Angiogenic response induced by acellular aortic matrix in vivo

In this study, we investigated the angiogenic response induced by acellular aortic matrices implanted in vivo onto the chick embryo chorioallantoic membrane (CAM), a useful model for such investigation. Results showed that acellular matrices were able to induce a strong angiogenic response comparable to that of fibroblast growth factor 2 (FGF-2), a well-known angiogenic cytokine. The angiogenic response was further increased when exogenous FGF-2 or transforming growth factor beta 1 (TGF-beta1) were added to the matrices and inhibited by the addition of an anti-FGF-2 or anti-TGF-beta1 antibodies. The response may be considered dependent on a direct angiogenic effect exerted by the matrices and in part also by the presence of FGF-2 and TGF-beta1 in the acellular matrices.     

Echocardiography and Doppler sonography in the evaluation of cardiac structure and function

Physical activity increases the work load of the heart. The adjustments of the heart depend on the quality and quantity of the work performed. These adjustments concern the function and the morphology of the cardiovascular system. It is important to underline that these adjustments are not permanent and can disappear when physical activity is stopped. In young subjects the risks are very few while the benefits may be shown on a better and more armonic body structure. In the elderly the benefits can be achieved with a lesser cost for submaximal activities, but the risks are of course more frequent due the possible onset of cardiovascular disease. It is important to correctly recognize the limits whitin which the physical activity can be allowed because beneficial. Echocardiography has given an important contribution to evaluate the morpho-functional adaptions of the athlete's heart. Similarly, it has proven useful in the detection of pathological cardiovascular modifications, asymptomatic or pausymptomatic, that do not allow certification to practise sport at agonistic levels.     

Bleeding patterns in recent postmenopausal outpatients with uterine myomas: comparison between two regimens of HRT

Objectives: To evaluate and to compare the bleeding patterns obtained with two regimens of hormone replacement therapy given to early postmenopausal women with asymptomatic uterine leiomyomas. Methods: In this randomised prospective 1-year study 50 early postmenopausal women with one to four asymptomatic uterine leiomyomas were enrolled into two study-groups to take two regimens of hormone replacement therapy for 12 28-day cycles: (A) Tibolone, 2.5 mg/day; (B) conjugated equine estrogens (CEE), 0.625 mg/day plus medroxyprogesterone acetate (MPA), 5 mg/day. The bleeding patterns and the changes in uterine volume of the 47 outpatients who completed the study were evaluated and compared. Results: Amenorrhea incidence was higher in group A (75.0% of the cycles) than in group B (65.6% of the cycles), while irregular bleeding and irregular spotting incidences were higher in group B (29.7 and 4.7% of the cycles, respectively) compared to group A (22.6 and 2.4% of the cycles, respectively). The mean bleeding and spotting lengths were not statistically different between patients in group A and those in group B. Finally, at the end of the study period transvaginal ultrasonography showed no significant change in leiomyoma size. Conclusions: The results demonstrate that, in early postmenopausal patients with asymptomatic uterine leiomyomas, Tibolone treatment seems to be preferable compared to CEE–MPA continuous combined treatment in relation to the lesser occurrence of irregular bleeding. Furthermore, neither Tibolone nor CEE–MPA therapy, at the doses used here, promote fibroid growth.     

Composition-properties relationships in propene-ethene random copolymers obtained with high-yield Ziegler-Natta supported catalysts

The crystallization and thermal behaviour of crystallizable random propene/ethene copolymers (P-co-E) was systematically investigated. Index of crystallinity and, index of γ-form, enthalpy and entropy of fusion, equilibrium melting temperature, spherulite growth rate, and overall kinetic rate constant were determined and correlated with the overall ethene content and with the concentration of specific chemical defects as determined by ¹³C NMR analysis (PEP, EPP, EPE triads). The samples of the copolymers, obtained with very-high-yield Ziegler-Natta catalysts, were characterized by IR, ¹³C NMR, wide angle X-ray scattering, and differential scanning calorimetry.     

Reduction of capsular polysaccharide production in Klebsiella pneumoniae by sodium salicylate.

Heavily encapsulated Klebsiella pneumoniae (serotypes 1 and 2) was cultured in the presence of sodium salicylate. The addition of salicylate (2 to 30 micrograms/ml) progressively decreased the amount of capsular polysaccharide produced by all strains without significantly inhibiting cell growth. Further addition of salicylate (50 to 200 micrograms/ml) was progressively inhibitory to cell growth and decreased the production of polysaccharide only slightly. The optimal concentration of salicylate that could reduce the polysaccharide production of heavily encapsulated, virulent strains by 50% or more was 30 micrograms/ml. Mutants of these bacteria that produced less capsule were affected by salicylate to a lesser degree. All concentrations of salicylate tested were physiologically achievable in humans and within the therapeutic range of aspirin. The addition of calcium and magnesium partially reversed the effects of salicylate on polysaccharide production. Chelating agents, particularly EGTA [ethylene-bis(oxyethylenenitrile)tetraacetic acid], reduce capsule production as salicylate did. Thus, the chelation of calcium and magnesium by salicylate could account, at least in part, for the reduction of capsule. Optical density measurements allowed for rapid monitoring of capsule production in various culture media because a large part of culture turbidity was apparently due to the capsule. Decreased production of the primary K. pneumoniae virulence factor with salicylate may have therapeutic potential.     

Genotype-phenotype relationship in human ATP6i-dependent autosomal recessive osteopetrosis

Autosomal-recessive osteopetrosis is a severe genetic disease caused by osteoclast failure. Approximately 50% of the patients harbor mutations of the ATP6i gene, encoding for the osteoclast-specific a3 subunit of V-ATPase. We found inactivating ATP6i mutations in four patients, and three of these were novel. Patients shared macrocephaly, growth retardation and optic nerve alteration, osteosclerotic and endobone patterns, and high alkaline phosphatase and parathyroid hormone levels. Bone biopsies revealed primary spongiosa lined with active osteoblasts and high numbers of tartrate-resistant acid phosphatase (TRAP)-positive, a3 subunit-negative, morphologically unremarkable osteoclasts, some of which located in shallow Howship lacunae. Scarce hematopoietic cells and abundant fibrous tissue containing TRAP-positive putative osteoclast precursors were noted. In vitro osteoclasts were a3-negative, morphologically normal, with prominent clear zones and actin rings, and TRAP activity more elevated than in control patients. Podosomes, alphaVbeta3 receptor, c-Src, and PYK2 were unremarkable. Consistent with the finding in the bone biopsies, these cells excavated pits faintly stained with toluidine blue, indicating inefficient bone resorption. Bone marrow transplantation was successful in all patients, and posttransplant osteoclasts showed rescue of a3 subunit immunoreactivity.     

Enhanced Phagocytosis and Bactericidal Activity of Hepatic Reticuloendothelial System During Endotoxin Tolerance

The effects of tolerance to Escherichia coli endotoxin on the phagocytic and bactericidal activity of the hepatic reticuloendothelial system against viable E. coli were examined using ex vivo perfused rat livers. Livers were isolated from control and endotoxin-tolerant rats and perfused with a medium containing 5% homologous serum from either control or tolerant rats. After the addition of the E. coli (2 × 10⁷ cells per ml) to the perfusate, the hepatic clearance of the bacteria was followed for 30 min. The highest activation of the hepatic reticuloendothelial system was observed when serum from tolerant animals was added to the perfusate. Under these conditions phagocytosis was 47% (12% in controls), and 37 to 38% of the bacteria were killed (5% in controls). This activation was less when livers obtained from tolerant rats were perfused with serum from controls or with saline only. The data suggests that, during endotoxin tolerance, humoral factors play an important role in the activation of the hepatic reticulendothelial system, although a direct stimulation of Kupffer cells also occurs. The enhancement of phagocytosis by tolerant serum did not require the presence of homologous antibodies and involved the activation of the alternative complement pathway, since it was lost after removal of factor B activity. On the other hand, stimulation of intracellular killing required both complement and specific antibodies. The data suggest a role of endotoxin in the activation of humoral and cellular mechanisms involved in the host resistance to gram-negative bacterial infection.     

The small subset of CD56brightCD16- natural killer cells is selectively responsible for both cell proliferation and interferon-gamma production upon interaction with dendritic cells

The encounter of NK cells with dendritic cells (DC) undergoing maturation may result in the induction of NK cell proliferation. Whether such proliferation involves most NK cells or just a subset has yet to be determined. In the present study we analyzed the nature of such proliferating NK cells by combining carboxyfluorescein succinimidyl ester staining and double-fluorescence cytofluorimetric analysis. Freshly isolated peripheral blood NK cells cultured with LPS and immature DC underwent proliferation; however, proliferating cells were confined to a minor NK cell subset. This subset is characterized by the CD56(bright)CD16(-)NKG2A(+)KIR(-) surface phenotype (KIR, killer Ig-like receptor). This was further confirmed by the fact that, after cell sorting, only the CD56(bright) NK cells were able to proliferate in response to the DC stimulus, whereas the CD56(dull) were not. We also provide evidence that the CD56(bright) subset is the main source of IFN-gamma-producing NK cells, upon interaction with DC. The CD56(bright)CD16(-) NK cells express a panel of surface molecules including CD62L, CCR7 and CXCR3 that may allow their homing either to secondary lymphoid compartments or to inflamed tissues. This implies that, in vivo, the interactions between DC undergoing maturation and CD56(bright) NK cells may occur in different tissues and have different functional implications.     

Use of the Roche LightCycler instrument in a real-time PCR for Trichomonas vaginalis in urine samples from females and males

Trichomonas vaginalis is the agent of a highly prevalent sexually transmitted infection (STI) that can result in vaginitis, urethritis, and preterm birth. Traditional methods of diagnosis, including wet preparation, can be unreliable. In this study, we describe the adaptation of an existing PCR method for specific detection of T. vaginalis DNA into a rapid real-time PCR assay based on fluorescence resonance energy transfer (FRET) probe chemistry. The FRET-based assay described demonstrated high sensitivity with a detection limit of 1.06 organisms, as well as a high specificity. A total of 253 urine samples collected prospectively from both men and women were tested for T. vaginalis DNA with both the FRET-based assay and a previously validated PCR assay. When the validated PCR assay was used as the "gold standard" and after discrepancies had been resolved, our FRET-based assay demonstrated an analytical sensitivity and specificity of 90.1 and 100%, respectively. Overall results suggest that FRET-based assays can provide rapid, accurate, and high-throughput detection of T. vaginalis and may prove useful in clinical settings and for large-scale screening programs.