RESUMEN
The Middle Pleistocene represents a period of critical importance in human evolution, marked by encephalisation and dental reduction, and increasing diversification of temporally and spatially distributed hominin lineages in Africa, Asia and Europe. New specimens, especially from areas less well represented in the fossil record, can inform the debate on morphological changes to the skeleton and teeth and the phylogenetic course of human evolution during this period. The mandible from the cave of Mala Balanica, Serbia has recently been re-dated to at least 400 ka, and its well-preserved dentition presents an excellent opportunity to characterize molar crown morphology at this time period, and re-examine claims for a lack of Neandertal affinities in the specimen. In this study we employ microtomography to image the internal structure of the mandibular molars (focusing on the morphology of the enamel-dentine junction, or EDJ) of the BH-1 specimen and a comparative sample (n = 141) of Homo erectus sensu lato, Homo neanderthalensis, Pleistocene Homo sapiens, and recent H. sapiens. We quantitatively assess EDJ morphology using 3D geometric morphometrics and examine the expression of discrete dental traits at the dentine surface. We also compare third molar enamel thickness in BH-1 to those of H. neanderthalensis and both Pleistocene and recent H. sapiens, and document previously unreported morphology of the BH-1 premolar and molar roots. Our results highlight the reliability of the EDJ surface for classifying hominin taxa, indicate a primitive dental morphology for BH-1 molars, and confirm a general lack of derived Neandertal features for the Balanica individual. The plesiomorphic character of BH-1 is consistent with several competing models of Middle Pleistocene hominin evolution and provides an important regional and temporal example for reconstructing morphological changes in the mandible and teeth during this time period.
Asunto(s)
Fósiles , Hominidae/anatomía & histología , Hominidae/clasificación , Mandíbula/anatomía & histología , Puntos Anatómicos de Referencia , Animales , Evolución Biológica , Esmalte Dental/anatomía & histología , Dentina/anatomía & histología , Humanos , Diente Molar/anatomía & histología , Raíz del Diente/anatomía & histologíaRESUMEN
Serum proteins are routinely used to diagnose diseases, but are hard to find due to low sensitivity in screening the serum proteome. Public repositories of microarray data, such as the Gene Expression Omnibus (GEO), contain RNA expression profiles for more than 16,000 biological conditions, covering more than 30% of United States mortality. We hypothesized that genes coding for serum- and urine-detectable proteins, and showing differential expression of RNA in disease-damaged tissues would make ideal diagnostic protein biomarkers for those diseases. We showed that predicted protein biomarkers are significantly enriched for known diagnostic protein biomarkers in 22 diseases, with enrichment significantly higher in diseases for which at least three datasets are available. We then used this strategy to search for new biomarkers indicating acute rejection (AR) across different types of transplanted solid organs. We integrated three biopsy-based microarray studies of AR from pediatric renal, adult renal and adult cardiac transplantation and identified 45 genes upregulated in all three. From this set, we chose 10 proteins for serum ELISA assays in 39 renal transplant patients, and discovered three that were significantly higher in AR. Interestingly, all three proteins were also significantly higher during AR in the 63 cardiac transplant recipients studied. Our best marker, serum PECAM1, identified renal AR with 89% sensitivity and 75% specificity, and also showed increased expression in AR by immunohistochemistry in renal, hepatic and cardiac transplant biopsies. Our results demonstrate that integrating gene expression microarray measurements from disease samples and even publicly-available data sets can be a powerful, fast, and cost-effective strategy for the discovery of new diagnostic serum protein biomarkers.
Asunto(s)
Proteínas Sanguíneas/genética , Biología Computacional/métodos , Rechazo de Injerto/metabolismo , Análisis de Secuencia por Matrices de Oligonucleótidos/métodos , ARN/análisis , Biomarcadores/sangre , Biomarcadores/metabolismo , Biomarcadores/orina , Proteínas Sanguíneas/análisis , Minería de Datos , Bases de Datos Genéticas , Ensayo de Inmunoadsorción Enzimática , Perfilación de la Expresión Génica , Rechazo de Injerto/sangre , Rechazo de Injerto/diagnóstico , Rechazo de Injerto/orina , Trasplante de Corazón/efectos adversos , Trasplante de Corazón/inmunología , Histocitoquímica , Humanos , Glomérulos Renales/metabolismo , Glomérulos Renales/patología , Trasplante de Riñón/efectos adversos , Trasplante de Riñón/inmunología , Proteinuria/sangre , Proteinuria/orina , ARN/biosíntesis , ARN/genética , Curva ROC , Reproducibilidad de los ResultadosRESUMEN
In environmental and human health protection, the role for geoscience may be expressed by how it enhances certainty in the hazard potential models that support risk assessment. For geochemical hazards, certainty reflects how well geoscience simplifies variability in the element concentrations and in the environmental conditions associated with exposure pathways. Through mineralogy, geoscience establishes natural geochemical background variability in terms of provenance, process, and past, and it links hazard potential to the physical and chemical transformation due to weathering and soil formation. The interpretation of hazard potential may be expressed by how analytical protocol, expressed by grain size and strength of acid decomposition, combines with geological factors, expressed by (1) mineralogy and mineral partitioning and (2) environmental cofactors, including moisture, pH, buffering capacity, and porosity. With this type of knowledge, geoscience enhances the potential to identify covariant relations between hazard indicators and disease, and to resolve potential causal factors.
Asunto(s)
Geología , Metales/efectos adversos , Metales/química , Oligoelementos/efectos adversos , Oligoelementos/química , Ambiente , Monitoreo del Ambiente , Humanos , Minerales , Modelos de Riesgos Proporcionales , Medición de Riesgo , Suelo/análisisRESUMEN
Publication of the human proteome has prompted efforts to develop high-throughput techniques that can catalogue and quantify proteins and peptides present in different tissue types. The field of proteomics aims to identify, quantify, analyze, and functionally define a large number of proteins in cellular processes in different disease states on a global scale. Peptidomics, a newer name in the -omics world, measures and identifies naturally occurring low molecular weight peptides, also providing an insight into enzymatic processes and molecular events occurring in the system of interest. One area of major interest is the use of proteomics to identify diagnostic and prognostic biomarkers for different diseases as well as for various clinical phenotypes in organ transplantation that can advance targeted therapy for various forms of graft injury. Outcomes in organ transplantation can be potentially improved by identifying noninvasive biomarkers that will serve as triggers that predate graft injury, and can offer a means to customize patient treatment by differentiating among causes of acute and chronic graft injury. Proteomic and peptidomic strategies can be harnessed for frequent noninvasive measurements in tissue fluids, allowing for serial monitoring of organ disease. In this review, we describe the basic techniques used in proteomic and peptidomic approaches, point out special considerations in using these methods, and discuss their applications in recently published studies in organ transplantation.
Asunto(s)
Péptidos/genética , Proteoma , Trasplante , HumanosRESUMEN
Pyrolytic carbon mechanical heart valves (MHVs) are widely used to replace dysfunctional and failed heart valves. As the human heart beats around 40 million times per year, fatigue is the prime mechanism of mechanical failure. In this study, a finite element approach is implemented to develop a model for fatigue analysis of MHVs due to the impact force between the leaflet and the stent and cavitation in the aortic position. A two-step method to predict crack propagation in the leaflets of MHVs has been developed. Stress intensity factors (SIFs) are computed at a small initiated crack located on the leaflet edge (the worst case) using the boundary element method (BEM). Static analysis of the crack is performed to analyse the stress distribution around the front crack zone when the crack is opened; this is followed by a dynamic crack analysis to consider crack propagation using the finite element approach. Two factors are taken into account in the calculation of the SIFs: first, the effect of microjet formation due to cavitation in the vicinity of leaflets, resulting in water hammer pressure; second, the effect of the impact force between the leaflet and the stent of the MHVs, both in the closing phase. The critical initial crack length, the SIFs, the water hammer pressure, and the maximum jet velocity due to cavitation have been calculated. With an initial crack length of 35 microm, the fatigue life of the heart valve is greater than 60 years (i.e. about 2.2 x 10(9) cycles) and, with an initial crack length of 170 microm, the fatigue life of the heart valve would be around 2.5 years (i.e. about 9.1 x 10(7) cycles). For an initial crack length greater than 170 microm, there is catastrophic failure and fatigue cracking no longer occurs. A finite element model of fatigue analysis using Patran command language (PCL custom code) in MSC software can be used to evaluate the useful lifespan of MHVs. Similar methodologies can be extended to other medical devices under cyclic loads.
Asunto(s)
Válvula Aórtica , Diseño Asistido por Computadora , Prótesis Valvulares Cardíacas , Modelos Teóricos , Falla de Prótesis , Simulación por Computador , Análisis de Elementos Finitos , Humanos , Estrés MecánicoRESUMEN
Radon has been identified as the second leading cause of lung cancer after tobacco smoking. Information on indoor radon concentrations is required to assess the lung cancer burden due to radon exposure. However, radon data in highly populated southern Ontario are very limited. Since radon in soil is believed to be the main source of radon in homes, measurements of soil gas radon concentrations can be used to estimate variations in radon potential of indoor environments. This study reports a transect survey of natural background variation in soil radon levels across southern Ontario. The results indicate that radon risk could be high in some areas of southern Ontario.
Asunto(s)
Contaminantes Radiactivos del Aire/análisis , Contaminación del Aire Interior/análisis , Radón/análisis , Contaminantes Radiactivos del Suelo/análisis , Radiación de Fondo , Canadá , HumanosRESUMEN
Megalin is a multiligand receptor heavily involved in protein endocytosis. We recently demonstrated that megalin binds and mediates internalization of ANG II. Although there is a strong structural resemblance between ANG II and ANG-(1-7), their physiological actions and their affinity for the angiotensin type 1 receptor (AT(1)R) are dissimilar. Therefore, the hypothesis of the present work was to test whether megalin binds and internalizes ANG-(1-7). The uptake of ANG-(1-7) was determined by exposure of confluent monolayers of BN/MSV cells (a model representative of the yolk sac epithelium) to fluorescently labeled ANG-(1-7) (100 nM) and measurement of the amount of cell-associated fluorescence after 4 h by flow cytometry. Anti-megalin antisera and an AT(1)R blocker (olmesartan) were used to interfere with uptake via megalin and the AT(1)R, respectively. ANG-(1-7) uptake was prevented by anti-megalin antisera (63%) to a higher degree than olmesartan (13%) (P < 0.001). In analysis by flow cytometry of binding experiments performed in brush-border membrane vesicles isolated from kidneys of CD-1 mice, anti-megalin antisera interfered with ANG-(1-7) binding more strongly than olmesartan (P < 0.05 against positive control). Interactions of megalin with ANG-(1-7) at a molecular level were studied by surface plasmon resonance, demonstrating that ANG-(1-7) binds megalin dose and time dependently and with an affinity similar to ANG II. These results show that the scavenger receptor megalin binds and internalizes ANG-(1-7).
Asunto(s)
Angiotensina I/metabolismo , Antihipertensivos/metabolismo , Proteína 2 Relacionada con Receptor de Lipoproteína de Baja Densidad/fisiología , Fragmentos de Péptidos/metabolismo , Técnicas de Cultivo de Célula , Epitelio , Citometría de Flujo , Humanos , Riñón/fisiología , Sistema Renina-Angiotensina/fisiología , Saco Vitelino/citologíaRESUMEN
Cubilin and megalin are giant glycoprotein receptors abundant on the luminal surface of proximal tubular cells of the kidney. We showed previously that light chains are a ligand for cubilin. As cubilin and megalin share a number of common ligands, we further investigated the ligand specificity of these receptors. Three lines of evidence suggest that light chains can also bind megalin: 1) anti-megalin antiserum largely displaces brush-border light chain binding and megalin-expressing BN-16 cell uptake more than anti-cubilin antiserum, 2) direct binding studies on isolated proteins using surface plasmon resonance techniques confirm that megalin binds light chains, and 3) light chains compete with known megalin ligands for brush-border membrane binding and BN-16 cell uptake. The megalin-light chain interaction is divalent ion dependent and similar for both kappa- and lambda-light chains. A fit of the data on light chain binding to megalin over a concentration range 0.078-2.5 mg/ml leads to an estimated dissociation constant of 6 x 10(-5) M, corresponding approximately to one light chain-binding site per megalin and in the same range for dissociation constants for cubilin binding. These data suggest that light chains bind the tandem megalin-cubilin complex. Megalin is the major mediator of light chain entry into megalin-expressing membrane such as the apical surface of proximal tubular epithelial cells.
Asunto(s)
Túbulos Renales Proximales/metabolismo , Receptores de Superficie Celular/metabolismo , Animales , Células Cultivadas , Túbulos Renales Proximales/ultraestructura , Ligandos , Masculino , Microvellosidades/metabolismo , Unión Proteica , Ratas , Ratas Sprague-DawleyRESUMEN
Megalin is an abundant membrane protein heavily involved in receptor-mediated endocytosis. The major functions of megalin in vivo remain incompletely defined as megalin typically faces specialized milieus such as glomerular filtrate, airways, epididymal fluid, thyroid colloid, and yolk sac fluid, which lack many of its known ligands. In the course of studies on ANG II internalization, we were surprised when only part of the uptake of labeled ANG II into immortalized yolk sac cells (BN-16 cells) was blocked by specific peptide inhibitors and direct competitors of the angiotensin type 1 receptor. This led us to test if megalin was a receptor for ANG II. Four lines of direct evidence demonstrate that megalin and, to a lesser extent, its chaperone protein cubilin are receptors for ANG II. First, in BN-16 cells anti-megalin and anti-cubilin antisera interfere with ANG II uptake. Second, also in BN-16 cells, pure ANG II competes for uptake of a known megalin ligand. Third, in proximal tubule cell brush-border membrane vesicles extracted from mice, anti-megalin antisera interfere with ANG II binding. Fourth, purified megalin binds ANG II directly in surface plasmon resonance experiments. The finding that megalin is a receptor for ANG II suggests a major new function for the megalin pathway in vivo. These results also indicate that ANG II internalization in some tissues is megalin dependent and that megalin may play a role in regulating proximal tubule ANG II levels.
Asunto(s)
Angiotensina II/metabolismo , Túbulos Renales Proximales/fisiología , Proteína 2 Relacionada con Receptor de Lipoproteína de Baja Densidad/fisiología , Vasoconstrictores/metabolismo , Animales , Ratones , Reacción en Cadena de la Polimerasa , Receptor de Angiotensina Tipo 1/biosíntesisRESUMEN
Although several heavy metal toxins are delivered to the kidney on the carrier protein metallothionein (MT), uncertainty as to how MT enters proximal tubular cells limits treatment strategies. Prompted by reports that MT-I interferes with renal uptake of the megalin ligand beta(2)-microglobulin in conscious rats, we tested the hypothesis that megalin binds MT and mediates its uptake. Three lines of evidence suggest that binding of MT to megalin is critical in renal proximal tubular uptake of MT-bound heavy metals. First, MT binds megalin, but not cubilin, in direct surface plasmon resonance studies. Binding of MT occurs at a single site with a K(d) approximately 10(-4) and, as with other megalin ligands, depends on divalent cations. Second, antisera and various known megalin ligands inhibit the uptake of fluorescently labeled MT in model cell systems. Anti-megalin antisera, but not control sera, displace >90% bound MT from rat renal brush-border membranes. Megalin ligands including beta(2)-microglobulin and also recombinant MT fragments compete for uptake by megalin-expressing rat yolk sac BN-16 cells. Third, megalin and fluorescently labeled MT colocalize in BN-16 cells, as shown by fluorescent microscopic techniques. Follow-up surface plasmon resonance and flow cytometry studies using overlapping MT peptides and recombinant MT fragments identify the hinge SCKKSCC region of MT as a critical site for megalin binding. These findings suggest that disruption of the SCKKSCC motif can inhibit proximal tubular MT uptake and thereby eliminate much of the renal accumulation and toxicity of heavy metals such as cadmium, gold, copper, and cisplatinum.
Asunto(s)
Cadmio/metabolismo , Proteína 2 Relacionada con Receptor de Lipoproteína de Baja Densidad/metabolismo , Metalotioneína/metabolismo , Secuencia de Aminoácidos , Animales , Anticuerpos , Sitios de Unión , Vesículas Citoplasmáticas/metabolismo , Citometría de Flujo , Ligandos , Proteína 2 Relacionada con Receptor de Lipoproteína de Baja Densidad/inmunología , Lisina/química , Masculino , Metalotioneína/química , Microscopía Fluorescente , Datos de Secuencia Molecular , Estructura Terciaria de Proteína , Ratas , Ratas Sprague-Dawley , Receptores de Superficie Celular/inmunología , Receptores de Superficie Celular/metabolismo , Resonancia por Plasmón de SuperficieRESUMEN
Strains of the yeast Pichia inositovora that carry the linear plasmids pPin1-1 (18 kb) and pPin1-3 (10 kb) display a killer activity towards Saccharomyces cerevisiae. Cloning and sequencing of the smaller plasmid, pPin1-3, revealed that it is 9683 bp long and has 154-bp terminal inverted repeats. Comparison of pPin1-3 with the only other completely sequenced killer plasmid, pGKL1 of Kluyveromyces lactis, revealed differences in genome organization. The Pichia element has four ORFs that account for 95% of the sequence. ORF1 is homologous to the putative immunity gene of the K. lactis system. A viral B-type DNA polymerase is encoded by ORF2. The predicted product of ORF3 displays similarities to the alpha- and beta-subunits of the heterotrimeric K. lactis killer toxin, also known as zymocin. A cysteine-rich chitin-binding site and a chitinase signature, characteristic for the alpha-subunit of zymocin were identified in Orf3p. Chitin affinity chromatography and Western analysis confirmed the plasmid specific expression and secretion of a protein that cross-reacts with an antibody raised against the alpha-subunit of K. lactis zymocin. Disruption of the major chitin synthase-gene ( CHS3) renders S. cerevisiae resistant to the toxin, providing further evidence that chitin is the cellular receptor for the P. inositovora toxin. Orf4p of pPin1-3 displays only weak similarities to the gamma-subunit of zymocin, which causes a G1 cell-cycle arrest in S. cerevisiae. However, disruption of the S. cerevisiae gene ELP3/TOT3, which encodes a histone-acetyltransferase that is essential for zymocin action, resulted in reduced sensitivity to the P. inositovora toxin also. Thus, despite obvious differences in genome organization and protein architecture, both killer systems very probably have similar modes of action.
Asunto(s)
Pichia/genética , Plásmidos/genética , Secuencia de Aminoácidos , Secuencia de Bases , Quitina/metabolismo , ADN de Hongos/genética , ADN Polimerasa Dirigida por ADN/genética , Factores Asesinos de Levadura , Datos de Secuencia Molecular , Micotoxinas/genética , Micotoxinas/metabolismo , Sistemas de Lectura Abierta , Fenotipo , Filogenia , Pichia/metabolismo , Pichia/patogenicidad , Saccharomyces cerevisiae/genética , Homología de Secuencia de Aminoácido , Virulencia/genéticaRESUMEN
The exozymocin secreted by Kluyveromyces lactis causes sensitive yeast cells, including Saccharomyces cerevisiae, to arrest growth in the G(1) phase of the cell cycle. Despite its heterotrimeric (alpha beta gamma) structure, intracellular expression of its smallest subunit, the gamma-toxin, is alone responsible for the G(1) arrest. The alpha subunit, however, has a chitinase activity that is essential for holozymocin action from the cell exterior. Here we show that sensitive yeast cells can be rescued from zymocin treatment by exogenously applying crude chitin preparations, supporting the idea that chitin polymers can compete for binding to zymocin with chitin present on the surface of sensitive yeast cells. Consistent with this, holozymocin can be purified by way of affinity chromatography using an immobilized chitin matrix. PCR-mediated deletions of chitin synthesis (CHS) genes show that most, if not all, genetic scenarios that lead to complete loss (chs3 Delta), blocked export (chs7 Delta) or reduced activation (chs4 Delta), combined with mislocalization (chs4 Delta chs5 Delta; chs4 Delta chs6 Delta; chs4 Delta chs5 Delta chs6 Delta) of chitin synthase III activity (CSIII), render cells refractory to the inhibitory effects of exozymocin. In contrast, deletions in CHS1 and CHS2, which code for CSI and CSII, respectively, have no effect on zymocin sensitivity. Thus, CSIII-polymerized chitin, which amounts to almost 90% of the cell's chitin resources, appears to be the carbohydrate receptor required for the initial interaction of zymocin with sensitive cells.
Asunto(s)
Pared Celular/metabolismo , Quitina/metabolismo , Kluyveromyces , Micotoxinas/metabolismo , Micotoxinas/farmacología , Saccharomyces cerevisiae/efectos de los fármacos , Secuencia de Aminoácidos , Quitina/genética , Quitina Sintasa/genética , Quitina Sintasa/metabolismo , Cromatografía de Afinidad , Eliminación de Gen , Factores Asesinos de Levadura , Datos de Secuencia Molecular , Micotoxinas/química , Micotoxinas/genética , Receptores de Superficie Celular/metabolismo , Saccharomyces cerevisiae/genéticaRESUMEN
The linear cytoplasmic element pPE1B from Pichia etchellsii CBS2011 (synonym Debaryomyces etchellsii) was totally sequenced. It consists of 12835 bp and has a remarkable high A+T content of 77.3%. The termini of pPE1B were found to consist of inversely orientated identical nucleotide repetitions 161 base pairs long, to which proteins are probably covalently linked at the 5' ends. Ten putative genes (open reading frames, ORFs) were identified, covering 96.5% of the total sequence. The predicted polypeptides correspond to proteins encoded by ORFs 2-11 of the linear plasmids pGKL2 of Kluyveromyces lactis and pSKL of Saccharomyces kluyveri. ORF1, existing on both latter elements, is lacking on pPE1B. An upstream conserved sequence motif (UCS) is located at the expected distance from the start codon of each of the 10 ORFs. As the arbitrarily chosen UCS6 was able to drive expression of a reporter gene in the heterologous pGKL-encoded killer system of K. lactis, extranuclear promoter function is probable. The almost congruent genome organization of pPE1B and other autonomous linear yeast plasmids sequenced so far, i.e. pGKL2 and pSKL, suggests a common, presumably viral, ancestor.
Asunto(s)
Citoplasma/genética , Genoma Fúngico , Pichia/genética , Plásmidos/genética , Secuencia de Bases , Clonación Molecular , Secuencia Conservada , Genes Reporteros , Kluyveromyces/genética , Datos de Secuencia Molecular , Sistemas de Lectura Abierta , Análisis de Secuencia de ADNRESUMEN
Twenty-one patients with Scheuermann's kyphosis had surgery for progressive kyphotic deformity of 50 degrees or greater. There were six adolescents, with a mean age of 15.6 years (range, 13-17 years) and 15 young adults, with a mean age of 25.4 years (range, 18-40 years). All patients had posterior spine arthrodesis with segmental compression instrumentation. Seven patients with rigid kyphosis had combined anterior and posterior spine arthrodesis. One patient died of superior mesenteric artery syndrome. In the group of 13 patients with posterior arthrodesis only, followup was 4.5 years. The mean preoperative thoracic kyphotic curve of 68.5 degrees improved to 40 degrees at latest review, with an average loss of correction of 5.75 degrees. Junctional kyphosis occurred in two patients with a short arthrodesis: one at the cephalad end and one at the caudal end of the fused kyphotic curve. In the second group of seven patients with combined anterior and posterior arthrodesis, followup was 6 years. The mean preoperative thoracic kyphotic curve of 86.3 degrees improved to 46.4 degrees at latest review, with an average loss of correction of 4.4 degrees. Overall, there was no postoperative neurologic deficit and no pseudarthrosis. Thus, posterior arthrodesis and segmental compression instrumentation seems to be effective for correcting and stabilizing kyphotic deformity in Scheuermann's disease. Despite a long operating time, this technique provided significant correction, avoiding the development of any secondary deformity in most patients. Combined anterior and posterior spine arthrodesis is recommended for rigid, more severe kyphotic deformities.
Asunto(s)
Artrodesis/instrumentación , Trasplante Óseo/métodos , Cifosis/cirugía , Enfermedad de Scheuermann/diagnóstico , Enfermedad de Scheuermann/cirugía , Adolescente , Adulto , Artrodesis/métodos , Terapia Combinada , Femenino , Estudios de Seguimiento , Humanos , Cifosis/diagnóstico , Cifosis/etiología , Masculino , Estudios Retrospectivos , Enfermedad de Scheuermann/complicaciones , Índice de Severidad de la Enfermedad , Resultado del TratamientoRESUMEN
Eight samples of 'processed food flavours' (PFFs), chosen from five different categories, were analysed for their mutagenic activity using the Ames Salmonella assay, and also for the presence of eight heterocyclic aromatic amines (HAAs), namely 2-amino-3-(trideuteromethyl)imidazo[4,5-f]quinoline (IQ), 2-amino-3,8-dimethyl-imidazo[4,5-f]quinoline (MeIQ), 2-amino-3,8-dimethylimidazo[4,5-f]quinoxaline (MeIQx), 2-amino-3,7,8-trimethylimidazo[4,5-f]quinoxaline (7,8-DiMeIQx), 2-amino-3,7,8-trimethylimidazo[4,5-f]quinoxaline (4,7,8-TriMeIQx), 2-amino-I-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP), 3-amino-1,4-dimethyl-5H-pyrido[4,3-b]indole (Trp-P-1) and 3-amino-1-methyl-5H-pyrido[4,3-b]indole (Trp-P-2) using liquid chromatography and mass spectrometry (LC/MS). The isolation of HAAs was based on sequential liquid-liquid extraction procedures of samples at both acidic and basic pH values. The recoveries and the clean-up were monitored by introduction of quality control samples and by spiking with three tri-deuterated standards of HAAs. Although the results for the mutagenicity assay were comparable by testing less-purified and highly-purified extracts, the analysis for identification and quantification of HAAs by LC/MS required highly purified concentrates. Four samples had little or no mutagenic activity and these results were in agreement with their LC/MS results: they had no detectable levels (detection limits 1-3 ppb) of any of the HAAs monitored. The mutagenic activity of one sample was in complete agreement with the quantification of HAAs by LC/MS. Two samples produced strong mutagenic responses (3115 and 2664 revertants/g). In one sample, LC/MS analysis revealed the presence of 9.6 ppb IQ, whereas LC/MS of the other could not confirm the presence of any of the eight HAAs monitored. Two samples produced mild mutagenic activity (204 and 160 revertants/g), but relatively elevated concentrations of IQ (6.7 and 6.8 ng/g) by LC/MS. The extracts from all samples were tested for their modifying effects on mutagenicity of four HAAs. The discrepancy between the Ames test and the LC/MS analysis of some samples indicates several possibilities, such as the presence of some other HAAs, of their isomers or of other mutagens. In addition, the presence of mutagen modifiers (inhibitors or synergists) was observed in most samples. The results indicate that although chemical tests (e.g. LC/MS) can provide quantitative data for the HAAs monitored, the Ames mutagenicity test should also be conducted to determine the mutagenic activities of PFFs, in order to assess their health risk potential.
Asunto(s)
Aminas/efectos adversos , Aromatizantes/efectos adversos , Aditivos Alimentarios/efectos adversos , Compuestos Heterocíclicos/efectos adversos , Aminas/análisis , Cromatografía Líquida de Alta Presión , Aromatizantes/análisis , Aditivos Alimentarios/análisis , Compuestos Heterocíclicos/análisis , Carne , Pruebas de Mutagenicidad , Control de CalidadRESUMEN
In an ongoing survey, the presence of heterocyclic aromatic amines (HAAs) was determined in processed, ready-to-eat meat products sold as 'meat cuts'. HAAs are a group of recently recognized mutagenic/carcinogenic contaminants in foods that are produced during the heat processing of meat. 16 samples of meat cuts (e.g. turkey breast, salami, chicken loaf, cooked ham, all beef meat, pepperoni, etc.), randomly purchased from supermarkets and specialty food stores in the Ottawa area, were analysed for the presence of eight HAAs. The isolation of HAAs was based on sequential liquid-liquid extraction procedures of the samples at both acidic and basic pH values. The mutagenic activity of these samples was determined using the Ames/Salmonella microsome assay with the strain TA98 plus rat liver S-9 metabolic activation. The mutagenicity of these samples ranged from undetectable to slightly active. The highest mutagenic activity, 141 induced revertants/g, was found in a smoked turkey breast sample. 11 samples were not mutagenic, including two that indicated a tendency for inhibition of the spontaneous revertants. The remaining four samples exhibited very low mutagenic activity. For chemical analysis, the extracts were purified with two solid phase extraction cartridges. Quantitative analysis was performed by using liquid chromatography for separation and mass spectrometry for detection. With the exception of trace amounts (0.4 ng/g) of 2-amino-3,8-dimethylimidazo[4,5-f]quinoxaline (MeIQx) in the sample with highest mutagenic activity, the chemical analysis did not detect the presence of any of the eight most frequently found HAAs in fried or broiled meat products. These data suggest that consumption of meat cuts does not present a serious health risk from HAA-type contaminants.
Asunto(s)
Aminas/análisis , Compuestos Heterocíclicos/análisis , Carne/análisis , Aminas/efectos adversos , Animales , Canadá , Manipulación de Alimentos , Compuestos Heterocíclicos/efectos adversos , Pruebas de Mutagenicidad , RatasAsunto(s)
Espectroscopía de Resonancia Magnética , Proteínas de la Membrana/química , Péptidos/química , Membrana Dobles de Lípidos/química , Espectroscopía de Resonancia Magnética/instrumentación , Espectroscopía de Resonancia Magnética/métodos , Micelas , Fosfolípidos/química , Fosfolípidos/metabolismo , Estructura Secundaria de ProteínaRESUMEN
Sixty-three consecutive total hip arthroplasties were performed with cement in fifty adolescent patients from 1972 through 1980, and the results were determined after a minimum of ten years. A polyethylene cup without a metal backing and a non-modular femoral component with a collar and a fixed neck length were inserted, with use of so-called first-generation cementing techniques, in each hip. Kaplan-Meier survival analysis of all sixty-three hips demonstrated that the probability of failure (defined as revision or symptomatic loosening) increased steadily over time and reached 45 per cent after fifteen years. A number of specific variables were associated with a significantly higher probability of failure: a history of more than one previous procedure involving the hip (p = 0.0002), unilateral arthroplasty (p = 0.006), previous trauma involving the hip (p = 0.01), the absence of other disease that limited function of the ipsilateral lower extremity (p = 0.03), a high postoperative level of activity (involving moderate or strenuous manual labor) (p = 0.03), and a preoperative weight of more than sixty kilograms (p = 0.03). The probability of failure in the patients who had inflammatory arthritis (11 per cent) was significantly lower than that in those who had previous trauma involving the hip (47 per cent) (p = 0.0006). Fifty-two hips (forty patients) were followed for a minimum of ten years or until revision. The mean duration of follow-up for these fifty-two hips was 12.6 years (range, 1.6 to 18.6 years). The result was evaluated clinically and radiographically with use of the Mayo hip-scoring system and was graded as excellent in ten hips (19 per cent), good in sixteen (31 per cent), fair in one (2 per cent), and poor in twenty-five (48 per cent). Most of the poor results were due to symptomatic loosening of the acetabular component. The probability of radiographic loosening after fifteen years was 60 per cent for the acetabular component and 20 per cent for the femoral component. Radiographic evidence of polyethylene wear was associated with probable loosening of the acetabular component (p = 0.03). The findings of the present study suggest that total hip arthroplasty in adolescents should be reserved for carefully selected patients for whom alternative procedures are contraindicated or unacceptable. Fixation of the acetabular component with cement is not recommended in this setting.
Asunto(s)
Prótesis de Cadera , Acetábulo , Adolescente , Adulto , Factores de Edad , Cementación , Niño , Interpretación Estadística de Datos , Estudios de Evaluación como Asunto , Femenino , Fémur , Estudios de Seguimiento , Humanos , Masculino , Esfuerzo Físico , Polietilenos , Falla de Prótesis , Factores de TiempoRESUMEN
Water extracts of eight brands (five types: 'green', 'black', 'oolong', decaffeinated and instant) of common teas (derived from Camellia sinensis) and infusions of six randomly selected herbal teas were examined for inhibitory or potentiating effects on the mutagenicity of eight heterocyclic aromatic amines (HAA) using the Ames Salmonella typhimurium TA98 and S-9 assay. HAA, produced in foods during regular heat processing of meat, exhibit mutagenic/carcinogenic activities. Tea extracts from C. sinensis displayed very potent antimutagenic effects against most HAA: total or substantial inhibition of mutagenic activity of the eight HAA was obtained with extracts equivalent to 50 mg tea leaves/plate (mgEq) and potent inhibition was frequently achieved even with 10 mgEq/plate. Decaffeinated tea produced the same effect as observed for 'regular' teas. However, lower concentrations of some tea extracts enhanced mutagenic activity of 2-amino-3,4,7,8-tetramethyl-3H-imidazo[4,5-f]quinoxaline (4,7,8-TriMeIQx) and 3-amino-1-methyl-5H-pyrido[4,3-b]indole (Trp-P-2). Herbal tea extracts displayed variable effects on the mutagenicity of different HAA. While some extracts had no effect, others exhibited a moderate inhibitory effect on the mutagenicity of IQ-type HAA. In contrast to common tea, herbal teas showed substantial potentiating effects on the mutagenicity of several HAA, especially Trp-P-2 and 4,7,8-TriMeIQx.