Search

The artificial urinary sphincter (AUS) is one of the most effective surgical treatments for male urinary incontinence regardless of its severity. Current knowledge comes from high-volume centers, but little is known about the performance of this surgery from community practices. This study aims to report contemporary AUS performance in a nationwide observational study in Colombia.Male patients who underwent AUS surgery with AMS 800™ between 2000 and 2020 in more than 17 centers and four cities were identified. Pre, intra, and postoperative characteristics were evaluated, mainly addressing patient reported outcomes measurements in the postoperative period. Retrospective and prospective data collection and descriptive analysis were completed. Kaplan-Meier analysis was used to determine AUS survival rate.Out of an initial 667 cases, a total of 215 patients met inclusion and exclusion criteria and were included. Mean age was 67 ± 9.4 years, and mean follow-up was 6.0 ± 4.4 years with maximum range of 14 years. The etiology of urinary incontinence was prostate cancer surgery in 141 (81%) of the cases. The rest of the cases were related to benign prostatic disease or spinal cord injury. It is noteworthy that out of 115 patients, only 59 (51.3%) reported previous formal pelvic floor rehabilitation. Subjective severity of urinary incontinence determined by a visual analog scale showed a decrease in 4.5 points after sphincter implantation. Sphincter removal was required in 50 (23.2%) cases. The main reasons for implant removal were urethral erosion and infection. The sphincter survival rate at 2, 5, 8, 10, and 14 years was 76%, 70%, 60%, 57%, and 17%, respectively. Of the subjects at the last follow-up with the device still in place, 80.7% defined their urinary condition as "does not cause or causes minor discomfort," and 99% would recommend the device to a friend or relative in the same condition.This series from a community-based practice shows the lack of adherence to clinical practice guidelines and the lack of standardized data collection. In contrast, this study provides real-world data on explantation and revision rates, allows physicians to inform patients and to have clear metrics for a shared decision-making process before the procedure.

CISD3 is a mitochondrial protein belonging to the NEET proteins family, bearing two [Fe

The H19X-encoded miR-424(322)/503 cluster regulates multiple cellular functions. Here, it is reported for the first time that it is also a critical linchpin of fat mass expansion. Deletion of this miRNA cluster in mice results in obesity, while increasing the pool of early adipocyte progenitors and hypertrophied adipocytes. Complementary loss and gain of function experiments and RNA sequencing demonstrate that miR-424(322)/503 regulates a conserved genetic program involved in the differentiation and commitment of white adipocytes. Mechanistically, it is demonstrated that miR-424(322)/503 targets gamma-Synuclein (SNCG), a factor that mediates this program rearrangement by controlling metabolic functions in fat cells, allowing adipocyte differentiation and adipose tissue enlargement. Accordingly, diminished miR-424(322) in mice and obese humans co-segregate with increased SNCG in fat and peripheral blood as mutually exclusive features of obesity, being normalized upon weight loss. The data unveil a previously unknown regulatory mechanism offat mass expansion tightly controlled by the miR-424(322)/503 through SNCG.

An increasing demand of consumers for natural bioactive compounds has boosted the interest of the scientific community for vegetables and fruits, with many efforts being made to identify new bioactive compounds and to develop sustainable extraction methods that preserve the compounds bioactivity. Such is the case of pressurized liquid extraction and supercritical fluid extraction using “green solvents” as supercritical CO2, ethanol, and ethyl acetate. The accurate design of these processes requires the knowledge of transport properties, namely, diffusion coefficients, D 12 . Astaxanthin and squalene are two bioactive compounds whose D 12 values were measured in compressed liquid ethyl acetate using a chromatographic technique. For astaxanthin the D 12 values were in the range 0.817 × 10 - 5 - 1.223 × 10 - 5 c m 2 s - 1 (at 303.15–333.15 K and 1–100 bar) and for squalene they were in the range 1.052 × 10 - 5 - 1.822 × 10 - 5 c m 2 s - 1 (at 303.15–333.15 K and 1–150 bar). The influence of temperature, pressure and Stokes-Einstein coordinate was studied for both systems and explained with the most relevant theories of the literature.

In this work the shape characteristics of a parabolic structure that consists of an initially flat circular membrane subject to solar radiation pressure in the space environment are examined. This problem arises in the analysis of space systems such as solar power collectors and solar sails. This paper presents parametric studies aimed at determining the power concentration capacity of a non-ideally reflecting flat circular membrane as a function of its radius size and thickness. The shape of the deformed membrane is close to parabolic. Both numerical simulations and analytical study show that the thin membrane subject solely to solar radiation pressure can concentrate the sunlight power several hundred times.

This paper addresses the preliminary design of a parabolic structure suitable for specific space applications, such as in solar power collector systems, which relies upon the sole use of an initially flat circular membrane rigidly constrained across its perimeter. In particular, the possibility for this concept to make use of solar pressure as a means of obtaining the intended parabolic curvature is explored. Assuming the membrane´s film as an ideally reflecting surface, parametric studies are conducted in order to determine several parameters of interest as functions of its radius and thickness. In order to do so, a set of numerical simulations are carried out using the finite element code ABAQUS®. It is shown that the shape of the deformed membrane is very close to parabolic, therefore being capable of concentrating sunlight power over a focal plane.

Aerospace components are characterized by having high strength to weight ratios in order to obtain lightweight structures. Recently, different types of sandwich components using composite materials have been developed with the purpose of combining the effect of reinforced face-sheets with low weight core materials, such as honeycombs and foams. However, these materials must combine damage tolerance characteristics with high resistance under both static and dynamic loads. Cork composites can be considered as an alternative material for sandwich components since cork is a natural material with some remarkable properties, such as high damage tolerance to impact loads, good thermal and acoustic insulation capacities and excellent damping characteristics for the suppression of vibrations. The experiments carried out in this investigation were oriented in order to optimize the specific strength of cork based composites for sandwich components. Static bending tests were performed in order to characterize the mechanical strength of different types of cork agglomerates which were obtained considering distinct production variables. The ability to withstand dynamic loads was also evaluated from a set of impact tests using carbon-cork sandwich specimens. The results from experimental tests showed that cork agglomerates performance depends on the cork granulate size, the type of reinforcing elements and the bonding procedure used for the cohesion with the matrix material.

It is well known that shot peening has a marked benefit on fatigue life for the majority of applications. This effect is attributed mainly due to the compressive residual stress state at the component’s surface due to shot peening. The present paper evaluates the ability of several fatigue life prediction models, commonly used for general analyses, to predict the behaviour of components with compressive residual stress due to shot peening. Advanced elastic-plastic finite element analyses were carried out in order to obtain stress, strain, strain energy and fracture mechanics parameters for cracks within a compressive residual stress field. With these results several total fatigue life prediction models (including critical distance methods) and fracture mechanics based models were applied in order to predict fatigue life. Fatigue life predictions were compared with several experimental fatigue tests carried out on specimens, representative of a critical region of a compressor disc in a gas turbine aero engine. The results obtained showed that total fatigue life methods, even if combined with critical distance methods, give conservative results when shot peening is considered. Fatigue life was successfully predicted using the method proposed by Cameron and Smith, by adding initiation life to crack propagation life. This last method was also successfully applied for the prediction of non-propagating cracks that were observed during the experimental tests.

O liberum arbitrium, para Santo Anselmo, não pode residir no poder permanente de pecar e de não pecar (posse peccare et non peccare), porque tal definiçao tomaria inútil «a graça, a predestinação e a presciencia de Deus» (DLA, I, 207, 1-2). Por outro lado, se nós não tivéssemos sempre essa potestas, o pecado não poderia ser-nos imputado porque nós pecaríamos «sine libero arbítrio». Procurando separar esta alternativa, que lhe parece armadilhada, Santo Anselmo busca uma definição de libertas arbitrii independente do poder negativo de pecar (potestas peccandi) e, a partir da distinção entre voluntas propter se (instrumentum volendi) e voluntas propter aliud (usus sive opus volendi), julga encontrá-la na seguinte: a libertas arbitrii é «o poder de guardar a rectitude da vontade pela própria rectitude» (DLA, III, 212, 19-20: «potestas servandi rectitudinem voluntatis propter ipsam rectitudinem»), poder que exprime a exacta e positiva noção da «justiça original». À luz de tal definição transcendental de libertas arbitrii, comum a Deus, aos anjos e aos homens, pode Anselmo avançar a tese teologicamente mais ousada do opúsculo: nem Deus, apesar de poder «reduzir a nada uma substância que Ele fez do nada», é capaz de «separar a rectidão de uma vontade que a possui» (DLA, VIII, 220, 13-15).

Metabolites of the potent DP antagonist, MK-0524, were generated using in vitro systems including hepatic microsomes and hepatocytes. Four metabolites (two hydroxylated diastereomers, a ketone and an acyl glucuronide) were characterized by LC-MS/MS and 1H NMR. Larger quantities of these metabolites were prepared by either organic synthesis or biosynthetically to be used as standards in other studies. The propensity for covalent binding was assessed and was found to be acceptable (50 pmol-equiv/mg protein).

During the last decade, some major improvements have been achieved concerning the evaluation of new types of materials suitable for aeronautical components exposed to severe operational conditions, such as turbine disks. Due to their outstanding mechanical properties, nickel base superalloys assumed a preferential position when compared with other conventional metallic alloys, benefiting from both their superior fatigue strength and high temperature behaviour. However, these alloys evince a high sensibility concerning possible defects that can arise due to certain types of loading, such as porosities and cavities associated with creep-fatigue at high temperatures. The present paper compiles some experimental results obtained for two types of recent nickel base superalloys. Some fatigue tests were performed using two configurations of these materials: a set of Udimet 720Li specimens (CT geometry) and a set of RR1000 specimens (CN geometry). A maximum temperature of 650°C was considered in both types of materials. The mechanical properties of these alloys were inferred via typical FCGR parameters, such as da/dN vs K curves, complemented with detailed analyses of the cracking mechanisms based on SEM observations. Finally, some metallographic characterization tests were carried out in order to determine the average grain size of these PM alloys and to confirm the presence of important microstructural constituents that can influence the overall fatigue performance of these materials.

L-454,560 is a potent phospodiesterase 4 (PDE4) inhibitor which was identified as a development candidate for the treatment of asthma and chronic obstructive pulmonary disease (COPD). As part of the discovery of this compound, interspecies in vitro metabolism data was generated using liver microsomes and hepatocytes in order to understand the metabolic fate of the compound. In microsomes, metabolism of the 3-methyl-1,2,4-oxadiazole ring was the predominant pathway observed, including ring cleavage. In rat hepatocytes, hydroxylation of the methyl group on the oxadiazole ring and double-bond isomerization were the most abundant metabolites observed. No major species differences were found in terms of microsomal metabolite profiles. The use of LC with UV and MS detection is highlighted, as well as information from tandem mass spectrometry and NMR. Copyright © 2006 John Wiley & Sons, Ltd.

The purpose of this investigation is to evaluate quantitatively and comparatively the effect of embedding optical fibers (OF) on the mechanical behavior of a carbon fiber-epoxy composite in order to verify whether their presence can possibly degrade the mechanical performance of the host material. The existing literature on this subject is not conclusive about the nature and intensity of this effect. Adding more reliable data to our systematic study contributes to this discussion favoring the conclusion about a harmful influence as a consequence of optical fiber embedment. Three kinds of mechanical tests have been performed in this work: impact tests, static flexural tests, and fatigue tests. The results of some experiments point to a possible detrimental influence related to the presence of the OF, being it different in nature and intensity for each of these tests. The mechanical behavior in static loading conditions seems to be not significantly affected as a consequence of the presence of the OF, while that in impact and fatigue tests are strongly affected, even though this influence being physically distinct from each other. Based on these results, some discussion is made about the possible failure mechanisms that can explain the detected differences.

The present paper is concerned with the fatigue behaviour ofcarbon-epoxy laminates with embedded optical fibres subjected to bending loads. The main goal of this investigation was to evaluate quantitatively the effect of the presence of optical fibres within the host structure on its whole fatigue behaviour. Two optical fibre positions were investigated: in the mid-plane of the laminate and near the surface subjected to loading. Two distinct geometries of the ply stacking sequence were also considered, namely unidirectional and crossply. In order to evaluate the fatigue life and the fatigue damage, two different loading levels were used, both at 6 Hz frequency, room temperature and R = 0.1. Fatigue damage was monitored using dynamic stiffness decay and acoustic emission techniques. Failure mechanisms were analysed by means of optical and scanning microscopy. The results obtained lead to the conclusion that the embedding of optical fibres markedly prejudices the fatigue performance of the material ...

The COX-2 inhibitor DFP [5,5-dimethyl-3-(2-propoxy)-4-methanesulfonylphenyl)-2(5H)-furanone] was found to have a long half-life in humans. Analogues have been characterized in order to optimize pharmacokinetics. This has lead to the discovery of 5(S)-(5-ethyl-5-methyl-3-(2-propoxy)-4-methanesulfonylphenyl)-2(5H)-furanone analogue 11 a potent and selective COX-2 inhibitor which is metabolized to a greater extent than DFP upon incubation with rat and human hepatocytes, suggesting a shorter half-life in humans.

Cytochrome P450 (P450) 2D6 was first identified as the polymorphic human debrisoquine hydroxylase and subsequently shown to catalyze the oxidation of a variety of drugs containing a basic nitrogen. Residue Asp301 has been characterized as being involved in electrostatic interactions with substrates on the basis of homology modeling and site-directed mutagenesis experiments [Ellis, S. W., Hayhurst, G. P., Smith, G., Lightfoot, T., Wong, M. M. S., Simula, A. P., Ackland, M. J., Sternberg, M. J. E., Lennard, M. S., Tucker, G. T., and Wolf, C. R. (1995) J. Biol. Chem. 270, 29055-29058]. However, pharmacophore models based on the role of Asp301 in substrate binding are compromised by reports of catalytic activity toward substrates devoid of a basic nitrogen, which have generally been ignored. We characterized a high-affinity ligand for P450 2D6, also devoid of a basic nitrogen atom, spirosulfonamide [4-[3-(4-fluorophenyl)-2-oxo-1-oxaspiro[4.4]non-3-en-4-yl]benzenesulfonamide], with K(s) 1.6 microM. Spirosulfonamide is a substrate for P450 2D6 (k(cat) 6.5 min(-)(1) for the formation of a syn spiromethylene carbinol, K(m) 7 microM). Mutation of Asp301 to neutral residues (Asn, Ser, Gly) did not substantially affect the binding of spirosulfonamide (K(s) 2.5-3.5 microM). However, the hydroxylation of spirosulfonamide was attenuated in these mutants to the same extent (90%) as for the classic nitrogenous substrate bufuralol, and the effect of the D301N substitution was manifested on k(cat) but not K(m). Analogues of spirosulfonamide were also evaluated as ligands and substrates. Analogues in which the sulfonamide moiety was modified to an amide, thioamide, methyl sulfone, or hydrogen were ligands with K(s) values of 1.7-32 microM. All were substrates, and the methyl sulfone analogue was oxidized to the syn spiromethylene carbinol analogue of the major spirosulfonamide product. The D301N mutation produced varying changes in the oxidation patterns of the spirosulfonamide analogues. The peptidometic ritonavir and the steroids progesterone and testosterone had been reported to be substrates for P450 2D6, but the affinities (K(s)) were unknown; these were estimated to be 1.2, 1.5, and 15 microM, respectively (cf. 6 microM for the classic substrate bufuralol). The results are consistent with a role of Asp301 other than electrostatic interaction with a positively charged ligand. H-Bonding or electrostatic interactions probably enhance binding of some substrates, but our results show that it is not required for all substrates and explain why predictive models fail to recognize the proclivity for many substrates, especially those containing no basic nitrogen.

A detailed study directed towards metabolic stability optimization of the alkoxy substituents on the catechol moiety of CDP-840 is reported. Replacement of the methoxy and cyclopentyloxy substituents by cyclobutyloxy and/or difluromethoxy groups resulted in the discovery of potent and selective PDE4 inhibitors where the formation of reactive metabolites that could covalently bind to microsomal protein was significantly reduced or eliminated.

Aircraft production is a complex and lengthy process which involves a judicious selection of materials in combination with the option of various production processes. The objective of this chapter is twofold: the first part encompasses the evolution of aircraft materials and related techniques, underlying their impact in the complete life cycle of an aircraft; the second part describes the industrialization process to allow manufacturing of airplanes. It essentially deals with the specific process where engineering is involved namely in terms of the various steps that need to be taken to generate the information essential for the process. It includes also the connection to specific ERP modules such as MRP II and CRP which are essential to the management of aircraft production.

Metabolites of the COX-2 inhibitor rofecoxib (MK-0966, Vioxx) were prepared by synthetic or biosynthetic methods. Metabolites include products of oxidation, glucuronidation, reduction and hydrolytic ring opening. Based on an in vitro whole blood assay, none of the known human metabolites of rofecoxib inhibits COX-1 nor contributes significantly to the inhibition of COX-2.

Our laboratory has been routinely using suspended and cultured human hepatocytes for predicting drug metabolism and enzyme induction by drug candidates to aid drug discovery. Increasing limitation and irregular availability of human tissue has indicated the need for maximizing the use of this valuable resource. Cryopreservation of surplus hepatocytes after isolation would greatly increase the potential of this model. However, cryopreservation of hepatocytes by various methods has resulted in cells with poor metabolic activity and unacceptably low survival rates in culture. Recently, Zaleski et al. (Biochem. Pharmacol. 46 (1993) 111-116) reported that cryopreserved rat hepatocytes retained metabolic capacity similar to fresh hepatocytes when the cells were preincubated for 30 min at 37 degrees C in Krebs Ringer bicarbonate buffer prior to freezing. To further explore this methodology, both the functional capacity of the cells in culture as well as their ability to retain CYP inducibility were investigated with thawed cryopreserved hepatocytes. Although human hepatocytes were used in this study the initial work focused on rat hepatocytes as a cell model. Our results showed that while the preincubation step did not appear to effect the initial viability of cryopreserved hepatocytes, survival of the cells in culture was greatly enhanced. Plating efficiencies for nonpreincubated cryopreserved hepatocytes were decreased to approximately 15% of fresh cells after 48 h in culture. In contrast, cells that had been preincubated prior to freezing had an excellent plating efficiency (approximately 60%) and responded to classical CYP inducers dexamethasone, beta-naphthoflavone and phenobarbital in a manner indistinguishable from that of fresh hepatocytes. Experiments with human hepatocytes have also demonstrated similar results. This is the first time to our knowledge that cryopreserved hepatocytes from both rat and human have been shown to reproducibly respond to CYP inducers in culture.

A series of 5,6-diarylthiazolo[3,2-b][1,2,4]triazoles was prepared for evaluation of potency and selectivity against human COX-1 and COX-2 enzymes. This lead to the discovery of L-768,277, a potent and selective COX-2 inhibitor that also demonstrated good in vivo activity.

Osteoporosis is a pathologic condition characterized by low bone mass and changes in the microarchitecture of the bone tissue. Although compromised bone strength and increased susceptibility to fracture have been established, little is known regarding the process of bone regeneration in osteoporotic conditions. Accordingly, this study sought to evaluate the intramembranous bone regeneration process in an ovariectomized rat model following the establishment of calvarial subcritical-size defects (sCSDs).Calvarial sCSDs were established in rats that had been ovariectomized (Ovx) or sham-operated 2 months previously and left to heal, unfilled, for 6 months. Bone regeneration was assessed by radiographic, densitometric, histologic, and histometric analyses.Radiologic and histologic analyses showed reduced new bone formation in calvarial sCSDs in Ovx animals in comparison to sham animals. Densitometric analysis of radiologic images and histometric analysis showed significant quantitative differences between groups that converged to substantiate reduced bone regeneration in Ovx animals.The intramembranous ossification process is impaired in the Ovx rat model. This may suggest an impairment of the bone regeneration process in clinical conditions of postmenopausal osteoporosis and highlight the requirement for selective bone regenerative strategies in affected patients.

Cork is a natural material with remarkable energy absorption properties, which make it an ideal candidate material when thermal insulation or vibration suppression are major design requirements. However, little work has been done towards the use of cork based materials in structural components. This paper envisages assessing the feasibility of using cork composites with improved specific strength and damage tolerant properties for aerospace applications. A review of some results and conclusions about the mechanical behavior of cork composites will be done based on existing literature and other previous works of the authors on this subject. In particular, two types of materials were considered: 1) a sandwich structure with a cork-epoxy agglomerate core; 2) a carbon-epoxy laminate with embedded cork granulates. In both cases, a set of static and dynamic tests were carried out to characterize the mechanical behavior of the material with a special emphasis on its damage tolerant properties. These tests were replicated using a benchmark core material commonly used in aerospace applications in order to confirm the comparative benefits of cork based composites. A final part of this work seeks to evaluate the advantages of combining the natural damping characteristics of cork with high performance composites aiming at improving the aeroelastic behavior of aerospace components. In particular, the critical flutter speed of a cork based sandwich plate was compared with other conventional materials through a computational analysis. Regardless the type of application, results are encouraging about the use of cork based materials in aerospace components due to their noticeable damage tolerant and high energy absorption properties under different loading scenarios.

Background The risk of coronary heart disease (CHD) is related to environmental factors and genetic variants. Apolipoprotein E (apoE) polymorphisms are heritable determinants of total and low-density lipoprotein cholesterol, with some authors suggesting an association between the ε4 allele and CHD. We investigated the relationship between apoE genotype and age at referral to a specialized lipid clinic by the primary care physician and whether the benefits of treatment with statin differed between genotypes. Methods We assessed individual apoE genotypes and lipid blood profile in a total of 463 patients followed at a specialized lipid clinic due to dyslipidemia, with a 3-year median follow-up time. The primary care physician at the time of the referral had no access to the apoE genotyping results. Carriers of apoE ε4/ε2 genotype were excluded. Results The frequencies of ε2, ε3 and ε4 alleles were 7.8, 78.9 and 13.3%, respectively. There were no significant differences between genders. Although with similar lipid profiles and antidyslipidemic drug usage at baseline, ε4-carriers were referred to the clinic at a younger age (44.2 ± 14.7 years) compared with non-ε4 carriers (50.6 ± 13.8 years) (p < 0.001), with a substantially younger age of referral for homozygous E4/4 and for all genotypes with at least one copy of the ε4 allele (p < 0.001 for trend). Although both ε4 and non-ε4 carriers achieved significant reductions in total cholesterol during follow-up (p < 0.001 vs. baseline), the mean relative decrease in total cholesterol levels was higher in non-ε4 carriers (-19.9 ± 2.3%) compared with ε4 carriers (-11.8 ± 2.3%), p = 0.003. Conclusion Our findings support the concept that there is a reduced response to anti-dyslipidemic treatment in ε4 carriers; this can be a contributing factor for the earlier referral of these patients to our specialized lipid clinic and reinforces the usefulness of apoE genotyping in predicting patients response to lipid lowering therapies.

A series of 5,6-diarylthiazolo[3,2-b][1,2,4]triazoles was prepared for evaluation of potency and selectivity against human COX-1 and COX-2 enzymes. This lead to the discovery of L-768,277, a potent and selective COX-2 inhibitor that also demonstrated good in vivo activity.

A detailed study directed towards metabolic stability optimization of the alkoxy substituents on the catechol moiety of CDP-840 is reported. Replacement of the methoxy and cyclopentyloxy substituents by cyclobutyloxy and/or difluromethoxy groups resulted in the discovery of potent and selective PDE4 inhibitors where the formation of reactive metabolites that could covalently bind to microsomal protein was significantly reduced or eliminated.

The experiments carried out in this investigation were oriented in order to optimize the properties of cork-based agglomerates as an ideal core material for sandwich components of lightweight structures, such as those used in aerospace applications. Static bending tests were performed in order to characterize the mechanical strength of different types of cork agglomerates which were obtained considering distinct production variables. The ability to withstand dynamic loads was also evaluated from a set of impact tests using carbon-cork sandwich specimens. The results got from experimental tests revealed that cork agglomerates performance essentially depends on the cork granule size, its density and the bonding procedure used for the cohesion of granulates, and these parameters can be adjusted in function of the final application intended for the sandwich component. These results also allow inferring that optimized cork agglomerates have some specific properties that confirm their superior ability as a core material of sandwich components when compared with other conventional materials.

Como é que a verdade da fé, sem se trair, pode ser formulada no espaço público de outras palavras concorrentes? Nesta condição agórica e agónica a convicção do fiel (fides qua) e a sua expressão comunicável (fides quae) têm de passar a mais dura das provas: inscrever-se no espaço não da verdade, mas da verosimilhança; não da realidade, mas da aparência; não do dogma, mas da dóxa. As Confissões de Agostinho de Hipona são uma das expressões mais bem conseguidas desta voluntária inscrição da fé no espaço plural da retórica. Consciente do carácter bífido da palavra, o próprio Agostinho confessa ter sido “um vendedor de palavras”; os oradores profissionais e seus aprendizes não passam de “vendedores e compradores de gramática”. Mas ao distinguir a “retórica ao serviço do erro e da mentira”, pura “feira de tagarelices”, da “retórica ao serviço da verdade”, Agostinho reconduz a retória ao lugar neutro de organon e aceita inscrever as razões da existência crente no espaço da persuasão e da verosimilhança, pois não basta à verdade ser verdadeira: importa que o pareça, que seja verosímil. Assim, a verdade íntima da palavra de fé pode e deve ser dita com eloquência e beleza, de tal modo “que ensine, que deleite, que comova”. E o resultado desta crítica retórica da retórica é a obra ímpar Confissões e suas inúmeras mises en scène adentro do ‘teatro do eu’, em busca de si e de Deus, pelos “vastos palácios da memória”. Mas é justamente aí que são denunciadas as pretensões idolátricas de anexar Deus às “experiências do eu”., Without betraying itself, how can the truth of the faith be formulated in the public forum of other competing words? In this agoric and agonic condition the conviction of the faithfull individual (fides qua) and its communicable expression (fides quae) have to passe the hardest of tests: to become involved in the forum not of truth, but of verisimilitude, not of reality, but of appearance; not of dogma, but of doxa. The Confessions of Augustine of Hippo constitute one of the most successful expressions of this willing involvement of faith in the pluralistic forum of rhetoric. Conscious of the divides character of the word, Augustine himself confesses having been “a word salesman”; professional orators and their apprentices are no more than “sellers and buyers of grammar”. But in distinguishing «rhetoric at the service of error and lies”, pure “bartering of chatter”, from “rhetoric in the service of truth”, Augustine redirects rhetoric to the neutral place of organon and agrees to involve the reasons of christian existence in the forum of persuasion and verisimilitude, for it is not sufficient for truth to be true; it must also seem true, be verisimilar. Thus, the intimate truth of the word a faith can and shoud be expressed with eloquence and beauty, so “that it teaches, that it delights, that it moves”. And the result of this rhetorical criticism of rhetoric is the unparalleled work Confessions and its innumerable mises en scène within the ‘theatre of self’, in search of itself and of God, through the “vast palaces of memory”. But it is precisely there that the idolatrous pretensions of annexing God to the “experiences of self” are denounced.

Some DP1 receptor antagonists from an indole-containing series were shown to cause in vitro covalent binding to protein in rat and human liver microsomes. Glutathione trapping experiments along with in vitro labeling assays confirmed that the presence of a strong electron withdrawing group was necessary to abrogate in vitro covalent binding, leading to the discovery of MK-0524. Hepatocyte incubations and in vivo studies showed that acyl-glucuronide formation did not translate into covalent binding.

The ZBRK1 protein is a member of the KRAB-ZFP family. It functions as a transcriptional repressor by binding to its recognition sequence within target genes and producing a nucleoprotein complex containing its co-repressor BRCA1 and also probably the co-repressor KAP-1. Alterations that affect the ZBRK1 gene have not been reported in human tumours. For this reason, possible alterations in the ZBRK1 gene have been studied by analysing mRNA expression using real-time polymerase chain reaction (PCR), and gene sequence using single strand conformation polymorphism (SSCP) analysis and DNA sequencing, in 61 patients with primary breast carcinomas. BRCA1 mRNA expression and allelic loss were also studied in 25 of the same patients. ZBRK1 was underexpressed in 28 (45.9%) and overexpressed in 18 (29.5%) of the 61 cases. No significant association was observed between ZBRK1 mRNA expression and clinicopathological parameters. Loss of heterozygosity of the BRCA1 gene was found in 3 of 23 (13%) informative cases and BRCA1 mRNA expression was altered in 11 of 25 (44%) cases. No significant correlation was found between altered BRCA1 expression and the different types of ZBRK1 mRNA expression. Nine polymorphisms were found in the ZBRK1 sequence, with no significant differences between patients and control subjects. Altered ZBRK1 expression correlated significantly with two of these polymorphisms. A point mutation in one polymorphism and allelic loss in one patient were also observed, findings that indicate that these inactivation mechanisms do not frequently affect this gene. Altered expression of the ZBRK1 gene is therefore frequent in primary breast carcinoma. The functional significance of the polymorphisms in this gene is unclear.

The INK4a/ARF locus encodes two tumour suppressor proteins, p16INK4a and p14ARF, which act in the two main cell-cycle control pathways, p16-Rb and p14-p53 respectively. The present study examined the mRNA expression of these genes by reverse transcription-polymerase chain reaction (RT-PCR), and the inactivation mechanisms that alter these levels, in 100 primary breast carcinomas. Furthermore, the interdependence of these mechanisms was examined, since it has been reported that p14ARF is altered in most tumours in concordance with p16INK4a. The results show that promoter hypermethylation, tested by methylation-specific PCR (MSP), was the major mechanism of inactivation of these genes and was present in 31 (31%) and 50 (50%) of the tumours that showed decreased p16INK4a and p14ARF expression, respectively. Hemizygous deletion was the second cause of down-regulation. Homozygous deletion was rare and mutation was absent. In most tumours overexpressing p16INK4a or p14ARF, no detectable inactivation mechanisms were observed. Finally, the results indicate that these proteins are often co-altered in primary breast tumours and that p16INK4a and p14ARF had non-independent behaviour, since they were silenced or overexpressed concomitantly with a significant correlation (p0.05).

Characterization of the metabolites of the COX-2 inhibitor etoricoxib (MK-0663 and L-791,456) produced in vitro indicate formation of an N-oxide pyridine and hydroxymethyl pyridine that can further be glucuronidated or oxidized to an acid. Significant turnover is observed in human hepatocytes. Several CYPs are involved in the oxidative biotranformations and, from in vitro studies, etoricoxib is not a potent CYP3A4 inducer or inhibitor. Based on an in vitro whole blood assay, none of the metabolites of etoricoxib inhibits COX-1 or contributes significantly to the inhibition of COX-2.

The standard method to evaluate CYP3A inhibition is to study the conversion of the specific CYP3A probe testosterone to its 6 beta-hydroxy metabolite in human liver microsomes, in the absence and presence of potential inhibitors. Quantification of the 6 beta-hydroxy metabolite is achieved by HPLC resulting in a tedious and time-consuming assay. In order to increase the P450 inhibition throughput, efforts were made to find a CYP3A probe that would produce a fluorescent metabolite. This paper reports the discovery of DFB as a potential CYP3A fluorescent probe. DFB was significantly metabolized in human microsomes (approximately 1-2 nmol/(min. mg protein)) to give the fluorescent compound DFH. The involvement of CYP3A in the metabolism of DFB was determined using multiple approaches. First, incubations conducted with microsomes made from cell lines expressing single CYPs (Gentest Supersomes) indicated that CYP3A played a major role in the metabolism of DFB. Secondly, immunoinhibition studies conducted with CYP3A antibody resulted in95% inhibition of DFB metabolism in HLM. Thirdly, inhibition studies with specific CYP1A1, 1A2, 2C8/9, 2C19, 2D6, and 2E1 chemical inhibitors did not suppress DFB activity in HLM. However, ketoconazole, miconazole, nicardipine, and nifedipine, all known CYP3A inhibitors, completely abolished the formation of DFH in HLM. The potency of several inhibitors determined using DFB and testosterone as CYP3A probes was consistent (R = 0.98). Finally, a good agreement was obtained for the formation of DFH and production of 6 beta-hydroxytestosterone when DFB and testosterone were incubated separately with various human liver microsome preparations (R = 0.94, N = 11). In order to use DFH as a fluorescent CYP3A marker in a 96-well plate format, it was important to remove the excess of NADPH at the end of the incubation because the fluorescence of NADPH interferes with DFH detection. This was achieved by adding oxidized glutathione and glutathione reductase to convert NADPH to NADP(+) which is not fluorescent. The liquid-handling steps were fully automated in a 96-well plate format and a template was designed to generate IC(50) curves and to address potential fluorescent interferences from the test compounds. The assay was found to be reproducible (intraday variability10% and interday variability indicated less than a 2-fold variation in the IC(50) values) and is now routinely used in our laboratory to evaluate CYP3A inhibition of NCEs.

Successful cryopreservation of freshly isolated hepatocytes would significantly decrease the need for freshly-procured livers for the preparation of hepatocytes for experimentation. Hepatocytes can be prepared, cryopreserved, and used for experimentation as needed at different times after isolation. Cryopreservation is especially important for research with human hepatocytes because of the limited availability of fresh human livers. Based on the cumulative experience of this international expert panel, a consensus was reached on the various aspects of hepatocyte cryopreservation, including cryopreservation and thawingprocedures and applications of the cryopreserved hepatocytes. Key to successful cryopreservation includes slow addition of cryopreservants, controlled-rate freezing with adjustment for the heat of crystallization, storage at -150 degrees C, and rapid thawing. There is a general consensus that cryopreserved hepatocytes are useful for short-term xenobiotic metabolism and cytotoxicity evaluation.

The cryopreservation of human liver slices is a promising way to enhance the ability to test the metabolism of drug candidates. This study demonstrates the use of a novel technique for the cryopreservation of both rat and human liver slices. In this technique the slices are treated with Me 2 SO and sandwiched between aluminum plates separated by a thin gasket. The device is then submerged in liquid nitrogen to freeze the slices, which can then be stored until use. To thaw the slices, the apparatus is submerged in a water bath at 37°C. Slices frozen and thawed in this manner were compared to those frozen in conventional cryovials. The viability of the slices was determined by incubating them in 12-well plates and measuring urea synthesis, ethoxycoumarin metabolism, and cytosolic enzyme leakage (LDH and ALT). The viability of rat slices frozen between plates approached that of fresh slices and was consistently higher than slices frozen in cryovials. Slices from two human samples gave similar results. The technique was found to work over a wide range of Me 2 SO concentrations (4.5 to 22% was tested) with an optimal concentration between 10 and 15%.