Discovery of potent carbonic anhydrase,acetylcholinesterase, and butyrylcholinesterase enzymes inhibitors: The new amides and thiazolidine‐4‐ones synthesized on an acetophenone base

Compounds containing nitrogen and sulfur atoms can be widely used in various fields, including industry, medicine, biotechnology, and chemical technology. Among them, amides of acids and heterocyclic compounds have an important place. These amides and thiazolidine‐4‐ones showed good inhibitory action against butyrylcholinesterase (BChE), acetylcholinesterase (AChE), and human carbonic anhydrase isoforms. AChE exists at high concentrations in the brain and red blood cells. BChE is an important enzyme that is plentiful in the liver, and it is released into the blood in a soluble form. They were demonstrated to have effective inhibition profiles with K_i values of 23.76–102.75 nM against hCA I, 58.92–136.64 nM against hCA II, 1.40–12.86 nM against AChE, and 9.82–52.77 nM against BChE. On the other hand, acetazolamide showed K_i value of 482.63 ± 56.20 nM against hCA I, and 1019.60 ± 163.70 nM against hCA II. Additionally, Tacrine inhibited AChE and BChE, showing K_i values of 397.03 ± 31.66 and 210.21 ± 15.98 nM, respectively.     

DNA photolyase of enterococci: possible explanation for its low sunlight inactivation rate

DNA photolyase is perhaps the most ancient and direct arsenal in curing the UV-induced dimers formed in the microbial genome. Out of two cofactors of the enzyme, catalytic and light harvesting, differences in the latter have provided basis for categorizing photolyases of prokaryotes as folate and deazaflavin types. In the present study, the homology modeling of DNA photolyase of Enterococcus faecalis was undertaken. The predicted models were structurally compared with the crystal structure coordinates of photolyases from Escherichia coli (folate type) and Anacystis nidulans (deazaflavin type). Discrepancies present in the multiple sequence alignment and tertiary structures, particularly at the light harvesting cofactor (methenyltetrahydrofolic acid, MTHF; 8-hydroxy-5-deazaflavin, 8-HDF) binding sites indicated the mechanistic nature of enterococcal photolyase. Concisely, despite the greater holistic homology with folate-type photolyase, enterococcal photolyase was characterized as deazaflavin-type. The presence of 8-HDF binding sites and groove architecture of substrate binding sites were also found supportive in this regard. The inter cofactor distance and/or orientation also implied to the efficient energy transfer in photolyase of Enterococcus in comparison with E. coli. In addition, we observed relatively high protein deformability in the enterococcal genome, which may favors the repair action of photolyase. The findings are expected to provide molecular insights into the difference in sunlight inactivation rate of two important fecal contamination indicators, namely Enterococcus and E. coli.     

Preparation and evaluation of an experimental iscom-based infectious bursal disease vaccine

The present study was conducted to develop and evaluate an experimental ISCOM-based infectious bursal disease (IBD) vaccine. The indigenous very virulent infectious bursal disease virus (vvIBDV) already attenuated and adapted to Vero cell line was used. After denaturation of viral proteins with sodium dodecyl sulphate (SDS), an IBD-ISCOM was constructed. The non-incorporated viral components were separated from ISCOM by centrifugation of dialysate. The pathogenicity and immunogenicity trials were conducted in 3-week-old broiler chicken. A commercial oil-emulsified vaccine (CEVAC IBD K) was used for comparison. There were no clinical signs of disease, gross or microscopic lesions in bursa of Fabricius in group G1 vaccinated with ISCOM-based vaccine and bursa to body weight ratio were comparable to un-vaccinated control group (G3). The virus-neutralizing antibody titers were significantly (P<0.05) higher in group G1 as compared with group G2 which was vaccinated with commercial vaccine. On challenge with vvIBDV, 100%, 75% and 0.00% protection was achieved in G1, G2 and G3, respectively. The results indicated that ISCOM-based IBD vaccine is safe and immunogenic.     

Clastogenic ROS and biophotonics in precancerous diagnosis

Background

Cancer is the leading cause of death worldwide. The application of biophotonics for diagnosing precancerous lesions is a major breakthrough in oncology and is associated with the expression of clastogenic bio-markers, such as reactive oxygen species (ROS), namely, superoxide anion radicals, hydrogen peroxide, hydroxyl radicals, and lipid peroxidation products. These ROS are the major sources of ultra-weak biophotons emission; in addition, biophotons are emitted from other biomolecules, which are not associated with ROS. The precancerous phase is diagnosed on the basis of biophoton emission from biomarkers. The type of biophotons emitted depends on the structure of the clastogenic ROS.

Methods

ROS-based emission of ultra-weak photons can be detected using charge coupled device (CCD) cameras and photomultiplier tubes. Furthermore, spectroscopic and microscopic analysis can yield more advanced and definite results.

Results

The frequency and intensity of biophoton emission associated with each ROS provides information regarding the precancerous phase. Previous have attempted to show an association between precancerous growth and biophoton emission; however, their results were not conclusive. In this review, we have addressed multiple aspects of the molecular environment, especially light- matter interactions, to derive a successful theoretical relationship which may have the ability to diaganose the tumor at precancerous stage and to give the solutions of previous failures. This can be a major quantum leap toward precancerous diagnosis therapy.

Conclusion

Biophotonics provides an advanced framework, for easily diagnosing cancer at its preliminary stage. The relationship between biophotons, clastogenic factors, and biochemical reactions in the cellular microenvironment can be understood successfully. The advancement in precancerous diagnosis will improve human health worldwide. The versatility of biophotonics can be used further for novel applications in biology, biochemistry, chemistry and social fields.

     

Small Interfering RNA-Mediated Control of Virus Replication in the CNS Is Therapeutic and Enables Natural Immunity to West Nile Virus

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Cloning, expression and characterization of two new IgE-binding proteins from the yeast Malassezia sympodialis with sequence similarities to heat shock proteins and manganese superoxide dismutase.

Malassezia sympodialis is an opportunistic yeast that colonizes human skin and may induce IgE and T cell reactivity in patients with atopic eczema/dermatitis syndrome (AEDS). Previously, we have cloned and expressed six recombinant allergens (rMala s 1 and rMala s 5 to rMala s 9) from this yeast. By combining high throughput screening and phage surface display techniques, 27 complete and partial IgE-binding clones of M. sympodialis have been identified. Here we enlarged the panel of recombinant M. sympodialis allergens by RACE-PCR, cloning and nucleotide sequencing to obtain the coding sequences of two new IgE-binding clones. The coding sequences of one of the clones showed similarity to the heat shock protein (HSP) family and the other to manganese superoxide dismutase (MnSOD), and both had a high degree of homology to human counterparts. The coding sequences were expressed in Escherichia coli as six-histidine tagged recombinant proteins and generated products with molecular masses of 86.1 kDa for HSP and 22.4 kDa for MnSOD. Their IgE-binding frequencies were shown to be 69% and 75%, respectively, to 28 sera from AEDS patients with serum IgE to M. sympodialis extract, indicating that HSP and MnSOD are major M. sympodialis allergens. In inhibition immunoblotting, M. sympodialis extract could inhibit the binding of serum IgE from AEDS patients to rHSP and rMnSOD in a concentration-dependent manner. The high frequency of sera from AEDS patients, showing IgE binding to both HSP and MnSOD, indicates that these allergens, designated Mala s 10 and Mala s 11, could play a role in AEDS.     

Effect of synthetic aβ peptide oligomers and fluorinated solvents on Kv1.3 channel properties and membrane conductance

The impact of synthetic amyloid β (1-42) (Aβ(1-42)) oligomers on biophysical properties of voltage-gated potassium channels Kv 1.3 and lipid bilayer membranes (BLMs) was quantified for protocols using hexafluoroisopropanol (HFIP) or sodium hydroxide (NaOH) as solvents prior to initiating the oligomer formation. Regardless of the solvent used Aβ(1-42) samples contained oligomers that reacted with the conformation-specific antibodies A11 and OC and had similar size distributions as determined by dynamic light scattering. Patch-clamp recordings of the potassium currents showed that synthetic Aβ(1-42) oligomers accelerate the activation and inactivation kinetics of Kv 1.3 current with no significant effect on current amplitude. In contrast to oligomeric samples, freshly prepared, presumably monomeric, Aβ(1-42) solutions had no effect on Kv 1.3 channel properties. Aβ(1-42) oligomers had no effect on the steady-state current (at -80 mV) recorded from Kv 1.3-expressing cells but increased the conductance of artificial BLMs in a dose-dependent fashion. Formation of amyloid channels, however, was not observed due to conditions of the experiments. To exclude the effects of HFIP (used to dissolve lyophilized Aβ(1-42) peptide), and trifluoroacetic acid (TFA) (used during Aβ(1-42) synthesis), we determined concentrations of these fluorinated compounds in the stock Aβ(1-42) solutions by (19)F NMR. After extensive evaporation, the concentration of HFIP in the 100× stock Aβ(1-42) solutions was ～1.7 μM. The concentration of residual TFA in the 70× stock Aβ(1-42) solutions was ～20 μM. Even at the stock concentrations neither HFIP nor TFA alone had any effect on potassium currents or BLMs. The Aβ(1-42) oligomers prepared with HFIP as solvent, however, were more potent in the electrophysiological tests, suggesting that fluorinated compounds, such as HFIP or structurally-related inhalational anesthetics, may affect Aβ(1-42) aggregation and potentially enhance ability of oligomers to modulate voltage-gated ion channels and biological membrane properties.