Novel fabrication of Cu(II)-incorporated chiral d-penicillamine-chitosan nanocomposites enantio-selectively inhibit the induced amyloid ß aggregation for Alzheimer's disease therapy.

It is well known that the chiral materials combined with metal ion's structure have been identified as promising candidate for the nursing Alzheimer Disease (AD) treatment, particularly to inhibit amyloid (Aß) due to their significant pharmacological effect on the living bodies. In the present study, Cu(II)/Chitosan nanocomposite caped with chiral penicillamine (Cu@D-PEN/Chitosan) have been synthesized and used as an effective amyloid-ß (Aß) inhibitor. The composite formations of the samples were confirmed from the FTIR and XRD, studies. FE-SEM, TEM and AFM studies have been carried out to depict the morphological analysis of the nanocomposites. The prepared samples have also been subjected to various in vitro studies such as encapsulation efficiency, drug loading capacity, drug release and biodegrading or compatibility of the nanocomposites to support the Aß aggregation inhibiting ability investigations. It was observed that the increase in the concentration of the Cu@D-PEN/Chitosan enhancing the Aß inhibiting ability. Thus, the Cu(II)@D-PEN/Chitosan showed improving memory effect suggesting that Cu(II)@D-PEN/Chitosan nanocomposites may be a potential candidate for inhibiting the Aß aggregation in nursing AD treatment.

Expression and significance of cyclin D1, cyclin-dependent kinase 4 and cyclin-dependent kinase inhibitor P27 in patients with non-neoplastic epithelial disorders of the vulva.

Non-neoplastic epithelial disorders of the vulva (NNEDV) are prevalent and refractory gynecological diseases. However, the underlying pathogenesis of these diseases remain unclear. The present study aimed to investigate the expression and significance of cyclin D1, cyclin-dependent kinase 4 (CDK4) and cyclin-dependent kinase inhibitor P27 (P27) in patients with NNEDV and provide a reference for clinical diagnosis and treatment. Normal vulvar skin samples from patients with perineum repair (control group, n=20) and skin samples from the vulvar lesions of patients with NNEDV (NNEDV group, n=36) were collected. Expression levels of cyclin D1, CDK4 and P27 were assessed in the samples using immunohistochemistry. The expression of each protein was evaluated based on the mean optical density (MOD). The MODs of cyclin D1 and CDK4 were significantly higher in samples of the three pathological types of NNEDV, namely squamous hyperplasia (SH), lichen sclerosus (LS) and mixed SH and LS lesions, compared with those of the control group. The MOD of P27 was lower in samples of the three pathological types of NNEDV than in the control group, although the difference was not statistically significant. No significant differences in the MOD of cyclin D1, CDK4 and P27 were detected among the three pathological types of NNEDV. The ratios of the MOD of cyclin D1 and CDK4 in the prickle cell layer to those in the basal cell layer were significantly higher in the NNEDV group than in the control group. However, the ratio of the MOD of P27 in the prickle cell layer to that in the basal cell layer exhibited no significant difference between the NNEDV and control groups. NNEDV has the potential for malignant transformation. The occurrence and development of NNEDV may be associated with the acceleration of cell proliferation, in which cyclin D1, CDK4 and P27 contribute to regulation of the cell cycle. Therefore, cyclin D1, CDK4 and P27 may be potential targets in the development of new clinical therapeutic drugs for patients with NNEDV.

Enhancement of antibacterial and growth-promoting effects of Paenibacillus polymyxa by optimizing its fermentation process.

AIMS: We aimed to enhance the antibacterial and growth-promoting effects of Paenibacillus polymyxa by improving the yield of spores, lipopeptides and indole-3-acetic acid (IAA) in the fermentation process. METHODS AND RESULTS: Through medium optimization by the response surface method and feeding fermentation, the number of spores reached 2.37 × 109  cfu ml-1 with an increase of 38%, the content of lipopeptides reached 60.8 mg L-1 with an increase of 89%, and the content of IAA reached 24.3 mg L-1 with an increase of 176%, respectively, comparing with the original (un-optimized) culture conditions. The fermentation culture of P. polymyxa from the optimized medium and feeding fermentation resulted in higher colonization of P. polymyxa in soils than that from the original culture during the 49 days for testing. Comparing with the supernatant of the original culture, the supernatant of the P. polymyxa culture from the optimized medium and feeding fermentation showed enhanced antibacterial effects and plant growth-promoting effects. The enhanced antibacterial effect was shown as the increase of the inhibition zone by 59%, 45% and 26% against Ralstonia solanacearum, Erwinia carotovora and Xanthomonas campestris. The enhanced growth-promoting effects on tomato and strawberry plants were the increase of plant height by 47% and 5%, root length by 23% and 15% and root weight by 65% and 110%. CONCLUSIONS: The combination of medium optimization and feeding fermentation effectively improved the yield of spores, lipopeptides and IAA. Lipopeptides and IAA lead to enhanced antibacterial and plant growth-promoting effects of the P. polymyxa product. SIGNIFICANCE AND IMPACT OF THIS STUDY: The optimized fermentation method significantly improved the yield of spores, lipopeptides and IAA, thus providing theoretical and technical support for enhancing the antibacterial and growth-promoting effects of P. polymyxa products in agriculture.

Consortium of Plant Growth-Promoting Rhizobacteria Strains Suppresses Sweet Pepper Disease by Altering the Rhizosphere Microbiota.

Beneficial microorganisms have been extensively used to make plants more resistant to abiotic and biotic stress. We previously identified a consortium of three plant growth-promoting rhizobacteria (PGPR) strains (Bacillus cereus AR156, Bacillus subtilis SM21, and Serratia sp. XY21; hereafter "BBS") as a promising and environmentally friendly biocontrol agent. In this study, the effect of BBS on a soil-borne disease of sweet pepper was evaluated. Application of BBS significantly reduced the prevalence of phytophthora blight and improved fruit quality and soil properties relative to the control. BBS was able to alter the soil bacterial community: it significantly increased the abundances of Burkholderia, Comamonas, and Ramlibacter, which were negatively associated with disease severity, relative to the control. A redundancy analysis suggested that BBS-treated soil samples were dominated by Burkholderia, Comamonas, Ramlibacter, Sporichthya, Achromobacter, and Pontibacter; abundance of these genera was related to total organic carbon (TOC), total nitrogen (TN), ammonium nitrogen (AN), total potassium (TP), and available phosphorus (AP) contents. This suggests that BBS treatment shifted the microbe community to one that suppressed soil-borne disease and improved the soil chemical properties.

Greater enhancement of Bacillus subtilis spore yields in submerged cultures by optimization of medium composition through statistical experimental designs.

Bacillus subtilis spore preparations are promising probiotics and biocontrol agents, which can be used in plants, animals, and humans. The aim of this work was to optimize the nutritional conditions using a statistical approach for the production of B. subtilis (WHK-Z12) spores. Our preliminary experiments show that corn starch, corn flour, and wheat bran were the best carbon sources. Using Plackett-Burman design, corn steep liquor, soybean flour, and yeast extract were found to be the best nitrogen source ingredients for enhancing spore production and were studied for further optimization using central composite design. The key medium components in our optimization medium were 16.18 g/l of corn steep liquor, 17.53 g/l of soybean flour, and 8.14 g/l of yeast extract. The improved medium produced spores as high as 1.52 +/- 0.06 x 10(10) spores/ml under flask cultivation conditions, and 1.56 +/- 0.07 x 10(10) spores/ml could be achieved in a 30-l fermenter after 40 h of cultivation. To the best of our knowledge, these results compared favorably to the documented spore yields produced by B. subtilis strains.

[Effect of Cry1Ca7 protein modified by site-directed mutagenesis on inhibiting Spodoptera exigua Hübner].

OBJECTIVE: To obtain the mutants with different toxicity from the wild-type CrylCa7. METHODS: Insecticidal crystal protein CrylCa7 from Bacillus thuringiensis which is highly toxic to Spodoptera exigua, an important agricultural pest in China, and we mutated this toxin by over-lapping extensive PCR method in different domains to obtain 11 chimeric mutants. RESULTS: The results of bioassays against Spodoptera exigua neonates showed that several conserved amino acid sites were crucial to insects. The pesticidal activities of most of mutated proteins were decreased, including Glycinel38 Serine, Threonine221 Aspartic acid, Threonine221 Argine, Asparagine251 Serine, 439GlycineGlycineThreonine440, Asparagine306 Argine, Tryp-tophan376 Phenylalanine, Argine522 Glutamic acid and Argine570 Glycine. The activity of those mutated proteins in the Domain II was 439GlycineGlycineThreonine440 < Asparagine306 Argine < Tryptophan376 Phenylalanine. In the Domain III, the mutant Argine522 Glutamic acid < Argine 570 Glycine, their toxicities reduced distinctly compared with CrylCa7. The toxicities of the mutant Argine148 Glycine in Domain I increased six-fold, nevertheless the activities of the mutants Glycine138 Serine, Threonine221 Argine and Asparagine251 Serine mutant reduced totally, even the mutant of Threonine221 Aspartic acid was not toxic entirely. CONCLUSION: It is relatively easier to obtain mutant with higher toxicity in Domain I of CrylCa7 protein than these in both DomainII and III. We can use the improved mutant genes as the potential resources to construct novel engineering bacteria and transgenic plant, meanwhile, to perform the study of interaction mechanism between insects and Cry proteins.