Nanomechanical properties of kidney stones, gallstones and oral stones compared with tap water scale by depth sensing indentation.

This article focuses on a description of research performed to identify structural and mechanical properties differences between calculi in stones, such as gallstones, kidney stones, dental tartar, and saliva gland sialolite, were analyzed and compared with tap water stone, in order to set interrelations. In this study, biological hard pebble-like structures were analyzed and compared among them using Scanning Electron Microscopy (SEM), X-Ray diffraction (XRD), and Atomic Force Microscopy (AFM). In addition, Nanoindentation was used to obtain values as example in kidney stones the in; stiffness S = 27,827 ± 620 N/nm elastic modulus E = 27.3 ± 4.5 GPa, hardness H = 1.5 ± 0.5 GPa. Samples with the highest amounts of calcium and magnesium oxides were; Tap water stone (39.60%), followed by dental tartar (39.40%), saliva gland sialolite (29.20%), kidney stones (27.70%), and lastly the gallstones (0.30%). Kidney stones showed in particular, whewellite and kaoulinite crystallographic phases, that confers characteristics of greater crystallization with respect to the other stones. Kidney stones positioned in the major hardness stone in human body with 1.5 GPa. In general, samples with the highest amount of calcium oxides, also showed the highest mechanical properties of H and E. Microstructural characteristics and nano-hardness of tap water stone from drinking water where similar to those of dental tartar and saliva gland sialolite, more research still required to associate health concerns and tap water scale derived from drinking water known as hardwater.

Comparative Evaluation of Bond Strength of Self-Etching and Total Etch Dental Adhesive Systems to Dentin of Deciduous and Permanent Teeth / Evaluación Comparativa de la Fuerza de Unión de los Sistemas de Adhesivo Dental Autograbado y Grabado Total para la Dentina de Dientes Deciduos y Permanentes

ABSTRACT: The aim of this study was to analyze the bond strength of total-etch and self-etch adhesive systems to dentin of primary and permanent teeth. Methods: Thirty sound naturally exfoliated primary molars deciduous teeth (DT) and thirty sound permanent bicuspids permanent teeth (PT) were randomly divided into six groups (n=10 per group) according to two commercial adhesive systems: Adper Single Bond 2; 3M ESPE (Total-etch) and Adper Easy Bond; 3M ESPE (selfetch and total-etch). Specimens submitted to cyclic loading in a universal Instron testing machine. Bond strength values (MPa) were analyzed by ANOVA test and Duncan post hoc test (a=0.05). Results: Mean values were higher in PT compared to DT. In deciduous teeth, no significantly differences observed. Total etch AdperTM Single Bond 2 showed significantly higher bond strength than self-etch AdperTM with additional acid etching in PT (p=0.031). Conclusion: Our findings suggest that the highest bond strength was found in dentin tissue of PT with total etch AdperTM employing the adhesive the Single Bond 2 of one step self-etch.

RESUMEN: El objetivo de este estudio fue analizar la fuerza de unión de los sistemas adhesivos de grabado total y autograbado a la dentina de los dientes primarios y permanentes. Métodos: treinta sonidos exfoliaron naturalmente los molares primarios dientes caducifolios (DT) y treinta sonidos. Los dientes permanentes de los premolares permanentes (PT) se dividieron aleatoriamente en seis grupos (n = 10 por grupo) de acuerdo con dos sistemas adhesivos comerciales: Adper Single Bond 2; 3M ESPE (Grabado total) y Adper Easy Bond; 3M ESPE (autograbado y grabado total). Muestras sometidas a carga cíclica en una máquina universal de pruebas Instron. Los valores de resistencia de la unión (MPa) se analizaron mediante la prueba ANOVA y la prueba post hoc de Duncan (a = 0.05). Resultados: Los valores medios fueron mayores en PT en comparación con DT. En dientes deciduos, no se observaron diferencias significativas. Total etch AdperTM Single Bond 2 mostró una fuerza de unión significativamente mayor que la autograbado AdperTM con grabado ácido adicional en PT (p = 0.031). Conclusión: Nuestros hallazgos sugieren que la mayor fuerza de unión se encontró en el tejido de dentina de PT con grabado total AdperTM empleando el adhesivo Single Bond 2 de autograbado de un solo paso.

Synthetic and natural protease inhibitors provide insights into parasite development, virulence and pathogenesis.

Protease function is essential to many biological systems and processes. In parasites, proteases are essential for host tissue degradation, immune evasion, and nutrition acquisition. Helminths (worms) depend on several classes of proteases for development, host tissue invasion and migration, and for degradation of host hemoglobin and serum proteins. The protozoa, which cause malaria, depend on both cysteine and aspartic proteases to initiate host hemoglobin digestion. Other types of proteases are involved in erythrocyte cell invasion and cell exit. Surface metalloproteases in kinetoplastids are implicated in the evasion of complement-mediated cell lysis and cell entry. Cysteine proteases in Entamoeba facilitate invasion of the host colon. Giardia utilizes a cysteine protease for both encystation and excystation. This review will summarize published data using protease inhibitors as tools to identify the function of parasite proteases in the development, virulence, and pathogenesis of parasites; as well as the role of endogenous parasite protease inhibitors in regulation.

Attention to rhotacism language problem by oral surgery and vibrostimulatory therapy: a case report / Tratamiento para el problema del lenguaje de rotacismo mediante cirugía oral y terapia vibroestimulatoria: reporte de un caso

Ankyloglossia may prevent the tongue from contacting the anterior palate, which promotes an infantile swallow and hamper the progression to an adult-like swallow. This can result in an open bite deformity and in some cases it can be associated to mandibular prognathism and language problems like rhotacism, described as the inability or difficulty in pronouncing the sound / r /. The surgical cut of the frenum tissue that connects the tongue to the jawbone (frenulectomy) and the language rehabilitation treatment with functional oral devices, represent an alternative treatment for this problem. An 11-year-old boy reported with language performance problems, dental malposition and diagnostic of ankyloglossia condition, received frenulectomy surgery and language rehabilitation treatment with functional oral devices during six months. Language diagnosis was carried out in three periods of time: prior to the surgery, four weeks after the surgery and six months later. Combined surgical and functional therapies proved to be a better alternative than only surgical therapy. Combined therapies increased the speech abilities as well as swallowing functions, therefore, the patient's self-esteem.

La anquiloglosia puede evitar que la lengua entre en contacto con la región anterior del paladar, lo que promueve una deglución infantil y dificultan la adecuada deglusión en el adulto. Esto también puede dar lugar a una mordida abierta y en algunos casos, estar asociada con prognatismo mandibular y problemas de lenguaje como el rotacismo, que se describen como la incapacidad o dificultad para pronunciar el sonido / r /. La exsición quirúrgica del tejido que conecta frenillo de la lengua a la mandíbula (frenectomía) y el tratamiento rehabilitador del lenguaje con dispositivos orales funcionales, representan una alternativa de tratamiento para este problema. Un niño de 11 años de edad, con problemas de lenguaje, malposición dental y diagnóstico de anquiloglosia, fue sometido a frenectomía y tratamiento de rehabilitación dellenguaje mediante dispositivos orales funcionales durante seis meses. El diagnóstico del lenguaje se llevó a cabo en tres periodos de tiempo: antes de la cirugía, cuatro semanas después de la cirugía y seis meses más tarde. La combinación de tratamiento quirúrgico y funcionales demostraron ser una alternativa mejor que la terapia quirúrgica por sí sola. Las terapias combinadas aumentaron la capacidad del habla, así como funciones de deglución, por lo tanto, la autoestima del paciente.

Al(2)O(3(w))-Al(2)O(3(n))-ZrO(2) (TZ-3Y)(n) multi-scale nanocomposite: an alternative for different dental applications?

The influence of the addition of Al(2)O(3) whiskers (2.5wt.% up to 30wt.%) on Vickers hardness and fracture toughness in an Al(2)O(3(n))+ZrO(2) (TZ-3Y)(n) (90, 80 and 70wt.%) composite was investigated. Green compacts were obtained by uniaxial pressing at 50MPa and pressureless sintering at 1500 degrees C in air for 2h. After sintering, relative densities ranging from 75% to 97% were reached. The whiskers resisted particle rearrangement owing to the extensive sliding distances along the whisker boundaries during sintering and the high length/diameter ratios. Sintering becomes more difficult with increasing whisker content, because whiskers come into contact with each other, forming a rigid network which hinders densification. The 2.5wt.% Al(2)O(3) whiskers+27.5wt.% Al(2)O(3) nanoparticles+70wt.% TZ-3Y composite showed a hardness>13GPa and a maximum fracture toughness of 6.9MPam(-1/2), with an average grain size of 0.4+/-0.17microm. The observed crack deflection was an important mechanism in the improved fracture toughness of the composite. In addition, the grain size and residual porosity also seem to be factors in obtaining a wide range of hardness as well as fracture toughness by varying the Al(2)O(3) whiskers and ZrO(2) (TZ-3Y) content. The use of alumina-whisker-reinforced composites in dental applications could be promising for increasing hardness and fracture toughness compared with other materials. The reported values for these composites can compete with those of commercially available materials in different dental applications.

Isoxazolo[3,4-d]pyridazinones and analogues as Leishmania mexicana PDE inhibitors.

A series of isoxazolopyridazinones and analogues has been prepared and evaluated as Leishmania mexicana phosphodiesterase (PDE) inhibitors. Some of the synthesized compounds showed a moderate PDE inhibitory activity at 100 microM and preliminary structure-activity relationships were discussed.

Characterization of cyclic AMP phosphodiesterases in Leishmania mexicana and purification of a soluble form.

The cyclic AMP phosphodiesterase (PDE) activity in Leishmania mexicana is mainly located (>95%) in the soluble fraction of the cell. The intact parasite, as well as plasma membranes, showed PDE activity, probably indicating that at least part of the activity in the particulate fraction resides on the parasite cell surface, with its catalytic domain facing the extracellular moiety. For the first time, a highly specific cAMP phosphodiesterase (PDE) was purified from the soluble fraction to apparent homogeneity after a single step 2239-fold purification using pseudo-affinity chromatography on Cibacron Blue 3GA agarose. The enzyme was identified as a 61-kDa protein on SDS-PAGE, with a K(m) of 277 microM at 30 degrees C (optimum temperature). The native enzyme protein showed an apparent molecular size of approximately 200000 estimated by molecular sieve chromatography on Sephacryl S-300. Further characterization of the PDE activity present in the soluble fraction shows that the enzyme requires Mg(2+) for maximal activity. Furthermore, no activity was detected when assayed at pHs below 6.0, but above this value it increased dramatically, reaching the optimum at pH 7.2. On the basis of the K(m) and PDE activity in presence of specific drugs or modulators such as rolipram, OPC-3911, cGMP, IBMX, zaprinast, theophylline, caffeine and Ca(2+)/calmodulin, this enzyme does not seem to conform to any of the ten previously described Class I PDE families but to the PDE class II (or non-mammalian PDEs) similar to the those found in Candida albicans, Dictyostelium discoideum, Saccharomyces cerevisiae or Vibrio fischeri.

Possible relationship between neurocysticercosis and hematological malignancies.

BACKGROUND: Previous studies have shown an increased frequency of chromosomal abnormalities in lymphocytes from animals and humans with cysticercosis. Some reports have suggested an association between cancer and cysticercosis. The aim of this study was to investigate the possible association between neurocysticercosis and cancer. METHODS: We designed a mortality rate study from the autopsy files of the Department of Pathology at the General Hospital of Mexico. A total of 1,271 autopsy files were reviewed. All files in which a malignant neoplasia was found during autopsy were selected as cases. Autopsies in which no malignant disease was found were used as controls. The odds ratio was determined between the frequency of neurocysticercosis in patients with any malignant neoplasia and that of the controls. RESULTS: Neurocysticercosis was more frequent in cases with malignant hematological diseases (MHD) than in controls (p = 0.01). The odds ratio for this association was 3.54, with 95% confidence interval from 1.17-9.79. CONCLUSIONS: Most human cancers arise from the interaction of a multiplicity of factors, including xenobiotics and endogenous constituents. Therefore, while it will be difficult to demonstrate that neurocysticercosis is a causal agent of malignant hematological diseases (MHD), it should be considered as a potential risk factor for cancer induction in countries where cysticercosis remains a public health problem.

[Mortality by poisoning in children]. / Mortalidad por envenenamiento en niños.

OBJECTIVE: To determine the standing of mortality by poisoning in children in the Mexican Republic, in the years from 1979-1994. MATERIAL AND METHODS: Secondary sources were employed. Analyzed variables were: age, sex, year, external cause of trauma and poisoning according to the 9th International Classification of Diseases: E850-E858, E860-E869 and E905. Tendencies by specific causes were analyzed with a Poisson regression model and relative risk by age, sex and district were obtained. RESULTS: A total of 11,272 children under 15 years of age were recorded. The main causes were poisoning and toxic reactions caused by venomous plants or animals (E905); accidental poisoning by household gas or carbon monoxide (E868); and accidental poisoning by other drugs (E858). The relative risk was highest in age group < 1 year; the values were RR 29.6, CI 95% 29.2-33.4; RR 3.47, CI 95% 2.86-4.22 and RR 31.86, CI 95% 24.8-40.9. Risk was similar for both sexes except for E905. The state of Aguascalientes consistently presented the highest risk values and the state of Nuevo Leon, the lowest. CONCLUSIONS: Poisoning is an important cause of child mortality. Considering that most of these deaths can potentially be prevented since they occur at home it is recommended that responsible adults can build protection into their environment and into the way society operates. Prevention should involve a multidisciplinary approach since the phenomenon has multiple causes and possible solutions.

[Cyclic nucleotide phosphodiesterases: diversity, classification, structure and function]. / Fosfodiesterasas de nucleótidos cíclicos: diversidad, clasificación, estructura y función.

During the last years there have been major advances in the knowledge of cyclic nucleotide phosphodiesterases. Particularly, referred to the presence of multiple different isozymes. Seven different phosphodiesterase gene families, have been described in mammalian tissues, containing several distinct genes, most of them expressed in different tissues as functionally unique splice variants. This article includes various aspects of the currently accepted nomenclature, structure and function of each family of phosphodiesterases. Finally, a brief discussion of the presence and role of these enzymes in the cell proliferation and differentiation processes, in parasites of the Trypanosmatidae family, is provided.

Identification of the phosphorylation site in vitro for cAMP-dependent protein kinase on the rat adipocyte cGMP-inhibited cAMP phosphodiesterase.

Rat adipocyte cGMP-inhibited cAMP phosphodiesterase (cGI-PDE) appears to be dually regulated in intact cells by serine phosphorylations induced by isoprenaline and insulin, respectively (Degerman, E., Smith, C. J., Tornqvist, H., Vasta, V., Belfrage, P., and Manganiello, V. C. (1990) Proc. Natl. Acad. Sci. U.S.A. 87, 533-537; Smith, C. J., Vasta, V., Degerman, E., Belfrage, P., and Manganiello, V. C. (1991) J. Biol. Chem. 266, 13385-13390). Since cAMP-dependent protein kinase (cAMP-PK) catalyzes the beta-adrenergic effects, the site in the isolated cGI-PDE phosphorylated by this kinase was explored. A peptide, LRRSSGASGLLTSEHHSR (P18), corresponding to the amino acid sequence Leu423-Arg440 in the putative regulatory domain of the rat adipocyte cGI-PDE was synthesized. It contains a consensus substrate sequence -RRXS- for cAMP-PK within two tryptic cleavage sites and was readily phosphorylated by cAMP-PK. Two phosphopeptides, identified as RS-[32P]SGASGLLTSEHHSR and S-[32P]SGASGLLTSEHHSR, were obtained after stoichiometric phosphorylation and trypsinization of the peptide. These two peptides and the two main tryptic phosphopeptides obtained from immunoisolated [32P]cGI-PDE phosphorylated with cAMP-PK in a solubilized crude adipocyte membrane fraction were immuno-precipitated by an affinity-purified polyclonal antibody raised against P18 and exhibited the same chromatographic and electrophoretic profiles in three different separation systems. Similar radiosequencing profiles indicated that the second most N-terminal serine, corresponding to Ser-427 in the intact cGI-PDE, was phosphorylated by cAMP-PK in both P18 and authentic cGI-PDE. It is concluded that serine 427 is the target for cAMP-PK phosphorylation of the rat adipocyte cGI-PDE in vitro.

Single-step affinity purification, partial structure and properties of human platelet cGMP inhibited cAMP phosphodiesterase.

The human platelet cilostamide- and cGMP-inhibited cAMP phosphodiesterase (cGI-PDE) was rapidly purified approximately 19,000-fold to apparent homogeneity using single step affinity chromatography on the isothiocyanate derivative of cilostamide coupled to aminoethyl agarose. Within 24 h, 30 micrograms of enzyme protein was obtained from 20 ml of packed platelets. Vmax for cAMP and cGMP was 6.1 and 0.9 mumol/min per mg protein, respectively. Several polypeptides (110/105, 79, 62, 55/53 kDa) were identified after SDS-PAGE, all of which were immunologically related to cGI-PDE and represented approx. 5, 20, 50 and 20% of the total protein, respectively. Limited proteolysis of the cGI-PDE with chymotrypsin produced a major fragment of approximately 47 kDa (and at least two smaller peptides) with catalytic activity and sensitivity to cGMP and OPC 3911 similar to controls. Phosphorylation of the cGI-PDE by cAMP-dependent protein kinase (A-kinase) resulted in maximal incorporation of 0.6-1.8 mol of 32P/mol 110/105 and 79 kDa polypeptides; much lower and variable amounts of phosphate were incorporated into the 62 and 55/53 kDa polypeptides. After digestion of cGI-PDE with several proteinases a number of peptides were isolated and sequenced. Most of the peptide sequences obtained could be aligned within the carboxy terminal domain of the deduced sequence of the human cardiac cGI-PDE. These and other results suggest that the subunit size of the intact platelet cGI-PDE is 110 kDa and that proteolytic fragments of 79, 62 and 55/53 kDa are produced during purification. The smaller fragments (62 and 55/53 kDa) contain the catalytic domain; the larger fragments (110 and 79 kDa) also contain the regulatory domain with phosphorylation sites for A-kinase.

Relationships between cell surface protease and acid phosphatase activities of Leishmania promastigote.

A correlation between the ratio of the cell surface protease activity to phosphatase activity and the complexity of the pattern of cell surface exposed polypeptides of Leishmania promastigotes was demonstrated for various strains grown under similar conditions. The ratio of the cell surface protease activity to acid phosphatase activity was high for L. major and L.b. panamensis and it correlates with the expression of a single polypeptide of 63 KDa on their cell surface. Intermediate and lower ratios of these enzymatic activities relate with more complex radio-iodinated patterns: two main bands in L.b. guyanensis (70 and 58 KDa) and L.b. braziliensis (72 and 60 KDa) and three main bands 65, 50, 27 KDa in all L.m. mexicana strains tested. Evidence is presented that the acid phosphatase located on the L.m. mexicana cell surface is not an artifact due to a secondary absorption of the secreted acid phosphatase from the culture medium. These results confirm the Leishmania antigen cell surface heterogeneity. The implications on the biology of Leishmania and the clinical manifestation of leishmaniasis are discussed.

Insulin induced phosphorylation and activation of the cGMP-inhibited cAMP phosphodiesterase in human platelets.

Insulin induced phosphorylation and activation of the cGMP inhibited cAMP phosphodiesterase (cGI-PDE) in human platelets were demonstrated after isolation of the enzyme with specific polyclonal cGI-PDE antibodies. The demonstration of this insulin effect required suppression of basal cGI-PDE phosphorylation, through the use of the protein kinase inhibitor H-7 (1-(5-isoquinolinylsulfonyl)-2-methylpiperazine). The human platelet insulin receptor beta-subunit, previously identified as a 97 kDa polypeptide, was detected with the use of wheat germ agglutinin chromatography and anti-phosphotyrosine antibodies. These results suggest that insulin, through phosphorylation/activation of cGI-PDE, could decrease cAMP/cAMP dependent protein kinase (cAMP-PK) activity and thereby make the platelets more sensitive towards aggregating agents.

Purification and properties of the cGMP-inhibited cAMP phosphodiesterase from bovine aortic smooth muscle.

Pure cGMP-inhibited cAMP phosphodiesterase (cGI-PDE) in micrograms quantities was isolated from bovine aortic smooth muscle after more than 5000-fold purification using DEAE ion-exchange and affinity chromatography with a derivative of the specific cGI-PDE inhibitor cilostamide conjugated as a ligand to aminoethyl agarose (CIT-agarose). The cGI-PDE, which constituted about half of the high affinity cAMP-PDE activity of a tissue homogenate, was identified with a 105-kDa protein on SDS-PAGE through use of antibodies towards the human platelet, bovine cardiac and bovine adipose tissue cGI-PDE in Western blot and immunoprecipitation/immunoinactivation analysis. As observed during purification of the enzyme from other tissues the enzyme protein was exquisitely sensitive to proteolytic nicking during purification, resulting in several 30-77-kDa polypeptide fragments. Rapid immunoprecipitation from fresh tissue extracts was the only was found to partially prevent the proteolysis. The native enzyme had apparent molecular sizes of approx. 100,000 or, mainly approx. 220,000 by gel chromatography, presumably indicating the presence of monomeric and dimeric forms. The enzyme hydrolyzed cAMP and cGMP with normal Michaelis-Menten kinetics with Km of 0.16 and 0.09 microM, respectively, with Vmax for hydrolysis of cAMP of 0.3 compared to 3.1 mumol/min per mg protein for cAMP. The enzyme was potently and selectively inhibited by cGMP (IC50 approximately 0.25 microM) and the cardiotonic/vasodilatory drugs OPC-3911 (a cilostamide derivative), milrinone and CI-930 (IC50 approximately 0.05, 0.40 and 0.25 microM, respectively). The cGI-PDE was phosphorylated by cAMP-dependent protein kinase as has been reported for the analogous enzymes in heart, adipose tissue and platelets. The identification of a cGI-PDE in the aortic smooth muscle and its inhibitor specificity is consistent with the hypothesis that inhibition of this enzyme is important in the mechanism through which these drugs produce vasorelaxation.

Selective inhibition of cGMP-inhibited and cGMP-noninhibited cyclic nucleotide phosphodiesterases and relaxation of rat aorta.

In the supernatant (50,000 g, 1 hr) fraction from rat aortic smooth muscle homogenates, approximately 50% of total cAMPE PDE activity was inhibited by OPC 3911 (3 microM), while approximately 20% was inhibited by rolipram (30 microM). A cGMP-inhibited cyclic nucleotide phosphodiesterase (cGI-PDE) was further purified using DEAE chromatography followed by affinity chromatography on the N-(2-isothiocyanato)ethyl derivative of cilostamide conjugated to aminoethyl agarose (CIT-agarose). OPC 3911, CI-930, and milrinone, but not rolipram, were potent and selective inhibitors of this enzyme. The PDE-activity in the CIT-agarose flow through fraction (RI-PDE), however, was inhibited potently by rolipram, but not by cGMP, OPC 3911, CI-930 or milrinone. Functional studies showed that OPC 3911, CI-930, and milrinone were potent relaxants of contracted rat aorta. Rolipram had little relaxant effect. When OPC 3911 or milrinone was combined with rolipram more than additive effects on aortic relaxation and cAMP content were obtained. OPC 3911 combined with milrinone had only additive effects. These results demonstrate the presence of a cGI-PDE in rat aortic smooth muscle, and that inhibition of this isozyme may be of primary importance for the relaxant effects of OPC 3911, CI-930, and milrinone. A RI-PDE activity was also found, but it appeared to be less important for modulation of vascular tone unless the cGI-PDE was already inhibited. This may explain the synergistic relaxant effects observed when both PDE-isozymes were inhibited.

Biochemical evidence of the antigenic cell surface heterogeneity of Leishmania mexicana.

In the present study, an enzymatical and structural analysis of Leishmania mexicana cell-surface components was carried out, demonstrating that protease and acid phosphatase activities were present at the L. mexicana cell surface. These findings correlate with the expression of the main components detected on the surface of L. mexicana promastigotes: the 50-kDa component is responsible for the acid phosphatase activity, whereas glycoprotein 65 (gp65) was characterized as the structural polypeptide of the surface protease. Furthermore, the 50- and 65-kDa antigens were found to be structurally different, inasmuch as no homology was observed in their peptide digestion profiles. The results presented in this communication confirm heterogeneity in the expression of the surface components of L. mexicana promastigotes at both the structural and the biochemical level.