, a 3‐fold increase (p&lt;0.001) in levels of calprotectin protein was observed in circulation of stressed animals. Last, we identified an approximate 2‐fold increase (p&lt;0.05) in calprotectin protein levels in circulation from human patients with PTSD with comorbid hypertension compared to PTSD alone. Taken together, our data suggest a new role for T‐lymphocytes in the regulation of stress‐induced inflammation and hypertension, and present an undescribed protein target for future investigation and potential therapeutic intervention.Support or Funding InformationNIH R00HL123471This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

Enhancing bioactivity, physicochemical, and pharmacokinetic properties of a nano-sized, anti-VEGFR2 Adnectin, through PASylation technology

The crucial role of VEGF receptor 2 (VEGFR2) signaling in the angiogenesis and metastasis of solid tumors has prompted the development of inhibitors with minimal bystander effects. Recently, Adnectin C has attracted attention for cancer treatment. To overcome the problematic properties of Adnectin, a novel form of Adnectin C has been designed by its fusion to a biodegradable polymeric peptide containing Pro/Ala/Ser (PAS) repetitive residues. E. coli-expressed recombinant fused and unfused proteins were compared in terms of bioactivity, physicochemical, and pharmacokinetic properties using standard methods. Dynamic light scattering (DLS) analysis of PASylated adnectin C revealed an approximate 2-fold increase in particle size with a slight change in the net charge. Additionally, fusion of the PAS sequence improved its stability against the growth of thermo-induced aggregated forms. The high receptor-binding and improved binding kinetic parameters of PASylated Adnectin C was confirmed by ELISA and surface plasmon resonance assays, respectively. Pharmacokinetic studies showed a noticeable increase in the terminal half-life of Adnectin C-PAS#1(200) by a factor of 4.57 after single dose by intravenous injection into female BALB/c mice. The results suggest that PASylation could offer a superior delivery strategy for developing Adnectin-derived drugs with improved patient compliance.

Prognostic Value of Nodal Response After Preoperative Treatment of Gastric Adenocarcinoma.

Background: Pathologically positive lymph nodes (ypN+) after preoperative chemotherapy are associated with poor survival in patients with gastric cancer. Little is known about the association between response to preoperative therapy and the benefit of postoperative therapy. Methods: This retrospective cohort study of the National Cancer Database included patients with clinically node-positive (cN+) gastric cancer treated with preoperative therapy followed by surgery (2006-2014). Preoperative treatment modality was categorized as the inclusion of radiation therapy (RT) or chemotherapy alone. Pretreatment clinical and pathologic stages were used to determine pathologic treatment response rates. The association between overall risk of death and preoperative treatment, disease response, and adjuvant therapy use was evaluated using multivariable Cox regression. Results: Preoperative RT was used in 53.6% of 1,976 patients with cN+ gastric cancer, (74.3% cardia and 10.1% noncardia). The nodal response rate was 38.9% and was higher with RT than with chemotherapy alone (cardia, 46.0% vs 29.1%; P<.001; noncardia, 43.8% vs 31.9%; P=.06). Preoperative RT was associated with an approximate 2-fold increase in the odds of pathologic response compared with chemotherapy. Overall, use of adjuvant therapy was not associated with a decreased risk of death. A primary tumor response with residual nodal disease was not associated with survival (hazard ratio [HR], 1.03; 95% CI, 0.66-1.60). However, a nodal response with residual primary disease was significantly associated with survival (HR, 0.54; 95% CI, 0.44-0.65). Conclusions: More than one-third of node-positive gastric cancers showed pathologic nodal response with preoperative treatment. RT is associated with a higher response than chemotherapy. Patients with ypN+ disease have worse survival, regardless of whether they receive postoperative therapy. Future gastric cancer trials should evaluate the role of preoperative RT and individualize postoperative therapy use.

Indocyanine green molecular angiography of choroidal neovascularization

Retinal diseases such as proliferative diabetic retinopathy and neovascular AMD are characterized by the formation of new blood vessels. Current imaging techniques such as fluorescein and ICG angiography help to identify areas of vascular leakage but are limited in their applicability due to their nonspecific nature. However, as new treatment paradigms emerge in an effort to have patient specific treatments, the development of new imaging techniques that are capable of identifying patient specific biomarkers will become crucial for the success of these approaches. In this study, we create and characterize an endoglin (CD105) targeted imaging probe that can be used for indocyanine green (ICG) molecular angiography. This anti-endoglin-ICG bioconjugate has a self-quenching “off-on” capacity to enable high contrast imaging of proliferative blood vessels at a molecular level in vivo. Using the laser CNV mouse model we demonstrate an approximate 3-fold increase in lesion visualization compared to non-targeting controls.

Comparative pathogenicity analysis of Cryptococcus neoformans RAPD types

Cryptococcus neoformans and Cryptococcus gattii are the major causes of Cryptococcosis, an invasive fungal infection of human and animals. Few data have been reported on the pathophysiology of Pattern I (PI) and Pattern IV (PIV) of C. neoformans with context of their clinical outcome. In this study an extensive comparison of PI and PIV was conducted to provide a more complete and reliable assessment of their biological and clinical behaviour. Representative strains of PI and PIV of C. neoformans were collected from Outreach Programme on Zoonotic Diseases (OPZD) at Department of Veterinary Public Health of West Bengal University of Animal and Fishery Sciences and infective inoculum was prepared by adjusting the number of Cryptococci at 3.25X 105 / ml in phosphate buffer saline (pH 7.4). Mice were challenged with 0.2 ml of inoculum at lateral tail vein and infected mice were examined for histopathological study and survivability study for 21 days. This present study showed that PIV produced more cystic structure in brain (c2 =4.22, P 0.05). PIV was associated with an approximate 4-fold increase in risk of death as compared with PI (P< 0.05, OR= 4.38, 95% CL= 1.32 to 14.50). Our study suggested that Pattern IV produced more severe Cryptococcosis in murine model due to its difference in genotype.

Engineering Bacillus pumilus alkaline serine protease to increase its low-temperature proteolytic activity by directed evolution

BackgroundMesophilic alkaline serine proteases from various bacteria have been commercially applied in a range of industries owing to their high catalytic efficiency and wide substrate specificity. However, these proteases have an optimal catalytic temperature of approximately 50 °C, and their activity decreases significantly at low temperature. Therefore, to enhance their cold activity, it is necessary to improve the catalytic performance of these proteases at low temperature. The alkaline serine protease (DHAP) from Bacillus pumilus BA06 is a typical mesophilic enzyme, which has demonstrated great potential in various industrial applications. Here we attempted to improve the cold activity of DHAP via directed evolution.ResultsSeven variants (P9S, A1G/K27Q, A38V, A116T, T162I, S182R, and T243S) of DHAP from B. pumilus were obtained via directed evolution. The results showed that all of the variants had increased proteolytic activity at 15 °C towards both the casein and synthetic peptide substrates. With the exception of variant T243S, the thermostability of these variants did not decrease in comparison with the wild-type enzyme. Kinetic analysis indicated that the increase in catalytic efficiency was largely attributed to the increase in turnover number (kcat). Furthermore, the combined variants generated by site-directed mutagenesis showed a further increase in specific caseinolytic activity and the kcat value for hydrolysis of the synthetic peptide. The combined variants of P9S/K27Q and P9S/T162I exhibited an approximate 5-fold increase in caseinolytic activity at 15 °C and almost no loss of thermostability. Finally, the possible mechanism responsible for the change in catalytic properties for these variants was interpreted based on structural modeling.ConclusionsDirected evolution and site-directed mutagenesis were combined to engineer variants of the DHAP from B. pumilus. All of the variants exhibited an increase in hydrolytic efficiency at low temperature towards both of the substrates, casein and synthetic peptide, without any loss of thermostability compared with the wild-type. These data suggest that engineering low-temperature activity for a bacterial protease is not always associated with the loss of thermostability. Furthermore, our findings demonstrate that enhanced cold activity and thermostability could be integrated into a single variant.

Using multi-echo simultaneous multi-slice (SMS) EPI to improve functional MRI of the subcortical nuclei of the basal ganglia at ultra-high field (7T)

The nuclei of the basal ganglia pose a special problem for functional MRI, especially at ultra-high field, because T2* variations between different regions result in suboptimal BOLD sensitivity when using gradient-echo echo-planar imaging (EPI). Specifically, the iron-rich lentiform nucleus of the basal ganglia, including the putamen and globus pallidus, suffers from substantial signal loss when imaging is performed using conventional single-echo EPI with echo times optimized for the cortex. Multi-echo EPI acquires several echoes at different echo times for every imaging slice, allowing images to be reconstructed with a weighting of echo times that is optimized individually for each voxel according to the underlying tissue or T2* properties. Here we show that multi-echo simultaneous multi-slice (SMS) EPI can improve functional activation of iron-rich subcortical regions while maintaining sensitivity within cortical areas. Functional imaging during a motor task known to elicit strong activations in the cortex and the subcortex (basal ganglia) was performed to compare the performance of multi-echo SMS EPI to single-echo SMS EPI. Notably within both the caudate nucleus and putamen of the basal ganglia, multi-echo SMS EPI yielded higher tSNR (an average 84% increase) and CNR (an average 58% increase), an approximate 3-fold increase in supra-threshold voxels, and higher t-values (an average 39% increase). The degree of improvement in the group level t-statistics was negatively correlated to the underlying T2* of the voxels, such that the shorter the T2*, as in the iron-rich nuclei of the basal ganglia, the higher the improvement of t-values in the activated region.

Abstract P133: Angiotensin-(1-7) Does Not Acutely Lower Blood Pressure in Patients With Essential Hypertension

Angiotensin (Ang)-(1-7) is a beneficial renin-angiotensin hormone that produces vasodilation to lower blood pressure (BP) in animal models of hypertension. There are limited clinical studies, however, and it is unclear if Ang-(1-7) contributes to BP regulation in human hypertension. We propose the difficulties in showing cardiovascular effects in previous clinical studies relates to the buffering capacity of the arterial baroreflex. In this study, we tested the hypothesis that Ang-(1-7) would produce negligible effects on BP with intact baroreceptors, and that the cardiovascular effects of this hormone would be unmasked following elimination of baroreflex buffering. To test this, we examined the effects of acute intravenous Ang-(1-7) infusion (ascending doses from 0.5 to 20 ng/kg/min) on supine BP in subjects with essential hypertension under intact conditions and following acute autonomic withdrawal with the ganglionic blocker trimethaphan in a randomized, open-label, crossover study. BP was restored to baseline levels following autonomic blockade with individually titrated phenylephrine doses. Seven subjects with essential hypertension completed this study (6 male; 48±4 years of age; 29±2 body mass index). All subjects were withdrawn from antihypertensive medications for at least two weeks, and were placed on a fixed sodium diet (150 mEq/day) for three days, prior to each study day. When comparing change from baseline to maximum dose, Ang-(1-7) did not alter systemic hemodynamics under intact conditions (systolic BP: 3±4 mmHg; diastolic BP: 3±1 mmHg; heart rate (0±1 bpm). In contrast to our hypothesis, Ang-(1-7) did not elicit a BP-lowering effect under autonomic blockade (systolic BP: 10±8 mmHg, p=0.299 vs. intact; diastolic BP: 7±3 mmHg, p=0.512; heart rate: -2±1 bpm, p=0.166). Plasma Ang-(1-7) was measured in 4 subjects and showed an approximate 3-fold increase with infusions [0.8±0.7 baseline vs. 2.1±1.2 ng/mL after Ang-(1-7)]. These findings suggest that infusion of Ang-(1-7) to levels within a physiologic range does not acutely induce vasodilation to lower blood pressure in essential hypertension. We cannot exclude that Ang-(1-7) lowers BP by sympathetic inhibition, since those pathways were excluded by our study design.

Oxidant production and SOD1 protein expression in single skeletal myofibers from Down syndrome mice

Down syndrome (DS) is a genetic condition caused by the triplication of chromosome 21. Persons with DS exhibit pronounced muscle weakness, which also occurs in the Ts65Dn mouse model of DS. Oxidative stress is thought to be an underlying factor in the development of DS-related pathologies including muscle dysfunction. High-levels of oxidative stress have been attributed to triplication and elevated expression of superoxide dismutase 1 (SOD1); a gene located on chromosome 21. The elevated expression of SOD1 is postulated to increase production of hydrogen peroxide and cause oxidative injury and cell death. However, it is unknown whether SOD1 protein expression is associated with greater oxidant production in skeletal muscle from Ts65Dn mice. Thus, our objective was to assess levels of SOD1 expression and oxidant production in skeletal myofibers from the flexor digitorum brevis obtained from Ts65Dn and control mice. Measurements of oxidant production were obtained from myofibers loaded with 2′,7′-dichlorodihydrofluorescein diacetate (DCFH2-DA) in the basal state and following 15min of stimulated unloaded contraction. Ts65Dn myofibers exhibited a significant decrease in basal DCF emissions (p < 0.05) that was associated with an approximate 3-fold increase in SOD1 (p < 0.05). DCF emissions were not affected by stimulating contraction of Ts65Dn or wild-type myofibers (p > 0.05). Myofibers from Ts65Dn mice tended to be smaller and myonuclear domain was lower (p < 0.05). In summary, myofibers from Ts65Dn mice exhibited decreased basal DCF emissions that were coupled with elevated protein expression of SOD1. Stimulated contraction in isolated myofibers did not affect DCF emissions in either group. These findings suggest the skeletal muscle dysfunction in the adult Ts65Dn mouse is not associated with skeletal muscle oxidative stress.

Fractionation of Interfacial Material Reveals a Continuum of Acidic Species That Contribute to Stable Emulsion Formation

The acidic components of Athabasca bitumen interfacial material (IM) were isolated and subsequently fractionated based on hydrophobicity by a modified aminopropyl silica (MAPS) method to determine whether low-molecular-weight IM acids are preferentially ionized in negative-ion electrospray ionization (ESI (−)) and, thus, bias the compositional information obtained by direct infusion (dilute and shoot) mass spectral analysis. Characterization by negative-ion electrospray ionization Fourier transform ion cyclotron resonance mass spectrometry (FT-ICR MS) revealed that MAPS fractionation of IM acids extends the detection of high-m/z (>600-Da) IM compounds by 2-fold, yields an approximate 10-fold increase in the number of assigned formulas, and exposes a continuum of acidic species that includes the first definitive identification of doubly charged acids in interfacial material. Comparison of the heteroatom contents of singly and doubly charged Ox species, combined with the acid-targeted extraction procedure, strongly suggests that the chemical functionalities are similar for the two ion types and are largely composed of mono- and dicarboxylic acids. Excitation emission matrix spectroscopy (EEMS) revealed that the most hydrophobic IM fractions approach the size and aromaticity of bitumen asphaltenes, but despite the increase in hydrophobicity, these asphaltene-like acids form the tightest emulsions, as revealed in simple bottle tests. Thus, the most surface-active material from Athabasca bitumen comprises low-molecular-weight, acidic, resin-like species, as well as larger (higher-molecular-weight) acidic asphaltene-like compounds.

The measurement of bisphenol A and its analogues, perfluorinated compounds in twenty species of freshwater and marine fishes, a time-trend comparison and human health based assessment

Our previous study in 2011 reported the detection of BPA and PFAAs in 20 species of marine and freshwater fishes. With an emerging evidence to suggest the metabolic-disrupting effects of BPA/PFAAs in animals, the present study was aimed to provide a time-trend analysis to determine the current concentrations of PFAAs and BPA in 20 commercially available Hong Kong species of fishes. Since the manufacture and use of BPA is being prohibited in most nations, the introduction of BPA alternatives has recently been incorporated in the markets. Therefore, the concentrations of BPB, BPF and BPS were determined. In the present study, all freshwater and seawater fish samples showed quantified concentrations [>Limit of Quantification (LOQ<0.5ng/g)] of BPA. BPF was detected in some marine (yellow seafin, bigeye, goldspotted rabbitfish, snubnose pompano, tongue sole, Bleeker's grouper and orange-spotted grouper) and freshwater fishes (mud carp, crucian carp, tilapia, catfish, mandarin fish, grass carp, grey mullet and spotted snakehead). Two of the compounds, BPS and BPB could only be identified in the marine fishes (snubnose pompano, yellow seafin). In PFAA analysis, PFOA, PFDA, PFOS, PFUdA and PFDoA were found in most of the marine and freshwater fishes. PFOS and PFOA were shown to be the two predominant PFAAs in fishes. On the basis of the measured concentrations of bisphenols, BPs (BPA, BPB, BPF, BPS) and PFAAs, the average daily intake for BPs (20.5–31.5ng/kgb.w./day) and PFAAs (1.17–1.83ng/kgb.w./day) were calculated and found to be lower than values of tolerable daily intake (TDI) established in Europe. However, as compared with our previous study in 2011, the present study revealed an approximate 10-fold increase in the concentrations of BPA in the fish samples. Although the hazard ratio of consuming fishes for BPA and PFAA exposure is expected to remain low, possible additive metabolic-disrupting effect of BPA and its analogues as well PFAAs should be taken into consideration for human health risk assessment.

Characterization of liquid state sulfur polymer/epoxy blend as asphalt pavement materials

Abstract In this study, a liquid state sulfur polymer (LSP) was employed as asphalt pavement materials. The LSP/epoxy asphalt (LSPA) was easily fabricated by mixing LSP and epoxy resin at room temperature. The workability of as-mixed LSP/epoxy mixture was explored by controlling the ratio of epoxy. The asphalt mixture with the epoxy content below 50 wt% exhibited reliable flowability above approximately 12.5 cm. The solidification of LSP was successfully accompanied by simultaneous curing of epoxy. For practical application of pavement on road, the Marshall stability, indirect tensile strength and wheel tracking test were performed. The Marshall stability and indirect tensile strength of the LSPA exhibited an approximate 2-fold increase compared to neat asphalt. Moreover, exceptional durability of the LSPA was achieved through the controllable crushed depth of the pavement under wheel passes of 25,000 cycles. For practical repairing test on the road, the LSPA was cast on a pothole prepared on asphalt road. Our as-cast asphalt materials were suitable to heal potholes with seamless interface and the hardening was accomplished within a very short time of 1 h. The hardened LSPA exhibited excellent adhesion strength of approximately 1.95 MPa, enabling the elimination of gaps arising from traffic cycles and climate variation.

Rosuvastatin Pharmacokinetics in Asian and White Subjects Wild Type for Both OATP1B1 and BCRP Under Control and Inhibited Conditions

The Food and Drug Administration recommends rosuvastatin dosage reductions in Asian patients because pharmacokinetic studies have demonstrated an approximate 2-fold increase in median exposure to rosuvastatin in Asian subjects compared with Caucasian controls. Yet, no explanation for this ethnic difference has been confirmed. Here we show that rosuvastatin exposure in Asians and Whites does not differ significantly when all subjects are wild-type carriers for both solute carrier organic anion transporter 1B1 *1a and ATP-binding cassette subfamily G member 2 c.421 transporters in a 2-arm, randomized, cross-over rosuvastatin pharmacokinetics study in healthy white and Asian volunteers. For single rosuvastatin doses, AUC0-48 were 92.5 (±36.2) and 83.5 (±32.2) ng/mL × h and Cmax were 10.0 (±4.1) and 7.6 (±3.0) ng/mL for Asians and Whites, respectively. When transporters were inhibited by intravenous rifampin, rosuvastatin AUC0-48 and Cmax also showed no ethnic differences. Our study suggests that both SLCO1B1 and ABCG2 polymorphisms are better predictors of rosuvastatin exposure than ethnicity alone and could be considered in precision medicine dosing of rosuvastatin.

Peroxisome proliferator-activated receptor-α expression induces alterations in cardiac myofilaments in a pressure-overload model of hypertrophy.

Although alterations in fatty acid (FA) metabolism have been shown to have a negative impact on contractility of the hypertrophied heart, the targets of action remain elusive. In this study we compared the function of skinned fiber bundles from transgenic (Tg) mice that overexpress a relatively low level of the peroxisome proliferator-activated receptor α (PPARα), and nontransgenic (NTg) littermates. The mice (NTg-T and Tg-T) were stressed by transverse aortic constriction (TAC) and compared with shams (NTg-S and Tg-S). There was an approximate 4-fold increase in PPARα expression in Tg-S compared with NTg-S, but Tg-T hearts showed the same PPARα expression as NTg-T. Expression of PPARα did not alter the hypertrophic response to TAC but did reduce ejection fraction (EF) in Tg-T hearts compared with other groups. The rate of actomyosin ATP hydrolysis was significantly higher in Tg-S skinned fiber bundles compared with all other groups. Tg-T hearts showed an increase in phosphorylation of specific sites on cardiac myosin binding protein-C (cMyBP-C) and β-myosin heavy chain isoform. These results advance our understanding of potential signaling to the myofilaments induced by altered FA metabolism under normal and pathological states. We demonstrate that chronic and transient PPARα activation during pathological stress alters myofilament response to Ca2+ through a mechanism that is possibly mediated by MyBP-C phosphorylation and myosin heavy chain isoforms.NEW & NOTEWORTHY Data presented here demonstrate novel signaling to sarcomeric proteins by chronic alterations in fatty acid metabolism induced by PPARα. The mechanism involves modifications of key myofilament regulatory proteins modifying cross-bridge dynamics with differential effects in controls and hearts stressed by pressure overload.

Second-Generation Antidepressants and Hyponatremia Risk: A Population-Based Cohort Study of Older Adults

Hyponatremia may occur after initiation of a second-generation antidepressant drug. However, the magnitude of this risk among older adults in routine care is not well characterized. Retrospective, population-based, matched-cohort study. In Ontario, Canada, 2003 to 2012, we compared older adults with a mood or anxiety disorder who were dispensed 1 of 9 second-generation antidepressant drugs with matched adults with comparable indicators of baseline health who were not dispensed an antidepressant drug (n=138,246 per group). A similar comparison was made in a subpopulation with available laboratory data (n=4,186 per group). Second-generation antidepressant prescription versus no antidepressant prescription. The primary outcome was hospitalization with hyponatremia. A secondary outcome was hospitalization with both hyponatremia and delirium. We assessed hospitalization with hyponatremia using a diagnosis code and, in the subpopulation, serum sodium values. We assessed hospitalization with hyponatremia and delirium using a combination of diagnosis codes. Second-generation antidepressant use versus nonuse was associated with higher 30-day risk for hospitalization with hyponatremia (450/138,246 [0.33%] vs 84/138,246 [0.06%]; relative risk [RR], 5.46 [95% CI, 4.32-6.91]). This association was consistent in the subpopulation with serum sodium values (73/4,186 [1.74%] vs 18/4,186 [0.43%]; RR, 4.23 [95% CI, 2.50-7.19]; absolute risk increase, 1.31% [95% CI, 0.87%-1.75%]). Second-generation antidepressant use versus nonuse was also associated with higher 30-day risk for hospitalization with both hyponatremia and delirium (28/138,246 [0.02%] vs 7/138,246 [0.005%]; RR, 4.00 [95% CI, 1.75-9.16]). Measures of serum sodium could be ascertained in only a subpopulation. Use of a second-generation antidepressant in routine care by older adults is associated with an approximate 5-fold increase in 30-day risk for hospitalization with hyponatremia compared to nonuse. However, the absolute increase in 30-day incidence is low.