Polydeoxyribonucleotides and nitric oxide release from guinea-pig hearts during ischaemia and reperfusion.

1. Two polydeoxyribonucleotides, produced by the controlled hydrolysis of DNA of mammalian lung (defibrotide and its lower molecular weight fraction, P.O. 085 DV), were studied for their ability to modify the release of nitrite and the coronary flow in perfusates collected from isolated, normally perfused hearts of guinea-pigs and from hearts subjected to regional ischaemia and reperfusion. 2. In guinea-pig normally perfused hearts, both defibrotide (DFT) and its fraction, P.O. 085 DV, increase the amount of nitrite appearing in perfusates in a concentration-dependent fashion. At the highest concentration studied (10(-6) M), P.O. 085 DV was more effective than DFT. A concomitant increase in the coronary flow was observed. 3. The increase in nitrite in perfusates and the increase in coronary flow induced by both DFT and P.O. 085 DV were significantly reduced by NG-monomethyl-L-arginine (L-NMMA, 10(-4) M), an inhibitor of nitric oxide synthase (NOS). 4. The endothelium-dependent vasodilator, acetylcholine (ACh), enhances the formation of nitrite and the coronary flow. Both the increase in coronary flow and in the formation of nitrite were significantly reduced by L-NMMA (10(-4) M). 5. In guinea-pig hearts subjected to ischaemia and reperfusion, the effect of both compounds in increasing the amount of nitrite in perfusates was more evident and more pronounced with P.O. 085 DV. 6. Reperfusion-induced arrhythmias were significantly reduced by both compounds to the extent of complete protection afforded by compound P.O. 085 DV. 7. The cardioprotective and antiarrhythmic effects of DFT and P.O. 085 DV are discussed.     

Nitric oxide modulates cardiac and mast cell anaphylaxis

Anaphylaxis in the isolated guinea-pig heart was associated with a sudden release of histamine with a long-lasting release of nitrite (NO ₂ ^– ), an oxidation product of NO.N ^G-monomethyl-l-arginine (MeArg, 300 M) increased the severity of cardiac anaphylaxis, as shown by the decrease in the coronary flow and by a prolonged duration of antigen-induced arrhythmias. Concomitantly, MeArg increased the release of histamine while decreasing the release of nitrite. Sodium nitroprusside (NaNP, 10^–5–10^–4 M) reduced the severity of cardiac anaphylaxis by increasing coronary flow and shortening the duration of antigen-induced arrhythmias. Concomitantly, NaNP decreased the release of histamine while increasing the release of nitrite. In mast cells isolated from actively sensitized guinea-pigs, the release of histamine elicited by specific antigen was increased by MeArg and decreased by NaNP.In conclusion, endogenous and exogenous NO antagonizes the effect of vasoconstrictor mediators released after antigen challenge and plays a protective role in anaphylactic reactions in vitro,     

Structural and functional macrophages alterations by ceramics of different composition

Biomaterials may initiate several and complex biological reactions in host tissues, and the cell-biomaterial interactions can determine the release of mediators including monocytes and lymphocytes chemotactic factors. The present work was aimed to investigate in vitro the macrophage biological reactions of a natural apatite obtained by heat treatment at 400 degrees C of bovine bone, compared to other ceramics usually used for dental and orthopedic applications, using synthetic apatite and three types of alumina as controls. Particles chemotactic activity and powders oxidative burst evidenced no monocyte macrophages sensitivity reaction for natural and synthetic hydroxyapatite powders at great granulometry (> 50 microm); data were confirmed by ultrastructural observations; electron microscopy analysis showed macrophages with the features of healthy cells in the presence of both natural and synthetic apatites while macrophages grown in the presence of alumina seemed to be negatively affected. In conclusion, among all ceramics tested, natural apatite displayed a good compatibility with living cells, being better tolerated than synthetic hydroxyapatite which in turn is better tolerated than alumina.     

Scanning electron microscopy (SEM) and X-ray dispersive spectrometry evaluation of direct laser metal sintering surface and human bone interface: a case series

Recent studies have shown that direct laser metal sintering (DLMS) produces structures with complex geometry and consequently that allow better osteoconductive properties. The aim of this patient report was to evaluate the early bone response to DLMS implant surface retrieved from human jaws. Four experimental DLMS implants were inserted in the posterior mandible of four patients during conventional dental implant surgery. After 8 weeks, the micro-implants and the surrounding tissue were removed and prepared for scanning electron microscopy (SEM) and histomorphometric analysis to evaluate the bone–implant interface. The SEM and EDX evaluations showed a newly formed tissue composed of calcium and phosphorus. The bone-to-implant contact presented a mean of 60.5 ± 11.6%. Within the limits of this patient report, data suggest that the DLMS surfaces presented a close contact with the human bone after a healing period of 8 weeks.     

Insulin resistance and glomerular hemodynamics in essential hypertension

BACKGROUND: Arterial hypertension is an important cause of end-stage renal failure. Insulin has been shown to modify glomerular hemodynamics in hypertensive subjects. The aim of this work, therefore, was to observe the relationships between renal hemodynamics and insulin resistance in arterial hypertension. METHODS: Sixty-two non-diabetic hypertensive patients and 25 healthy normal subjects were studied. Renal plasma flow and the glomerular filtration fraction were determined by renoscintigraphy and the insulin sensitivity by an oral glucose test. RESULTS: Renal plasma flow in hypertensive subjects was lower than expected and was related to pressure values, whereas the mean glomerular filtration rates were not different in the two groups. In most patients the filtration fraction was higher than expected. A lower glomerular filtration rate and lower filtration fraction were found in patients with higher insulin resistance. CONCLUSIONS: The progressive decrease of glomerular function in subjects with hypertension is linked with insulin-resistance.     

Structural aspects of the extracellular matrix of the tendon: an atomic force and scanning electron microscopy study

The mutual interactions of small proteoglycans with collagen fibrils in the extracellular matrix remain to be completely understood. The present research investigated the extracellular matrix of the rat tail tendon by atomic force microscopy (AFM) as well as by scanning electron microscopy (SEM). Observations showed simply dehydrated specimens made of large heterogeneous fibrils, tightly packed in mutual contact with no visible interfibrillar spaces. Proteoglycans usually extended onto neighboring fibrils, forming an intricate interfibrillar weaving highly sensitive to chondroitinase digestion. Pre-treatment with cupromeronic blue only affected the proteoglycans side chains, which appeared better preserved but somewhat thickened. Observation of hydrated specimens by AFM confirmed the close packing of collagen fibrils and the abundance of collagen-bound proteoglycans. Interfibrillar bridges were only occasionally observed in this tissue, whose fibrils are instead tightly bound together by proteoglycans in a structure quite consistent with its functional requirements. The molecular machinery responsible for these interactions is the subject of ongoing research.     

Efficacy of immunization against bovine mastitis using a Staphylococcus aureus avirulent mutant vaccine

Bovine mastitis is responsible of major economic losses on dairy farms worldwide. In Argentine dairy herds, Staphylococcus aureus is the main causative agent of the disease.     

Two distinct types of depolarizing afterpotentials are differentially expressed in stellate and pyramidal‐like neurons of entorhinal‐cortex layer II

Two types of principal neurons, stellate cells and pyramidal‐like cells, are found in medial entorhinal‐cortex (mEC) layer II, and are believed to represent two distinct channels of information processing and transmission in the entorhinal cortex–hippocampus network. In this study, we found that depolarizing afterpotentials (DAPs) that follow single action potentials (APs) evoked from various levels of holding membrane voltage (V_h) show distinct properties in the two cells types. In both, an evident DAP followed the AP at near‐threshold V_h levels, and was accompanied by an enhancement of excitability and spike‐timing precision. This DAP was sensitive to voltage‐gated Na⁺‐channel block with TTx, but not to partial removal of extracellular Ca²⁺. Application of 5‐μM anandamide, which inhibited the resurgent and persistent Na⁺‐current components in a relatively selective way, significantly reduced the amplitude of this particular DAP while exerting poor effects on the foregoing AP. In the presence of background hyperpolarization, DAPs showed an opposite behavior in the two cell types, as in stellate cells they became even more prominent, whereas in pyramidal‐like cells their amplitude was markedly reduced. The DAP observed in stellate cells under this condition was strongly inhibited by partial extracellular‐Ca²⁺ removal, and was sensitive to the low‐voltage‐activated Ca²⁺‐channel blocker, NNC55‐0396. This Ca²⁺ dependence was not observed in the residual DAP evoked in pyramidal‐like cells from likewise negative V_h levels. These results demonstrate that two distinct mechanism of DAP generation operate in mEC layer‐II neurons, one Na⁺‐dependent and active at near‐threshold V_h levels in both stellate and‐pyramidal‐like cells, the other Ca²⁺‐dependent and only expressed by stellate cells in the presence of background membrane hyperpolarization. © 2015 Wiley Periodicals, Inc.     

Characterization of Staphylococcus aureus strains isolated from humans in Argentina

Humans are a natural reservoir of Staphylococcus aureus and asymptomatic colonization is far more common than infection. The aim of this work was to characterize genotypically 68 S. aureus strains isolated from nasal swabs of healthy people and from human clinical infections. A total of fourteen (20%) strains were susceptible to all the antimicrobials tested. The strains isolated from nasal swabs showed the lowest percentages of resistance. Resistance to one or more than one antibiotics tested was detected in 83% and 70% of the S. aureus strains isolated from clinical infections and nasal swabs, respectively. All of the 68 S. aureus strains were subject to RAPD-PCR analysis. Cluster A-I grouped 42 (87%) clinical infection strains and cluster A-II grouped 13 (65%) strains isolated from nasal swabs suggesting a genetic relationship among S. aureus strains. Cluster A-II grouped 65% of the S. aureus strains associated with the anterior nares, suggesting that these strains may be adapted to this site. Furthermore, five RAPD profiles isolated from nasal swabs, belonged to clusters B to F, were similar to strains isolated from clinical infection, suggesting that they might have a high propensity to cause disease. The results of the present study allow a characterization of S. aureus strains isolated from humans and shows that some S. aureus genotypes from nasal swabs are similar to the genotypes obtained from clinical infections, suggesting that clinical isolates may be originated from human normal flora.