Paediatric head and neck malignant neoplasms: A brazilian retrospective study.

BACKGROUND: To assess the prevalence of oral and maxillofacial malignant neoplasias in children and adolescents diagnosed through biopsies sent to the Oral Pathology Laboratory at the University of Sao Paulo School of Dentistry. MATERIAL AND METHODS: A retrospective analysis of anatomopathological reports on patients between 1 and 18 years old issued by the oral and maxillofacial pathology laboratory between 1997 and 2021 was performed for demographic data, lesion site, type of biopsy, diagnostic hypothesis and final diagnosis. RESULTS: The laboratory issued 76,194 anatomopathological reports during this period, of which 10.77% were of children and adolescents. Of this total, only 32 biopsies (32/8.204; 0.39%) were neoplasias in children and adolescents. Sarcomas were the most prevalent malignant neoplasms (19/32; 59%), followed by carcinomas (7/32; 22%), lymphomas (5/32; 16%) and ganglioneuroblastomas (1/32; 3%). Of these 32 patients, the most affected individuals were aged between 4 and 11 years old (47%), 18 (56%) were male, and the mandible was the main anatomical site involved (28%). In 41% of the cases (13/32), the diagnostic hypothesis of the biopsied lesion was mistakenly considered benign and there was no diagnostic hypothesis in 18% of the cases. CONCLUSIONS: Oral and maxillofacial malignant neoplasms in children and adolescents are uncommon and the accuracy of provisional diagnoses is low in these cases. Better knowledge on oral and maxillofacial malignant lesions in this population would help professionals to reduce the diagnostic time and consequently improve the patient's prognosis.

Accuracy of CT and MRI to assess resection margins in primary malignant bone tumours having histology as the reference standard.

AIM: To evaluate the accuracy of magnetic resonance imaging (MRI) and computed tomography (CT) in assessing the resection margins of primary malignant bone tumours. MATERIALS AND METHODS: Resected primary malignant bone tumour specimens removed from 46 patients (27 male; mean age: 48±22 years) were imaged using MRI (fat-saturated proton density-weighted and three-dimensional fat-suppressed T1-weighted gradient-recalled-echo) and CT immediately after surgery. A radiologist and an orthopaedist evaluated bone and soft-tissue margins of the specimens on both examinations. Histological evaluation was performed by a senior orthopaedic oncology pathologist. Margins were classified as R0 (safe margins), R1 (residuals between 0 and 1 mm), and R2 (macroscopic residuals). Cohen's k, chi-square, and McNemar's statistics were used. RESULTS: Having histology as the reference standard, reproducibility of the radiologist ranged from moderate (k=0.544) to substantial (k=0.741) for bone and soft-tissue margins on CT, respectively, while that of the orthopaedist ranged from fair (k=0.316) to moderate (k=0.548). When comparing R2 and R0+R1 scores, reproducibility of readers' evaluation of bone margins increased ranging from substantial (k=0.655) to perfect (k=1.000). Inter-reader agreement ranged from fair (k=0.308) to substantial (k=0.633). Accuracy of the radiologist and orthopaedist ranged from 76% to 83% and from 68% to 72%, respectively. When comparing R2 and R0+R1 scores, the accuracy of both readers ranged from 83% to 100%. There was no association between local recurrence and margin scores of histology, MRI, and CT (p≥0.058). CONCLUSIONS: MRI and CT may be useful for extemporaneous analysis of resection margins of primary malignant bone tumours, although wide accuracy variability between the different imaging methods was observed.

METABOLIC BONE CHANGES IN OSTEOARTHRITIS: THE ROLE OF IMAGING AND PATHOGENETIC INTERPRETATION.

Osteoarthritis (OA) is the most prevalent chronic joint disease and one of the major causes of disability in the adult population. Although OA is considered a progressive degenerative process which involves the whole joint, articular cartilage and subchondral bone play a determinant role in its pathogenesis. In particular, metabolic-triggered subchondral bone damage, together with biochemical markers, are referred as important indicators of the disease. Magnetic resonance (MR) is the best imaging technique to detect and characterize such bone abnormalities. It represents an effective method through which to not only diagnose, describe and follow the course of OA but also to deepen our understanding of the natural history of the disease, with the ultimate purpose of attaining improved outcome in terms of therapy and prognosis. Even though MR has enormous potential, some diagnostic pitfalls may occur in clinical practice, hence an accurate clinical assessment of the patient is mandatory in combination with optimal imaging evaluation.

Phototherapy during treadmill training improves quadriceps performance in postmenopausal women.

OBJECTIVE: To evaluate the effects of infrared-light-emitting diode (LED) during treadmill training on functional performance. METHODS: Thirty postmenopausal women aged 50-60 years were randomly assigned to one of three groups and successfully completed the full study. The three groups were: (1) the LED group, which performed treadmill training associated with phototherapy (n = 10); (2) the exercise group, which carried out treadmill training only (n = 10); and (3) the sedentary group, which neither performed physical training nor underwent phototherapy (n = 10). Training was performed over a period of 6 months, twice a week for 45 min per session at 85-90% of maximal heart rate, which was obtained during progressive exercise testing. The irradiation parameters were 100 mW, 39 mW/cm(2) and 108 J/cm(2) for 45 min. Quadriceps performance was measured during isokinetic exercise testing at 60°/s and 300°/s. RESULTS: Peak torque did not differ amongst the groups. However, the results showed significantly higher values of power and total work for the LED group (∆ = 21 ± 6 W and ∆ = 634 ± 156 J, p < 0.05) when compared to both the exercise group (∆ = 13 ± 10 W and = 410 ± 270 J) and the sedentary group (∆ = 10 ± 9 W and ∆ = 357 ± 327 J). Fatigue was also significantly lower in the LED group (∆ = -7 ± 4%, p < 0.05) compared to both the exercise group (∆ = 3 ± 8%) and the sedentary group (∆ = -2 ± 6%). CONCLUSIONS: Infrared-LED during treadmill training may improve quadriceps power and reduce peripheral fatigue in postmenopausal women.

Patient dose management solution directly integrated in the RIS: "Gray Detector" software.

On X-ray modalities, the information concerning the dose delivered to the patient is usually available in image headers or in structured reports stored in the picture archiving and communication system (PACS). Sometimes this information is sent in the Modality Performed Procedure Step message. By saving the information inside the Radiological Information System, it can be linked to the patient and to his/her episode/request. A software, "Gray Detector," implementing different and complementary extraction methods was developed. Query/retrieve on images header, Modality Performed Procedure Step message analysis, or the combination of the two methods were used. In order to avoid erroneous dose-protocol association, every accession number is linked to its unique report code, allowing multiple-protocols exam recognition. The adoption of different methods to extract dosimetric information makes it possible to integrate any kind of modality in a vendor/version neutral way. Linking the dosimetric information received from a modality to the patient and to the unique report code solves, for example, common problems in computed tomography exams, where the dosimetric value related to multiple segments/studies on the modality can be associated by the technician who performs the exam only to one accession number corresponding to a single study/segment. Analyses of dosimetric indexes' dependence on modality type, patient age, technician, and radiologist were performed. Linking dosimetric information to radiological information system data allows a contextualization of the former and helps to optimize the image-quality/dose ratio, thereby making it possible to take a clinical decision that is "patient-centered."

Autotransplantation with tooth replica: Technical note.

INTRODUCTION: Dental autotransplantation is a surgical procedure that involves the implantation of a tooth with incomplete root development to replace an injured or missing tooth. An initial surgery was performed to attempt orthodontic traction without success after 6 months. TECHNIQUE: Preoperative assessment included mandibular cone beam computed tomography to visualise two canines. The double autotransplantation technique was performed under general anaesthesia using three-dimensional resin-printed dental replicas. The extra-alveolar time for each tooth was reportedly <1 min, and pulp sensibility was observed 3 months later. CONCLUSION: In the digital era, dental autotransplantation has become an essential part of orthodontic-surgical treatment in children and adolescents with dental agenesis or non-preservable teeth. The use of three-dimensional dental replicas printed from radiological data offers a reliable, reproducible, and valid therapeutic solution with an advantageous benefit/risk balance.

Effect of polyphosphates on the activity of amine oxidases.

The interaction between polyphosphates and polyamines was investigated by 31P-NMR spectroscopy and by amine oxidase activity measurements. An apparent competition between negatively charged polyphosphates (ATP, ADP, AMP, tripolyphosphate and pyrophosphate) and positively charged polyamine, for the active site of bovine serum and soybean seedling amine oxidases, was observed by activity measurements. This behavior was explained by formation of polyamine-polyphosphate complexes and the stability constants of these complexes were calculated by 31P NMR. However, at a given concentration of polyphosphate, the amine oxidase activity was found higher than that expected on the basis of the free amine concentration calculated according to the NMR stability constant. This fact, and the different extent of inhibition of the spermidine oxidase activity of soybean seedling and of bovine serum amine oxidases observed in the presence of a given polyphosphate, suggest that amine oxidases may be active also on the polyamine-polyphosphate complexes. This hypothesis was supported by the strong dependence of the kcat/Km of bovine serum amine oxidase on ionic strength, indicating an electrostatic interaction between the charged amine and the active site, while no effect of ionic strength on kcat/Km was observed in the presence of ATP. A kinetic model of this behavior was found to fit the experimental data.

beta2-microglobulin H31Y variant 3D structure highlights the protein natural propensity towards intermolecular aggregation.

beta2-Microglobulin (beta2m) is the non-covalently bound light chain of the human class I major histocompatibility complex (MHC-I). The natural turnover of MHC-I gives rise to the release of beta2m into plasmatic fluids and to its catabolism in the kidney. beta2m dissociation from the heavy chain of the complex is a severe complication in patients receiving prolonged hemodialysis. As a consequence of renal failure, the increasing beta2m concentrations can lead to deposition of the protein as amyloid fibrils. Here we characterize the His31-->Tyr human beta2m mutant, a non-natural form of beta2m that is more stable than the wild-type protein, displaying a ten-fold acceleration of the slow phase of folding. We report the 2.9A resolution crystal structure and the NMR characterization of the mutant beta2m, focussing on selected structural features and on the molecular packing observed in the crystals. Juxtaposition of the four mutant beta2m molecules contained in the crystal asymmetric unit, and specific hydrogen bonds, stabilize a compact protein assembly. Conformational heterogeneity of the four independent molecules, some of their mutual interactions and partial unpairing of the N-terminal beta-strand in one protomer are in keeping with the amyloidogenic properties displayed by the mutant beta2m.

Deuterium nuclear magnetic resonance for evaluating the metabolic status of livers subjected to warm ischemia.

BACKGROUND: The development of reliable methods for assessing the viability of currently available livers is expected to increase the number of successful transplantations. METHODS: 2 H nuclear magnetic resonance (NMR) was used to search for metabolic markers of ischemia in explanted rat livers. Deuterium oxide (2 H2O) was used as a source of 2 H. A total of 10-80% v/v 2 H2O was added to homogenates obtained from a liver biopsy and the formation of 2 H-labeled metabolites was monitored. RESULTS: Some well-resolved 2 H resonances were found in the homogenates from biopsies of warm ischemic liver. Two of these were identified as [3-2 H] lactate and [2-2 H] lactate, and a linear relationship was found between the ratio of [[2-2 H] lactate] to [[3-2 H] lactate] and the warm ischemia time. The deuterium incorporation into lactate was explained on the basis of the metabolic events occurring under hypoxic conditions. CONCLUSIONS: The experimental results support the application of 2 H NMR for a reliable evaluation of the metabolic status of a liver harvested from non-heart-beating donors.

Biphasic behavior of the kinetics of 31P-containing metabolites in ischemic porcine kidneys.

The assessment of kidney viability before transplantation (with a view of discarding nonviable organs) remains an obstacle to confidently extending organ harvesting to marginal donors. In the present study phosphorus magnetic resonance spectroscopy was used to monitor metabolic changes in (31)P-containing metabolites in isolated porcine kidneys. After various warm ischemia times, the organs were stored at 0 degrees C. Time-dependent changes in the phosphomonoester/inorganic-phosphate ratio were recorded at 0 degrees C were shown to follow a biexponential decay. The first-order kinetic rate constant of the short-time decay was strongly dependent on the warm ischemia time, a result that was discreted in terms of the underlying biochemistry. The metabolic events responsible for the dramatic decrease in phosphomonoester/inorganic phosphate ratio that occur immediately after organ perfusion and storage, suggest that any procedure to minimize organ damage must occur immediately after harvesting.

Oligomeric states along the folding pathways of ß2-microglobulin: kinetics, thermodynamics, and structure.

The transition of proteins from their soluble functional state to amyloid fibrils and aggregates is associated with the onset of several human diseases. Protein aggregation often requires some structural reshaping and the subsequent formation of intermolecular contacts. Therefore, the study of the conformation of excited protein states and their ability to form oligomers is of primary importance for understanding the molecular basis of amyloid fibril formation. Here, we investigated the oligomerization processes that occur along the folding of the amyloidogenic human protein ß2-microglobulin. The combination of real-time two-dimensional NMR data with real-time small-angle X-ray scattering measurements allowed us to derive thermodynamic and kinetic information on protein oligomerization of different conformational states populated along the folding pathways. In particular, we could demonstrate that a long-lived folding intermediate (I-state) has a higher propensity to oligomerize compared to the native state. Our data agree well with a simple five-state kinetic model that involves only monomeric and dimeric species. The dimers have an elongated shape with the dimerization interface located at the apical side of ß2-microglobulin close to Pro32, the residue that has a trans conformation in the I-state and a cis conformation in the native (N) state. Our experimental data suggest that partial unfolding in the apical half of the protein close to Pro32 leads to an excited state conformation with enhanced propensity for oligomerization. This excited state becomes more populated in the transient I-state due to the destabilization of the native conformation by the trans-Pro32 configuration.

1H,13C-NMR and X-ray absorption studies of copper(I) glutathione complexes.

The tripeptide glutathione (gamma-L-Glu-L-Cys-Gly, GSH) is an important intracellular reducing agent for Cu(II) and complexation agent for Cu(I). We have studied the complexation of Cu(I) to GSH in aqueous solution at a range of pH values and Cu(I):GSH molar ratios by 1H-NMR and 13C-NMR spectroscopy and X-ray absorption spectroscopy. The NMR data are consistent with formation of a complex with approximate 1:1 stoichiometry [Cu(SG)] as the major species with only thiolate sulfur of GSH binding to Cu(I). The rate of exchange of GSH with GS-Cu was determined to 13 s(-1) at 283 K, pH 6.8. X-ray absorption spectroscopic measurements showed that Cu(I) is coordinated to 3.1+/-0.3 sulfur atoms at approximately 0.222 nm in solutions (and solids) containing GSH:Cu in 1:1 and 2:1 mol ratios. The possible structures of polymeric Cu(I)-glutathione complexes are discussed. The high thermodynamic stability of Cu(I)-S bonds in Cu(I)-glutathione complexes coupled with their kinetic lability may provide efficient and specific pathways for the transport of copper in cells.

Effect of phosphate ion on the activity of bovine plasma amine oxidase.

The system bovine plasma amine oxidase-polyamine-phosphate ion was investigated by activity measurements and 31P NMR spectroscopy. Lineweaver-Burk plots showed that phosphate ion, under physiological conditions, is an apparent competitive inhibitor of bovine plasma amine oxidase. While NMR measurements of the T1 of 31P do not suggest the binding of phosphate to/or near the paramagnetic Cu(II) sites of bovine plasma amine oxidase, the chemical shift dependence of 31P on spermidine concentration indicates the formation of a spermidine-phosphate complex. The value of the dissociation constant of this complex was found 18.5 +/- 1.4 mM, at pH 7.2, by NMR, in good agreement with the value 17.0 +/- 0.8 mM calculated from activity measurements, assuming the enzyme activity is proportional to the free amine concentration, under second order conditions. Our data suggest that the decrease of the free spermidine, due to the binding of phosphate ion, is responsible of the observed inhibition of bovine plasma amine oxidase.

Platination of a GG site on single-stranded and double-stranded forms of a 14-base oligonucleotide with diaqua cisplatin followed by NMR and HPLC -- influence of the platinum ligands and base sequence on 5'-G versus 3'-G platination selectivity.

Detailed studies of the kinetics of platination of the single-stranded 14-base DNA oligonucleotide d(ATACATGGTACATA) and the corresponding duplex by cis-[Pt(NH3)2(H2O)2]2+ show that HPLC and NMR are complementary methods which provide similar results. The 5'-G and 3'-G monofunctional intermediates were trapped, separated and characterized by NMR (via 15NH3 labeling) and enzymatic digestion followed by mass spectrometry. The kinetic data are compared with those for the corresponding reactions of cis-[PtCl2(NH3)2] (cisplatin) and its monohydrolysed analogue. For both single and double strands of the oligonucleotide, the aqua complex shows little selectivity for the 5'-G or the 3'-G in the initial platination step, whereas the chloro-complex preferentially platinates the 3'-G. The base on the 3' side of the GG sequence appears to play an important role in controlling this selectivity; replacement of T by C increases the selectivity of duplex platination by the diaqua complex by a factor of about 6, and the selectivity of chelation of the 3'-G monofunctional adduct by a factor of about 3. In general the reactivity of the 5'-G in a GG sequence appears to be enhanced in a duplex compared with a single-strand. For both the aqua-monoadduct and chloro-monoadduct, cis-[Pt(NH3)2(N7G)(H2O or Cl)], the 5'-G monoadduct is much longer lived (t1/2 approximately 4 h at 288 K for aqua, 80 h at 298 K for chloro) than the 3'-G monoadduct (t1/2 < or = 45 min at 288 K for aqua, 6 h at 298 K for chloro). Inspection of molecular mechanics models of the end states of various monofunctional adducts provided insight into H-bonding and destacking interactions in these adducts and the sequence selectivity observed in their formation. Such adducts may play an important role in the mechanism of action of platinum anticancer drugs.

Enzyme mimics complexing Cu(II) ion: structure-function relationships.

Five peptides containing (His-X2)-His or (His-X3)-His motifs have been designed and synthesized to coordinate Cu(II). Structural information was obtained by various spectroscopic techniques and was used as constraint to search for local conformational energy minima by molecular mechanics. Thermodynamic stability constants of the Cu(II) chelates was obtained by 19F-NMR. The synthesized Cu(II)-peptide chelates were tested as catalysts of some important red-ox processes occuring in biological systems, in particular oxidation of ascorbate and dismutation of superoxide ion. The catalytic efficiency of the five chelates was much lower than that of ascorbate oxidase. On the contrary, two of them showed kinetic constants for superoxide dismutation about one order of magnitude lower than that of the enzyme Cu,Zn superoxide dismutase. In both cases, the catalytic properties were dependent on the peptide sequence. The relationships between structure and activity are discussed to find the structural parameters crucial for catalytic activity that can be modulated by appropriate design and synthesis of the peptides.

Structural transitions of a GG-platinated DNA duplex induced by pH, temperature and box A of high-mobility-group protein 1.

[1H, 15N] and 1H NMR, and CD spectroscopy are used to show that the duplex d(A-T-A-C-A-T-Pt 7G-Pt7G-T-A-C-A-T-A).d(T-A-T-G-T-A-C-C-A-T-G-T-A-T), where Pt7G is platinated guanine, containing the cis-[Pt(NH3)2]2+ adduct, undergoes reversible temperature-induced (T0.5 310 K) and pH-induced (pKa approximately 4.8) transitions between kinked-duplex and distorted forms, with the latter forms predominating at high temperature and low pH. A related pH-induced structural change was observed for the unplatinated duplex (pKa 4.69, Hill coefficient n = 1.4) but was less cooperative than for the platinated duplex (n = 2). The pH-induced transition is attributed to protonation of cytosine residues and has wider implications, since many reported NMR studies of DNA are carried out near pH 5 to minimize NH-exchange rates. The [Pt(en)]2+ (where en is 1,2-ethanediamine) GG chelate of the same duplex is shown to exist in kinked and distorted forms, and the [1H,15N]-NMR shifts for the kinked form are indicative of the presence of highly stereospecific interactions with the Pt-NH protons. On binding of the duplex platinated with [Pt(NH3)2]2+ to high-mobility-group protein 1 (HMG1) box A, similar changes in shifts of the Pt-NH3 resonances to those induced by raising the temperature or lowering the pH were observed. The specific changes in 1H-NMR chemical shifts of HMG1 box A are consistent with binding of the platinated duplex (intermediate exchange rate on the 1H-NMR time-scale) to the concave face of the protein via helices I and II and the intervening loop.