Lipid-induced recognition of a conformational determinant (residues 65 to 83) in myelin basic protein

The precipitation by antibodies to intact myelin basic protein (BP) and to synthetic peptides containing a sequence based on the region 65 to 83 of bovine BP, S82, S81, S79, and S24, of intact BP in solution or bound to lipid vesicles was compared, using 125I-BP or 14C-DPPC-labeled lipid-BP vesicles. The antipeptide antibodies were shown earlier to recognize conformational determinants which are not expressed in the intact protein in solution. Several anti-BP antibodies precipitated more of the BP free in solution than when bound to lipid vesicles, suggesting that some of the determinants recognized by these antibodies were either sequestered in the bilayer or were altered in conformation. In contrast, one anti-peptide antisera, which had a high titer for the conformational determinant in two of these peptides, S82 and S81, precipitated the protein to a significant degree when it was bound to PG vesicles, even though it did not react with the intact protein in solution. These results indicated that PG was able to confer on the protein the unique peptide conformation recognized by this antibody. PS was less effective, and other lipids were ineffective at conferring this conformation on the protein, supporting earlier results which showed that the conformation of the protein is influenced by the lipid composition of its environment. None of the other anti-peptide antibodies studied bound to the protein either in solution or in lipid vesicles. These results indicate that the lipid environment can sequester or alter the conformation of some antigenic determinants, preventing recognition by some anti-BP antibodies, and can expose or generate other conformational determinants, allowing recognition by an anti-peptide antiserum.     

Prevention of experimental allergic encephalomyelitis by nonencephalitogenic basic peptides

The biological properties of the chymotrypsin-treated encephalitogenic basic protein are described. The basic protein, isolated from bovine spinal cord, was digested with chymotrypsin and filtered through Sephadex gel resulting in three distinct well-separated peaks starting at the void volume of the column. Tubes common to each peak were combined into Fractions I, II, and III, respectively. More than 90% of the original protein was recovered in the three fractions. Fraction II, representing 76% of the original protein, was nonencephalitogenic when tested in guinea pigs at 0.010-, 0.025-, and 0.500-mg doses emulsified in complete Freund's adjuvant. Guinea pigs immunized with Fraction II were protected from EAE when challenged with encephalitogenic emulsions. A critical dose of 1.0 mg completely protected the animals from disease, while partial protection was obtained with lower doses. In addition to producing circulating antibodies, animals sensitized with Fraction II, the encephalitogenic tryptophan peptide or the basic protein displayed a delayed-type hypersensitivity response when skin tested with either of the three antigens. The positive skin reactivity in animals sensitized with Fraction II was not followed by EAE during 5 months of observation. In contrast, animals sensitized with extracts from bovine tissues other than the central nervous system were not protected from disease when challenged with encephalitogenic emulsions.The main finding here reported is the prevention of EAE with nonencephalitogenic peptides derived from the parent EAE-producing protein. The peptides retain the ability to induce delayed-type hypersensitivity and provide antigens to study the role of delayed hypersensitivity in experimental allergic encephalomyelitis.     

How I Met Nico M. van Gelder: A Commentary

Neurochemical Research -     

Adaptive Neuro-Fuzzy Methodology for Noise Assessment of Wind Turbine

Wind turbine noise is one of the major obstacles for the widespread use of wind energy. Noise tone can greatly increase the annoyance factor and the negative impact on human health. Noise annoyance caused by wind turbines has become an emerging problem in recent years, due to the rapid increase in number of wind turbines, triggered by sustainable energy goals set forward at the national and international level. Up to now, not all aspects of the generation, propagation and perception of wind turbine noise are well understood. For a modern large wind turbine, aerodynamic noise from the blades is generally considered to be the dominant noise source, provided that mechanical noise is adequately eliminated. The sources of aerodynamic noise can be divided into tonal noise, inflow turbulence noise, and airfoil self-noise. Many analytical and experimental acoustical studies performed the wind turbines. Since the wind turbine noise level analyzing by numerical methods or computational fluid dynamics (CFD) could be very challenging and time consuming, soft computing techniques are preferred. To estimate noise level of wind turbine, this paper constructed a process which simulates the wind turbine noise levels in regard to wind speed and sound frequency with adaptive neuro-fuzzy inference system (ANFIS). This intelligent estimator is implemented using Matlab/Simulink and the performances are investigated. The simulation results presented in this paper show the effectiveness of the developed method.     

Artonin E Induces Apoptosis via Mitochondrial Dysregulation in SKOV-3 Ovarian Cancer Cells

Artonin E is a prenylated flavonoid isolated from the stem bark of Artocarpus elasticus Reinw.(Moraceae). This study aimed to investigate the apoptotic mechanisms induced by artonin E in a metastatic human ovarian cancer cell line SKOV-3 in vitro. MTT assay, clonogenic assay, acridine orange and propidium iodide double staining, cell cycle and annexin V analyses were performed to explore the mode of artonin E-induced cell death at different time points. DNA laddering, activation of caspases-3, -8, and -9, multi-parametric cytotoxicity-3analysis by high-content screening, measurement of reactive oxygen species generation, and Western blot were employed to study the pathways involved in the apoptosis. MTT results showed that artonin E inhibited the growth of SKOV-3 cells, with IC₅₀ values of 6.5±0.5μg/mL after 72 h treatment, and showed less toxicity toward a normal human ovarian cell lineT1074, with IC₅₀ value of 32.5±0.5μg/mL. Results showed that artonin E induced apoptosis and cell cycle arrest at the S phase. This compound also promoted the activation of caspases-3, -8, and -9. Further investigation into the depletion of mitochondrial membrane potential and release of cytochrome c revealed that artonin E treatment induced apoptosis via regulation of the expression of pro-survival and pro-apoptotic Bcl-2 family members. The expression levels of survivin and HSP70 proteins were also down regulated in SKOV-3 cells treated with artonin E. We propose that artonin E induced an antiproliferative effect that led to S phase cell cycle arrest and apoptosis through dysregulation of mitochondrial pathways, particularly the pro- and anti-apoptosis signaling pathways.     

Estimation of linear and mass attenuation coefficients of soy–lignin bonded Rhizophora spp. particleboard as a potential phantom material using caesium-137 and cobalt-60

In this study, linear and mass attenuation coefficients of fabricated particleboards intended for use as phantom material were estimated using ¹³⁷Cs and ⁶⁰Co radiation sources. Particleboards made of Rhizophora spp. wood trunk bonded with soy flour and lignin were fabricated at a target density of 1.0 g cm^?3, with and without gloss finish coating. Elemental composition of the particleboards was obtained by means of energy dispersive X-ray (EDX) spectroscopy. Experimental setups were simulated via the GATE Monte Carlo (MC) package, with particle histories of 1?×?10⁶–1?×?10⁷. Linear and mass attenuation coefficients obtained from measurements and GATE simulations were compared and discussed. The percentage differences between the measured and simulated linear and mass attenuation coefficients of the samples were reasonably small (2.05–4.88% for ¹³⁷Cs and 3.24–5.38% for ⁶⁰Co). It is shown that all the particleboards have the potential to be used as phantom materials as the attenuation coefficients measured were in good agreement with those of water (calculated with XCOM) and with those simulated with the GATE toolkit. The use of gloss finish coating also did not show any significant effect on the attenuation coefficient of the phantom material. Verification of experimental results via GATE simulations has been shown crucial in providing reliable data for energy transmission studies. Based on the results achieved in this study, it is concluded that the studied material—Rhizophora spp. wood trunk bonded with soy flour and lignin including gloss finish coating—can be used in radiation dosimetry studies.

     

Design and Evaluation of the Highly Concentrated Human IgG Formulation Using Cyclodextrin Polypseudorotaxane Hydrogels

To achieve the potent therapeutic effects of human immunoglobulin G (IgG), highly concentrated formulations are required. However, the stabilization for highly concentrated human IgG is laborious work. In the present study, to investigate the potentials of polypseudorotaxane (PPRX) hydrogels consisting of polyethylene glycol (PEG) and α- or γ-cyclodextrin (α- or γ-CyD) as pharmaceutical materials for highly concentrated human IgG, we designed the PPRX hydrogels including human IgG and evaluated their pharmaceutical properties. The α- and γ-CyDs formed PPRX hydrogels with PEG (M.W. 20,000) even in the presence of highly concentrated human IgG (>100 mg/mL). According to the results of ¹H-NMR, powder X-ray diffraction, and Raman microscopy, the formation of human IgG/CyD PPRX hydrogels was based on physical cross-linking arising from their columnar structures. The release profiles of human IgG from the hydrogels were in accordance with the non-Fickian diffusion model. Importantly, the stabilities of human IgG included into the hydrogels against thermal and shaking stresses were markedly improved. These findings suggest that PEG/CyD PPRX hydrogels are useful to prepare the formulation for highly concentrated human IgG.KEY WORDS: cyclodextrin, human IgG, hydrogel, polypseudorotaxane, stability     

Antisecretory,Gastroprotective, Antioxidant and Anti-Helicobcter Pylori Activity of Zerumbone from Zingiber Zerumbet (L.) Smith

Background

Zingiber zerumbet Smith is a perennial herb, broadly distributed in many tropical areas. In Malaysia, it’s locally known among the Malay people as “lempoyang” and its rhizomes, particularly, is widely used in traditional medicine for the treatment of peptic ulcer disease beyond other gastric disorders.

Aim of the study

The aim of the current study is to evaluate the gastroprotective effect of zerumbone, the main bioactive compound of Zingiber zerumbet rhizome, against ethanol-induced gastric ulcer model in rats.

Materials and Methods

Rats were pre-treated with zerumbone and subsequently exposed to acute gastric ulcer induced by absolute ethanol administration. Following treatment, gastric juice acidity, ulcer index, mucus content, histological analysis (HE and PAS), immunohistochemical localization for HSP-70, prostaglandin E2 synthesis (PGE₂), non-protein sulfhydryl gastric content (NP-SH), reduced glutathione level (GSH), and malondialdehyde level (MDA) were evaluated in ethanol-induced ulcer in vivo. Ferric reducing antioxidant power assay (FRAP) and anti-H. pylori activity were investigated in vitro.

Results

The results showed that the intragastric administration of zerumbone protected the gastric mucosa from the aggressive effect of ethanol-induced gastric ulcer, coincided with reduced submucosal edema and leukocyte infiltration. This observed gastroprotective effect of zerumbone was accompanied with a significant (p <0.05) effect of the compound to restore the lowered NP-SH and GSH levels, and to reduce the elevated MDA level into the gastric homogenate. Moreover, the compound induced HSP-70 up-regulation into the gastric tissue. Furthermore, zerumbone significantly (p <0.05) enhanced mucus production, showed intense PAS stain and maintained PG content near to the normal level. The compound exhibited antisecretory activity and an interesting minimum inhibitory concentration (MIC) against H. pylori strain.

Conclusion

The results of the present study revealed that zerumbone promotes ulcer protection, which might be attributed to the maintenance of mucus integrity, antioxidant activity, and HSP-70 induction. Zerumbone also exhibited antibacterial action against H. pylori.