OBJECTIVES/GOALS: Allopregnanolone (ALLO), a modulator of GABAA receptors, may be useful as a treatment for human and canine benzodiazepine-refractory status epilepticus (SE). Our objective was to develop a phamacokinetic-pharmacodynamic (PKPD) model relating ALLO plasma concentrations to electroencephalographic (EEG) effects in dogs. METHODS/STUDY POPULATION: Four healthy dogs and one dog with epilepsy that had implanted intracranial electrodes were utilized. ALLO doses ranging from 1-6 mg/kg were administered IV over 5 min. EEG data were collected during four IM doses (1-2 mg/kg). Blood samples were collected up to 6 hr following dosing. ALLO concentrations were measured using HPLC-MS/MS. Power density was determined in EEG bands using a custom algorithm. A two-compartment link PKPD model was developed to describe the relation between ALLO plasma concentration and change in EEG power in the alpha, beta, delta and theta bands. RESULTS/ANTICIPATED RESULTS: ALLO caused a rapid increase in absolute power density in all EEG bands measured (1-4, >4 – 8, >8 – 12, >12 – 25, and >25 – 100 Hz). The onset of effect was rapid (1-3 min) and demonstrated by frequency band and dose analysis. Concentration-EEG data were best fit by a two-compartment PK model and sigmoidal Emax PD indirect link model. The beta frequency band was most sensitive, showing increases in power at the lowest ALLO concentrations. The EC50 concentration for the beta frequency was ~270 ng/mL. The EC50 values for effects on the other frequency bands were ~500-700 ng /mL. In conclusion, IV ALLO causes a rapid effect on EEG that can be used to determine minimal plasma concentrations associated with target engagement. DISCUSSION/SIGNIFICANCE OF IMPACT: Dose selection for future clinical trials will use the effective concentrations determined here in conjunction with studies in animal status epilepticus models. Studies are planned in client owned dogs with epilepsy to evaluate clinical efficacy in dogs and as nonclinical proof-of-concept evidence supporting translational studies in people. CONFLICT OF INTEREST DESCRIPTION: Michael Rogawski and Dorota Zolkowska are named as inventors on patent applications claiming use of neuroactive steroids including allopregnanolone and ganaxolone in the treatment of status epilepticus.

The present study was undertaken to investigate the antiparasitic activity of extracellular products of Streptomyces albus. Bioactivity-guided isolation of chloroform extracts affording a compound showing potent activity. The structure of the compound was elucidated as salinomycin (SAL) by EI-MS, 1H NMR and 13C NMR. In vitro test showed that SAL has potent anti-parasitic efficacy against theronts of Ichthyophthirius multifiliis with 10 min, 1, 2, 3 and 4 h (effective concentration) EC50 (95% confidence intervals) of 2.12 (2.22–2.02), 1.93 (1.98–1.88), 1.42 (1.47–1.37), 1.35 (1.41–1.31) and 1.11 (1.21–1.01) mg L−1. In vitro antiparasitic assays revealed that SAL could be 100% effective against I. multifiliis encysted tomonts at a concentration of 8.0 mg L−1. In vivo test demonstrated that the number of I. multifiliis trophonts on Erythroculter ilishaeformis treated with SAL was markedly lower than that of control group at 10 days after exposed to theronts (P < 0.05). In the control group, 80% mortality was observed owing to heavy I. multifiliis infection at 10 days. On the other hand, only 30.0% mortality was recorded in the group treated with 8.0 mg L−1 SAL. The median lethal dose (LD50) of SAL for E. ilishaeformis was 32.9 mg L−1.

Morphological evolution and phase transformation of metastable intermediate precipitates are critical to their mechanical properties for the non-isothermal processing. During the non-isothermal precipitation, the formation of the new phases usually couples with structural evolution. Traditional structural characterization has limitation to resolve comprehensive changes simultaneously. In this study, we report direct observation, precipitation sequence, and the details of concurrent morphological and structural changes of various intermediate precipitates during non-isothermal heating in the Al–Cu systems with different pretreatments. The structural heterogeneity during the non-isothermal precipitation processes is resolved into coexistence of two different precipitate phases and quantitatively studied in terms of the phase transition and the morphological evolution. This paper presents the in situ small- and wide-angle synchrotron x-ray scattering (SAXS and WAXS) to refine and to identify the mixed structural information during multiple precipitation stages. The WAXS results show that the precipitation sequence is θ″ → (θ″ + θ′) → θ′ → (θ′ + θ) → θ upon heating. Due to the fact of the specifically oriented SAXS intensity, the evolution of the aforementioned phase transformation is resolved by the refinement of the SAXS intensity integrated over the selected area. These methods reveal multiscale information that is not trivial comparing to the traditional characterization methods.

In order to determine the transference of Bacillus thuringiensis Berliner (Bacillaceae) (Bt) insecticidal protein in the food chain, enzyme-linked immunosorbent assay was used to detect Bt insecticidal protein levels in transgenic Bt cotton (GK12, New variety 33B and SGK321), Spodoptera exigua (Hübner) (Lepidoptera: Noctuidae) fed on the Bt cotton varieties, and two natural enemies of S. exigua, Chrysoperla carnea (Stephens) (Neuroptera: Chrysopidae) and Microplitis pallidipes Szépligeti (Hymenoptera: Braconidae). The results showed that Bt insecticidal protein was found not only in cotton leaves, but also in the body and excrement of S. exigua and the bodies of both C. carnea and M. pallidipes. Bt toxin was detected in S. exigua larvae of all the examined instars (second, third, fourth, and fifth) that fed on transgenic cotton varieties and the Bt toxin level was the highest in the body of the second instar. In addition, the Bt toxin content in the excrement of the second instar was lower than that in the older ones. After the natural enemies C. carnea and M. pallidipes preyed/parasitised the S. exigua larvae that fed on transgenic cotton, Bt toxin was found in both the predator and parasite. This research indicates that Bt protein can be transferred through the food chain and to natural enemies of various predatory habits.

The electrode materials for VRFB should possess higher electric conductivity, corrosion resistance and hydrophilic properties in sulfuric acid. The characteristics of the electrode materials affect the stability and the energy efficiency of VRFB. Carbon materials are the best suited for VRFB applications. In this study, the calcined treatment, acid treatment and ozone treatment were used to modify the surface of carbon papers. The redox reaction of [VO]2+/[VO2]+ on the modified carbon papers was evaluated by cyclic voltammetry (CV). The surface compositions of carbon materials were analyzed by X-ray photoelectron spectrometry (XPS). The experimental results reveal that three oxidative methods enhance the redox reaction of [VO]2+/[VO2]+. The calcined treatments and acid treatments also enhanced hydrolysis reaction. The mole ratio of O/C apparently increased, but the binding energy of C1s and O1s were not chemically shifted in the acid treatment. The intensity of binding energy of O1s, between 532 eV and 534 eV, apparently increased in the ozone and calcined treatments. The Ox treated samples were more hydrophilic than the Oz treated samples. In the Ox treated samples, the decrease of Rct value indicates that was contributed from the redox reaction of [VO]2+/[VO2]+ and hydrolysis reaction. It does not completely benefit the energy efficiency of VRFB. The 5 x 5 cm2 modified carbon papers were used as electrode materials in the VRFB. The voltage efficiency, coulomb efficiency and energy efficiency reached 93 %, 90 % and 83 %, respectively, at a current density of 12 mA．cm-2 at 0.8-1.8 V.

The stability of the negative electrode electrolyte affects the efficiency and capacity of energy storage in the vanadium redox flow battery (VRFB) system. To explore the stability of vanadium electrolytes, the study prepared five types of V(II) electrolytes that were exposed to air in a fixed open area and monitored the charge state of vanadium ions over time by UV/Visible spectrophotometer. This study succeeded in preparing pure V(II) electrolytes. Five characteristics are found in the UV/Visible spectra, respectively, during the oxidation process from V(II) electrolytes to V(III) electrolytes and V(III) electrolytes to V(IV) electrolytes. The experimental results show that the oxidation rate of a solution of 1 M V(II) electrolytes to V(III) electrolytes and 1 M V(III) electrolytes to V(IV) electrolytes under an atmosphere of air is 4.79 and 0.0089 mol/h per square meter. The oxidation rates of 0.05-1 M V(II) electrolytes to V(III) electrolytes are approximately 96-538 times than that of V(III) electrolytes to V(IV) electrolytes.

Fluorescent nanodiamonds (FNDs) with a size in the range of 10 – 100 nm have been produced by ion irradiation and annealing, and isolated by differential centrifugation. Single particle spectroscopic characterization with confocal fluorescence microscopy and fluorescence correlation spectroscopy indicates that they are photostable and useful as an alternative to far-red fluorescent proteins for bioimaging applications. We demonstrate the application by performing in vivo imaging of bare and bioconjugated FND particles (100 nm in diameter) in C. elegans and zebrafishes and exploring the interactions between this novel nanomaterial and the model organisms. Our results indicate that FNDs can be delivered to the embryos of both organisms by microinjection and eventually into the hatched larvae in the next generation. No deleterious effects have been observed for the carbon-based nanoparticles in vivo. The high fluorescence brightness, excellent photostability, and nontoxic nature of the nanomaterial have allowed long-term imaging and tracking of embryogenesis in the organisms.

The outcome of Plasmodium yoelii 17XL-infected BALB/c and DBA/2 mice, ranging from death to spontaneous cure, respectively, depends largely on the establishment of effective pro-inflammatory type 1 responses during the early stages of infection and associates with CD4+CD25+Foxp3+regulatory T cells (Tregs). Here, effects of Tregs were analysed on early P. yoelii 17XL infection in BALB/c and DBA/2 mice. In vivo depletion of Tregs significantly reversed the inhibited establishment of effective pro-inflammatory type 1 responses in BALB/c mice, indicating that this cell population contributed to the suppression of T-cell function in malaria. Moreover, the proportion and absolute numbers of IL-10-secreting Tregs in BALB/c mice were significantly higher than that found in DBA/2 mice by intracytoplasmic staining, and IL-10 production was correlated with the Tregs population. In addition, in vivo Tregs depletion decreased the production of IL-10 and the apoptosis of CD4+ T cells. Consistently, IL-10R blockade also had the same effect as that of Tregs depletion in P. yoelii 17XL-infected BALB/c mice. Our data demonstrate that Tregs perhaps have an important role in regulating pro-inflammatory type 1 responses in an IL-10-dependent manner and induce CD4+ T cell apoptosis during the early stage of P. yoelii 17XL infection.

The ZnO thin film was successfully deposited on a glass substrate at RT by a RF reactive magnetron sputtering method. Structural, chemical, optical, and hydrophilic/hydrophobic properties are measured by using a surface profilometer, an x-ray diffractometry (XRD), an x-ray photoelectron spectroscopy (XPS), a UV-VIS spectrophotometer, and a contact angle system, respectively. Results show that the deposition rate decreases with increasing O2/(Ar+O2) ratio. Otherwise, the best stoichiometric and quality of ZnO thin film was observed at 0.30 of O2/(Ar+O2) ratio by the smallest FWHM and the strong O-Zn bonds. Regardless of O2/(Ar+O2) ratio effect or thickness effect, high transmittance (> 86%) in the visible region is observed, while the UV-shielding characteristics depend upon both the magnitude of film thickness. The film thickness plays a more prominent role in controlling optical properties, especially in the UV-shielding characteristics, than the O2/(Ar+O2) ratio. However, the hydrophobic characteristics can be obtained when the glass coating with ZnO thin films. In general, with properly coated ZnO thin film, we can obtain a glass substrate which is highly transparent in the visible region, has good UV-shielding characteristics, and possesses highly hydrophobic characteristics (self-clean capability), which is highly suitable for applications in the glass industries.

Genetic diversity and genetic variation of 240 adult plants of four tea populations in Hunan – Camellia sinensis var. sinensis, C. sinensis var. assamica cv. Duntsa, C. ptilophylla and C. sinensis var. assamica cv. Jianghua – were studied by rapid amplification of polymorphic DNA (RAPD) markers. The results showed 226 loci using 21 random primers (10 bp), of which 201 (88.9%) were polymorphic. The genetic diversity analysis indicated that Shannon's index was 0.43; 74.7% of which was within-population genetic diversity while 25.3% was among-population variation. The gene diversity indexes of total populations (HT), within populations (HS) and among populations (HST) were, respectively, 0.37, 0.28 and 0.09. The coefficient of gene differentiation (GST) among populations was 0.23, indicating a 76.7% variation within populations and 23.3% among populations. These results displayed a rich within-population genetic variation, as in Shannon's diversity index. Interpopulation gene flow (Nm) was 0.74, which indicates the limited genetic exchange between populations.

We investigated a cluster of postoperative febrile episodes and episodes of Acinetobacter baumannii bacteremia in obstetrics and gynecology wards after an electrical blackout and loss of the water supply. The use of patient-controlled analgesia was the only independent risk factor associated with postoperative fever, and A. baumannii isolates recovered from the blood of patients who had received patient-controlled analgesia were genetically related to an isolate recovered from the diluted morphine solution used for this procedure. After inappropriate preparation of the morphine solution was identified and stopped, the outbreak ended.