Targeting Serotonin 2A and Adrenergic α1 Receptors for Ocular Antihypertensive Agents: Discovery of 3,4‐Dihydropyrazino[1,2‐b]indazol‐1(2H)‐one Derivatives

Glaucoma affects millions of people worldwide and causes optic nerve damage and blindness. The elevation of the intraocular pressure (IOP) is the main risk factor associated with this pathology, and decreasing IOP is the key therapeutic target of current pharmacological treatments. As potential ocular hypotensive agents, we studied compounds that act on two receptors (serotonin 2A and adrenergic α₁) linked to the regulation of aqueous humour dynamics. Herein we describe the design, synthesis, and pharmacological profiling of a series of novel bicyclic and tricyclic N2‐alkyl‐indazole‐amide derivatives. This study identified a 3,4‐dihydropyrazino[1,2‐b]indazol‐1(2H)‐one derivative with potent serotonin 2A receptor antagonism, >100‐fold selectivity over other serotonin subtype receptors, and high affinity for the α₁ receptor. Moreover, upon local administration, this compound showed superior ocular hypotensive action in vivo relative to the clinically used reference compound timolol.     

Cigarette characteristics,smoker characteristics,and the relationship to cigarette fires

Data were collected from eight cities on a wide range of cigarette and smoker characteristics for a sample of smokers. Of these, 564 smokers had had fires and were identified through fire department response to those fires, while the other 1,611 smokers had not had fires and were identified through a telephone sample survey of the communities. The characteristics analyzed included those that had shown evidence of a relationship to the risk of a cigarette-initiated fire, either in laboratory studies or in previous statistical analyses of fire experience.The smoker characteristics analyzed were household income, education, age, gender, and race. The cigarette characteristics analyzed were filter, tobacco column length, filter length, circumference, density, amount of tobacco, menthol, citrate, porosity, and pack type. In addition, a variable was used to control for the smoker's city.After controlling for all smoker characteristics and city, logistic regression modeling showed four cigarette characteristics to be significant: filter, filter length, porosity, and type of pack. Filter, filter length, and porosity all affect air intake, which, therefore, appears to be an important physical element in the combustion process associated with risk. Analysis limited to filtered cigarettes only showed the same characteristics to be significant, plus tobacco column length. Extension of the analysis to two-way interaction terms did not change any of the conclusions on which cigarette characteristics are important, but it did indicate that the role of pack type was different for men and women.Sensitivity analyses, shown in the appendix, supported the main conclusions that cigarette characteristics are significant after controlling for smoker characteristics and that the four specific cigarette characteristics—filter, filter length, porosity, and pack—are the ones that are significant. These analyses checked the impact of cluster sampling, sensitivity to missing data on smoker characteristics, and sensitivity to nonfire smoker cases with responses by people other than the smokers themselves.All this means that there are already cigarettes commercially available that exhibit a reduced propensity for ignition when one controls for smoker characteristics.This report was originally written as the final report in June 1993 as part of CPSC-P-91-1147 Contract Cigarette Fire Incident Study that the National Fire Protection Association conducted for the U.S. Consumer Product Safety Commission.Harwood, formerly with the Consumer Product Safety Commission, is retired.     

Demulsifier performance at low temperature in a low permeability reservoir

The crude oil in Longdong area is produced in the form of emulsion containing associated oil and water, which needs to be separated before dispatch to end user. Chemical demulsification under high temperature is the most widely used technology to break the emulsions. In this study a rheological method was used to determine the curve of viscosity-temperature and lower limit of temperature was determined. A series of experiments on low-temperature commercial demulsifies were implemented for studying demulsification performance by bottle test method. Mechanism of low-temperature demulsifier was studied by using spinning drop interfacial tensiometer to determine interfacial tension between the crude oil and demulsifier solution by considering the concentration. Turbiscan stability analyzer was used to study the effect of water content, temperature, and demulsifier concentration on emulsion stability. The corresponding relationship between interfacial tension and demulsification was verified through the study of low-temperature demulsifier effect on interfacial tension. Efficient low-temperature demulsifiers AR102, AR901, PR929, and PRC06 were selected. PRC06 was chosen to be the best at 40°C, and when the optimal concentration was 200 mg/L, dehydration rate was 99.51%.     

Physicochemical characteristics and bronchial epithelial cell cytotoxicity of Folpan 80 WG® and Myco 500®, two commercial forms of folpet

Background

Pesticides, in particular folpet, have been found in rural and urban air in France in the past few years. Folpet is a contact fungicide and has been widely used for the past 50 years in vineyards in France. Slightly water-soluble and mostly present as particles in the environment, it has been measured at average concentration of 40.1 μg/m³ during its spraying, 0.16–1.2 μg/m³ in rural air and around 0.01 μg/m³ in urban air, potentially exposing both the workers and the general population. However, no study on its penetration by inhalation and on its respiratory toxicity has been published. The objective of this study was to determine the physicochemical characteristics of folpet particles (morphology, granulometry, stability) in its commercial forms under their typical application conditions. Moreover, the cytotoxic effect of these particles and the generation of reactive oxygen species were assessed in vitro on respiratory cells.

Results

Granulometry of two commercial forms of folpet (Folpan 80WG^® and Myco 500^®) under their typical application conditions showed that the majority of the particles (>75%) had a size under 5 μm, and therefore could be inhaled by humans. These particles were relatively stable over time: more than 75% of folpet remained in the particle suspension after 30 days under the typical application conditions. The inhibitory concentration (IC₅₀) on human bronchial epithelial cells (16HBE14o-) was found to be between 2.89 and 5.11 μg/cm² for folpet commercial products after 24 h of exposure. Folpet degradation products and vehicles of Folpan 80 WG^® did not show any cytotoxicity at tested concentrations. At non-cytotoxic and subtoxic concentrations, Folpan 80 WG^® was found to increase DCFH-DA fluorescence.

Conclusion

These results show that the particles of commercial forms of folpet are relatively stable over time. Particles could be easily inhaled by humans, could reach the conducting airways and are cytotoxic to respiratory cells in vitro. Folpet particles may mediate its toxicity directly or indirectly through ROS-mediated alterations. These data constitute the first step towards the risk assessment of folpet particles by inhalation for human health. This work confirms the need for further studies on the effect of environmental pesticides on the respiratory system.     

A study of electrochemically treated PAN based carbon fibres by IGC and XPS

Inverse gas chromatography (IGC) has been used with conventional and novel probes in conjunction with X-ray photoelectron spectroscopy (XPS) to assess the surface chemistry of a range of aged polyacrylonitrile carbon fibres. Using conventional probes the thermodynamic surface properties were investigated at 100 °C. The dispersive component of the surface free energy, , was determined and ranged from 73.1 mJ m⁻² to 92.2 mJ m⁻². Unconventional probes, chosen as analogues of the functionalities present in a potential matrix material, were also eluted through the IGC columns. Surface compositions and peak fitting of C 1s spectra from XPS analysis helped identify groups on the fibre surface. Using these results, mechanisms of interactions occurring at the interface of a carbon fibre reinforced polymer composite have been proposed. Hydrogen bonding and stronger acid-base interactions are the main mechanisms of interactions. Although, the carbon fibres are aged, oxygen and nitrogen functionalities introduced from surface treatment have remained active.     

Nanocrystal-encoded fluorescent microbeads for proteomics: antibody profiling and diagnostics of autoimmune diseases

The first application of nanocrystal (NC)-encoded microbeads to clinical proteomics is demonstrated by multiplexed detection of circulating autoantibodies, markers of systemic sclerosis. Two-color complexes, consisting of NC-encoded, antigen-covered beads, anti-antigen antibody or clinical serum samples, and dye-tagged detecting antibodies, were observed using flow cytometry assays and on the surface of single beads. The results of flow cytometry assays correlated with the ELISA technique and provided clear discrimination between the sera samples of healthy donors and patients with autoimmune disease. Microbead fluorescence signals exhibited narrow distribution regardless of their surface antigen staining, without the need of any fluorescence compensation-a parameter determining the limit of sensitivity of flow cytometry assays. In single bead measurements, less than 30 dye-labeled antibodies interacting with the topoI-specific antibodies at the surface of a bead have been detected by the emission of dye excited through the FRET from NCs. In this format, the antibody-bead interaction reaction turns specifically the fluorescence signal from dye label off and on, additionally increasing autoantibody detection sensitivity.     

Phenotypic Characterization of Immortalized Chondrocytes from a Desbuquois Dysplasia Type 1 Mouse Model: A Tool for Studying Defects in Glycosaminoglycan Biosynthesis

The complexity of skeletal pathologies makes use of in vivo models essential to elucidate the pathogenesis of the diseases; nevertheless, chondrocyte and osteoblast cell lines provide relevant information on the underlying disease mechanisms. Due to the limitations of primary chondrocytes, immortalized cells represent a unique tool to overcome this problem since they grow very easily for several passages. However, in the immortalization procedure the cells might lose the original phenotype; thus, these cell lines should be deeply characterized before their use. We immortalized primary chondrocytes from a Cant1 knock-out mouse, an animal model of Desbuquois dysplasia type 1, with a plasmid expressing the SV40 large and small T antigen. This cell line, based on morphological and biochemical parameters, showed preservation of the chondrocyte phenotype. In addition reduced proteoglycan synthesis and oversulfation of glycosaminoglycan chains were demonstrated, as already observed in primary chondrocytes from the Cant1 knock-out mouse. In conclusion, immortalized Cant1 knock-out chondrocytes maintained the disease phenotype observed in primary cells validating the in vitro model and providing an additional tool to further study the proteoglycan biosynthesis defect. The same approach might be extended to other cartilage disorders.     

Lipid Cubic Mesophases Combined with Superparamagnetic Iron Oxide Nanoparticles: A Hybrid Multifunctional Platform with Tunable Magnetic Properties for Nanomedical Applications

Hybrid materials composed of superparamagnetic iron oxide nanoparticles (SPIONs) and lipid self-assemblies possess considerable applicative potential in the biomedical field, specifically, for drug/nutrient delivery. Recently, we showed that SPIONs-doped lipid cubic liquid crystals undergo a cubic-to-hexagonal phase transition under the action of temperature or of an alternating magnetic field (AMF). This transition triggers the release of drugs embedded in the lipid scaffold or in the water channels. In this contribution, we address this phenomenon in depth, to fully elucidate the structural details and optimize the design of hybrid multifunctional carriers for drug delivery. Combining small-angle X-ray scattering (SAXS) with a magnetic characterization, we find that, in bulk lipid cubic phases, the cubic-to-hexagonal transition determines the magnetic response of SPIONs. We then extend the investigation from bulk liquid-crystalline phases to colloidal dispersions, i.e., to lipid/SPIONs nanoparticles with cubic internal structure (“magnetocubosomes”). Through Synchrotron SAXS, we monitor the structural response of magnetocubosomes while exposed to an AMF: the magnetic energy, converted into heat by SPIONs, activates the cubic-to-hexagonal transition, and can thus be used as a remote stimulus to spike drug release “on-demand”. In addition, we show that the AMF-induced phase transition in magnetocubosomes steers the realignment of SPIONs into linear string assemblies and connect this effect with the change in their magnetic properties, observed at the bulk level. Finally, we assess the internalization ability and cytotoxicity of magnetocubosomes in vitro on HT29 adenocarcinoma cancer cells, in order to test the applicability of these smart carriers in drug delivery applications.     

Efficacy of electrolyzed water in the inactivation of planktonic and biofilm Listeria monocytogenes in the presence of organic matter

The ability of electrolyzed (EO) water to inactivate Listeria monocytogenes in suspension and biofilms on stainless steel in the presence of organic matter (sterile filtered chicken serum) was investigated. A five-strain mixture of L. monocytogenes was treated with deionized, alkaline EO, and acidic EO water containing chicken serum (0, 5, and 10 ml/liter) for 1 and 5 min. Coupons containing L. monocytogenes biofilms were also overlaid with chicken serum (0, 2.5, 5.0, and 7.5 ml/liter) and then treated with deionized water, alkaline EO water, acidic EO water, alkaline EO water followed by acidic EO water, and a sodium hypochlorite solution for 30 and 60 s. Chicken serum decreased the oxidation-reduction potential and chlorine concentration of acidic EO water but did not significantly affect its pH. In the absence of serum, acidic EO water containing chlorine at a concentration of 44 mg/liter produced a > 6-log reduction in L. monocytogenes in suspension, but its bactericidal activity decreased with increasing serum concentration. Acidic EO water and acidified sodium hypochlorite solution inactivated L. monocytogenes biofilms to similar levels, and their bactericidal effect decreased with increasing serum concentration and increased with increasing time of exposure. The sequential 30-s treatment of alkaline EO water followed by acidic EO water produced 4- to 5-log reductions in L. monocytogenes biofilms, even in the presence of organic matter.     

Are car drivers holding a motorcycle licence less responsible for motorcycle--car crash occurrence? A non-parametric approach

The purpose of this work is to evaluate the effect of a specific motorcycle licence, held by car drivers, in responsibility for motorcycle-car crashes. The data were provided by a multicentric case-control study (MAIDS) regarding the risk of crash and serious injuries of motorcyclists. A non-parametric method, classification and regression tree (CART), was used to accomplish the objective, and then compared to standard unconditional logistic regression. Drivers owning a motorcycle licence turned out to be less responsible for motorcycle-car crashes than drivers who do not have one; both types of analysis are consistent with this result. It is reasonable to assume that car drivers who hold a motorcycle licence have acquired more ability in riding and controlling two wheeled vehicles than drivers without a licence, and this may help them in predicting motorcycles manoeuvres.