Chitosan, bio-polyaminosacharide, is derived from chitin. Two sources (shrimp wastes and fungus biomass) were used to produce chitosan. And then the chitosan was produced in the nano-form followed by characterization by transmission electron microscopy. The images obtained clearly showed that the size of nano-chitosan ranged between 7 and 13 and 3-6 nm with spherical shape for shrimp and fungal sources, respectively. The antimicrobial activities of the tested concentrations of chitosan and nano-chitosan were examined and found to have high activity against the tested pathogens. The evaluation of the toxicity of the tested concentrations of the produced chitosan and its nano-size were performed using brine shrimp and rat bioassay. Toxicity examination of chitosan and their nano derivatives is an essential procedure to assess the possibility of using these concentrations as food ingredient. Nine groups of rats were treated with either chitosan or nano-chitosan of both sources at 100 and 200 mg kg-1 bw. Adding chitosan in the diet of all groups showed no significant changes in both the blood biochemical and oxidative stress parameters when compared with control group. The histopathology of liver, kidney and stomach confirmed the results of the previous parameters. No signs of inflammation, fibrosis or cirrhosis were found in examined organs. It is concluded that chitosan and nano-chitosan of shrimp and Rhizopus stolonifer had high antimicrobial activity and are not toxic in the same time and it can be used as food ingredients.

Fruit juices are amongst the most non-alcoholic beverages appreciated and consumed in European countries, including Portugal. These beverages contain minerals, nutrients, trace elements, vitamins and phytochemicals, which are essential for a healthy life. However, fruit juices may also contain high levels of metals, posing a health risk to humans, especially to children, since they consume more fruit juice per body weight unit, and have a less varied diet than adults. Thus, in order to guarantee food safety and to make sound nutritional considerations, fruit juices require careful investigation. The main purpose of this study was to determine arsenic (As), cadmium (Cd), chromium (Cr), lead (Pb), manganese (Mn) and nickel (Ni) concentrations in 21 fruit juices from 4 different brands, previously selected by the ASAE (Portuguese Food and Economic Safety Authority), and available in the Portuguese market. Results obtained were compared with permissible levels set out by WHO (World Health Organization), USEPA (United States Environmental Protection Agency), by the Portuguese law, and with similar studies performed in other countries. A validation process, including linearity, range, analytical thresholds, precision, accuracy and specificity/selectivity was conducted in order to guarantee reliable analytical data. The results showed that As levels in four samples, Ni in thirteen samples and Mn in all the twenty-one samples, were above the maximal permissible values specified by Decree-Law 306/2007 from 27th August of the Portuguese Legislation. These data establish the need for reduction of metal concentrations in consumed juices.

Chromium is considered as one of the most common ubiquitous pollutants in the aquatic environment, but the pure metallic form is absent naturally. There are three oxidation states in case of Chromium viz., Cr (II), Cr (III), Cr (VI). Among which Cr (II) is most unstable. Cr (III) and Cr (VI) are the stable oxidation state of Chromium in the environment. Being one of the commonly used metals Chromium and its particulates enter the aquatic medium through effluents discharged from different industries like textiles, tanneries, electroplating workshops, ore mining, dyeing, printing-photographic and medical industries. Among these, hexavalent chromium is considered as the most toxic form because it readily passes cellular membranes and then reduced to trivalent form. This trivalent chromium combines with several macromolecules including genetic material inside the cytosol, and is ultimately exposes the toxic and mutagenic alterations due of chromium toxicity. Chromium is taken up either through gastrointestinal tract or respiratory tract. The amount varies depending upon the medium and the form of chromium. In this review, an attempt has been made to accumulate the mammoth available data regarding impact of chromium on fresh water fishes into a systematic representation. The main objective of the review is to provide a future guideline for the scientific community and public officials involved in health risk assessment and management ensuring a better environmental condition for human health.

Application of ZnO nanoparticles in sunscreens exposes human skin with their adverse effects, which correlates to dissolution/translocation of free Zn+2 ions. The possibility of decreasing solubility and therefore, reducing toxicity, by structural modifications have been discussed as a solution. The present investigation has developed new metallic lattices of ZnO to reduce cytotoxicity of ZnO nanoparticles. Novel metal-promoted Zn-based nanocomposites ([Zn(O)/M], M = Mg, Al, Ca, Ti) were synthesized and their physicochemical properties and their cytotoxicity were evaluated. Solubility and release studies showed that modification of ZnO structure decreases release of Zn+2 into culture medium. XRD and UV absorbance analyses showed that metallic-dopants percolate into crystalline lattice of ZnO. This phenomenon is basic reason for stability of Zn-based network. Cell culture studies and MTT assay on human skin cells (HFF-1) exposed to UVA radiation showed that the level of protection of [Zn(O)/M] compounds were more than of [ZnO]. Dichlorofluoroscein diacetate-ROS assay and Zn+2 release experiments indicated that [Zn(O)/M] nanocomposites decreased the level of ROS generation and Zn+2 release in compared to ZnO, indicating higher safety of nanocomposites. This study shows that the synthesized Zn-based nanocomposites have potential to be used as safer and more effective sunscreens than ZnO.

The objective was to evaluate the effect of black bean protein fraction (PFRA), and its derived peptides on glucose uptake, SGLT1 and GLUT2 expression and translocation on Caco-2 cells. The effect of treatments was evaluated on glucose uptake, protein expression and localization and gene expression on Caco-2 cells. PFRA (10 mg/mL) lowered glucose uptake from 27.4% after 30 min to 33.9% after 180 min of treatment compared to untreated control (p < 0.05). All treatments lowered GLUT2 expression after 30 min of treatment compared to untreated control (31.4 to 48.6%, p < 0.05). Similarly, after 24 h of treatment, GLUT2 was decreased in all treatments (23.5% to 48.9%) (p < 0.05). SGLT1 protein expression decreased 18.3% for LSVSVL (100 μM) to 45.1% for PFRA (10 mg/mL) after 24 h. Immunofluorescence microscopy showed a decrease in expression and membrane translocation of GLUT2 and SGLT1 for all treatments compared to untreated control (p < 0.05). Relative gene expression of SLC2A2 (GLUT2) and SLC5A1 (SGLT1) was downregulated significantly up to two-fold change compared to the untreated control after 24 h treatment. Black bean protein fractions are an inexpensive, functional ingredient with significant biological potential to reduce glucose uptake and could be used as an adjuvant in the treatment of colorectal cancer.

Thromboxane is a key clinical risk endpoint of smoking-induced inflammation which has been associated in the pathogenesis of cardiovascular disease. The goal of this review is to quantify the effect of smoking and smoking cessation on one of its urinary metabolites, 11-dehydrothromboxaneB2.

The objective of this study was to evaluate the anti-obesity activity and the action mechanism of Cissus quadrangularis extracts (CQR-300) in 3T3-L1 adipocytes. Cissus quadrangularis was extracted with hot water, resulting in CQR-300. The anti-obesity activity of CQR-300 in 3T3-L1 adipocytes was examined by Oil-red O staining. Possible mechanisms of CQR-300 in 3T3-L1 adipocytes were determined by real-time PCR and western blot. Treatment with CQR-300 inhibited lipid accumulation without showing cytotoxicity to 3T3-L1 adipocytes. Furthermore, CQR-300 decreased adipogenesis/lipogenesis-related mRNA expression levels of fatty acid binding protein (aP2), fatty acid synthase (FAS), lipoprotein lipase (LPL), stearoyl-CoA desaturase-1 (SCD-1), and acetyl-CoA carboxylase (ACC). CQR-300 also down-regulated expression levels of adipogenesis/lipogenesis-associated proteins, including peroxisome proliferator-activated receptor γ (PPARγ), CCAAT/enhancer-binding protein α (C/EBPα), sterol regulatory element binding protein-1c (SREBP-1c), and FAS. It's also up-regulated the expression level of phosphorylated-AMPK (p-AMPK). Collectively, these results suggested that CQR-300 might have an anti-obesity effect by its ability to decrease expression levels of adipogenesis/lipogenesis-related genes and proteins.

Chemotherapeutic agents for cancer are highly toxic to healthy tissues at therapeutic doses and hence alternative medicine avenues are widely researched. Most of the studies on alternative medicine have suggested that Euphorbia plant possesses considerable antitumor and antibacterial properties. The present study was designed to evaluate the in vivo genotoxic effects of Euphorbia triaculeata extract on mice bone marrow cells using chromosomal aberration test and micronucleus assay. This study also deals with the effect of E. triaculeata on the standard drug cyclophosphamide (CP) treatment in mice. Three different doses 250, 500 and 1000 mg/kg body weight were selected. In micronucleus assay, single oral dose administration of Euphorbia triaculeata extract at the three doses did not increase the number of micronucleated polychromatic erythrocytes. Similarly, a single oral administration of Euphorbia triaculeata extract showed no significant changes on mitotic indices or in induction of chromosomal aberrations in mice bone marrow cells. Pretreatment with E. triaculeata extract significantly reduced the clastogenicity of CP. Hence it can be concluded that, E. triaculeata extract showed no significant genotoxic effect on mice bone marrow cells. Under the conditions of this study, it has been demonstrated that the Euphorbia triaculeata extract is not genotoxic and not clastogenic at the concentrations used.

Although silicone implants are widely used in breast and other reconstructive surgeries, the limited biocompatibility of these materials leads to severe complications, including capsular contracture. Here, we aimed to clarify the relationship between periostin and the process of capsule formation after in vivo implantation. Seven-week-old wild-type (WT) C57BL/6 mice and periostin-deficient mice were used. Round silicone implants were inserted into a subcutaneous pocket on the dorsum of the mice. After 8 weeks, the fibrous capsule around the implant was harvested and histologically examined to estimate capsular thickness and the number of inflammatory cells. Additionally, immunohistochemical analysis (periostin, α-SMA, and collagen type I) and western blotting (CTGF, TGF-β, VEGF, and MPO) were performed for a more detailed analysis of capsule formation. The capsules in periostin-knockout mice (PN-KO) were significantly thinner than those in WT mice. PN-KO mice showed significantly lower numbers of inflammatory cells than WT mice. Fibrous tissue formation markers (α-SMA, periostin, collagen type I, and CTGF) were significantly reduced in PN-KO mice. We also confirmed that inflammatory reaction and angiogenesis indicators (TGF-β, MPO, and VEGF) had lower expression in PN-KO mice. Inhibition of periostin could be important for suppressing capsule formation on silicone implants after in vivo implantation.

IL-18 is a proinflammatory cytokine with multiple immunoregulatory properties. We studied the effect of IL-18 gene therapy on the development of murine collagen-induced arthritis (CIA).

Upregulated meteorin-like (Metrnl) protein in peripheral tissues because of exercise-induced increases in intramuscular Metrnl may effectively alleviate obesity by improving metabolism in whole-body tissues. The objective was to analyse the effects of regular treadmill exercise on Metrnl levels in muscle and peripheral tissues of chronic high-fat diet- (HFD-) induced obese mice. Forty-eight-week-old male C57BL/6 mice were first divided equally into normal-diet (CO) and high-fat diet (HF) groups. Following 16 weeks of a HFD, each group was again split equally into control (CO, HF) and training groups (COT, HFT). The HFT group expressed significantly higher phospho-AMP-activated protein kinase (AMPK), AMPK activity, and peroxisome proliferator-activated receptor gamma coactivator-1α (PGC-1α) in muscle tissue than the HF group (p < 0.05). Similar to muscle energy sensing network protein levels, the HFT group also expressed significantly higher muscle, plasma, and adipose tissue Metrnl (p < 0.05). Moreover, regular exercise increased acyl-CoA oxidase 1 (ACOX-1) and monoglyceride lipase (MGL) expression in adipose tissue (p < 0.05) and significantly decreased abdominal fat mass (p < 0.05). This study suggests that exercise-induced muscle Metrnl effectively reduces fat accumulation through the increase of Metrnl in adipose tissue, which may be a therapeutic target for chronic obesity.

The present study tested the effect of a bacterial lactone N-(3-oxododecanoyl)-homoserine lactone (C12-HSL) on the cytoskeletal and transcriptional genes and proteins in prostate adenocarcinoma (PA) cells (DU145 and LNCaP) and prostate small cell neuroendocrine carcinoma (SCNC) PC3 cells including their cellular viability and apoptosis. Our data indicate that cell migration and colony formation were affected in the presence of C12-HSL. C12-HSL induced apoptosis and altered viability of both PA and SCNC cells in a concentration dependent manner as measured by fluorescence and chemiluminescence assays. Compared to PCa cells, noncancerous prostate epithelial cells (RWPE1) were resistant to modification by C12-HSL. Further, the viability of PC3 cells in 3D matrix was suppressed by C12-HSL treatment as detected using calcein AM fluorescence in situ. C12-HSL treatment induced cytoskeletal associated protein expression of vinculin and RhoC, which may have implications in cancer cell motility, adhesion, and metastasis. IQGAP protein expression was reduced in DU145 and RWPE1 cells in the presence of C12-HSL. C12-HSL decreased STAT3 phosphorylation in DU145 cells but increased STAT1 protein phosphorylation in PC3 and LNCaP cells. Overall, these studies indicate that C12-HSL can trigger changes in transcription factors and cytoskeletal proteins and thereby modulate growth and migration properties of PCa cells.

Sepsis, an infection-induced systemic disease, leads to pathological, physiological, and biochemical abnormalities in the body. Organ dysfunction is caused by a dysregulated host response to infection during sepsis which is a major contributing factor to acute kidney injury (AKI) and the mortality rate for sepsis doubles due to coincidence of AKI. Sepsis-induced AKI is strongly associated with increased mortality and other adverse outcomes. More timely diagnosis would allow for earlier intervention and could improve patient outcomes. Sepsis-induced AKI is characterized by a distinct pathophysiology compared with other diseases and may also have unique patterns of plasma and urinary biomarkers. This concise review summarizes properties and perspectives of the biomarkers for their individual clinical utilization.

Dunaliella salina is the popular microalga for β-carotene production. There is still a growing demand for the best strain identification and growth conditions optimization for maximum carotenoids production. Some strains are noncarotenogenic while other strains may respond differently to applied growth conditions and produce enhanced carotenoid levels. This study tested the carotenogenic ability of Dunaliella salina CCAP 19/20 under sixteen stress conditions and certain biochemical changes in response to specific stress were investigated. This study identified the above strain as carotenogenic, which produces maximum carotenoids under high light (240 μmol photons m-2 sec-1) when combined nitrogen and micronutrients (Cu or CuMn) were limited. Based on the intensity of extracted ions chromatograms, lutein (m/z 568.4357) appears as the major carotenoid followed by β-carotene (m/z 536.4446) and α-carotene (m/z 536.4435). A polypeptide of 28.3 kDa appeared while another polypeptide of 25.5 kDa disappeared in stress cells as compared to noncarotenogenic cells. Expression levels of antioxidative-enzyme superoxide dismutase-1 (SOD1, H2O2-resistant) remained identical, while the prominent H2O2-sensitive isoforms SOD2 and SOD3 were downregulated during carotenogenic conditions. Overall, increased carotenoids levels might be due to the response of differential expression of specific polypeptides and retention of H2O2-resistant SOD, which eventually might help the organism to thrive in the tested stress conditions.

It is well established that, during development of malignancies, metabolic changes occur, including alterations of enzyme activities and isoenzyme expression. Arginase and nitric oxide (NO) synthase (NOS) are two of those enzymes considered to be involved in tumorigenesis. The goal of this article was to study the involvement of arginase and NOS in the development of different stages of breast cancer. Our results have shown that human serum arginase activity and NO (resp., and NOS activity) and polyamines quantities increased in parallel with cancer stage progression and decreased after neoadjuvant chemotherapy. For breast cancer, the only isoenzyme of arginase expressed in serum before and after chemotherapy was in a cationic form. The data of Lineweaver-Burk plot with a Km value of 2 mM was calculated, which is characteristic for human liver type isoform of arginase. During electrophoresis at pH 8.9, the enzyme exhibited high electrophoretic mobility and was detected near the anode. The presented results demonstrated that arginase in human serum with breast cancer and after chemotherapy is not polymorphic. We suggest that arginase and NOS inhibition has antitumor effects on cancer development, as it can inhibit polyamines and NO levels, a precursor of cancer cell proliferation, metastasis, and tumor angiogenesis.

The osteopontin has been involved in therapeutic resistance in a variety of cancers. But, the significance of osteopontin expression on the prognosis of patients with locally advanced esophageal squamous cell carcinoma (ESCC) receiving chemoradiotherapy is unclear.

Nasopharyngeal carcinoma (NPC) is a head and neck tumor with high degree of malignancy and with high incidence especially in southern China. AIMP2-DX2, one isoform of the aminoacyl-tRNA synthetase interacting multifunctional proteins (AIMPs), is shown to be a potential target in many cancers. However, the detailed mechanisms of AIMP2-DX2 in NPC development remain to be elucidated. Here, we found that the mRNA expression level of AIMP2-DX2 was significantly increased in NPC specimens, compared with normal nasopharyngeal tissues. Microarray immunohistochemical analysis of NPC specimens and Kaplan-Meier analysis showed that patients with high AIMP2-DX2 protein expression had shorter overall survival than those with low AIMP2-DX2 level. Furthermore, mRNA and protein expression levels of AIMP2-DX2 were both increased in cultured NPC cell lines (5-8F, CNE-2Z, and CNE-1), by being compared with normal nasopharyngeal cell line NP69. Overexpression of AIMP2-DX2 remarkably promoted the cell viability, cell migration, and invasion of cultured NPC cells. Genetic knockdown of AIMP2-DX2 by shRNA lentiviruses significantly suppressed the proliferation, migration, and invasion and induced apoptosis of NPC cells. Inhibition of AIMP2-DX2 decreased the highly expressed level of matrix metalloproteinase- (MMP-) 2 and MMP-9, further suppressed proliferation, migration, and invasion in cultured NPC cells in vitro, and inhibited tumor growth in a xenograft mouse model in vivo. Taken together, these results suggest that AIMP2-DX2 plays an important role in the regulation of NPC and could be a potential therapeutic target and prognostic indicator for the treatment of NPC.

To evaluate the association between mean platelet volume (MPV) and gestational diabetes mellitus (GDM).

Loss of first-phase insulin release is an early pathogenic feature of type 2 diabetes (T2D). Various mouse models exist to study T2D; however, few recapitulate the early β-cell defects seen in humans. We sought to develop a nongenetic mouse model of T2D that exhibits reduced first-phase insulin secretion without a significant deficit in pancreatic insulin content. C57BL/6J mice were fed 10% or 60% fat diet for three weeks, followed by three consecutive, once-daily intraperitoneal injections of the β-cell toxin streptozotocin (STZ; 30, 50, or 75 mg/kg) or vehicle. Four weeks after injections, the first-phase insulin response to glucose was reduced in mice when high-fat diet was combined with 30, 50, or 75 mg/kg STZ. This was accompanied by diminished second-phase insulin release and elevated fed glucose levels. Further, body weight gain, pancreatic insulin content, and β-cell area were decreased in high fat-fed mice treated with 50 and 75 mg/kg STZ, but not 30 mg/kg STZ. Low fat-fed mice were relatively resistant to STZ, with the exception of reduced pancreatic insulin content and β-cell area. Together, these data demonstrate that in high fat-fed mice, three once-daily injections of 30 mg/kg STZ produces a model of β-cell failure without insulin deficiency that may be useful in studies investigating the etiology and progression of human T2D.

Models of Parkinson's disease with neurotoxins have shown that microglial activation does not evoke a typical inflammatory response in the substantia nigra, questioning whether neuroinflammation leads to neurodegeneration. To address this issue, the archetypal inflammatory stimulus, lipopolysaccharide (LPS), was injected into the rat substantia nigra. LPS induced fever, sickness behavior, and microglial activation (OX42 immunoreactivity), followed by astrocyte activation and leukocyte infiltration (GFAP and CD45 immunoreactivities). During the acute phase of neuroinflammation, pro- and anti-inflammatory cytokines (TNF-α, IL-1β, IL-6, IL-4, and IL-10) responded differentially at mRNA and protein level. Increased NO production and lipid peroxidation occurred at 168 h after LPS injection. At this time, evidence of neurodegeneration could be seen, entailing decreased tyrosine hydroxylase (TH) immunoreactivity, irregular body contour, and prolongation discontinuity of TH+ cells, as well as apparent phagocytosis of TH+ cells by OX42+ cells. Altogether, these results show that LPS evokes a typical inflammatory response in the substantia nigra that is followed by dopaminergic neurodegeneration.