Moringa oleifera Protects SH-SY5YCells from DEHP-Induced Endoplasmic Reticulum Stress and Apoptosis

Moringa oleifera (MO) is a medicinal plant that has been shown to possess antioxidant, anticarcinogenic and antibiotic activities. In a rat model, MO extract (MOe) has been shown to have a protective effect against brain damage and memory decline. As an extending study, here, we have examined the protective effect of MOe against oxidative stress and apoptosis caused in human neuroblastome (SH-SY5Y) cells by di-(2-ethylhexyl) phthalate (DEHP), a plasticizer known to induce neurotoxicity. Our data show that MOe prevents oxidative damage by lowering reactive oxygen species (ROS) formation, restoring mitochondrial respiratory chain complex activities, and, in addition, by modulating the expression of vitagenes, i.e., antioxidant proteins Nrf2 and HO-1. Moreover, MOe prevented neuronal damage by partly inhibiting endoplasmic reticulum (ER) stress response, as indicated by decreased expression of CCAAT-enhancer-binding protein homologous protein (CHOP) and Glucose-regulated protein 78 (GRP78) proteins. MOe also protected SH-SY5Y cells from DEHP-induced apoptosis, preserving mitochondrial membrane permeability and caspase-3 activation. Our findings provide insight into understanding of molecular mechanisms involved in neuroprotective effects by MOe against DEHP damag

Activating mutations of STAT5B and STAT3 in lymphomas derived from gamma delta-T or NK cells

Peer reviewe

Spent grain as a sustainable and low-cost carrier for laccase immobilization

Spent grain is promising lignocellulosic by-product support for laccase immobilization. The waste digestion with two different approaches (HCl/NaOH and H2SO4/NaOH) was performed. Different procedures (soaking and dropping), based on chemical and physical reactions, were also used to obtain the highest immobilized activity. Results showed that H2SO4/NaOH digestion guaranteed an immobilized activity five times higher than HCl/NaOH digestion. The best immobilization conditions with physical dropping procedure resulted in the highest immobilized activity on digested spent grain (2500 U/Kg). Good reusability (42% of activity retained after four cycles), and lower catalytic efficiency (Vmax/Km) of 0.053 min−1 than free laccase (0.14 min−1) with ABTS as substrate, were also obtained. Besides, when 20 mg of biocatalyst (0.02 U) were tested for syringic acid removal, complete oxidation of the phenol was achieved in just 4 h

Eggshell membrane as feedstock in enzyme immobilization

Eggshell membrane, an eco-compatible, safe and cheap by-product was employed as carrier for the laccase from Trametes versicolor immobilization. In order to evaluate the best protocol to apply for the syringic acid degradation, two different types of laccase loading on eggshell membrane were used by incubation in solution or by enzyme-dropping. Chemicals (covalent) and physicals (adsorption) immobilizations were tested for both procedure using native or periodate-oxidized laccase. It is shown that immobilization of periodate-oxidized laccase on NiCl2-pretreated eggshell membrane was the best method for the first procedure (immobilized activity 1300 U/Kg, a residual activity of 30 % for 6 reuse). For the enzyme-dropping protocol a covalent method with the bifunctional cross linker (glutaraldehyde) was the best method (immobilized activity 3500 U/Kg, a residual activity of 45 % for 6 reuse

Fabrication of a novel biocatalyst with Laccase immobilized on eggshell membrane

The research provides the suitability of a not-lignocellulosic agro-industrial wastes, such as egg shell membrane, for laccase from Trametes versicolor immobilization. This result is of significant interest due to the low cost and great availability of such waste, which actually is underused and causes significant environmental problem for improper storage. In addition, the development of economic biocatalysts more sustainable, besides reduce environmental impacts, improve the application of enzymatic technology in industry. Laccase is a eco-friendly oxidoreductase enzyme and has been used in different industrial fields for different purposes, from food additive and beverage processing to biomedical diagnosis and production of biofuels. It catalyzes the oxidation of a great number of substrates which can be widened to higher‐redox potential compounds than laccase one with the help of laccase-mediator-system . A very simple method was based on the cross-linking reaction between the amino functional groups of the protein and of the ESM solid surface. So, glutaraldehyde, as cross linker, was dropped onto the ESM surface in order to form the Shiff bases .The performance of biocatalyst was evaluated taking into account the operational stability expressed as reuse and the enzyme recovery. The goal of this research is to apply the biocatalyst in the removal of contaminants from waste water

Design of a heterogeneous enzymatic catalyst on chitosan: investigation of the role of conjugation chemistry in the catalytic activity of a laccase from trametes versicolor

BACKGROUND: Three protocols are presented in order to immobilise a laccase (EC 1.10.3.2) from Trametes versicolor on chitosan. In particular, chitosan is functionalized with glutaraldehyde and epichlorohydrin to explore to what extent the conjugation of the enzyme is affected by a carbonyl- or epoxy-modified surface, respectively. In addition, an oxidation procedure is tested to modify, for the first time, the carbohydrate moiety of the enzyme and exploit it for linking with the amine group of chitosan. RESULTS: The system in which laccase is directly conjugated to chitosan seems to be the best-performing since it is able to maintain 100% of initial activity over a 30 day period and at least 50% of the starting activity after 3 catalytic cycles. CONCLUSION: The methods show the capability to develop three biocatalysts suitable for the immobilization of laccase. All of them provide covalent bonds that don't inactivate the enzyme through any conformational change and any distortion of the active pocket. Specifically, the oxidation procedure proves to be feasible for the preparation of stabilized glycoproteins without the introduction of foreign molecules, suggesting that the carbohydrate moiety of laccase does not appear to be closely related to the catalytic site of the protein. © 2017 Society of Chemical Industry

Agro-industrial wastes as potential carriers for enzyme immobilization: a review

This review provides a general overview of the suitability of different agro-industrial wastes for enzymeimmobilization. For the purposes of this literary study, the support materials are divided into two maingroups, called lignocellulosic (coconutfiber, corn cob, spent grain, spent coffee, husk, husk ash, and strawrice, soybean and wheat bran) and not lignocellulosic by-products (eggshell and eggshell membranes).The study pointed out that all of these wastes are materials of great potentiality for enzyme immobili-zation even if coconutfiber is preferred. This result is of significant interest due to the low cost and greatavailability of such wastes, which actually are underused and cause significant environmental problemsfor improper storage. In addition, the development of economic biocatalysts more sustainable, besidesreduce environmental impacts, improve the application of enzymatic technology in industry. Therefore,the enzyme immobilization reaction and the application of biocatalysts are reviewed and discussed

A new Laccase-Mediator system facing the biodegradation challenge: insight into the NSAIDs removal

Non-steroidal anti-inflammatory drugs (NSAIDs) are widely found pollutants in the aquatic environment and the currently available treatments for their removal are usually associated with some drawbacks. The aim of this research was to apply a laccase-mediator system for the degradation of some commonly used NSAIDs, namely diclofenac (DCF), naproxen (NAP) and ketoprofen (KP). The biocatalyst was obtained by direct immobilization on chitosan beads of a periodate-oxided laccase from Trametes versicolor. A preliminary study aimed to optimize DCF degradation in the presence of 2,2-azinobis (3-ethylbenzothiazoline-6-sulfonicacid) diammonium salt (ABTS) as mediator. It turned out that pH 3 and a 1:1 M ratio for ABTS:drug were the best experimental conditions under which DCF was degraded at 90% after 3 h. In addition, an efficient reuse of the biocatalyst for up to 5 cycles emerged. DCF was further mixed with naproxen and ketoprofen to test whether laccase was still able to eliminate DCF and eventually act on the other compounds. At just 0.02 U of laccase activity, diclofenac was completely degraded within 3 h, while an almost complete removal for naproxen (∼90%) and a partial removal for ketoprofen (30%) occurred in 7 d when drugs were added at high concentrations (78.5 μM, 98 μM and 108 μM, respectively). After 7 d of degradation, transformation products of diclofenac, identified as hydroxylated compounds, disappeared. Naproxen products were, instead, reduced to very small amounts

Interferon-gamma, tumor necrosis factor-alpha, and lipopolysaccharide promote chitotriosidase gene expression in human macrophages

Human chitotriosidase (Chit), a chitinolytic enzyme, is a member of the chitinase family. In human plasma, Chit activity has been proposed as a biochemical marker of macrophage activation in several lysosomal diseases. Recently we found that Chit activity is higher in patients affected by Plasmodium falciparum malaria infection, suggesting that Chit may reflect induction of an immunological response. To assess this hypothesis, we evaluated the CHIT1 mRNA levels in human monocytes/macrophages (HMMs) following treatment with interferon-gamma (IFNϒ), tumor necrosis factor-alpha (TNFα), and lipopolysaccharide (LPS). Stimulation of macrophages with INF-ϒ, TNF-α, and LPS resulted in an increase in Chit activity as well as the levels of CHIT1 mRNA as measured by quantitative real-time PCR. The data presented in this article show that Chit plays a role in the response to the activation of INF-ϒ-, TNF-α-, and LPS-driven macrophages, suggesting that the production of Chit by macrophages could have biological relevance in the immune-response