Bimanes (1,5-diazabicyclo[3.3.0]octadienediones): Laser activity in syn-bimanes

The conversion of 3-methyl-4-benzyl-4-chloro-2-pyrazolin-5-one 10b was catalyzed by a mixture of potassium fluoride and alumina to give syn-(methyl, benzyl)bimane 6 (62%) without detectable formation of the anti isomer, A6 [a 1 : 1 mixture (87%) of the isomers 6 and A6 was obtained when the catalyst was potassium carbonate]. In a similar reaction syn-(methyl,carboethoxymethyl)bimane 7 (15%) with the anti isomer A7 (36%) was obtained from 3-methyl-4-carboethoxymethyl-4-chloro-2-pyrazolin-5-one 10c . syn-(Methyl, β-acetoxyethyl)bimane 8 (70%) was obtained from 3-methyl-4-β-acetoxyethyl-4-chloro-2-pyrazolin-5-one 10d (potassium carbonate catalysis) and was converted by hydrolysis to syn-(methyl, β-hydroxyethyl)bimane 9 (40%). Acetyl nitrate (nitric acid in acetic anhydride) converted anti-(amino,hydrogen)bimane 11 to anti-(amino,nitro)bimane 15 (91%), anti-(methyl,hydrogen)bimane 13 to anti-(methyl,nitro)(methyl,hydrogen)bimane 16 (57%), and degraded syn-(methyl,hydrogen)bimane 12 to an intractable mixture. Treatment with trimethyl phosphite converted syn-(bromomethyl,methyl)bimane 17 to syn-(dimethoxyphosphinylmethyl,methyl)bimane 18 (78%) that was further converted to syn-(styryl,methyl)bimane 19 (29%) in a condensation reaction with benzaldehyde. Treatment with acryloyl chloride converted syn-(hydroxymethyl,methyl)bimane 20 to its acrylate ester 21 (22%). Stoichiometric bromination of syn-(methyl,methyl)bimane 1 gave a monobromo derivative that was converted in situ by treatment with potassium acetate to syn-(acetoxymethyl,methyl)(methyl,methyl)bimane 47 . N-Amino-μ-amino-syn-(methylene,methyl)bimane 24 (68%) was obtained from a reaction between the dibromide 17 and hydrazine. Derivatives of the hydrazine 24 included a perchlorate salt and a hydrazone 25 derived from acetone. Dehydrogenation of syn-(tetramethylene)bimane 26 by treatment with dichlorodicyanobenzoquinone (DDQ) gave syn-(benzo,tetramethylene)bimane 27 (58%) and syn-(benzo)bimane 28 (29%). Bromination of the bimane 26 gave a dibromide 29 (92%) that was also converted by treatment with DDQ to syn-(benzo)bimane 28 . Treatment with palladium (10%) on charcoal dehydrogenated 5, 6, 10, 11-tetrahydro-7H,9H-benz [6, 7] indazol [1, 2a]benz[g]indazol-7,9-dione 35 to syn-(α-naphtho)bimane 36 (71%). The bimane 35 was prepared from 1,2,3,4-tetrahydro-1-oxo-2-naphthoate 37 by stepwise treatment with hydrazine to give 1,2,4,5-tetrahydro-3H-benz[g]indazol-3-one 38 , followed by chlorine to give 3a-chloro-2,3a,4,5-tetrahydro-3H-benz[g]indazol-3-one 39 , and base. Dehydrogenation over palladium converted the indazolone 34 to 1H-benz[g] indazol-3-ol 36 . Helicity for the hexacyclic syn-(α-naphtho)bimane 36 was confirmed by an analysis based on molecular modeling. The relative efficiencies (RE) for laser activity in the spectral region 500–530 nm were obtained for 37 syn-bimanes by reference to coumarin 30 (RE 100): RE > 80 for syn-bimanes 3, 5, 18 , and μ-(dicarbomethoxy)methylene-syn-(methylene,methyl)bimane 22 : RE 20–80: for syn-bimanes 1,2,4,20,24,26 , and μ-thia-syn-(methylene,methyl)bimane 50 : and RE 0-20 for 26 syn-bimanes. The bimane dyes tended to be more photostable and more water-soluble than coumarin 30. The diphosphonate 18 in dioxane showed laser activity at 438 nm and in water at 514 nm. Presumably helicity, that was demonstrated by molecular modeling, brought about a low fluorescence intensity for syn-(α-naphtho)bimane 36 , Φ0.1, considerably lower than obtained for syn-(benzo)bimane 28 , Φ0.9.     

The optimisation of ultrasonic cleaning procedures for dairy fouled ultrafiltration membranes

Ultrafiltration (UF) of whey is a major membrane based process in the dairy industry. However, commercialization of this application has been limited by membrane fouling, which has a detrimental influence on the permeation rate. There are a number of different chemical and physical cleaning methods currently used for cleaning a fouled membrane. It has been suggested that the cleaning frequency and the severity of such cleaning procedures control the membrane lifetime. The development of an optimal cleaning strategy should therefore have a direct implication on the process economics. Recently, the use of ultrasound has attracted considerable interest as an alternative approach to the conventional methods. In the present study, we have studied the ultrasonic cleaning of polysulfone ultrafiltration membranes fouled with dairy whey solutions. The effects of a number of cleaning process parameters have been examined in the presence of ultrasound and results compared with the conventional operation. Experiments were conducted using a small single sheet membrane unit that was immersed totally within an ultrasonic bath. Results show that ultrasonic cleaning improves the cleaning efficiency under all experimental conditions. The ultrasonic effect is more significant in the absence of surfactant, but is less influenced by temperature and transmembrane pressure. Our results suggest that the ultrasonic energy acts primarily by increasing the turbulence within the cleaning solution.     

Effects of degassing on the long-range attractive force between hydrophobic surfaces in water

The long-ranged attractions between hydrophobic amorphous fluoropolymer surfaces are measured in water with and without dissolved air. An atomic force microscope is used to obtain more than 500 measured jump-in distances, which yields statistically reliable results. It is found that the range of the attraction and its variability is generally significantly decreased in deaerated water as compared to normal, aerated water. However, the range and strength of the attraction in deaerated water remain significantly greater than the van der Waals attraction for this system. The experimental observations are consistent with (1) nanobubbles being primarily responsible for the long-ranged attraction in normal water, (2) nanobubbles not being present in deaerated water when the surfaces are not in contact, and (3) the attraction in the absence of nanobubbles being most probably due to the approach to the separation-induced spinodal cavitation of the type identified by Bérard et al. [J. Chem. Phys. 1993, 98, 7236]. It is argued that the measurements in deaerated water reveal the bare or pristine hydrophobic attraction unobscured by nanobubbles.     

Identification of tyrosine nitration in UCH-L1 and GAPDH

Protein tyrosine nitration is a post-translational modification commonly used as a marker of cellular oxidative stress associated with numerous pathophysiological conditions. We focused on ubiquitin carboxyl terminal hydrolase-L1 (UCH-L1) and glyceraldehyde-3-phosphate (GAPDH) which are high-abundant brain proteins that have been identified to be highly susceptible to oxidative modification. Both UCH-L1 and GAPDH have been linked to the pathogenesis of Alzheimer's and Parkinson's disease, however specific nitration sites have not been elucidated. Identification of specific nitration sites and quantitation of endogenous nitrated proteins are important in correlating this modification to disease pathology. In this study, purified UCH-L1 and GAPDH were nitrated in vitro with peroxynitrite and the presence of nitrated proteins was confirmed by anti-3-nitrotyrosine Western blots. Data-dependent LC-MS/MS analysis identified several distinct tyrosine nitration sites in UCH-L1 (Tyr-80) and GAPDH (Tyr-47, Tyr-92, and Tyr-312). Subsequent validation with synthetic peptides was conducted for selected nitropeptides. An LC-MS/MS method was developed for semi-quantitative determination of the synthetic nitropeptides: KGQEVSPKVY(*) (UCH-L1) and mFQY(*) DSTHGKF (GAPDH). The nitropeptides were detectable in the mid-attomole range and the peak area response was linear over three orders of magnitude. Targeted analysis of endogenous UCH-L1 and GAPDH nitration was then conducted in an in vivo second-hand smoke rat model to evaluate the utility of this approach.     

Highly sensitive non‐isotopic restriction endonuclease fingerprinting of nucleotide variability in the gp60 gene within Cryptosporidium species,genotypes and subgenotypes infective to humans,and its implications

The high‐resolution analysis of genetic variation has major implications for the identification of parasites and micro‐organisms to species and subspecies as well as for population genetic and epidemiological studies. In this study, we critically assessed the effectiveness of a PCR‐based restriction endonuclease fingerprinting (REF) method for the detection of mutations in the 60 kDa glycoprotein gene (gp60) of Cryptosporidium, a genus of parasitic protists of major human and animal health importance globally. This gene displays substantial intraspecific variability in sequence, particularly in a TCA (perfect and imperfect) microsatellite region, is present as a single copy in the nuclear genome and is used widely as a marker in molecular epidemiological studies of Cryptosporidium hominis and C. parvum, the two predominant species that infect humans. The results of this study demonstrated an exquisite capacity of REF to detect nucleotide variability in the gp60 gene within each of the two species. The differentiation of genotypes/subgenotypes based on REF analysis was supported by targeted sequencing, allowing the detection of levels of variation as low as a single‐nucleotide transversion for amplicons of ∼1 kb in size. The high‐throughput potential and relatively low‐cost of REF make it a particularly useful tool for large‐scale genetic analyses of C. hominis and C. parvum. REF could also be utilized for comparative surveys of genetic variability across large nuclear genomic regions. Such analyses of Cryptosporidium in clinical and environmental samples by REF have important implications for identifying sources of infection, modes of transmission and/or possible infectivity to humans, thus assisting in the surveillance and control of cryptosporidiosis. Given its excellent mutation detection capacity, REF should find broad applicability to various single‐copy genes as well as a wide range of other protozoan and metazoan parasites. (The nucleotide sequences reported in this article are available in the GenBank database under accession numbers GU214343–GU214371).     

DANGLE: A Bayesian inferential method for predicting protein backbone dihedral angles and secondary structure

This paper introduces DANGLE, a new algorithm that employs Bayesian inference to estimate the likelihood of all possible values of the backbone dihedral angles ? and ψ for each residue in a query protein, based on observed chemical shifts and the conformational preferences of each amino acid type. The method provides robust estimates of ? and ψ within realistic boundary ranges, an indication of the degeneracy in the relationship between shift measurements and conformation at each site, and faithful secondary structure state assignments. When a simple degeneracy-based filtering procedure is applied, DANGLE offers an ideal compromise between accuracy and coverage when compared with other shift-based dihedral angle prediction methods. In addition, per residue analysis of shift/structure degeneracy has potential to be a useful new approach for studying the properties of unfolded proteins, with sufficient sensitivity to identify regions of residual structure in the acid denatured state of apomyoglobin.     

Relativistic effective potential SCF calculations of AgH and AuH

Relativistic effective potentials (REP) are now widely used in molecular electronic structure calculations. Tests of these REP are needed to assess their accuracy. This can now be done for AgH and AuH since Lee and McLean have published Dirac-Fock calculations for these molecules. Comparative SCF calculations have been performed for two types of effective potential. Satisfactory agreement between the effective potential results and Dirac-Fock values is found for spectroscopic constants and dipole moments, which supports the use of these potentials for heavy atom containing molecules.     

Heat capacity calorimetry

Experimental heat capacity measurements of α-ZrW₂O₈, and zeolitic polymorphs of SiO₂, BEA and MFI, have been made from 0.6 to 400 K. Measurements on β-ZrMo₂O₈ have been made from 8 to 400 K. Analysis of the results yields evidence for very low frequency modes in all four materials. These modes are responsible for negative thermal expansion behavior in α-ZrW₂O₈ and β-ZrMo₂O₈. Negative thermal expansion has been observed in some pure SiO₂ zeolites, but no studies have been made to look for it in BEA and MFI. The appearance of low frequency modes in these two zeolites suggests that temperature dependent structural investigations would be worthwhile. These modes are lower in energy than the Boson peak in vitreous silica. This revised version was published online in August 2006 with corrections to the Cover Date.     

Cubosomes: The Next Generation of Smart Lipid Nanoparticles?

Cubosomes are highly stable nanoparticles formed from the lipid cubic phase and stabilized by a polymer based outer corona. Bicontinuous lipid cubic phases consist of a single lipid bilayer that forms a continuous periodic membrane lattice structure with pores formed by two interwoven water channels. Cubosome composition can be tuned to engineer pore sizes or include bioactive lipids, the polymer outer corona can be used for targeting and they are highly stable under physiological conditions. Compared to liposomes, the structure provides a significantly higher membrane surface area for loading of membrane proteins and small drug molecules. Owing to recent advances, they can be engineered in vitro in both bulk and nanoparticle formats with applications including drug delivery, membrane bioreactors, artificial cells, and biosensors. This review outlines recent advances in cubosome technology enabling their application and provides guidelines for the rational design of new systems for biomedical applications.