Loredana Cerroni

Simultaneous separation of malondialdehyde, ascorbic acid, and adenine nucleotide derivatives from biological samples by ion-pairing high-performance liquid chromatography

A method for a simultaneous separation of malondialdehyde (MDA), ascorbic acid and adenine nucleotide derivatives in biological samples by ion-pairing high-performance liquid chromatography is presented. The separation is obtained by an LC-18-T 15 cm x 4.6 mm 3 microns particle size column using tetrabutylammonium as the pairing ion. The starting buffer consists of 10 mM tetrabutylammonium hydroxide, 10 mM KH2PO4 plus 1% methanol, pH 7.00. A step gradient is formed using a second buffer consisting of 2.8 mM tetrabutylammonium hydroxide, 100 mM KH2PO4 plus 30% methanol, pH 5.5. Under these chromatographic conditions a highly resolved separation of MDA, ATP, ADP, AMP, adenosine, ascorbic acid, GTP, GDP, IMP, inosine, Hypoxanthine, Xanthine, uric acid, NAD, and NADP can be performed in about 36 min. In addition, the separation of NADH and NADPH can also be obtained; this renders the present method suitable for the detection of these reduced coenzymes in alkaline extracts from tissue samples. Data referring to PCA extracts from ischemic and reperfused isolated rat hearts and from human erythrocytes peroxidized in vitro by a challenge with 1 mM NaN3 and various concentrations of H2O2 are reported. The relevance of this chromatographic method lies in the possibility to determine directly MDA concentrations avoiding the unspecific thiobarbituric acid colorimetric test, any other manipulation of the sample out of the PCA extraction, and any possible coelution of other acid soluble compounds. The simultaneous determination of MDA, ascorbic acid, and of ATP and its degradation products gives the opportunity to correlate, by a single chromatographic run, peroxidative damages with the energy state of the cell which is of great importance in studies of ischemic and reperfused tissues.

A comparative study of smear layer removal and erosion in apical intraradicular dentine with three irrigating solutions: a scanning electron microscopy evaluation.

INTRODUCTION This study compared the efficacy of BioPure MTAD (Dentsply Tulsa, Tulsa, OK), 17% EDTA, and 42% citric acid in endodontic smear layer removal and degree of erosion in the apical third of endodontic canals. METHODS Ninety-six extracted single-rooted human teeth were randomized into four groups (n = 24) and instrumented using System GT nickel-titanium rotary instruments (Dentsply Tulsa, Tulsa, OK). Each canal was irrigated with one of the following solutions: BioPure MTAD, 17% EDTA, 42% citric acid, or 5.25% NaOCl (control). Next, all specimens were irrigated with 5.25% NaOCl. RESULTS Evaluation by scanning electron microscopy showed no significant differences among test irrigants in removing the smear layer. However, the efficacy of BioPure MTAD and 17% EDTA in removing the smear layer was significantly greater than 5.25% NaOCl (control). The erosive effects of irrigating solutions could not be evaluated. CONCLUSIONS In conclusion, the protocols used in this study were not sufficient to completely remove the smear layer in the apical third of prepared root canals.

Preserving effect of fructose-1,6-bisphosphate on high-energy phosphate compounds during anoxia and reperfusion in isolated langendorff-perfused rat hearts

Isolated Langendorff-perfused rat hearts after 10 min pre-perfusion, were subjected to a substrate-free anoxic perfusion (20 min) followed by 20 min reperfusion with a glucose-containing oxygen-balanced medium. A similar experimental protocol was repeated in the presence either of 5 mM fructose or of 5 mM fructose-1,6-bisphosphate throughout the different perfusion conditions. High-energy phosphate compounds (adenosine triphosphate, creatine phosphate), adenine nucleotides, nicotinic coenzymes, lactate, pyruvate and glycogen content in the tissue were determined at the end of each perfusion period, while coronary flow, heart rate and lactate and pyruvate output were monitored throughout the whole duration of the experiments. On the whole, the results indicate that exogenous fructose-1,6-bisphosphate preserves high-energy metabolites during anoxia and restores myocardial metabolism and contractility during reperfusion, that a prolonged period of substrate-free anoxic perfusion renders the heart unable to normalize its metabolism during re-oxygenation and that fructose is not utilized by the heart for its energy demand. A possible hypothesis concerning the mechanism of action of fructose-1,6-bisphosphate is presented.

Smear layer removal and canal cleanliness using different irrigation systems (EndoActivator, EndoVac, and passive ultrasonic irrigation): field emission scanning electron microscopic evaluation in an in vitro study.

INTRODUCTION The purpose of this study was to evaluate the effectiveness of different irrigating methods in removing the smear layer at 1, 3, 5, and 8 mm from the apex of endodontic canals. METHODS Sixty-five extracted single-rooted human mandibular premolars were decoronated to a standardized length of 16 mm. Specimens were shaped to ProTaper F4 (Dentsply Maillefer, Ballaigues, Switzerland) and irrigated with 5.25% NaOCl at 37°C. Teeth were divided into 5 groups (2 control groups [n = 10] and 3 test groups [n = 15]) according to the final irrigant activation/delivering technique (ie, sonic irrigation, passive ultrasonic irrigation [PUI], or apical negative pressure). Root canals were then split longitudinally and observed by field emission scanning electron microscopy. The presence of debris and a smear layer at 1, 3, 5, and 8 mm from the apex was evaluated. Scores were analyzed by Kruskal-Wallis and Mann-Whitney U tests. RESULTS The EndoActivator System (Dentsply Tulsa Dental Specialties, Tulsa, OK) was significantly more efficient than PUI and the control groups in removing the smear layer at 3, 5, and 8 mm from the apex. The EndoVac System (Discus Dental, Culver City, CA) removed statistically significantly more smear layer than all groups at 1, 3, 5, and 8 mm from the apex. At 5 and 8 mm from the apex, PUI and the EndoVac did not differ statistically significantly, but both performed statistically better than the control groups. CONCLUSIONS In our study, none of the activation/delivery systems completely removed the smear layer from the endodontic dentine walls; nevertheless, the EndoActivator and EndoVac showed the best results at 3, 5, and 8 mm (EndoActivator) and 1, 3, 5, and 8 mm (EndoVac) from the apex.

Oxygen radical injury and loss of high-energy compounds in anoxic and reperfused rat heart: Prevention by exogenous fructose-1,6-bisphosphate

Isolated Langendorff-perfused rat hearts after 10 minutes preperfusion, were subjected to a substrate-free anoxic perfusion (20 minutes) followed by 20 minutes reperfusion with a glucose-containing oxygen-balanced medium. Under the same perfusion conditions, the effect of exogenous 5mM fructose-1,6-bisphosphate has been investigated. The xanthine dehydrogenase to xanthine oxidase ratio, concentrations of high-energy phosphates and of TBA-reactive material (TBARS) were determined at the end of each perfusion period in both control and fructose-1,6-bisphosphate-treated hearts. Results indicate that anoxia induces the irreversible transformation of xanthine dehydrogenase into oxidase as a consequence of the sharp decrease of the myocardial energy metabolism. This finding is supported by the protective effect exerted by exogenous fructose-1,6-bisphosphate which is able to maintain the correct xanthine dehydrogenase/oxidase ratio by preventing the depletion of phosphorylated compounds during anoxia. Moreover, in control hearts, the release of lactate dehydrogenase during reperfusion, is paralleled by a 50% increase in the concentration of tissue TBARS. On the contrary, in fructose-1,6-bisphosphate-treated hearts this concentration does not significantly change after reoxygenation, while a slight but significant increase of lactate dehydrogenase activity in the perfusates is observed. On the whole these data indicate a direct contribution of oxygen-derived free radicals to the worsening of post-anoxic hearts. A hypothesis on the mechanism of action of fructose-1,6-bisphosphate in anoxic and reperfused rat heart and its possible application in the clinical therapy of myocardial infarction are presented.

Reduced sensitivity of O2 transport to allosteric effectors and temperature in loggerhead sea turtle hemoglobin: functional and spectroscopic study

The functional and spectroscopic (EPR and absorbance) properties of the adult loggerhead sea turtle (Caretta caretta) hemoglobin have been studied with special reference to the action of allosteric effectors and temperature. Present results indicate that turtle Hb displays a very low O2 affinity and a very small sensitivity to allosteric effectors and temperature. Furthermore, the amplitude of the Bohr effect for O2 binding is strongly reduced. In parallel, EPR and absorbance spectroscopic properties of the nitrosylated derivative of turtle Hb suggest that the hemoprotein is in a low-affinity conformation, even in the absence of allosteric effectors. These findings suggest the existence of unusual molecular mechanisms modulating the basic reaction of Hb with O2, which may be linked to specific physiological needs related to the diving behavior of the turtle.

Selective Binding of Met-Hemoglobin to Erythrocytic Membrane: A Possible Involvement in Red Blood Cell Aging

It is well known that in vivo and under normal physiological conditions intraerythrocytic hemoglobin may exist in three different forms represented by oxygenated, deoxygenated and partially oxidized hemoglobin (1–4). Apart from the first two derivatives whose relative proportions are continuously changing during the oxygenation deoxygenation cycle, met-hemoglobin is normally present at a steady-state level of about 1%.

Periodontal bacteria in the genital tract: are they related to adverse pregnancy outcome?

One of the most important factors implicated in preterm birth (PTB) is acute genitourinary tract infection. The bacteria causing chronic periodontal inflammation include Gram-negative rods and anaerobes similar to those found in women with bacterial vaginosis. The aim of this prospective study is to investigate the relationship between oral and vaginal microflora and preterm low birth weight. Real-time polymerase chain reaction was used to detect both the presence and level of six periodontitis-related species: Aggregatibacter actinomycetemcomitans (Aa), Porphyromonas gingivalis (Pg), Tannerella forsythia (Tf), Treponema denticola (Td), Fusobacterium nucleatum ssp (Fn), and Prevotella intermedia (Pi) for both oral samples of subgingival plaque and cervical samples, obtained from 80 patients, during gynaecological examinations. The more representative oral pathogen (>60%) species in oral samples of preterm and term group were Tf, Td, and Fn. 24.4% of pregnant women presented periodontal pathogens in vaginal swab; the most representative species with a percentage over 0.1% of total bacteria in genital tract of preterm group were Tf, Td, and Pi with a positive correlation (>0.5). The presence of the bacterium T. denticola in the vagina, regardless of the amount, adversely affects preterm delivery.

Root canal irrigation: chemical agents and plant extracts against Enterococcus faecalis

Background: There are various microorganisms related to intra and extra-radicular infections and many of these are involved in persistent infections. Bacterial elimination from the root canal is achieved by means of the mechanical action of instruments and irrigation as well as the antibacterial effects of the irrigating solutions. Enterococcus faecalis can frequently be isolated from root canals in cases of failed root canal treatments. Antimicrobial agents have often been developed and optimized for their activity against endodontic bacteria. An ideal root canal irrigant should be biocompatible, because of its close contact with the periodontal tissues during endodontic treatment. Sodium hypoclorite (NaOCl) is one of the most widely recommended and used endodontic irrigants but it is highly toxic to periapical tissues. Objectives: To analyze the literature on the chemotherapeutic agent and plant extracts studied as root canal irrigants. In particularly, the study is focused on their effect on Enterococcus faecalis. Method: Literature search was performed electronically in PubMed (PubMed Central, MEDLINE) for articles published in English from 1982 to April 2015. The searched keywords were “endodontic irrigants” and “Enterococcus faecalis” and “essential oil” and “plant extracts”. Results: Many of the studied chemotherapeutic agents and plant extracts have shown promising results in vitro. Conclusion: Some of the considered phytotherapic substances, could be a potential alternative to NaOCl for the biomechanical treatment of the endodontic space.

Autogenous Tooth Fragment Adhesive Reattachment for a Complicated Crown Root Fracture: Two Interdisciplinary Case Reports

Trauma of anterior teeth is quite a common occurrence in both children and adults. Various degrees of trauma leading to fracture may affect teeth in different ways depending on the age of the patient and extent of fracture and other factors that will be discussed. Guidelines have been given as to how each of these situations should be treated. In the past, often more aggressive restorations were performed to restore fractured teeth. However improved and more efficient adhesion may affect the type of treatment we decide to carry out, leading to more conservative therapies through an increased preservation of tooth structures.