Rutile nano-bio-interactions mediate dissimilar intracellular destiny in human skin cells.

The use of nanoparticles (NPs) in the healthcare market is growing exponentially, due to their unique physicochemical properties. Titanium dioxide nanoparticles (TiO2 NPs) are used in the formulation of sunscreens, due to their photoprotective capacity, but interactions of these particles with skin cells on the nanoscale are still unexplored. In the present study we aimed to determine whether the initial nano-biological interactions, namely the formation of a nano-bio-complex (other than the protein corona), can predict rutile internalization and intracellular trafficking in primary human fibroblasts and keratinocytes. Results showed no significant effect of NPs on fibroblast and keratinocyte viability, but cell proliferation was possibly compromised due to nano-bio-interactions. The bio-complex formation is dependent upon the chemistry of the biological media and NPs' physicochemical properties, facilitating NP internalization and triggering autophagy in both cell types. For the first time, we observed that the intracellular traffic of NPs is different when comparing the two skin cell models, and we detected NPs within multivesicular bodies (MVBs) of keratinocytes. These structures grant selected input of molecules involved in the biogenesis of exosomes, responsible for cell communication and, potentially, structural equilibrium in human tissues. Nanoparticle-mediated alterations of exosome quality, quantity and function can be another major source of nanotoxicity.

Use of triethylcitrate plasticizer in the production of poly-L-lactic acid implants with different degradation times.

Bioabsorbable materials have been widely used in the repair of damaged tissue as well as in the controlled release of drugs and as a supports for cultured cells. The degradation time of poly-L-(lactic acid) (PLLA) may be controlled by altering the polymer porosity through the addition of the plasticizer triethylcitrate. This in turn influences the extent cellular infiltration. In this study, we examined the degradation of PLLA membranes containing different concentrations of plasticizer. PLLA discs were implanted subcutaneouly in rats and withdrawn 2, 14 and 60 days after implantation. The samples were processed for light microscopy and scanning electron microscopy (SEM). Polymer degradation was proportional to the concentration of plasticizer, indicating that triethylcitrate could affect the degradation time of the implants, without damaging the polymer biocompatibility.

In vivo interaction of cells on poly L-(lactic acid) membranes containing plasticizer.

The development of biodegradable materials has lead to renewed interest in the study of their interactions with the host organism in order to make the resulting products appropriate for use as temporary materials in protheses. Poly L-(lactic acid)(PLLA)-based biodegradable devices have been used for several purposes. The physical properties of these materials can be modified by the addition of a plasticizer, such as the triethylcitrate, to provide flexibility and porosity to the implants and enhance control of the polymer degradation time. In this work we examined the biological properties of a PLLA porous membrane containing 7% triethylcitrate, by assessing the process of degradation and the interaction with dermal tissue. Samples of skin obtained from female Wistar rats 2-180 days after implantation with PLLA-based membrane were processed for light microscopy and scanning electron microscopy. The membranes became surrounded by a delicate network of connective tissue which gradually invaded the membrane structure. Polymer degradation began with the appearance of radial fractures in the globular units of the biodegradable membrane, especially by 90 and 180 days after implantation.

Glycogen phosphorylase activity in the liver of the frog Rana esculenta.

1. Phosphorylase activity has been assayed in liver extracts of the frog, Rana esculenta, during the winter period. In native conditions, most of the phosphorylase is present as AMP-independent activity and shows properties similar to those of the a form of the liver enzyme from other vertebrates. 2. It is suggested that regulation of phosphorylase activity is through interconversion between a and b forms operated by endogenous phosphorylase kinase and phosphatase. 3. Kinetic studies show hyperbolic saturation curves for glycogen with apparent Km of 2.91 mM and 9.67 mM for a and b forms, respectively. 4. A hyperbolic saturation curve is also observed for glucose 1-P in the case of phosphorylase a, with an apparent Km of 3.95 mM, whereas a sigmoidal kinetic is shown by the b form for the same substrate; from Hill plots an S0.5 of 24.2 mM was derived. 5. Hyperbolic responses were observed in the case of AMP, and Ka of 70 microM and 0.31 mM were calculated for phosphorylase a and b, respectively.

Annual variations in the binding of insulin to hepatic membranes of the frog Rana esculenta.

Amphibia undergo regular annual cycles of metabolic activity that are influenced by both exogenous factors and hormones. Insulin binding to crude frog hepatic membranes was studied throughout the year. The general character of insulin binding was similar to that in other vertebrates; the maximum specific binding was achieved after 4 hr at 4 degrees, the optimum pH was 7.8, half-maximal displacement of bound insulin was from 9 x 10(-10) to 1 x 10(-9) M, and insulin analogs competed for the insulin receptor in line with their relative biological potencies. A biphasic Scatchard plot and negative cooperativity of the receptor were also observed in frog liver membranes. Affinity constants from Scatchard plots revealed high and low affinity binding sites which were unchanged during the year. The seasonal cycle, however, markedly affected the binding capacity for both sites. Maximum binding occurred in May-June and the minimum in November-December for both classes of receptors. Binding capacities ranged from 1.71 to 11.33 fmol/mg protein for the high affinity sites and from 432 to 3171 fmol/mg protein for the low affinity sites. It is concluded that annual cycles of insulin binding reflect modulation of receptor number rather than receptor affinity.

Cholesterol metabolism in frog (Rana esculenta) liver: seasonal and sex-related variations.

Many aspects of lipid metabolism have been studied in amphibians, but seasonal lipid modulation in male and female frogs has not been investigated. We describe here the yearlong patterns of hepatic lipid content and enzyme activities related to cholesterol homeostasis, 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase activity and acyl coenzyme A:cholesterol acyltransferase (ACAT) activity in liver of the male and female frog, Rana esculenta. Lipid storage follows distinct seasonal patterns, with an increase in June that is more pronounced in the female than in the male frog. Cholesterol content and cholesterol storage as cholesteryl ester in male liver are consistent with the activity of HMG-CoA reductase and of ACAT enzymes. HMG-CoA reductase activity of the female frog shows an extra peak in fall unrelated to cholesterol storage and probably related to the production of essential compound for oogenesis.

Tyrosine aminotransferase activity of frog (Rana esculenta) liver. III. A circannual study.

A circannual study of tyrosine aminotransferase and other metabolic enzymes in frog liver is reported. The subcellular distribution of all enzymatic activities under investigation was also studied. Results show significant oscillations of all enzymatic activities throughout the year; in particular tyrosine aminotransferase has a marked summer maximum. The subcellular distribution of tyrosine aminotransferase shows significant variations: the soluble activity of the enzyme presents a bimodal circannual distribution, which has its counterpart in an increased activity of heavier fractions.

Glucocorticoid receptor of frog (Rana esculenta) liver.

The presence of a glucocorticoid soluble receptor is demonstrated in frog liver cytosol. The kinetic characterization of frog liver cytosolic receptor for glucocorticoids is reported and its steroid specificity assessed. Results indicate a gross similarity between frog liver and mammalian glucocorticoid receptor, being a major difference the reduced binding capacity.

Tyrosine aminotransferase activity of frog (Rana esculenta) liver.

1. The presence of tyrosine aminotransferase is reported both in particulate and soluble fractions of frog liver. 2. The activity of the soluble enzyme of frog liver was investigated with regard to its dose and time dependence, its substrate specificity and concentration dependence, its thermal sensitivity as well as pH and temperature dependence. 3. It appears that the properties of the soluble tyrosine aminotransferase of frog liver are in close agreement with those reported for the mammalian liver enzyme.

Seasonal fluctuations of pancreatic activities and cyclic nucleotide levels in the frog Rana esculenta.

1. Seasonal influence on pancreatic activity has been determined in the frog Rana esculenta. 2. Protein content, amylase activity and cyclic nucleotide levels, show significant changes throughout the year, being higher in the active period (spring, summer), with respect to a constant pancreas weight. 3. A significant correlation between protein content and cyclic AMP levels has been observed. 4. Results suggest that feeding habits exert a control on the pancreatic activity even if stimuli other than food intake cannot be ruled out.

Tyrosine aminotransferase activity of frog (Rana esculenta) liver--II. Comparative aspects of intracellular distribution.

1. A subcellular fractionation procedure for frog liver is reported and validated by the distribution pattern of several marker enzymes, also in comparison with rat liver. 2. The subcellular distribution of tyrosine aminotransferase was investigated in frog liver as compared to rat liver: a different distribution of the enzyme was observed, being the activity mostly recovered in mitochondrial and cytosolic compartments. 3. Results indicate that mitochondrial tyrosine aminotransferase of both frog and rat liver is a matrix enzyme, even if differences are observed concerning its release from the organelles upon detergent treatment.

Insulin-sensitive adenosine 3',5'-cyclic monophosphate phosphodiesterase of hepatocyte plasma membrane.

Adenosine 3',5'-cyclic monophosphate phosphodiesterase (EC 3.1.4.17) has been investigated in rat liver as to its insulin sensitivity. Hormone action has been assayed in vitro on a liver homogenate purified by DEAE-cellulose column chromatography, on isolated hepatocytes, on isolated plasma membranes. The DEAE-cellulose chromatography purified homogenate showed no sensitivity to insulin, whereas isolated hepatocytes incubated in presence of insulin showed increased phosphodiesterase activity in a plasma membrane-containing fraction. The plasma membrane-bound enzyme, which shows both high and low affinity components, was significantly stimulated after hormonal treatment; this effect being dependent on a V increase of the low Km form.

Ultrastructural analysis of some functional aspects of Xenopus laevis pancreas during development and metamorphosis.

Morphological studies using both light and electron microscope were carried out with the aim of characterizing cells present in the larval and adult pancreas of Xenopus laevis. The following cell types have been seen: (1) exocrine cells, with a very well developed r.e.r. (rough endoplasmic reticulum), well defined Golgi complexes and numerous large secretory granules (A cells); (2) cells without either r.e.r. or secretory granules but with a large number of well developed mitochondria (B cells); (3) endocrine cells often clustered in the typical islets and with small membrane-coated granules showing a very dense central core surrounded by a light halo (C cells). During development, the aspect is seen to change from an unorganized tissue in which the acinar structures are still not clearly visible (stage 42), to a more organized form in which the exocrine cells (A cells) are seen to be arranged around the lumen of the acinus together with some B cells. At the stages 54-56, an increasing number of acini surrounded both by A and B cells was observed. At about stage 61, large quantities of necrotic cells were seen and it became more difficult to individualize the acinar organization found in the preceding stages. Finally, there are no necrotic cells in the adult but only A, B cells which are organized in well developed acinar structures and C cells. The investigation also included a study of some pancreatic enzymes (lipase and amylase) synthesized during larval life. Lipase activity shows a peak at stage 54-56 in which the most well organized tissue of the entire larval life was observed. The activity then decreases, reaching a minimum at stage 66, after which it rapidly rises. Maximum amylase activity occurs at stage 51 after which there is a decrease, to a minimum at stage 66. The activity then remains at constant level.

[Rapeseed oil: nutritional aspects and safety considerations (author's transl)]. / Il problema dell'olio di colza: aspetti nutrizionali e di sicurezza

In this review data are reported on the effects of rapeseed oil feeding to several animal species (mice, pigs, rats, guinea pigs, ducklings, chickens, turkeys) with particular regard to its nutritional value as compared to other vegetable fats, and to physiological and pathological modifications on different organs and tissues, namely: gonads, adrenals, thyroid, liver, myocardium. Finally, the results obtained after administration of regular rapeseed oil (i.e. with high levels of erucic acid) are compared with those obtained using the modern types Canbra, Span, Oro, Zephir (i.e. with low levels of erucic acid). From this comparison it could be inferred that erucic acid could be the major responsible for the observed modifications, but also that other long chain monoenes and the altered ratio between saturated and monounsaturated fatty acids might have a certain degree of pathogenic effects, although in this case the lesions appear later, are less frequent and less severe.