Carboxymethyl Starch Films as Enteric Coatings: Processing and Mechanistic Insights.

This study proposes the application of carboxymethyl starch derivatives as tablet coatings affording gastro-protection. Carboxymethyl starch (CMS) films were obtained by casting of aqueous filmogenic starch solutions with or without plasticizers and their structural organization was followed using Fourier transform infrared (FTIR), Thermogravimetric analysis (TGA), X-ray diffraction (XRD). Together with data from mechanical tests (tensile strength, elongation, Young's modulus) the results were used to select filmogenic formulations adapted for coatings of tablets. The behaviour of these films was evaluated in simulated gastric and intestinal fluids. The effect of plasticizers (glycerol and sorbitol) on the starch organization, on the rate of drying of the films and on the water vapor absorption was also analyzed. Various types of starch have been compared and the best results were found with high amylose starch (HAS) that was carboxymethylated in an aqueous phase to obtain carboxymethyl high amylose starch (CMHAS). The CMHAS coating solutions containing sorbitol or glycerol as plasticizers have been applied with an industrial pan coater and the final tablets exhibited a good gastro-resistance (up to 2h) in simulated gastric fluid followed by disintegration in simulated intestinal fluid (SIF). The CMHAS derivatives present a high potential as coatings for nutraceutical and pharmaceutical solid dosage forms.

Anionic and Ampholytic High-Amylose Starch Derivatives as Excipients for Pharmaceutical and Biopharmaceutical Applications: Structure-Properties Correlations.

Many chemical modifications of starch are realized in organic (mostly methanol) phase, allowing high degrees of substitution (DS). Some of these materials are used as disintegrants. To expand the usage of starch derivative biopolymers as drug delivery system, various starch derivatives obtained in aqueous phase were evaluated with the aim to identify materials and procedures which would generate multifunctional excipients providing gastro-protection for controlled drug delivery. Chemical, structural and thermal characteristics of anionic and ampholytic High Amylose Starch (HAS) derivatives under powder (P), tablet (T) and film (F) forms were evaluated by X-ray Diffraction (XRD), Fourier Transformed Infrared (FTIR) and thermogravimetric analysis (TGA) methods and correlated with the behavior of tablets and films in simulated gastric and intestinal media. At low DS, the HAS carboxymethylation (CMHAS) in aqueous phase, generated tablets and films that were insoluble at ambient conditions. The CMHAS filmogenic solutions, with a lower viscosity, were easier to cast and gave smooth films without the use of plasticizer. Correlations were found between structural parameters and the properties of starch excipients. Compared to other starch modification procedures, the aqueous modification of HAS generated tunable multifunctional excipients that may be recommended for tablets and functional coatings for colon-targeted formulations.

Characterization of a KDM5 small molecule inhibitor with antiviral activity against hepatitis B virus.

Chronic hepatitis B (CHB) is a global health care challenge and a major cause of liver disease. To find new therapeutic avenues with a potential to functionally cure chronic Hepatitis B virus (HBV) infection, we performed a focused screen of epigenetic modifiers to identify potential inhibitors of replication or gene expression. From this work we identified isonicotinic acid inhibitors of the histone lysine demethylase 5 (KDM5) with potent anti-HBV activity. To enhance the cellular permeability and liver accumulation of the most potent KDM5 inhibitor identified (GS-080) an ester prodrug was developed (GS-5801) that resulted in improved bioavailability and liver exposure as well as an increased H3K4me3:H3 ratio on chromatin. GS-5801 treatment of HBV-infected primary human hepatocytes reduced the levels of HBV RNA, DNA and antigen. Evaluation of GS-5801 antiviral activity in a humanized mouse model of HBV infection, however, did not result in antiviral efficacy, despite achieving pharmacodynamic levels of H3K4me3:H3 predicted to be efficacious from the in vitro model. Here we discuss potential reasons for the disconnect between in vitro and in vivo efficacy, which highlight the translational difficulties of epigenetic targets for viral diseases.

Synthesis of Metal-Loaded Carboxylated Biopolymers with Antibacterial Activity through Metal Subnanoparticle Incorporation.

Carboxymethyl starch (CMS) and carboxymethyl cellulose (CMC) loaded by highly dispersed metal subnanoparticles (MSNPs) showed antibacterial activity against E. coli and B. subtilis strains. Copper and silver were found to act in both cationic and zero-valence forms. The antibacterial activity depends on the metal species content but only up to a certain level. Silver cation (Ag+) showed higher antibacterial activity as compared to Ag0, which was, however, more effective than Cu0, due to weaker retention. The number of carboxyl groups of the biopolymers was found to govern the material dispersion in aqueous media, the metal retention strength and dispersion in the host-matrices. Cation and metal retention in both biopolymers was found to involve interactions with the oxygen atoms of both hydroxyl and carboxyl groups. There exists a ternary interdependence between the Zeta potential (ZP), pH induced by the biocidal agent and its particle size (PS). This interdependence is a key factor in the exchange processes with the surrounding species, including bacteria. Clay mineral incorporation was found to mitigate material dispersion, due to detrimental competitive clay:polymer interaction. This knowledge advancement opens promising prospects for manufacturing metal-loaded materials for biomedical applications.

In vivo evaluation of targeted delivery of biological agents using barium sulfate.

This study was aimed to monitor the transit through the intestine by X-ray imaging using barium sulfate (BS) as tracer. The in vitro features of monolithic tablets were correlated with their in vivo behavior in order to provide a tool for the development of targeted formulations containing macromolecular bioactive agents. The impact of BS on various matrices (neutral, ionic) was studied in simulated fluids using the disintegration time (DT) as main parameter. Dry tablets were characterized by spectroscopic methods (X-ray diffraction and Infra-Red) and scanning electron microscopy (SEM). The selected formulations were followed in a beagle dog model. The in vivo and in vitro DT of tablets formulated with BS were compared. Results: anionic excipients carboxymethylcellulose (CMC) and carboxymethylstarch (CMS) protected the active ingredient from the gastric acidity, ensuring its targeted delivery in the intestine. The SEM analysis, before and after transit in simulated fluids, showed that BS remained in the tablets allowing their good follow-up in vivo. The incorporation of 30% protein in tablets with 40% BS had no impact on their behavior. In conclusion, BS and X-ray imagery could be a good alternative to scintigraphy for development of targeted formulations containing high molecular weight bioactive agents.

Effect of ozonation on anaerobic digestion sludge activity and viability.

The effect of ozonation of anaerobic digested sludge on methane production was studied as a means of increasing the capacity of municipal anaerobic digesters. Ozone doses ranging from 0 to 192 mg O3/g sludge COD were evaluated in batch tests with a bench scale ozonation unit. Ozonation initially, and temporarily, reduced biomass viability and acetoclastic methanogenic activity, resulting in an initial lag phase ranging from 0.8 to 10 days. Following this lag phase, ozonation enhanced methane production with an optimal methane yield attained at 86 mg O3/g COD. Under these conditions, the yield of methane and the rate of its formation were 52% and 95% higher, respectively, than those factors measured without ozonation. A required optimal ozone dose could be feasible to improve the anaerobic digestion performance by increasing the methane production potential with a minimum impact on microbial activity; thus, an optimal ozone dose would enable an increase in the capacity of anaerobic digesters.

Activated Sludge Production Parameters and Nutrient Content of Organic Sludge Components.

An important part of biological treatment system design is quantifying the sludge production and the nutrient removal capacity. Influent wastewater COD fractionation, biomass growth and endogenous respiration directly impacts the composition of the mixed liquor solids in activated sludge systems. The objectives of this project were to determine the model kinetic and stoichiometric parameters associated with activated sludge production and the nutrient content (N and P) of unbiodegradable organic matter components. A complete sludge retention experiment was conducted over 70 days in a pilot-scale membrane bioreactor fed with a real municipal wastewater, and operated with alternating growth and famine periods. Experimental results were simulated and compared using the default values from two well-accepted model parameter sets. The General ASDM parameter set was found to better fit the experimental data than the Metcalf and Eddy parameter set, mainly to characterize endogenous respiration and the heterotrophic biomass concentration. An influent unbiodegradable organic particulate fraction (fXU,Inf) value of 0.16 g COD/g COD was determined by calibration of the accumulated sludge total COD, suspended solids and heterotrophic biomass concentrations. The nutrient content of the accumulated endogenous residue (XE) and influent unbiodegradable organic particulate (XU,Inf) components were calibrated to 0.030 and 0.100 g N/g COD and 0.035 and 0.008 g P/g COD, respectively. These values are in the range of those reported in the literature except for the high P content found in the endogenous residue, possibly due to the presence of coagulants added for P removal in the accumulated sludge. These results were consistent under the wide range of dynamic conditions tested and could improve model prediction of sludge production and composition.

Compact secondary treatment train combining a lab-scale moving bed biofilm reactor and enhanced flotation processes.

High-rate wastewater processes are receiving a renewed interest to obtain energy positive/efficient water resource recovery facilities. An innovative treatment train combining a high-rate moving bed biofilm reactor (HR-MBBR) with an enhanced flotation process was studied. The two objectives of this work were 1) to maximize the conversion of soluble organics to particulate matter in an HR-MBBR and 2) to maximize the particulate matter recovery from the HR-MBBR effluent by green chemicals to enhance biogas production by anaerobic digestion. To achieve these objectives, lab-scale MBBRs fed with synthetic soluble wastewater were operated at organic loading rates (OLRs) between 4 and 34 kg COD m-3 reactor d-1 corresponding to hydraulic retention times (HRTs) between 6 and 54 min. Colloidal and soluble chemical oxygen demand (COD) removal efficiency in the HR-MBBR increased with HRT to reach a plateau of 85% at an HRT longer than 27 min. Carrier clogging observed at an OLR higher than 16 kg COD m-3 d-1 (HRT < 13 min) resulted in about 23% loss in colloidal and soluble COD removal efficiency. Thus, the recommended parameters were between 22 and 37 min and between 6 and 10 kg COD m-3 d-1 for the HRT and the OLR, respectively, to maximize the conversion of soluble organics to particulate matter. Total suspended solids (TSS) recovery of 58-85% and 90-97% were achieved by enhanced flotation using green and unbiodegradable chemicals, respectively, corresponding to a TSS effluent concentration below 14 and 7 mg TSS/L. Among the synthetic polymers tested, a high molecular weight and low charge density cationic polyacrylamide was found to give the best results with less than 2 mg TSS/L in the clarified effluent (97% TSS recovery). Green chemicals, although performing slightly less for solids separation than unbiodegradable chemicals, achieved a mean TSS concentration of 10 ± 3 mg/L in the clarified effluent.

Sleep Deprivation Reveals Altered Brain Perfusion Patterns in Somnambulism.

BACKGROUND: Despite its high prevalence, relatively little is known about the pathophysiology of somnambulism. Increasing evidence indicates that somnambulism is associated with functional abnormalities during wakefulness and that sleep deprivation constitutes an important drive that facilitates sleepwalking in predisposed patients. Here, we studied the neural mechanisms associated with somnambulism using Single Photon Emission Computed Tomography (SPECT) with 99mTc-Ethylene Cysteinate Dimer (ECD), during wakefulness and after sleep deprivation. METHODS: Ten adult sleepwalkers and twelve controls with normal sleep were scanned using 99mTc-ECD SPECT in morning wakefulness after a full night of sleep. Eight of the sleepwalkers and nine of the controls were also scanned during wakefulness after a night of total sleep deprivation. Between-group comparisons of regional cerebral blood flow (rCBF) were performed to characterize brain activity patterns during wakefulness in sleepwalkers. RESULTS: During wakefulness following a night of total sleep deprivation, rCBF was decreased bilaterally in the inferior temporal gyrus in sleepwalkers compared to controls. CONCLUSIONS: Functional neural abnormalities can be observed during wakefulness in somnambulism, particularly after sleep deprivation and in the inferior temporal cortex. Sleep deprivation thus not only facilitates the occurrence of sleepwalking episodes, but also uncovers patterns of neural dysfunction that characterize sleepwalkers during wakefulness.

Mechanisms for Reduced Excess Sludge Production in the Cannibal Process.

Reducing excess sludge production is increasingly attractive as a result of rising costs and constraints with respect to sludge treatment and disposal. A technology in which the mechanisms remain not well understood is the Cannibal process, for which very low sludge yields have been reported. The objective of this work was to use modeling as a means to characterize excess sludge production at a full-scale Cannibal facility by providing a long sludge retention time and removing trash and grit by physical processes. The facility was characterized by using its historical data, from discussion with the staff and by conducting a sampling campaign to prepare a solids inventory and an overall mass balance. At the evaluated sludge retention time of 400 days, the sum of the daily loss of suspended solids to the effluent and of the waste activated sludge solids contributed approximately equally to the sum of solids that are wasted daily as trash and grit from the solids separation module. The overall sludge production was estimated to be 0.14 g total suspended solids produced/g chemical oxygen demand removed. The essential functions of the Cannibal process for the reduction of sludge production appear to be to remove trash and grit from the sludge by physical processes of microscreening and hydrocycloning, respectively, and to provide a long sludge retention time, which allows the slow degradation of the "unbiodegradable" influent particulate organics (XU,Inf) and the endogenous residue (XE). The high energy demand of 1.6 kWh/m³ of treated wastewater at the studied facility limits the niche of the Cannibal process to small- to medium-sized facilities in which sludge disposal costs are high but electricity costs are low.

Sleep deprivation impairs inhibitory control during wakefulness in adult sleepwalkers.

Sleepwalkers often complain of excessive daytime somnolence. Although excessive daytime somnolence has been associated with cognitive impairment in several sleep disorders, very few data exist concerning sleepwalking. This study aimed to investigate daytime cognitive functioning in adults diagnosed with idiopathic sleepwalking. Fifteen sleepwalkers and 15 matched controls were administered the Continuous Performance Test and Stroop Colour-Word Test in the morning after an overnight polysomnographic assessment. Participants were tested a week later on the same neuropsychological battery, but after 25 h of sleep deprivation, a procedure known to precipitate sleepwalking episodes during subsequent recovery sleep. There were no significant differences between sleepwalkers and controls on any of the cognitive tests administered under normal waking conditions. Testing following sleep deprivation revealed significant impairment in sleepwalkers' executive functions related to inhibitory control, as they made more errors than controls on the Stroop Colour-Word Test and more commission errors on the Continuous Performance Test. Sleepwalkers' scores on measures of executive functions were not associated with self-reported sleepiness or indices of sleep fragmentation from baseline polysomnographic recordings. The results support the idea that sleepwalking involves daytime consequences and suggest that these may also include cognitive impairments in the form of disrupted inhibitory control following sleep deprivation. These disruptions may represent a daytime expression of sleepwalking's pathophysiological mechanisms.

Discovery of 6-phenylpyrimido[4,5-b][1,4]oxazines as potent and selective acyl CoA:diacylglycerol acyltransferase 1 (DGAT1) inhibitors with in vivo efficacy in rodents.

The discovery and optimization of a series of acyl CoA:diacylglycerol acyltransferase 1 (DGAT1) inhibitors based on a pyrimido[4,5-b][1,4]oxazine scaffold is described. The SAR of a moderately potent HTS hit was investigated resulting in the discovery of phenylcyclohexylacetic acid 1, which displayed good DGAT1 inhibitory activity, selectivity, and PK properties. During preclinical toxicity studies a metabolite of 1 was observed that was responsible for elevating the levels of liver enzymes ALT and AST. Subsequently, analogues were synthesized to preclude the formation of the toxic metabolite. This effort resulted in the discovery of spiroindane 42, which displayed significantly improved DGAT1 inhibition compared to 1. Spiroindane 42 was well tolerated in rodents in vivo, demonstrated efficacy in an oral triglyceride uptake study in mice, and had an acceptable safety profile in preclinical toxicity studies.

Psychopathologic correlates of adult sleepwalking.

OBJECTIVES: Sleepwalking (SW) often has been associated with psychopathology, but the nature and magnitude of this relation remains unclear. The aim of our study was to investigate the presence of psychopathology in a large cohort of sleepwalkers and to determine if levels of psychopathology showed differential relations to specific characteristics of the disorder, including clinical history. METHODS: One-hundred and five sleepwalkers (39 men, 66 women; mean age, 32.4±9.5years) referred to our sleep disorders clinic for chronic SW underwent a comprehensive clinical investigation that included an overnight polysomnography (PSG) assessment in 90% of cases. All participants also completed a series of questionnaires, including the Beck Depression Inventory, Second Revision (BDI-II), the Beck Anxiety Inventory (BAI), and the Symptom Checklist 90-Revised (SCL-90-R). RESULTS: The proportion of sleepwalkers who scored above the minimal clinical threshold on the BDI-II, BAI, and SCL-90-R was 27%, 40%, and 28%, respectively. Only 15% of sleepwalkers showed moderate to severe symptoms on the BDI-II and 19% on the BAI. Taken as a whole, these profiles are similar to those observed in the general adult population. The presence of psychopathology in sleepwalkers was associated with a negative family history for SW, a higher frequency of nightmares, and with potentially injurious behaviors enacted during somnambulistic episodes. CONCLUSIONS: A majority of adult sleepwalkers consulting for the disorder do not report clinically significant levels of depression or anxiety. Overall, sleepwalkers with and without psychopathology appear more similar than dissimilar.

Daytime somnolence in adult sleepwalkers.

OBJECTIVES: Sleepwalkers often complain of excessive daytime somnolence (EDS). Our retrospective study aimed to document the presence of EDS in a substantial sample of sleepwalkers and to explore the contribution of other sleep disorders, nocturnal sleep disruption, and sleep depth to the alteration of their daytime vigilance. METHODS: Seventy adult sleepwalkers and 70 control subjects completed the Epworth Sleepiness Scale (ESS). Sleepwalkers also were studied for one night in the sleep laboratory. We compared the sleep profiles of 32 somnolent vs 38 nonsomnolent sleepwalkers and investigated the relationship between ESS scores and sleep-related variables. RESULTS: No differences were found in polysomnographic (PSG) parameters. Slow-wave activity (SWA) also was similar in the two subgroups. Sleepwalkers' ESS scores were not correlated with their body mass index (BMI) or periodic limb movements during sleep (PLMS) index, but they tended to be negatively correlated with indices of respiratory events. CONCLUSIONS: The EDS reported by adult sleepwalkers does not appear to be explained by the presence of concomitant sleep disorders or PSG signs of nocturnal sleep disruption. These results raise the possibility that EDS is part of the sleepwalking phenotype and that it is linked to its underlying pathophysiology.

Modelling the degradation of endogenous residue and 'unbiodegradable' influent organic suspended solids to predict sludge production.

Activated sludge models have assumed that a portion of organic solids in municipal wastewater influent is unbiodegradable. Also, it is assumed that solids from biomass decay cannot be degraded further. The paper evaluates these assumptions based on data from systems operating at higher than typical sludge retention times (SRTs), including membrane bioreactor systems with total solids retention (no intentional sludge wastage). Data from over 30 references and with SRTs of up to 400 d were analysed. A modified model that considers the possible degradation of the two components is proposed. First order degradation rates of approximately 0.007 d(-1) for both components appear to improve sludge production estimates. Factors possibly influencing these degradation rates such as wastewater characteristics and bioavailability are discussed.

Discovery and optimization of a series of liver X receptor antagonists.

The present report describes our efforts to convert an existing LXR agonist into an LXR antagonist using a structure-based approach. A series of benzenesulfonamides was synthesized based on structural modification of a known LXR agonist and was determined to be potent dual liver X receptor (LXR α/ß) ligands. Herein we report the identification of compound 54 as the first reported LXR antagonist that is suitable for pharmacological in vivo evaluation in rodents.

Discovery of a new binding mode for a series of liver X receptor agonists.

Structural modification of a series of dual LXRα/ß agonists led to the identification of a new class of LXRß partial agonists. An X-ray co-crystal structure shows that a representative member of this series, pyrrole 5, binds to LXRß with a reversed orientation compared to 1.

A novel series of IKKß inhibitors part II: description of a potent and pharmacologically active series of analogs.

A novel series of (E)-1-((2-(1-methyl-1H-imidazol-5-yl) quinolin-4-yl) methylene) thiosemicarbazides was discovered as potent inhibitors of IKKß. In this Letter we document our efforts at further optimization of this series, culminating in 2 with submicromolar potency in a HWB assay and efficacy in a CIA mouse model.

A novel series of IKKß inhibitors part I: Initial SAR studies of a HTS hit.

A novel series of (E)-1-((2-(1-methyl-1H-imidazol-5-yl) quinolin-4-yl) methylene) thiosemicarbazides was discovered as potent inhibitors of IKKß. In this Letter we document our early efforts at optimization of the quinoline core, the imidazole and the semithiocarbazone moiety. Most potency gains came from substitution around the 6- and 7-positions of the quinoline ring. Replacement of the semithiocarbazone with a semicarbazone decreased potency but led to some measurable exposure.