Effect of TiO2 Microparticles in Lettuce (Lactuca sativa L.) Seeds and Seedlings.

The particle size reduction technology is used in several segments, including sunscreens and new techniques and product improvement. One of the main particles used in the sunscreens formulation is titanium dioxide (TiO2). This formulation allows for better characteristics of these products. Perspectives like incorporation of the particles by other biological systems beyond humans and their effects should be observed. This work aimed to evaluate the titanium dioxide microparticles phytotoxicity on Lactuca sativa L. plants through tests of germination, growth, and weight analysis using microscopy techniques: optical microscopy (OM) and scanning electron microscopy (SEM). Some of the results showed cellular and morphological damage, mainly in the roots and 50 mg L-1 TiO2 concentration, confirmed by SEM. Additionally, anatomical damages like vascular bundle disruption and irregularity in the cortex cells were confirmed by SEM. Additionally, anatomical damages were observed on the three main organs (root, hypocotyl, and leaves) evidenced by the OM. Perspectives to confirm new hypotheses of the interaction of nanomaterials with biological systems are necessary.

Acute toxicity of titanium dioxide microparticles in Artemia sp. nauplii instar I and II.

In this study, the toxicity effects of titanium dioxide (MTiO2 ) microparticles on Artemia sp. nauplii instar I and II between 24 and 48 h was evaluated. The MTiO2 were characterized using different microscopy techniques. MTiO2 rutile was used in toxicity tests at concentration of 12.5, 25, 50, and 100 ppm. No toxicity was observed in Artemia sp. nauplii instar I at the time of 24 and 48 h. However, Artemia sp. nauplii instar II toxicity was observed within 48 h of exposure. MTiO2 at concentrations of 25, 50 and 100 ppm was lethal for Artemia sp. with a significant difference (p ≤ .05) in relation to the control artificial sea water with LC50 value at 50 ppm. Analysis of optical and scanning electron microscopy revealed tissue damage and morphological changes in Artemia sp. nauplii instar II. By using confocal laser scanning microscopy, cell damage was observed due to the toxicity of MTiO2 at a concentration of 20, 50, and 100 ppm. The high mortality rate is related to the filtration of MTiO2 by Artemia sp. nauplii instar II due to the complete development of the digestive tract.

A new bioinspired peptide on defensin from C. annuum fruits: Antimicrobial activity, mechanisms of action and therapeutical potential.

BACKGROUND: Antimicrobial peptides, natural or synthetic, appear as promising molecules for antimicrobial therapy because of their both broad antimicrobial activity and mechanism of action. Herein, we determine the anti-Candida and antimycobacterial activities, mechanism of action on yeasts, and cytotoxicity on mammalian cells in the presence of the bioinspired peptide CaDef2.1G27-K44. METHODS: CaDef2.1G27-K44 was designed to attain the following criteria: high positive net charge; low molecular weight (<3000 Da); Boman index ≤2.5; and total hydrophobic ratio ≥ 40%. The mechanism of action was studied by growth inhibition, plasma membrane permeabilization, ROS induction, mitochondrial functionality, and metacaspase activity assays. The cytotoxicity on macrophages, monocytes, and erythrocytes were also determined. RESULTS: CaDef2.1G27-K44 showed inhibitory activity against Candida spp. with MIC100 values ranging from 25 to 50 µM and the standard and clinical isolate of Mycobacterium tuberculosis with MIC50 of 33.2 and 55.4 µM, respectively. We demonstrate that CaDef2.1G27-K44 is active against yeasts at different salt concentrations, induced morphological alterations, caused membrane permeabilization, increased ROS, causes loss of mitochondrial functionality, and activation of metacaspases. CaDef2.1G27-K44 has low cytotoxicity against mammalian cells. CONCLUSIONS: The results obtained showed that CaDef2.1G27-K44 has great antimicrobial activity against Candida spp. and M. tuberculosis with low toxicity to host cells. For Candida spp., the treatment with CaDef2.1G27-K44 induces a process of regulated cell death with apoptosis-like features. GENERAL SIGNIFICANCE: We show a new AMP bioinspired with physicochemical characteristics important for selectivity and antimicrobial activity, which is a promising candidate for drug development, mainly to control Candida infections.

Effect of preservation solution and distension pressure on saphenous vein's endothelium.

OBJECTIVES: Approaches to improve saphenous vein (SV) patency in coronary artery bypass graft (CABG) surgery remain relevant. This study aimed to evaluate the effects of different preservation solutions and different pressures of intraluminal distention on the endothelium of SV segments in CABG. METHODS: Forty-two SV segments obtained from 12 patients undergoing CABG were divided into 7 groups. Group 1 (control) was prepared without preservation or intraluminal distension, while the other 6 groups were preserved in autologous heparinized autologous arterial blood or normal saline (NS), with distention pressures 30, 100 and 300 mmHg. To assess the effects of using these solutions and pressures on the endothelium, the grafts were analysed by scanning electron microscopy, with the measurement of endothelial damage degree. RESULTS: Segments in group 1 showed minimal endothelial damage. SV grafts preserved with NS had significantly greater endothelial damage both compared to the control group and compared to groups preserved with autologous arterial blood (P < 0.001). Segments distended with pressures up to 100 mmHg showed less damage when compared to those distended at 300 mmHg, with the ones subjected to higher pressures presenting a maximum degree of damage, with considerable loss and separation of endothelial cells, extensive foci of exposure of the basement membrane and numerous fractures of the intimate layer, without differences regarding the solution used. CONCLUSIONS: Preparation of SV using NS and with intraluminal distension pressures above 100 mmHg is factors related to increased damage to the venous endothelium.

Silver nanoparticles (AgNPs) internalization and passage through the Lactuca sativa (Asteraceae) outer cell wall.

Silver nanoparticle (AgNPs) toxicity is related to nanoparticle interaction with the cell wall of microorganisms and plants. This interaction alters cell wall conformation with increased reactive oxygen species (ROS) in the cell. With the increase of ROS in the cell, the dissolution of zero silver (Ag0) to ionic silver (Ag+) occurs, which is a strong oxidant agent to the cellular wall. AgNP interaction was evaluated by transmission electron microscopy (TEM) on Lactuca sativa roots, and the mechanism of passage through the outer cell wall (OCW) was also proposed. The results suggest that Ag+ binds to the hydroxyls (OH) present in the cellulose structure, thus causing the breakdown of the hydrogen bonds. Changes in cell wall structure facilitate the passage of AgNPs, reaching the plasma membrane. According to the literature, silver nanoparticles with an average diameter of 15nm are transported across the membrane into the cells by caveolines. This work describes the interaction between AgNPs and the cell wall and proposes a transport model through the outer cell wall.

Morphological and ultrastructural changes in seminal coagulum of the squirrel monkey (Saimiri collinsi Osgood, 1916) before and after liquefaction.

Studies with squirrel monkey semen are of special interest due to the large amount of coagulation that is a component of the semen, which is a problem that has to be overcome when the objective is harvesting of gametes. In the present study, there was characterization of the seminal coagulum of captive S. collinsi. Four samples of ejaculates were collected using electroejaculation procedures from four animals. The aim in conducting this study was to evaluate seminal coagulum of S. collinsi using histological and scanning electron microscopy (SEM) procedures before and after semen liquefaction in an ACP-118® extender. Seminal coagulum of S. collinsi was composed of a superficial plate (external), which coats the spongy seminal plasma matrix of S. collinsi. Additionally, there were sperm in the external and internal components of the coagulum with these gametes being isolated or grouped and with there being a heterogeneous distribution of gametes. The supplementation of semen with ACP-118® resulted in a partial dissolution of the seminal plate and spongy matrix portions of the seminal coagulum within the first hour of incubation.

H2O2 priming promotes salt tolerance in maize by protecting chloroplasts ultrastructure and primary metabolites modulation.

Hydrogen peroxide priming has emerged as a powerful strategy to trigger multiple responses involved in plant acclimation that reinforce tolerance to abiotic stresses, including salt stress. Thus, this study aimed to investigate the impact of foliar H2O2 priming on the physiological, biochemical, and ultrastructural traits related to photosynthesis of salt-stressed plants. Besides, we provided comparative leaf metabolomic profiles of Zea mays plants under such conditions. For this, H2O or H2O2 pretreated plants were grown under saline conditions for 12-days. Salinity drastically affected photosynthetic parameters and structural chloroplasts integrity, also increased reactive oxygen species contents promoting disturbance in the plant metabolism when compared to non-saline conditions. Our results suggest that H2O2-pretreated plants improved photosynthetic performance avoiding salinity-induced energy excess and ultrastructural damage by preserving stacking thylakoids. It displayed modulation of some metabolites, as arabitol, glucose, asparagine, and tyrosine, which may contribute to the maintenance of osmotic balance and reduced oxidative stress. Hence, our study brings new insights into an understanding of plant acclimation to salinity by H2O2 priming based on photosynthesis maintenance and metabolite modulation.

Toxicity evaluation of process water from hydrothermal carbonization of sugarcane industry by-products.

Hydrothermal carbonization (HTC) is a thermochemical process carried out in an aqueous medium. It is capable of converting biomass into a solid, carbon-rich material (hydrochar), and producing a liquid phase (process water) which contains the unreactive feedstock and/or chemical intermediates from the carbonization reaction. The aim of this study was to evaluate the characteristics of process water generated by HTC from vinasse and sugarcane bagasse produced by sugarcane industry and to evaluate its toxicity to both marine (using Artemia salina as a model organism) and the terrestrial environment (through seed germination studies of maize, lettuce, and tomato). The experiments showed that concentrated process water completely inhibited germination of maize, lettuce, and tomato seeds. On the other hand, diluted process water was able to stimulate seedlings of maize and tomato and enhance root and shoot growth. For Artemia, the LC50 indicated that the process water is practically non-toxic; however, morphological changes, especially damages to the digestive tube and antennas of Artemia, were observed for the concentration of 1000 mg C L-1.

Spherical neutral gold nanoparticles improve anti-inflammatory response, oxidative stress and fibrosis in alcohol-methamphetamine-induced liver injury in rats.

This study aimed to elucidate the anti-inflammatory, anti-oxidant and antifibrotic effects of gold nanoparticles (GNPs) in rats subjected to liver injury with ethanol and Methamphetamine (METH). The liver injury was induced by gavage administrations of 30% alcoholic solution (7 g/kg) once a day during 28 days, followed by METH (10 mg/kg) on the 20th and 28th days of treatment. GNPs treatment (724.96 µg/kg) during the ethanol and METH exposure was associated with reduced steatosis, hepatic cord degeneration, fibrosis and necrosis. Furthermore, there was a reduction in biochemical markers of liver damage and oxidative stress, and pro-inflammatory cytokines IL-1ß and TNF-α, compared to ethanol + METH group alone. A decrease of FGF, SOD-1 and GPx-1 expression was also observed. GNPs down-regulated the activity of Kupffer cells and hepatic stellate cells affecting the profile of their pro-inflammatory cytokines, oxidative stress and fibrosis through modulation of signaling pathways AKT/PI3K and MAPK in ethanol + METH-induced liver injury in a rat model.

Apoptosis in human liver carcinoma caused by gold nanoparticles in combination with carvedilol is mediated via modulation of MAPK/Akt/mTOR pathway and EGFR/FAAD proteins.

In cancers, apoptosis signaling pathways and cell survival and growth pathways responsible for resistance to conventional treatments, such as Pi3K/Akt/mTOR and mitogen-activated protein kinase (MAPK) become dysregulated. Recently, alternative treatments to promote tumor cell death have become important. The present study reports on the antitumor and cytoprotective action of gold nanoparticles (GNPs) and carvedilol in combination and in isolated application. Apoptosis was analyzed by FITC/propidium iodide staining flow cytometry; caspase-3, caspase-8, Bcl-2 and MAPK/ERK activity by immunofluorescence microscopy; gene expression of proteins related to cell death as Akt, mTOR, EGFR, MDR1, survivin, FADD and Apaf, by the real-time PCR; and western blot analysis for MAPK/ERK, Akt and mTOR. Oxidative stress evaluation was performed by reduced glutathione (GSH) and malondialdehyde (MDA) levels. Intracellular GNPs targets were identified by transmission electron microscopy. After exposure to a combination of GNPs (6.25 µg/ml) and carvedilol (3 µM), death as promoted by apoptosis was detected using flow cytometry, for expression of pro-apoptotic proteins FADD, caspase-3, caspase-8 and sub-regulation of anti-apoptotic MAPK/ERK, Akt, mTOR, EGFR and MDR1 resistance. Non-tumor cell cytoprotection with GSH elevation and MDA reduction levels was detected. GNPs were identified within the cell near to the nucleus when combined with carvedilol. The combination of GNP and carvedilol promoted downregulation of anti-apoptotic and drug resistance genes, over-regulation of pro-apoptotic proteins in tumor cells, as well as cytoprotection of non-tumor cells with reduction of apoptosis and oxidative stress.

Tributyltin impairs the coronary vasodilation induced by 17ß-estradiol in isolated rat heart.

Triorganotins, such as tributyltin (TBT), are environmental contaminants that are commonly used as antifouling agents for boats. However, TBT is also known to alter mammalian reproductive functions. Although the female sex hormones are primarily involved in the regulation of reproductive functions, 17ß-estradiol also protects against cardiovascular diseases, in that this hormone reduces the incidence of coronary artery disease via coronary vasodilation. The aim of this study was to examine the influence of 100 ng/kg TBT administered daily by oral gavage for 15 d on coronary functions in female Wistar rats. Findings were correlated with changes in sex steroids concentrations. Tributyltin significantly increased the baseline coronary perfusion pressure and impaired vasodilation induced by 17ß-estradiol. In addition, TBT markedly decreased serum 17ß-estradiol levels accompanied by a significant rise in serum progesterone levels. Tributyltin elevated collagen deposition in the heart interstitium and number of mast cells proximate to the cardiac vessels. There was a positive correlation between the increase in coronary perfusion pressure and incidence of cardiac hypertrophy. In addition, TBT induced endothelium denudation (scanning electron microscopy) and accumulation of platelets. Moreover, TBT impaired coronary vascular reactivity to estradiol (at least in part), resulting in endothelial denudation, enhanced collagen deposition and elevated number of mast cells. Taken together, the present results demonstrate that TBT exposure may be a potential risk factor for cardiovascular disorders in rats.

Tributyltin impairs the reproductive cycle in female rats.

Triorganotins are environmental contaminants, commonly used in antifouling agents for boats, that bioaccumulate and thus are found in mammals and humans due to ingestion of contaminated seafood diets. The importance of triorganotins as environmental endocrine disruptors and consequent reproductive toxicity in different animal models is well known; however, the adverse effects on reproductive cycle are less well understood. The potential reproductive toxicity of tributyltin (TBT) on regular reproductive cycling of female rats was examined. Wistar female rats (12 wk old, weighing approximately 230 g) were divided into two groups: control (vehicle, ethanol 0.4%) and tributyltin (100 ng/kg/d, 7 d/wk, for 16 d by gavage). Tributyltin significantly decreased the cycle regularity (%), duration of the reproductive cycle, the proestrus and diestrus phases, and number of epithelial cell in proestrus phase. TBT also increased the duration of metestrus and the number of cornified cells in this phase. Ovary weight and serum 17ß-estradiol levels decreased markedly, accompanied by a significant increase in progesterone levels. Histological analysis showed apoptotic cells in corpus luteum and granulosa cells layer, with cystic follicles after TBT exposure. Tributyltin also elevated number of atretic follicles and corpoa lutea. The micronucleus (MN) test, using Chinese hamster ovary cells, demonstrated a concentration-dependent mutagenic effect of TBT, and at 2.0 × 10(-2)ng/ml most of the cells were nonviable. The toxic potential of TBT over the reproductive cycle may be attributed to changes found in the ovarian weight, unbalanced levels of sexual female hormones, and number of ovarian follicles and corpora lutea.