Evaluation of the mutagenic, antimutagenic and antiproliferative potential of Croton lechleri (Muell. Arg.) latex.

Sangre de Drago is a red viscous latex extracted from Croton lechleri (Euphorbiaceae) cortex, renowned in South American popular medicine for its wound-healing properties. The in vitro antiproliferative effects were determined on the human myelogenous leukemia K562 cells line (IC50 = 2.5 +/- 0.3 microg ml(-1)). The mutagenic and antimutagenic activity of C. lechleri sap was examined by means of the Ames/Salmonella test. No mutagenic activity was found on the Salmonella typhimurium strains T98 and T100, either with or without S9 activation. On the other hand, the sap showed an inhibitory effect against the mutagenic activity of the indirectly acting mutagen 2-Aminoanthracene in presence of S9 and a moderate protective activity against directly acting mutagens Sodium Azide and 2-Nitrofluorene. Therefore we suggest that C. lechleri sap interacts with the enzymes of the S9 mix, thereby inhibiting the transformation of 2-Aminoantracene into its active forms.

17beta-estradiol modulates prostaglandin E2 release from human amnion-derived wish cells.

In human amnion-derived WISH cells [(3)H]estradiol-17beta binding sites are not detectable, but they become measurable in cells exposed to cAMP elevating agents such as forskolin or Ro 20-1724. In cells unexposed to these drugs, 17beta-estradiol stimulates prostaglandin (PG)E(2) release but exerts an evident inhibitory effect in cells exposed to Ro 20-1724. Both stimulatory and inhibitory actions are inhibited by the estrogen receptor antagonist, tamoxifen, by cell pretreatment with cycloheximide, or when the hormone is bound to BSA. Our data demonstrate for the first time that 1) 17beta-estradiol modulates PGE(2) release from WISH cells, interacting with specific intracellular receptors and probably evoking new protein synthesis, and 2) WISH cell responsiveness to 17beta-estradiol seems to be modulated by cAMP, whose levels are significantly increased by the steroid hormone in the presence of Ro 20-1724. The nucleotide is presumably responsible for the enhacement of hormone receptor availability and for the inhibition of PGE(2) release observed in the presence of Ro 20-1724.

Phthalic acid mimics 17beta-estradiol actions in WISH cells.

The object of this study was to evaluate whether phthalic acid, which is one of the metabolites of phthalic acid esters, exerts estrogenic actions in WISH cells, an immortalized cell line derived from human amniotic tissue. Our data demonstrate that phthalic acid (i) displaces [3H]estradiol from its binding sites, (ii) enhances the intracellular cyclic AMP concentration, without influencing adenylyl cyclase activity, (iii) stimulates or inhibits prostaglandin output, probably depending on the intracellular nucleotide level. The effects on prostanoid release are counteracted by addition of the protein-synthesis inhibitor cycloheximide, or when the diffusion of phthalic acid through the cell membrane is prevented. On the basis of our previous demonstration, that 17beta-estradiol exerts similar effects in WISH cells, we suggest that the molecular mechanisms underlying phthalic acid and steroid-hormone responses in this cell line are the same. This is the first demonstration that phthalic acid binds to the estrogen receptor with high affinity and mimics the hormone physiological actions.

Endocrine-disrupting agents on healthy human tissues.

A vast number of substances have been suggested as possibly contributing to perturbation of the endocrine system. Several have been tested with different approaches ranging from yeast expression system of human oestrogenic receptors to human breast cancer cells assays. Surprisingly, no inhibition-binding experiments to steroid receptors on healthy human tissue have been performed so far. Our study provides inhibition binding experiments to oestrogens, progesterone, testosterone and retinoic acid receptors in prostate and uterine human tissue of organochlorine pesticides, phthalate esters, oestrogenic constituents derived from plants and phenol derivates. Affinities of significant extent of phthalates on oestrogenic, progestinic and androgenic receptors have not been detected. As for retinoic acid receptors, mono(2-ethylexyl)phthalate provokes a notable reduction of the binding of the tritiated retinoic acid, phtalic acid ethyl-n-butyl ester and 4-octylphenol show an affinity comparable to that of isoflavonoid genistein, whereas 4-nonylphenol reduces the binding of retinoic acid in prostate.

Fat and water photon scattering data for in vivo lean and fatty tissue composition studies.

Photon scattering cross-section data for freshly excised and filtered liquid pig fat was measured in the interval chi = 0.02 to 0.64 A-1 (chi = E*[sin(theta/2)]/12.4; E being the photon incident energy (keV) and theta the scattering angle). The experimental results demonstrate that the marked intermolecular effects of coherent scattering in the forward direction can be exploited as a tool for characterizing lean and fatty tissue.

X-ray diffraction of bone at the interface with hydroxyapatite-coated versus uncoated metal implants.

The microstructural characteristics of the newly formed bone tissue at the interface with hydroxyapatite-coated and uncoated stainless steel pins used in an external fracture fixation system have been evaluated. The bone far from the interface was used as a control. Pins were transversally inserted into the diaphyses of sheep tibiae and were loaded in for six weeks. Three sheep received coated pins and two received uncoated pins. Crystallographic habit and mineralization of the implant-facing bone were evaluated. Moreover, lattice parameters of bone apatite were measured and hydroxyapatite (HA) coating degradation was investigated, by means of conventional and microbeam X-ray diffraction (XRD). In coated pins, six weeks after the implantation the newly formed bone tissue at the interface did not reach complete maturation, but the presence of the implant did not alter the apatite lattice structure; the lattice parameters did not show statistically significant variations with respect to those observed in the control bone. In uncoated pins, bone tissue rarely appeared totally mineralized and lattice parameters were significantly different with respect to those observed in the bone far from the implant. HA particles were observed spreading in the bone-facing coated pins; the XRD pattern of bone apatite surrounding HA particles was unmodified. It was concluded that HA coatings improved the bone remodelling process during pin fixation in comparison to uncoated pins and did not alter the crystallographic habit of apatite.

Effects of orally administered bisphosphonates on bone loss in a disuse osteopenia model involving the rat.

OBJECTIVE: This study was carried out to evaluate the effectiveness of two newly synthesized bisphosphonates (BPs) [Alendronate (4-amino-1-hydroxybutylidene-1, 1-bisphosphonic acid (AHBuBP)) and Neridronate (6-amino-1-hydroxyhexylidene-1,1-bisphosphonic acid (AHHexBP)], administered orally, in reducing experimentally induced bone loss. METHODS: Unilateral sciatic nerve section was performed on the Sprague-Dawley rat to induce osteopenia in one of the hind limbs. Histomorphometric measurements of the tibial trabecular bone and femur ash content determinations were effected to assess the degree of osteopenia. For comparison Chlodronate (dichloromethylene-1-bisphosphonic acid (Cl2MBP) was employed as the reference drug. CONCLUSIONS: The results of this investigation show that both BPs were significantly active in reducing the osteopenic process in the involved limb and were more active than Chlodronate.

High-performance liquid chromatographic determination of some of the hydrolytic decomposition products of poly(alpha-hydroxyacid)s.

An HPLC procedure is described for the separation and identification of some hydrosoluble by-products resulting from the hydrolytic degradation of poly(alpha-hydroxyacid)s having biomedical interest: poly(L-lactide), poly(DL-lactide), poly-(glycolide) and poly(lactide-co-glycolide). Peak identification was performed by comparing the respective retention times with those of pure standards. It was observed that optimum shape and separation of peaks are considerably affected by the composition of the mobile phase, consisting of acetonitrile (A) and a 0.006 M K2HPO4 buffer (B), and, in particular, its pH and A:B ratio, which had to be adjusted to around 5.8 and 75:25 (v/v), respectively. Under the investigated experimental conditions (aqueous suspension, 100 degrees C for 12 h under stirring), poly(L-lactide) is quite stable, poly(glycolide) degrades easily to glycolic acid, whereas poly(DL-lactide) and poly(DL-lactide-co-glycolide) exhibit intermediate behaviour. Upon hydrolytic decomposition, these poly(alpha-hydroxyacid)s yield not only the corresponding acids, but also their linear dimers and, possibly, trimers, tetramers and higher oligomers.

Microstructural investigation of bone-cement interface.

We evaluated the microstructural characteristics of newly formed bone tissue at the interface with cement. The bone-cement interfaces of the femoral components of nine hip prostheses retrieved after loosening were investigated by means of X-ray diffraction on microareas and microhardness. The bone far from the interface of two stable prostheses was used as a control. The newly formed bone adjacent to cement in the loosened prostheses showed a maturity degree lower than that of bone adjacent to cement in stable prostheses. The lattice parameters of bone apatite did not show significant variations as compared to the reference values. Bone trabeculae at the interface with loosened prostheses often showed an osteoid lining characterized by a strongly demineralized lamellar and haversian structure. Radioopaque cement particles are sometimes found in the trabecular bone tissue around the prosthesis.

X-ray diffraction of newly formed bone close to alumina- or hydroxyapatite-coated femoral stem.

The study was focused on the bone tissue response to two types of ceramic coating [alumina and hydroxyapatite (HA)] obtained with a plasma-spray technique. The HA coating was performed on titanium stems, while the alumina coating was on chromium-cobalt-molybdenum stems. The investigation was carried out by means of micro-area X-ray diffractometric analyses and microdiffractometric analyses at the bone-implant interface. Moreover, the coating before and after implantation was analysed by means of the conventional powder X-ray diffractometric technique. This study demonstrated that 1 yr after implantation in animals the newly formed bone adjacent to the HA-coated hip prosthesis stems was mineralized as much as the pre-existing bone, while 24-64 months after surgery in humans the newly formed bone close to the alumina-coated hip prosthesis stems showed a lamellar and Haversian structure strongly demineralized, probably due to the release of aluminum ions from the alumina covering. The authors conclude that the X-ray diffraction allows the evaluation of the structural modifications of the ceramic coating, the bone formation rate close to the coating and the chemical nature of the particles released from the coating.

Some theoretical aspects of the mechanism of action of benzhydrazone, an inhibitor of glycosylation in herpes simplex virus.

Some hypotheses of the mechanism of action of benzhydrazone, a selective inhibitor of herpes simplex virus glycoprotein biosynthesis, are presented in this paper. Comparative analysis of different models indicates that the drug may interfere with herpes simplex virus DNA. Thus, benzhydrazone seems to act like a bifunctional alkylating agent.