Lobe variation effects of experimental diabetes and insulin replacement on rat prostate

We investigated the impact of diabetes with simultaneous and late insulin replacement on rat prostate growth during puberty, paying special attention to different prostatic lobes. Diabetes was induced by administration of streptozotocin (STZ) in 40-day-old male Wistar rats. A subset of diabetic rats underwent simultaneous insulin replacement (3 days after STZ administration), and another subset underwent a late insulin replacement (20 days after STZ administration). The ventral, dorsolateral, and anterior prostatic lobes were weighed and processed for histological, immunohistochemical, and morphometric analyses. Both diabetic and insulin-treated animals maintained low plasma testosterone (T) concentrations, whereas dihydrotestostenore (DHT) levels were normal. Diabetic animals had a decreased gain in absolute prostatic weight when compared to age-matched controls and insulin replacement animals. However, prostatic lobe weight in the diabetic animals was ～100% higher, even at the beginning of the experiment. Among the lobes, the anterior lobe showed the highest weight gain in diabetic and insulin replacement conditions. Epithelial cell proliferation in all lobes was significantly reduced in diabetic animals and significantly increased in insulin replacement animals, although apoptosis was unaltered. In conclusion, diabetes diminishes, but does not abolish, prostate growth during puberty. Even late insulin administration reduces the adverse effects of this disease on the prostate. In a scenario with both low insulin and T levels, DHT and other factors may play an important role in pubertal prostate growth. The adverse effects of diabetes on the rat prostate show a variation in lobe response, suggesting that diabetes may affect human prostate zones differently.     

Influence of nanostructure composition on its morphometric characterization by different techniques

Morphometric characterization of nanoparticles is crucial to determine their biological effects and to obtain a formulation pattern. Determining the best technique requires knowledge of the particles being analyzed, the intended application of the particles, and the limitations of the techniques being considered. The aim of this article was to present transmission (TEM) and scanning (SEM) electron microscopy protocols for the analysis of two different nanostructures, namely polymeric nanoemulsion and poly(lactic‐co‐glycolic acid) (PLGA) nanoparticles, and to compare these results with conventional dynamic light scattering (DLS) measurements. The mean hydrodynamic diameter, the polydispersity index, and zeta potential of the nanostructures of polymeric nanoemulsion were 370.5 ± 0.8 nm, 0.133 ± 0.01, and ?36.1 ± 0.15 mV, respectively, and for PLGA nanoparticles were 246.79 ± 5.03 nm, 0.096 ± 0.025, and ?4.94 ± 0.86 mV, respectively. TEM analysis of polymeric nanoemulsion revealed a mean diameter of 374 ± 117 nm. SEM analysis showed a mean diameter of 368 ± 69 nm prior to gold coating and 448 ± 70 nm after gold coating. PLGA nanoparticles had a diameter of 131 ± 41.18 nm in TEM and 193 ± 101 nm in SEM. Morphologically, in TEM analysis, the polymeric nanoemulsions were spherical, with variable electron density, very few showing an electron‐dense core and others an electron‐dense surface. PLGA nanoparticles were round, with an electron‐lucent core and electron‐dense surface. In SEM, polymeric nanoemulsions were also spherical with a rough surface, and PLGA nanoparticles were round with a smooth surface. The results show that the “gold standards” for morphometric characterization of polymeric nanoemulsion and PLGA nanoparticles were, respectively, SEM without gold coating and TEM with negative staining. Microsc. Res. Tech. 77:691–696, 2014. © 2014 Wiley Periodicals, Inc.     

Disruption of Redox Homeostasis by Alterations in Nitric Oxide Synthase Activity and Tetrahydrobiopterin along with Melanoma Progression

Cutaneous melanoma emerges from the malignant transformation of melanocytes and is the most aggressive type of skin cancer. The progression can occur in different stages: radial growth phase (RGP), vertical growth phase (VGP), and metastasis. Reactive oxygen species contribute to all phases of melanomagenesis through the modulation of oncogenic signaling pathways. Tetrahydrobiopterin (BH4) is an important cofactor for NOS coupling, and an uncoupled enzyme is a source of superoxide anion (O₂^•−) rather than nitric oxide (NO), altering the redox homeostasis and contributing to melanoma progression. In the present work, we showed that the BH4 amount varies between different cell lines corresponding to distinct stages of melanoma progression; however, they all presented higher O₂^•− levels and lower NO levels compared to melanocytes. Our results showed increased NOS expression in melanoma cells, contributing to NOS uncoupling. BH4 supplementation of RGP cells, and the DAHP treatment of metastatic melanoma cells reduced cell growth. Finally, Western blot analysis indicated that both treatments act on the PI3K/AKT and MAPK pathways of these melanoma cells in different ways. Disruption of cellular redox homeostasis by the altered BH4 concentration can be explored as a therapeutic strategy according to the stage of melanoma.     

Active Compounds in Zingiber officinale as Possible Redox Inhibitors of 5-Lipoxygenase Using an In Silico Approach

5-Lipoxygenase (5-LOX) converts arachidonic acid to lipidic inflammatory mediators such as leukotrienes (LTs). In diseases such as asthma, LTs contribute to a physiopathology that could be reverted by blocking 5-LOX. Natural products with anti-inflammatory potential such as ginger have been used as nutraceuticals since ancient times. 6-Gingerol and 6-shogaol are the most abundant compounds in the ginger rhizome; they possess anti-inflammatory, antioxidant, and chemopreventive properties. In the present study, 6-gingerol and 6-shogaol structures were analyzed and compared with two commercial 5-LOX inhibitors (zileuton and atreleuton) and with other inhibitor candidates (3f, NDGA, CP 209, caffeic acid, and caffeic acid phenethyl ester (CAPE)). The pharmacokinetics and toxicological properties of 6-gingerol, 6-shogaol, and the other compounds were evaluated. Targeted molecular coupling was performed to identify the optimal catalytic pocket for 5-LOX inhibition. The results showed that 6-gingerol and 6-shogaol follow all of the recommended pharmacokinetic parameters. These compounds could be inhibitors of 5-LOX because they present specific interactions with the residues involved in molecular inhibition. The current study demonstrated the potential of 6-gingerol and 6-shogaol as anti-inflammatory agents that inhibit 5-LOX, as they present a high level of performance in the toxicological analysis and could be catabolized by the cytochrome p450 enzymatic complex; however, 6-gingerol was superior in safety compared to 6-shogaol.     

Fibrotic Idiopathic Interstitial Lung Disease: The Molecular and Cellular Key Players

Interstitial lung diseases (ILDs) that are known as diffuse parenchymal lung diseases (DPLDs) lead to the damage of alveolar epithelium and lung parenchyma, culminating in inflammation and widespread fibrosis. ILDs that account for more than 200 different pathologies can be divided into two groups: ILDs that have a known cause and those where the cause is unknown, classified as idiopathic interstitial pneumonia (IIP). IIPs include idiopathic pulmonary fibrosis (IPF), non-specific interstitial pneumonia (NSIP), cryptogenic organizing pneumonia (COP) known also as bronchiolitis obliterans organizing pneumonia (BOOP), acute interstitial pneumonia (AIP), desquamative interstitial pneumonia (DIP), respiratory bronchiolitis-associated interstitial lung disease (RB-ILD), and lymphocytic interstitial pneumonia (LIP). In this review, our aim is to describe the pathogenic mechanisms that lead to the onset and progression of the different IIPs, starting from IPF as the most studied, in order to find both the common and standalone molecular and cellular key players among them. Finally, a deeper molecular and cellular characterization of different interstitial lung diseases without a known cause would contribute to giving a more accurate diagnosis to the patients, which would translate to a more effective treatment decision.     

Study of oxygen reduction on copper applied to the peroxidase-mediated oxidation of methylene blue

The kinetics of oxygen reduction was studied on copper in tartarate solutions using a rotating disc electrode. The effect of pH on the cathodic reduction was examined. AC impedance and cathodic polarization curves showed that the reaction is partially controlled by mass transport. At high current densities, the limiting current values showed that O₂ is reduced mainly through the four electron pathway. From measurements of density and viscosity and data taken from Levich and Tafel plots, diffusion coefficients for oxygen in tartarate medium were calculated. Peroxidase-mediated oxidation of methylene blue (MB) results using electrogenerated H₂O₂ on copper electrode showed that O₂ reduction reaction occurs in two steps producing the adsorbed intermediate species H₂O₂ by k ₂. In the absence of peroxidase, the hydrogen peroxide elimination has to be rapid with k ₃ ≫ k _M. The MB oxidation by lignin peroxidase using electrogenerated H₂O₂ was studied. After 24 h LiP is able to oxide the MB, producing different oxidized forms: azure C and thionine, with yield of 23% and 66%, respectively.     

Development of a green integrated process for biodiesel esters production: Use of fermented macaúba cake as biocatalyst for macaúba acid oil transesterification

In this study, a new green integrated process for biodiesel ester production was developed, consisting of the enzymatic transesterification of acid oil from macaúba palm tree (Acrocomia aculeata) catalyzed by dry fermented solid (DFS) with lipase activity, which was obtained from solid-state fermentation (SSF) of macaúba cake. Analysis of statistical experimental designs showed that nitrogen supplementation of macaúba cake improved production of lipases displaying esterification capacity by Rhizomucor miehei. The conditions chosen to produce R. miehei DFS were cake supplementation with 2 wt% urea, 60 wt% of initial moisture during 48 h of fermentation at 40°C. DFS showed, by zymography assays, at least one enzyme that was different from those that are present in the commercial lipase of R. miehei. Furthermore, DFS was used in transesterification reactions between macaúba acid oil and alcohol (methanol or ethanol), and the ester content obtained was 92%. This is the first time that this integrated process using low-cost biocatalysts and a low-value oil derived from the processing of the same fruit was used for biodiesel ester production, reducing costs and logistic constraints, and optimizing the raw material utilization.     

Assessment of exposure of broiler chicken in Brazil to mycotoxins through naturally contaminated feed

The objective of the present study was to assess the degree of exposure of broiler chicken to mycotoxins through naturally contaminated feed and the hygienic quality of feeds. For this purpose the total fungal count and occurrence of fumonisins and aflatoxins were evaluated in four feed types intended for broilers (n?=?158), collected from a poultry breeding farm in northern Paraná State, Brazil. In most feed samples (94 % pre-starter, 91 % starter, 99 % grower and 97 % finisher feeds) the total mould and yeast counts were below 1.0?×?10⁴ CFU/g, the maximum limit established to assure a good hygienic quality of the product. Fumonisins were detected in 94.9 % feed samples at mean levels ranging from 0.52 mg/kg (finisher) to 0.68 mg/kg (pre-starter and grower), while aflatoxins were detected in 72.1 % feed samples at mean levels ranging from 0.0022 mg/kg (pre-starter) to 0.0064 mg/kg (grower). The maximum estimated daily intake of fumonisin B₁ for broilers (0.057 mg/kg body weight/day) was below the Lowest Observed Adverse Effect Level (2 mg/kg bw/day). Concerning aflatoxins, most of the positive samples (97 %) showed aflatoxin levels below the maximum limit allowed by the European Commission 0.02 mg aflatoxin B₁/kg. Nevertheless, a rigorous monitoring program of the feed producing chain is essential in order to minimize human health hazards. To our knowledge, this is the first report on the degree of exposure of broilers to mycotoxins through naturally contaminated feed in Brazil.