Amelioration of experimental rheumatoid arthritis by selected ultra-diluted preparations by down regulating increased expression of TNF-α & IL-6

956-964The current work explored the inhibitory effect of selected homeopathic drugs, in experimental models of inflammation and CFA-induced arthritis. Twelve groups of animals were made, each containing 6 animals. The selected homeopathic drugs (causticum, calcarea, medorrhinum, mercurius, formica, proteus, silica, sulphur, thuja), placebo and standard drug, Indomethacin. In CFA model, treatment groups and the reference drug were administered daily for a period of 21 days. Dysfunction in joints was evaluated by parameters such as joint diameter, expression of inflammatory markers (TNF-α, IL-6). Findings of the study revealed that on CFA administration, there is a significant (p<0.01) increase in joint diameter in all the tested animals. On day 3, we found highest increase in the joint diameter in all treatment groups. Medorrhinum, silica, sulphur showed significant (p<0.01) decrease in joint diameter on day 21. Significant (p<0.05) reduction in paw edema was observed at 5 h post carrageenan administration. IHC of NF-kB in CFA treated group revealed presence of vacuoles, infiltration of inflammatory cells. However, prominent reversal of joint damage was seen in homeopathic drugs (medorrhinum, silica, sulphur) and indomethacin. Study inferred that the homeopathic drugs (medorrhinum, silica, sulphur) and indomethacin were found to be potent in ameliorating inflammation

Amelioration of experimental rheumatoid arthritis by selected ultra-diluted preparations by down regulating increased expression of TNF-α &amp; IL-6

The current work explored the inhibitory effect of selected homeopathic drugs, in experimental models of inflammation and CFA-induced arthritis. Twelve groups of animals were made, each containing 6 animals. The selected homeopathic drugs (causticum, calcarea, medorrhinum, mercurius, formica, proteus, silica, sulphur, thuja), placebo and standard drug, Indomethacin. In CFA model, treatment groups and the reference drug were administered daily for a period of 21 days. Dysfunction in joints was evaluated by parameters such as joint diameter, expression of inflammatory markers (TNF-α, IL-6). Findings of the study revealed that on CFA administration, there is a significant (p&lt;0.01) increase in joint diameter in all the tested animals. On day 3, we found highest increase in the joint diameter in all treatment groups. Medorrhinum, silica, sulphur showed significant (p&lt;0.01) decrease in joint diameter on day 21. Significant (p&lt;0.05) reduction in paw edema was observed at 5 hours post carrageenan administration. IHC of NF-kB in CFA treated group revealed presence of vacuoles, infiltration of inflammatory cells. However, prominent reversal of joint damage was seen in homeopathic drugs (medorrhinum, silica, sulphur) and indomethacin. Study inferred that the homeopathic drugs (medorrhinum, silica, sulphur) and indomethacin were found to be potent in ameliorating inflammation

Do Parallel β-Helix Proteins Have a Unique Fourier Transform Infrared Spectrum?

AbstractSeveral polypeptides have been found to adopt an unusual domain structure known as the parallel β-helix. These domains are characterized by parallel β-strands, three of which form a single parallel β-helix coil, and lead to long, extended β-sheets. We have used ATR-FTIR (attenuated total reflectance-fourier transform infrared spectroscopy) to analyze the secondary structure of representative examples of this class of protein. Because the three-dimensional structures of parallel β-helix proteins are unique, we initiated this study to determine if there was a corresponding unique FTIR signal associated with the parallel β-helix conformation. Analysis of the amide I region, emanating from the carbonyl stretch vibration, reveals a strong absorbance band at 1638cm−1 in each of the parallel β-helix proteins. This band is assigned to the parallel β-sheet structure. However, components at this frequency are also commonly observed for β-sheets in many classes of globular proteins. Thus we conclude that there is no unique infrared signature for parallel β-helix structure. Additional contributions in the 1638cm−1 region, and at lower frequencies, were ascribed to hydrogen bonding between the coils in the loop/turn regions and amide side-chain interactions, respectively. A 13-residue peptide that forms fibrils and has been proposed to form β-helical structure was also examined, and its FTIR spectrum was compared to that of the parallel β-helix proteins

Equilibrium unfolding studies of barstar: evidence for an alternative conformation which resembles a molten globule

The folding of the small protein barstar, which is the intracellular inhibitor to barnase in Bacillus amyloliquefaciens, has been studied by equillibrium unfolding methods. Barstar is shown to exist in two conformations; the A form, which exists at pH values lower than 4, and the N state, which exists at pH values above 5. The transition between the A form and the N state is completely reversible. UV absorbance spectroscopy, fluorescence spectroscopy, and circular dichroism spectroscopy were used to study the two conformation. The mean residue ellipticity measured at 220 nm of the A form is 60% that of the N state, and the A form has some of the properties expected for a molten globule confirmation, Flourescence energy transfer experiments using 1-anilino-8-napthalenesulfonate indicate that at least one of the three tryptophan residues in the A form is accessible to water. Surprisingly, high concentrations of denaturant are required to unfold the A form. For denaturation by guanidine hydrochloride, the midpoint of teh cooperative unfolding transition measured by circular dichroism for the A form at pH 3 is 3.7±0.1M, which is significantly higher than the value of 2.0±0.1M observed for the N state at pH 7. The unfolding of the A form by guanidine hydrochloride or urea is complex and cannot be satisfactorily fit to a two-state (A⇌U) model for unfolding, Fluorescence-monitored tertiary structure melts before circular dichrosim-monitored secondary structure, and an equilibrium unfolding intermediate must be present on the unfolding pathway of A

Is Congo Red an Amyloid-specific Dye?

Congo red (CR) binding, monitored by characteristic yellow-green birefringence under crossed polarization has been used as a diagnostic test for the presence of amyloid in tissue sections for several decades. This assay is also widely used for the characterization of in vitro amyloid fibrils. In order to probe the structural specificity of Congo red binding to amyloid fibrils we have used an induced circular dichroism (CD) assay. Amyloid fibrils from insulin and the variable domain of Ig light chain demonstrate induced CD spectra upon binding to Congo red. Surprisingly, the native conformations of insulin and Ig light chain also induced Congo red circular dichroism, but with different spectral shapes than those from fibrils. In fact, a wide variety of native proteins exhibited induced CR circular dichroism indicating that CR bound to representative proteins from different classes of secondary structure such as α (citrate synthase), α + β (lysozyme), β (concavalin A), and parallel β-helical proteins (pectate lyase). Partially folded intermediates of apomyoglobin induced different Congo red CD bands than the corresponding native conformation, however, no induced CD bands were observed with unfolded protein. Congo red was also found to induce oligomerization of native proteins, as demonstrated by covalent cross-linking and small angle x-ray scattering. Our data suggest that Congo red is sandwiched between two protein molecules causing protein oligomerization. The fact that Congo red binds to native, partially folded conformations and amyloid fibrils of several proteins shows that it must be used with caution as a diagnostic test for the presence of amyloid fibrils in vitro