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Urban areas are facing challenges for the provision of public services, with water scarcity arising as one of the main problems. A twin track approach of supply and demand management is essential, and water-loss management contributes to reducing water demand. However, small municipalities from developing countries have technical, information, and financial limitations to locate and monitor water losses. This paper presents the estimation of real and apparent losses in a small municipality from a developing country in a data-scarce situation. For this, several tools were used, allowing data integration that resulted in a water balance, from which water losses were estimated at 46%, and four alternatives for water-loss reduction were developed. A cost-benefit analysis and financial indicators were estimated for the proposed alternatives, resulting in a water savings of 19%, a payback period of 3 years and an internal rate of return of 39%. The proposed strategies have potential to improve water quantity and quality, the technical stability of the system, utility performance, and water security.

Our aim was to identify the potential risk factors for developing post-traumatic endophthalmitis (PTE) and the possible measures of prevention. Retrospective case-control study, with 15 cases of PTE and 2 matched controls. We reviewed the medical records of the cases and their respective controls during the period 1996-2008 at a Spanish Hospital. We collected demographic data and information about the type of trauma, the potential risk factors, comorbidities, microbial isolations, antimicrobial susceptibility, administered treatments, and the visual outcome. The independent predictor factors identified for PTE were intraocular foreign body (IOFB) (OR 5.48; CI 95 % 1.05-28.7), dirty wound (OR 4.91; CI 95 % 0.96-25.3), and wound closure delays of 24 h or more (OR 5.48; CI 95 % 1.05-28.7). The probability of endophthalmitis in patients without these risk factors was 5.9 %, but ascended to 65.3 % and 90.3 %, in those patients with two and three risk factors, respectively. Infected patients presented a complication rate of 80 %, with an evisceration rate of 53 %; both were significantly associated with infection. The visual outcome was poor and related to the presence of IOFB and virulent microorganisms (Bacillus sp., filamentous fungus), visual acuity at presentation, and retinal detachment. Patients who presented an IOFB, dirty wound, and delayed wound closure were 15 times more likely to develop infection, and when infected, patients fared much worse than those non-infected. We thus recommend aggressive prophylactic measures in patients with these risk factors, adding antifungal prophylaxis when the injury is contaminated with vegetable matter.

While they account for one-fifth of new cancer cases, rare cancers are difficult to study. A higher than average degree of uncertainty should be accommodated for clinical as well as for population-based decision making. Rules of rational decision making in conditions of uncertainty should be rigorously followed and would need widely informative clinical trials. In principle, any piece of new evidence would need to be exploited in rare cancers. Methodologies to explicitly weigh and combine all the available evidence should be refined, and the Bayesian logic can be instrumental to this end. Likewise, Bayesian-design trials may help optimize the low number of patients liable to be enrolled in clinical studies on rare cancers, as well as adaptive trials in general, with their inherent potential of flexibility when properly applied. While clinical studies are the mainstay to test hypotheses, the potential of electronic patient records should be exploited to generate new hypotheses, to create external controls for future studies (when internal controls are unpractical), to study effectiveness of new treatments in real conditions. Framework study protocols in specific rare cancers to stepwisely test sets of new agents, as from the early post-phase I development stage, should be encouraged. Also the compassionate and the off-label settings should be exploited to generate new evidence, and flexible regulatory innovations such as adaptive licensing could convey new agents early to rare cancer patients, while generating evidence. Though validation of surrogate end points is problematic in rare cancers, the use of an updated notion of tumor response may be of great value in the single patient to optimize the use of therapies, all the more the new ones. Disease-based communities, involving clinicians and patients, should be regularly consulted by regulatory bodies when setting their policies on drug approval and reimbursement in specific rare cancers ., While they account for one-fifth of new cancer cases, rare cancers are difficult to study. A higher than average degree of uncertainty should be accommodated for clinical as well as for population-based decision making. Rules of rational decision making in conditions of uncertainty should be rigorously followed and would need widely informative clinical trials. In principle, any piece of new evidence would need to be exploited in rare cancers. Methodologies to explicitly weigh and combine all the available evidence should be refined, and the Bayesian logic can be instrumental to this end. Likewise, Bayesian-design trials may help optimize the low number of patients liable to be enrolled in clinical studies on rare cancers, as well as adaptive trials in general, with their inherent potential of flexibility when properly applied. While clinical studies are the mainstay to test hypotheses, the potential of electronic patient records should be exploited to generate new hypotheses, to create external controls for future studies (when internal controls are unpractical), to study effectiveness of new treatments in real conditions. Framework study protocols in specific rare cancers to sequentially test sets of new agents, as from the early post-phase I development stage, should be encouraged. Also the compassionate and the off-label settings should be exploited to generate new evidence, and flexible regulatory innovations such as adaptive licensing could convey new agents early to rare cancer patients, while generating evidence. Though validation of surrogate end points is problematic in rare cancers, the use of an updated notion of tumor response may be of great value in the single patient to optimize the use of therapies, all the more the new ones. Disease-based communities, involving clinicians and patients, should be regularly consulted by regulatory bodies when setting their policies on drug approval and reimbursement in specific rare cancers.

Background and Objectives: Endoscopic ultraendosonography (UES) is a sensitive technique for the diagnosis of choledocholithiasis. Our study aimed to determine the positive predictive value (PPV) of UES for the detection of choledocholithiasis. Methods: We performed a descriptive, retrospective, prospective observational, analytical study, with 150 patients who met the inclusion criteria. Endosonography and endoscopic retrograde cholangiopancreatography (ERCP) were performed in the Union of Surgeons Clinical Presentation of the city of Manizales; statistical analysis was performed using the SPSS version 21 program. Results: Fifty-five patients (36.7%) were male and 95 (63.3%) were female. Of the 150 patients positive for choledocholithiasis by UES, 29 of them (19.3%) had images compatible with the presence of bile clay, with 28 patients corroborated in ERCP, with a PPV of 96.6%. As for choledocholithiasis, it was corroborated in 139 patients in ERCP for a PPV of 92.7%. Conclusion: Endosonography has a high valor preditivo positivo for the detection of calculi and mud in the bile duct, being useful in the diagnosis of choledocholithiasis, due to its sensitivity for the detection of microlithiasis (

Our objectives are to describe the microbial spectrum and antimicrobial susceptibility of isolates from patients with culture-proven endophthalmitis.Retrospective study of patients with microbiological diagnosis of endophthalmitis treated at the Ophthalmology Department of the General Hospital La Mancha Centro in the period 1996-2008. The identification of isolates was performed using the automated VITEK-2® and Api galleries (bioMérieux, Spain SA). The antimicrobial susceptibility was performed by the VITEK-2® system (bioMérieux, Spain SA), E-test strips (MIC Test Strip, Liofilchem, Italy) and Sensititre® YeastOne trading system (Trek Diagnostic Systems, Ohio, USA) for Candida species.Forty four (70%) of 63 cases of endophthalmitis were culture positive. Gram-positive bacteria were much more common than gram-negative bacteria in both postoperative endophthalmitis (POE) and post-traumatic endophthalmitis (PTE). Staphylococcus epidermidis was predominant in POE, while Bacillus sp. predominated in the PTE; furthermore, the 75% of total fungal isolates corresponded to postraumatic cases. The isolated strains showed 100% susceptibility to vancomycin, ceftazidime and amikacin, while resistance to ciprofloxacin was greater than 15%. The empirical antifungal therapy failed in 50% of cases. The visual prognosis was significantly less favorable in the PTE.Based on the susceptibility of our isolates, vancomycin, ceftazidime and amikacin are good choices for empirical treatment of endophthalmitis, unlike ciprofloxacin. We recommend conducting antifungal prophylaxis after penetrating ocular trauma in a rural environment.

The search for techniques that allow a more efficient use of water is a global task, since this resource is increasingly scarce and its quality is constantly deteriorating. The objective of this study was to determine the effect of applying coconut fiber CF on physical-hydric parameters of the soil and its water use efficiency (WUE). The effects of coconut fiber (CF) were assessed on water consumption, water use efficiency and aerial and root biomass of Italian ryegrass (Lolium multiflorumL.). In addition, it was also evaluated whether CF causes toxic effects on lettuce seeds (Lactuca sativaL.). Treatments were established according to the proportion of CF and soil in g g-1soil CF: T1 = 1:80, T2 = 1:40 and T3 = 1:20, and a control treatment (T0 = no substrate). Additionally, 3 charges of water (100, 75, 50 % of field capacity) were established. Results showed that the best treatment for water consumption was T3-50 (377.8 ml), for WUE the best treatment was T3-100 (8.52 kg DM m-3), for aerial biomass it was T3-100 (4.59 g MS) and in the case of root biomass the best treatment was T3-100 (1.92 g MS). It was concluded that the addition of coconut fiber substrate to the mix improves the physical and hydrological characteristics of degraded soil. There were no toxic effects on lettuce among the applied rates of CF. The germination index (IG) ranged from 124.82 to 156.03 % and the best treatments were T3 and T2. &nbsp, La búsqueda de alternativas que permitan un uso más eficiente del agua es una tarea mundial, puesto que este recurso es cada vez más escaso y su calidad se deteriora. De ahí que la presente investigación tuvo como objetivo determinar el efecto de la aplicación de fibra de coco (FC) sobre parámetros físico-hídricos del suelo y su eficiencia en el uso del agua (EUA). Se probaron los efectos de la FC sobre el gasto de agua, sobre la EUA, biomasa aérea y radical en ballica italiana (Lolium multiflorumL.). Además se evaluó si la FC presenta efectos tóxicos sobre las semillas de lechuga (Lactuca sativaL.). Los tratamientos fueron establecidos de acuerdo a la proporción de FC y suelo, en gramos FC g-1suelo: T1 = 1:80, T2 = 1:40 y T3 = 1:20, y un testigo (T0 = Sin sustrato). Adicionalmente, se establecieron 3 cargas de agua (100, 75, 50 % de capacidad de campo). Los resultados muestran que el menor consumo de agua se dio en el tratamiento T3-50 (377,8 ml), para la EUA el mejor tratamiento fue T3-100 (8,52 kg MS m-3), para biomasa aérea, el mejor fue el tratamiento T3-100 (4,59 g MS) y en el caso de la biomasa radical su mejor tratamiento fue el T3-100 (1,92 g MS). Por lo tanto, al adicionar el sustrato de fibra de coco en la mezcla, este permite un mejoramiento en las características físico-hídricas del suelo degradado. Las pruebas con lechuga indicaron que no existen efectos tóxicos entre las dosis aplicadas de FC. Los índices de germinación (IG) variaron entre 124,82 a 156,03 % y los mejores tratamientos fueron para T3 y T2. &nbsp

International audience

Poster presented at the Fourth International Conference from Nanoparticles and Nanomaterials to Nanodevices and Nanosystems that took place in Corfu Island (Greece) during 16-20th June 2013., This study [1] addresses the growth dynamics of nanocrystalline diamond (NCD) films deposited by hot-filament chemical vapor deposition (HFCVD). It is based on the statistical analysis of the evolution of the surface morphology of the films at the mesoscale using atomic force microscopy (AFM). Concepts of the Dynamic Scaling Theory are used focusing on the changes of the growing surface roughness and its lateral correlations with time. In conclusion, the NCD growth dynamics displays two growth regimes., JGB would like to thank the Executive Research Agency of the EU for funding under Marie Curie grant 272448 support by Comunidad Autónoma de Madrid (S2009/PPQ-1642) and Ministerio de Economía y Competitividad (FIS2012-38866-C05-05).

Ni-TiO2nanocomposites coatings with a layer thickness of 21 μm have been obtained by electrocodeposition. Incorporation in nickel matrix of TiO2nanoparticles (10 nm), affects the morphology of the nickel matrix as indicated from scanning electron microscope (SEM) and atomic force microscope (AFM) results. The chemical composition was studied by energy dispersive X-ray spectroscopy (EDS). Analysis by X-ray diffraction (XRD) revealed the inclusion of nickel particles in the matrix and their effect on nickel matrix crystal size changes., This work was financially supported by the research Project IFA-CEA C2-02/01/03/2012.

Even though concentrations of most chemical elements in Antarctic ecosystems are very low as compared to other world's areas, their increasing time trends, as a consequence of the strong population growth and industrial development in countries of the Southern Hemisphere, could be affecting some vulnerable endemic species such as penguins. Concentrations (dry weight) of arsenic (As), cadmium (Cd), mercury (Hg) and lead (Pb) and porphyrins (copro-, uro- and proto-) were determined in feces of Gentoo penguins (Pygoscelis papua), collected at two locations of the Antarctic Peninsula (O'Higgins and Videla). We found higher (p< 0.05) levels of Hg in excreta of penguin colonies at O'Higgins (7.55 ± 1.28 μg g-1), whereas penguins at Videla showed the highest concentrations of Cd (1.68 ± 0.71 μg g-1). Gentoo penguins from O'Higgins site showed higher levels of copro- (1.81 ± 0.61 nmol g-1), uro- (1.74 ± 0.72 nmol g-1) and protoporphyrins (1.24 ± 0.46 nmol g-1), directly related to higher levels of Hg and Pb measured. These findings imply that some Gentoo penguin colonies of the Antarctic Peninsula are likely to develop some biochemical effects through trace metal contamination at some level. These preliminary results raise concerns about anthropogenic pollutants in Antarctica. Aunque las concentraciones de la mayoría de los elementos químicos en la Antártica son bajas en comparación con otras áreas del planeta, éstas se han ido incrementando con el tiempo debido al gran crecimiento de la población y desarrollo industrial de los países del Hemisferio Sur, lo cual podría estar afectando a especies vulnerables como los pingüinos. Se determinaron las concentraciones (peso seco) de arsénico (As), cadmio (Cd), mercurio (Hg) y plomo (Pb) y porfirinas (copro-, uro- y proto-) en heces de colonias de pingüinos Papua (Pygoscelis papua), colectadas en dos lugares de la península Antártica (O'Higgins y Videla). Se encontraron mayores (P < 0,05) niveles de Hg (7,55 ± 1,28 μg g-1) en las heces de colonias de pingüinos en O'Higgins, mientras que las colonias de pingüinos en Videla exhibieron mayores concentraciones de Cd (1,68 ± 0,71 μg g-1). Los pingüinos Papua de O'Higgins mostraron mayores (P < 0,05) niveles de copro- (1,81 ± 0,61 nmol g-1), uro- (1,74 ± 0,72 nmol g-1) y protoporfirinas (1,24 ± 0,46 nmol g-1), en directa relación con los mayores niveles de Hg y Pb medidos. Esto podría implicar que algunas colonias de pingüinos Papua de la península Antártica están propensas a desarrollar algunos efectos bioquímicos asociados a contaminación por metales traza. Estos resultados preliminares levantan inquietantes interrogantes acerca de la contaminación de origen antropogénico que estaría afectando áreas tan remotas como la Antártica.

The biocompatible metals made available for powder injection moulding (PIM) are increasing, and as a result, the PIM process is becoming attractive for manufacturers of medical implants and surgical instruments. In addition, friction and wear properties play an important role in biomaterial applications. This study aimed to systematically investigate the fretting wear properties of 316L stainless steel parts produced by PIM under dry contact conditions. The fretting experiments were performed under gross-slip regime. The influence of a number of fretting test parameters on the friction and wear behaviour was investigated, as the contact frequency, the normal load, and number of reciprocating sliding cycles. Fretting tests were performed against corundum counter bodies. After the tests, the wear scars were examined by laser surface profilometry, scanning electron microscopy (SEM) and energy dispersive X-ray microanalysis. The results indicate that wear occurred predominantly by abrasion, plastic deformation, and cracking.

OBJETIVO: Determinar la frecuencia de litiasis vesicular en pacientes post-operados con gastrectomía por adenocarcinoma gástrico, determinar el tiempo de aparición de litiasis vesicular en pacientes gastrectomizados por Adenocarcinoma Gástrico. MÉTODOS: Se realizo un estudio observacional, descriptivo retrospectivo. Se revisó un total de 148 historias clínicas de pacientes sometidos a gastrectomía por adenocarcinoma gástrico en el Instituto Nacional de Enfermedades Neoplásicas en el periodo de 1990 al 2000. RESULTADOS: De los 148 casos de pacientes post gastrectomizados por adenocarcinoma gástrico que fueron enrolados, 29(19.6%) presentaron litiasis vesicular versus 119(80.9%) que no presentaron litiasis vesicular como complicación post gastrectomía. La media de edad de 29 pacientes con litiasis vesicular post gastrectomía por adenocarcinoma gástrico fue de 59.9años (min. 39años máx.74años). La distribución de la litiasis vesicular en relación al sexo fue, femenino 18(62.1%) casos y masculino 11(37.9%) casos. La media del tiempo de aparición de la litiasis vesicular post gastrectomía fue de 3.1 años, para el sexo masculino 2.7años y para el sexo femenino de 3.3 años .Según el tipo de cirugía se presento litiasis vesicular en 14 pacientes que fueron sometidos a Gastrectomía Subtotal Distal y a 15 pacientes a los que se les practicó Gastrectomía Total. CONCLUSIONES: la Frecuencia de litiasis vesicular post gastrectomía fue de 19.6% . El periodo de aparición de colelitiasis fue de 3.1 años. La colecistectomía podría ser una toma de decisión importante en pacientes con alto riesgo de litiasis y cáncer vesicular post gastrectomía. El presente trabajo de investigación invita a realizar otros estudios de tipo cohorte en el cual se determine el factor de riesgo principal para la aparición de esta complicación. SUBJECT: To determinate the frecuency and the time of development of the gallbladder stones in gastrectomy post-operated patients with stomach adenocarcinoma at The National Cancer Institute in Lima, Peru. METHODS: In an observational, descriptive and retrospective case series design, 148 patiens' files who underwent gastrectomy for stomach adenocarcinoma in the National Cancer Institue of Lima during 1990 and 2000, have been reviewed looking for the development. RESULTS: A total de 148 patients were involved in this study. 29 of them (19.6%) develop gallbladder stones during the (x years of) follow up vs 119 (80.9%). The mean age ot the 29 patients with gallbladder stones were 59.9 years ans 18 of them were female and 11 male.\The mean time of develop gallbladder stones was 3.1 years. According to the type of surgery, 14 patient wiht gallbladder stones underwent to subtotal gastrectomy and 15 to total gastrectomy. CONCLUSIONS The frecuency of gallbladder stones post gastrectomy in this study was 19.6%. The mean time of the develop and diagnosis of litiasis was 3.1 years. To perform the colecistectomy at the same time of the gastrectomy could be an important decision in patients with high risk of gallstones and gallbladder cancer. We need furthermore studies to have conclusions about the risk factors.

Even though titanium (Ti) has the ability to develop a highly stable passive oxide layer (TiO2) that provides an excellent corrosion resistance; it’s not inert to corrosive attack. Under in vivo conditions, such as in orthopedic and dental implants, a tribological contact may be formed in association with a chemical aggressive environment. Consequently, wear debris and corrosion products or ions can lead to inflammatory reactions, such as osteolysis (bone resorption), provoking peri-prosthetic bone loss and subsequent loosening of the implant/prosthesis [1]. The microbial effect on the corrosion of metallic biomaterials remains unknown. Chang et al. [2] reported an increase on the corrosion behaviour of dental metallic materials in the presence of Streptococcus mutans and its growth byproducts. Moreover, brushing can lead to the production of wear debris, which together with the attachment of microbes can disturb the passivity of the metal oxide. In addition, the organic acids produced by bacteria may reduce the pH, favoring the corrosion and tribocorrosion of implants. The aim of this work was to study the influence of mixed biofilms on the biotribocorrosion behaviour of commercially pure (cp) Ti for dental implants, and the consequent effect of fluoride.

The use of dental implants has been increasing even though failures do occur. The presence of wear debris and oral microorganisms can contribute to infections and jeopardize implant integration. The aim of this work was to study the influence of mixed biofilms in the tribological behaviour of commercially pure titanium for dental implants under different concentrations of fluoride. Samples of titanium with two different surface topographies were used. Mixed biofilms of Candida albicans and Streptococcus mutans were formed on both surfaces at 37 °C in a tryptic soy broth containing mucin, peptone, yeast extract and sucrose. After 8 days, biofilm biomass was analysed by crystal violet staining method. Biofilm biomass was significantly higher for the samples with higher roughness. Some samples with biofilms were analysed under friction (using a force of 100 mN) in an artificial saliva solution (Fusayama) without or with different concentrations of fluoride (30 and 227 ppm). It was verified that the coefficient of friction (COF) decreased in the presence of biofilms. Moreover, samples with more biomass (0.4 μm of roughness) presented the lowest values of COF. Concerning the effect of the presence of fluoride, although there were no significant differences on the COF for 30 ppm, for 227 ppm a transition regimen was observed. These results were confirmed by sample observation under scanning electron microscopy. In conclusion, it can be highlighted that biofilm formation on dental implants can significantly affect the tribological behaviour of titanium, namely, the presence of biofilms reduces the release of wear debris.

Pollution of aquatic fresh water and soil is of great concern in the salmon farming industry. Water is the most important vehicle for the transmission of organic polluting agents, and for that reason the sediments (sludge) near to salmon facilities could become enriched reservoirs of pathogens and chemicals. A solution to this problem could be to incorporate the sludge into the soil under sustainable procedures. In order to investigate the addition of salmon sludge to soils, sediment samples from a pisciculture and a lake were applied to a degraded in the soil southern Chilean Patagonia. Microbial biological activity was evaluated under different salmon sludge addition rates: 25, 50, 75, 100 y 150 t ha-1by using respirometry tests (C-CO2) in samples under closed incubation in laboratory. The results showed that microorganisms increased their respiratory activity along with time and increased sludge concentrations, indicating the amount of sludge that is possible to incorporate into a soil without causing adverse environmental effects. &nbsp, La contaminación de los suelos y de los ecosistemas acuáticos de agua dulce es una preocupación constante para la industria de la salmonicultura. El agua es el vehículo más importante para la transmisión de contaminantes orgánicos, y por ello los sedimentos (lodos) próximos a las instalaciones de una piscicultura o salmonicultura pueden convertirse en reservorios de patógenos y químicos. Una solución es enmendar estos lodos en suelos de manera sustentable. Para investigar la adición de lodos residuales de salmonicultura en suelos, muestras de sedimentos obtenidos de piscicultura y de lago fueron aplicadas a un suelo patagónico degradado del sur de Chile. Se evaluó la actividad biológica de los microorganismos bajo diferentes dosis de lodos enmendados al suelo: 25, 50, 75, 100 y 150 t ha–1. La determinación se realizó mediante pruebas de respirometría (C-CO2) en muestras sometidas a incubación cerrada en laboratorio por60 días. Los resultados mostraron que los microorganismos aumentan su actividad respiratoria a medida que transcurre el tiempo y con mayores tasas de aplicación de lodo, indicando con ello la cantidad de lodo residual que es posible disponer o verter en un suelo sin provocar efectos ambientales adversos. &nbsp

The oral cavity is a complex environment where corrosive substances from dietary, human saliva and oral biofilms may accumulate in retentive areas of implant-supported prostheses promoting a corrosion of their surfaces. On the other side, during mastication, micro-movements may occur in artificial joints causing a relative contact motion between surfaces. Both processes result in a tribocorrosion system. The objective of this work is to investigate the in-vitro tribocorrosion behavior of titanium covered by biofilms and immersed in artificial saliva. Reciprocating sliding wear tests coupled with electrochemical techniques were performed at 25 o C on titanium samples covered or not with biofilms. Mixed biofilms of Candida albicans and Streptococcus mutans were grown at 37 oC on titanium surfaces for 9 days in Tryptic Soy Broth medium, containing mucin, yeast extract, peptone and sucrose. The open-circuit potential (OCP) was measured until stabilization in artificial saliva. After that, electrochemical impedance spectroscopy (EIS) measurements were carried out at open circuit potential. Subsequently, sliding tests were performed at normal loads of 0.1, 0.2 and 3 N, at a sliding frequency of 1 Hz, and linear displacement amplitudes of 0.5 and 2 mm using a tribometer equipped with a test viewer software. During the sliding tests, the OCP was constantly monitored. At the end of the sliding test, the OCP was further monitored until stabilization, and another series of EIS measurements was made. Worn and unworn surfaces were inspected by SEM. The detachment of the biofilms was not noticed when low loads were applied, and biofilms appear to have some lubricating effect. However, it was found that acidic substances released from Streptococcus mutans and external substances accumulated into the biofilm might corrode surfaces located below and around the biofilms. At a normal load of 3 N, the biofilms were removed from the contact region by the sliding action, and it was observed that the repassivation rate of titanium covered with biofilms was lower than without biofilms., Fundação para a Ciência e a Tecnologia (FCT), ALBAN, Flemish Science Foundation. Scientific Research Community on Surface Modification of Materials, Programa Alban - Bolsas de Alto Nível da União Europeia para a América Latina

The oral cavity can be considered as an aggressive environment to restorative materials considering that several acidic substances can be retained in the saliva and biofilms as well as loads from mastication and abrasive particles take place in the wear of oral surfaces. The objective of this work is to investigate the in-vitro corrosion and wear in fluoridated artificial saliva and in presence of biofilms. Electrochemical tests of titanium surfaces were performed in artificial saliva containing different fluoride concentrations at 37 oC. Another group of titanium surfaces were prepared to the growth of oral mixed biofilms for 9 days in a specific culture medium. Then, electrochemical tests were carried out with titanium surfaces covered with biofilms in artificial saliva without fluorides. After that, wear sliding tests were performed in the same mediums using a tribometer equipped with a test viewer software. Also, electrochemical measurements were carried during the wear sliding tests. After corrosion-wear tests, worn and unworn surfaces were analyzed by AFM and SEM. A localized corrosion of titanium was only noticed in high fluoride concentration. Therefore, there was a decrease of the corrosion resistance of titanium when the fluoride concentration was increased. Moreover, the presence of biofilms affected harmfully the corrosion resistance of titanium probably due to acids release from the biofilms. However, the wear was decreased when low loads were applied on the biofilms., Fundação para a Ciência e a Tecnologia (FCT) - PTDC/CTM/67500/2006, Programa Alban - Bolsas de Alto Nível da União Europeia para a América Latina - E06D103407BR, Flemish Science Foundation. Scientific Research Community on Surface Modification of Materials

In the oral cavity, restorative surfaces such as of titanium-based implant systems are in contact with several acidic substances originated from dietary intake and microbial metabolism. During the mastication, a simultaneous corrosion-wear process known by Biotribocorrosion can take place on the surfaces which are in relative contact motion. The objective of this work is to investigate the in-vitro simultaneous corrosion-wear behavior of titanium in the presence of biofilms. Electrochemical tests of titanium surfaces covered or not by mixed oral biofilms (Streptococcus mutans and Candida albicans) were carried out in artificial saliva. After stabilization of open circuit potential of titanium in artificial saliva, reciprocating wear sliding tests were performed against an alumina ball at different loads using a tribometer equipped with a test viewer software. Moreover, electrochemical measurements were carried during the wear sliding tests. Worn and unworn surfaces were analyzed by different SEM techniques. Biofilms decreased significantly the friction of titanium surfaces. However, the presence of biofilms affected negatively the corrosion resistance of titanium probably due to acids release from the biofilms., Comissão Europeia - programa Alban (cod. E06D103407BR), Fundação para a Ciência e a Tecnologia (FCT) - PTDC/CTM/67500/2006), Flemish Science Foundation. Scientific Research Community on Surface Modification of Materials (WOG-FWO Vlaanderen)

Titanium and titanium alloys have been widely used in orthopeadics and dental implants because of their excellent properties such as low modulus, good fatigue strength, good corrosion resistance and biocompatibility. However, their poor wear resistance still limits their application. When inserted in the oral environment dental implants are under a complex degradation phenomenon as a result of the combined action of chemical and mechanical solicitations, which can result in its failure. The relatively poor wear resistance of titanium may be improved by surface modification, namely by anodic oxidation. Also, as suggested by recent works carried out using animal models, bone regeneration may be enhanced by acting on the surface topography of the implant, essentially if the an oxide is present at the surface. The present study aims to investigate the surface properties of the anodic oxide films created on c.p. Ti. Oxide films were prepared by anodization (potentiostatic mode) in citric acid with different molar concentration and different applied voltages. The thickness, composition, structure and topography of the prepared films were investigated by XRD, RBS, and AFM. Also electrochemical impedance spectroscopy was used to characterize the original native oxide film before anodization, in comparison with the oxide film after anodic oxidation treatment. AFM results demonstrate substantial differences on film topography, with an enhancement on surface roughness. Also, changes on structure, thickness and chemical composition of the anodic oxide layers influences the electrochemical characteristics of the films.

International audience; As-plated and annealed NiP coatings and composite NiP-SiC coatings were investigated in uni-directional ball-on-disc sliding tests. Abrasive wear was noticed in the case of composite NiP coatings containing submicron SiC particles, whereas in NiP coatings oxidational wear was active. The addition of submicron SiC particles not only increases the hardness of these electrolytic coatings but also hinders the formation of an oxide film in the sliding wear track. As a consequence, the wear loss on as-plated NiP coatings is not markedly reduced by the addition of SiC particles. On the contrary, a heat treatment at 420 °C for 1 h decreases the wear loss on both pure NiP and composite NiP-SiC coatings. During that heat treatment, Ni3P precipitates are formed in the NiP matrix and owing to this fact, the hardness of both pure NiP and composite NiP-SiC coatings increases. However, the heat treatment of composite NiP-SiC coatings induces the sensitivity for crack formation in the NiP matrix around these SiC particles. As a result, the pull out of SiC particles in the wear track occurs easily during sliding, and the wear loss of composite NiP-SiC coatings remains above the wear loss on NiP coatings.

International audience; As-plated and heat-treated electrodeposited NiP and composite NiP-SiC coatings were investigated in bi-directional ball-on-disc sliding tests. All tests were performed under gross slip conditions. Heat treatment decreases the wear volume loss during fretting in ambient air for all coatings investigated. Heat-treated NiP coating has a lower wear volume loss compared to composite NiP-SiC coatings for all sliding tests. The wear rate at the bi-directional sliding test was found to be lower relative to the wear rate at uni-directional sliding test.

Titanium has been widely used for dental implant applications because their attractive properties such as low Young modulus, good fatigue strength, excellent corrosion resistance and biocompatibility. However it cannot meet all clinical requirements. 1-2 When inserted in the oral environment, dental implants are submitted to a complex degradation phenomena occurring due to the interaction of tribological and corrosive phenomena which can result on its failure. In fact, most of the failures are a result of the combined action of mechanical solicitations (sliding or abrasive wear, erosion, impact, fretting or fatigue processes) and chemical solicitations as a consequence of contact with saliva, cells or bacteria. Consequently, dental implants become part of a tribocorrosion system, and the investigation of the tribocorrosion mechanisms in such systems becomes essential. 3 Recent work has shown that wear resistance of titanium might be improved selectively using the appropriate surface treatment, such as plasma oxidation, which is also often used to enhance the osseointegration process. 4-5 The main aim of this paper is to study the influence of the characteristics of the oxide layers obtained by plasma oxidation on the tribocorrosion behaviour of c.p. Ti intended for the fabrication of dental implants.The plasma oxidation treatment were performed at three different temperatures (300, 450 and 530ºC) and a detailed study of surface topography (Fig. 1), thickness, structure and chemical composition of the oxidized layers was performed. Tribocorrosion experiments were performed using a reciprocating motion pin-on-plate tribometer with an alumina pin as a counterface material in the presence of Fusayama solution. The applied normal force used was 3N, the stroke length 1mm and the displacement frequency was 1Hz. During rubbing the friction coefficient was monitored. Electrochemical techniques were used to follow the evolution of the system during sliding. Each experiment involved an in situ surface cleaning by a cathodic polarization at -900mv during 3 minutes, and after that a potentiostat was used to impose a passive potential to titanium samples. This experiment allows in situ and in real time following of the corrosion kinetics through current measurements. It is important to refer that electrochemical impedance spectroscopy was also used before and after rubbing in order to evaluate the protective character of the oxide film. The influence of the thickness of the oxide layer and of surface roughness on the wear-corrosion behaviour of the material is discussed in detail. Results clearly showed that plasma oxidation treatments have a strong influence on the tribocorrosion behaviour of titanium. Both the corrosion and wear performance of the samples are improved by the increase of the processing temperature. In relation to friction profile no significant differences were found between all samples tested (μ remains between 0.35 and 0.45). Results show that electrochemical techniques yield useful results.

The demand for metallic materials in medical and dental devices is large; metals and alloys are widely used as biomedical materials and are indispensable in the medical field. In particular, toughness, elasticity, rigidity, and, some times, electrical conductivity are essential properties for metallic materials used in medical devices. In dentistry, titanium is used for restorations, orthodontic wires, and dental implants. Materials implanted in the human body are intermittently stressed with loads, which are repeated in a tremendous amount of cycles. During mastication, dental implants are simultaneously subjected to corrosion by the oral environment and to the sliding-wear process by bite forces. Dental wear can be attributed to mastication where the teeth are worn by attrition and abrasion. The main aim of the present study was to investigate the tribocorrosion behaviour of titanium grade 2 in reciprocating sliding conditions in contact with artificial saliva solutions. Particular attention is given to the relationship between mechanical damage and the electrochemical behaviour of the material. Sliding-corrosion experiments were performed using a reciprocating ball-on-flat contact configuration and the open circuit potential was monitored during the test. Also electrochemical impedance spectroscopy was used to obtain a more detailed description of the corrosion mechanisms and to characterise the passive layer that forms on the titanium surface. To reproduce the oral environment around the dental implant, some additives (citric acid, anodic, cathodic and organic inhibitors) were added to simple artificial saliva constituted mainly by NaCl and KCl, and with a pH between 5 and 7. A corundum ball was loaded on the top of the flat rectangular sample at a normal force of 10N. Linear peak-to-peak displacement amplitude of 3 mm at vibrating frequencies of 1 Hz was applied to the tribocorrosion system. The tests were performed for 11160 cycles. The surface roughness and topography of the titanium samples were evaluated prior to testing. In relation to the friction profile, for all solutions, no significant differences were observed. In fact, the aqueous environment used doesn’t have sufficient viscosity to cause changes in the friction behavior. Regarding the weight loss results it is possible to conclude that titanium in artificial saliva solution with citric acid present the high weight lost. Additionally, cathodic and organic inhibitors solutions exposed a higher tendency for corrosion, forming an oxide passive film with lower protection characteristics.

Tribocorrosion refers to a complex degradation of materials occurring due to the interaction between a tribological process and a corrosive environment. From the tribological point of view, the interaction with the material may include sliding or abrasive wear, erosion, impact, fretting or fatigue processes. The environmental solicitation may result from temperature, humidity, liquids or gases in permanent or intermittent contact with the materials. Failure in the metallic part or of the metal/ceramic interfaces existing in dental restorations, such as crowns and bridges, are still relatively frequent. Also, excessive degradation of dental materials is sometimes observed in other dental materials. Most of the failures are due to the simultaneous action of mechanical (wear, fretting and fatigue) and chemical (saliva) solicitations. As a consequence, the investigation of the tribocorrosion mechanisms in such systems becomes essential. Regarding this aspect, in last years particular focus is being given to the exploitation of electrochemical methods as a tool for the investigation of the combined corrosion-wear degradation of the materials. Also, it is known that the mechanical contact geometry is of crucial importance regarding the degradation behaviour. In this work the tribocorrosion behaviour of commercially pure titanium in contact with artificial saliva solutions was investigated. Tests were conducted in a reciprocating sliding geometry with movement amplitudes varying from 200 m (fretting) up to 6 mm (wear), and normal loads between 2 and 10 N. The electrochemical noise technique was used in order to follow the evolution of both the corrosion current and of the open circuit potential of the system during the wear tests. The pH of the artificial saliva solution was varied between 4 and 7. Results show that the behaviour of the material is strongly influenced by the pH of the solution. In fact, acidification of the solution improves the electrochemical response of the material, in particular the repassivation kinetics after sliding, in all the test conditions used in this work. However, the characteristics of the passive film formed on the material surface appear to be less protective than that formed at neutral pH. Also, an abnormal amount of titanium is removed from the material at low pH, when the normal load applied during the wear test is increased.

Sol-gel hybrid organic-inorganic and inorganic SiO2-based protective coatings with and without added 3 μm glass particles were developed and tested for their corrosion and wear behavior of an stainless steel substrate (AISI316L). The corrosion resistance greatly increases by incorporating glass particles in the sols. The incorporation of particles in the coatings allows the synthesis of thicker crack-free coatings. On the other hand, the corrosion resistance increases for coatings with a higher organic content obtained at lower sintering temperature. These coatings are also highly stable in saline aqueous solutions. However, the wear resistance is badly affected by the hybrid character of the SiO2 matrix. The optimum coating process in terms of corrosion and wear resistance, appears to be a hybrid system with a dense SiO2 network achieved at intermediate sintering temperatures.

The fretting wear of sulphur-deficient MoSx coatings with different crystallographic orientations has been investigated in ambient air of controlled relative humidity. The coefficient of friction and the wear rate of MoSx coatings sliding against corundum depend not only on fretting parameters like contact stress, fretting frequency, and relative humidity, but also strongly on the crystallographic orientation of the coatings. For randomly oriented MoSx coatings, the coefficient of friction and the wear rate increased significantly with increasing relative humidity. In contrast, basal-oriented MoSx coatings were less sensitive to relative humidity. The coefficient of friction of both types of MoSx coatings decreased on sliding against corundum with increasing contact stress and decreasing fretting frequency. A correlation between dissipated energy and wear volume is proposed. This approach allows detection in a simple way of differences in fretting wear resistance between random- and basal-oriented MoSx coatings tested in ambient air of different relative humidity.

In our laboratories we are exploring the possibility of using proteome expression profiles of fresh bladder tumors (transitional cell carcinomas, TCCs; squamous cell carcinomas, SCCs) and random biopsies as fingerprints to subclassify histopathological types and as a starting point to search for protein markers that may form the basis for diagnosis, prognosis, and treatment. Ultimately, the goal of these studies is to identify signaling pathways and components that are affected at various stages of bladder cancer progression and that may provide novel leads in drug discovery. Here we present our ongoing efforts to establish comprehensive two-dimensional polyacrylamide gel electrophoresis (2-D PAGE) databases of TCCs and SCCs which are being constructed based on the proteomic and immunohistochemical analysis of hundreds of fresh tumors, random biopsies and cystectomies received shortly after operation (http://biobase.dk/cgi-bin/celis).

To better understand the process of multistage carcinogenesis in Schwann cells, we have attempted to isolate novel candidate genes involved in neoplastic progression of mouse malignant Schwannoma cells. The semi-differentiated Schwannoma cell line 56-24 and the less differentiated Schwannoma cell line 64-39 were established from peripheral nerve sheath tumors arising in transgenic mice of the MBP/SV40 large T strain Tg29. By using the chemical cross-linking subtraction technique, we have cloned a novel murine cDNA that detects pronounced expression in 56-24 cells but not in 64-39 cells. The longest open reading frame of the cDNA predicts a peptide showing 95% amino acid sequence homology to the recorded sequence of the human immunophilin homolog huFKBPr38, one of a family of proteins that are thought to interface with a wide range of intracellular signal transduction systems. The predicted open reading frame of the corresponding gene, named muFKBP38, encodes a 38 kDa protein that harbors an FK-binding protein (FKBP) domain that is 36% identical to that of muFKBP52, a three-unit tetratricopeptide repeat and a consensus leucine-zipper repeat. Although muFKBP38 mRNA was detected in both neurons and glial cells, pronounced expression of the immunophilin homolog appeared in various classes of neurons associated with the hippocampal formation, as shown by in situ hybridization analysis of adult mouse brains. Taken together, these data indicate that muFKBP38 is (i) a novel potential marker for semi-differentiated Schwannomas, (ii) may form homomultimers and/or interact with other proteins, and (iii) may have a role in neurons associated with memory function.

Members of the phospholipase D (PLD) superfamily are defined by the conserved HXKXXXXD motif, which is essential for the catalytic function of mammalian PLD. PLD enzymes are thought to play roles in signal transduction and membrane vesicular trafficking in mammalian cells. Here we describe a 54-kDa novel murine polypeptide (designated SAM-9) that is predicted to be a membrane-associated member of the PLD superfamily. SAM-9 shares 40, 30, and 29% amino acid identity with potential orthologs, in vaccinia virus, Caenorhabditis elegans, and Dictyostelium discoideum, respectively, and belongs to a subclass of PLD homologs in which the second HXKXXXXD motif is imperfect and harbors a conserved Asp to Glu substitution. The sam-9 gene has more than eight exons, and the two HXKXXXXD motifs are encoded by two highly conserved exons. The expression of the sam-9 gene is greater in the brain than in non-nervous tissue and appears to be predominantly of neuronal origin. sam-9 expression is pronounced in mature neurons of the forebrain and appears to be turned on at late stages of neurogenesis as revealed by in situ hybridization analysis of sam-9 expression during postnatal development of the hippocampal formation and the primary somatosensory cortex.

Human 2-D PAGE Databases established at the Danish Centre for Human Genome Research are now available on the World Wide Web (http://biobase.dk/cgi-bin/celis). The databanks, which offer a comprehensive approach to the analysis of the human proteome both in health and disease, contain data on known and unknown proteins recorded in various IEF and NEPHGE 2-D PAGE reference maps (non-cultured keratinocytes, non-cultured transitional cell carcinomas, MRC-5 fibroblasts and urine). One can display names and information on specific protein spots by clicking on the image of the gel representing the 2-D gel map in which one is interested. In addition, the database can be searched by protein name, keywords or organelle or cellular component. The entry files contain links to other databases such as Medline, Swiss-Prot, PIR, PDB, CySPID, OMIM, Methabolic pathways, etc. The on-line information is updated regularly.

We report a novel approach for identifying monomeric GTP-binding proteins that is based on probing cDNA expression libraries with [alpha-32P]GTP. In short, a nitrocellulose replica from a plated cDNA expression library is treated with 2% SDS to block the GTP-binding activity of various G proteins expressed by E. coli, thus allowing the direct identification of positive clones. Using this procedure we have cloned several small GTP-binding proteins from human keratinocytes including the human homolog of rheb, a novel member of the ras-related GTP-binding proteins. Human rheb cDNA shares 90% identity with the rat counterpart and it is highly upregulated in transformed human cells of various origin. Northern analysis showed that human rheb is ubiquitously expressed, with the highest levels observed in skeletal and cardiac muscle, and not in brain, as it is the case for rat rheb. The human RHEB gene was mapped to chromosome 10q11.

Udgivelsesdato: 1995-Nov Psoriasin is a low molecular weight protein of the S100 family, which is highly upregulated in psoriatic epidermis, and whose function is related to skin inflammatory responses. We have applied a cDNA probe from the corresponding psoriasin gene S100A7 in a refined localisation analysis. S100A7 was mapped physically by human/rodent somatic cell hybrid analysis, and more precisely genetically by multilocus linkage analysis of 40 CEPH (Centre d'Etude du Polymorphisme Humain) families. The resulting 12-point linkage map was supported by odds of at least 1000:1, where S100A7 could be placed with a multipoint lodscore of 27.4 in an approximately 7-cM interval. The order of the 12 loci was as follows (with the best estimates of recombination frequencies given in between): AMY2B-0.091-D1S73(0.039)-D1S11(-0.053)-D1S189(-0 .017)-D1S1252 (-0.017)-D1S13(-0.078)-D1Z5(-0.051)-S100A7(-0.022)- MUC1(-0.026)-SPTA1 (-0.066)-ATP1A2(-0.014)-APOA2. Furthermore, from this map S100A7 could be assigned to the regional position of chromosome 1cen-q21. The linkage information presented should be of great value in association and linkage studies of diseases where psoriasin, or some of the several other very closely linked and functionally related genes, are seen as candidate genes, e.g. in psoriasis.

The master two-dimensional (2-D) gel database of human keratinocytes currently lists 3087 cellular proteins (2168 isoelectric focusing, IEF; and 919 none-quilibrium pH gradient electrophoresis, NEPHGE), many of which correspond to posttranslational modifications, 890 polypeptides have been identified (protein name, organelle components, etc.) using one or a combination of procedures that include (i) comigration with known human proteins, (ii) 2-D gel immunoblotting using specific antibodies (iii) microsequencing of Coomassie Brilliant Blue stained proteins, (iv) mass spectrometry and (v) vaccinia virus expression of full length cDNAs. These are listed both in alphabetical order and with increasing SSP number, together with their M(r), pI, cellular localization and credit to the investigator(s) that aided in the identification. Furthermore, we list 239 microsequenced proteins recorded in the database. We also report a database of proteins recovered from the medium of noncultured, unfractionated keratinocytes. This database lists 398 polypeptides (309 IEF; 89 NEPHGE) of which 76 have been identified. The aim of the comprehensive databases is to gather, through a systematic study of keratinocytes, qualitative and quantitative information on proteins and their genes that may allow us to identify abnormal patterns of gene expression and, ultimately, to pinpoint signaling pathways and components affected in various skin diseases, cancer included. Udgivelsesdato: 1994-Nov

Analysis of cellular protein patterns by computer-aided two-dimensional gel electrophoresis together with recent advances in protein sequence analysis and expression systems have made possible the establishment of comprehensive two-dimensional gel protein databases that may link protein and DNA mapping and sequence information and that offer an integrated approach to the study of gene expression. With the integrated approach offered by two-dimensional gel protein databases it is now possible to reveal phenotype-specific protein(s), to microsequence them, to search for homology with previous identified proteins, to clone the cDNAs, to assign partial protein sequences to genes for which the full DNA sequence and the chromosome location are known, and to study the regulatory properties and function of groups of proteins that are coordinately expressed in a given biological process. Comprehensive two-dimensional gel protein databases will provide an integrated picture of the expression levels and properties of the thousands of protein components of organelles, pathways, and cytoskeletal systems, both under physiological and abnormal conditions, and are expected to lead to the identification of new regulatory networks. So far, about 20% (600 out of 2,980) of the total number of proteins recorded in the human keratinocyte protein database have been identified and we are actively gathering qualitative and quantitative biological data on all resolved proteins. Given the current improvements on microsequencing as well as the availability of specific antibodies, it seems feasible to expect that most known keratinocyte proteins will be identified in the very near future. This feast will reveal a wealth of new proteins that will become amenable to experimentation both at the biochemical and molecular biology level. Udgivelsesdato: 1993-null

The master two-dimensional gel database of human keratinocytes currently lists 2980 cellular proteins (2098 isoelectric focusing, IEF; and 882 nonequilibrium pH gradient electrophoresis, NEPHGE) many of which correspond to posttranslational modifications. About 20% of all recorded proteins have been identified (protein name, organelle components, etc.) and they are listed in alphabetical order together with their M(r), pI, cellular localization and credit to the investigator(s) that aided in the identification. Also, we have listed 145 microsequenced proteins that are recorded in this database. As an aid in localizing the polypeptides we have included blow-ups of the master images (IEF, NEPHGE) displaying all the protein numbers. In the long run, the master keratinocyte database is expected to link protein and DNA sequencing and mapping information (Human Genome Program) and to provide an integrated picture of the expression levels and properties of the thousands of proteins that orchestrate various keratinocyte functions both in health and disease.

Neuronal protein gene product 9.5 (PGP 9.5) most likely identical to ubiquitin carboxyl-terminal hydrolase isozyme L1 (UCH-L1) has been reported to be expressed almost exclusively in neuronal and neuroendocrine tissues. By two-dimensional (2D) immunoblotting, comigration and microsequencing of proteins recovered from 2D gels we have identified PGP 9.5 UCH-L1 as polypeptide IEF SSP 6104 (Mr = 27,000, pI = 5.49) in the comprehensive 2D gel cellular protein database of human embryonal lung MRC-5 fibroblasts [(1989) Electrophoresis 10, 76 115; (1990) Electrophoresis 11, 1072 1113]. This protein is expressed at high levels in quiescent and proliferating cultured normal fibroblasts and is strongly down-regulated (about 10 times) in their transformed counterparts. Udgivelsesdato: 1991-Mar-25

Elongation factor 2 was identified in the two-dimensional gel patterns of asynchronous human amnion cells (AMA) by comigration with purified rabbit reticulocyte elongation factor 2 and by two-dimensional gel immunoblot analysis using a specific rabbit polyclonal antibody. In all, four polypeptides were identified corresponding to isoelectric focusing polypeptides 2713 (95.0 kDa), 2714 (94.8 kDa), 3727 (94.8 kDa), and 3702 (93.6 kDa) (listed in order of decreasing pI values) in the computerized comprehensive two-dimensional gel data base of human AMA proteins. The relative proportion of two of these variants (isoelectric focusing polypeptides 3727 and 3702), which are phosphorylated, increased dramatically during mitosis. Since phosphorylation is known to render elongation factor 2 inactive in translation, this observation may partly explain the decline in the rate of protein synthesis observed during cell division.

Udgivelsesdato: 1990-Feb

PCNA autoantibodies specific for cyclin/PCNA were used to determine the nuclear distribution of this protein in transformed human amnion cells (AMA) irradiated with ultraviolet light (254 nm) under conditions that induced nucleotide excision DNA repair synthesis. The results showed a striking increase in nuclear cyclin/PCNA antigen staining of non S-phase cells that was not abolished by cycloheximide (20 micrograms/ml, added 2 h before irradiation), and that is most likely due to a redistribution of pre-existing cyclin. These observations raise the possibility that cyclin/PCNA may play a role in nucleotide excision DNA repair synthesis in addition to its putative role in replicative DNA synthesis.

Databases of protein information derived from the analysis of two-dimensional gels have been established from transformed human amnion cells (AMA) and peripheral blood mononuclear cells (PBMCs). A total of 1781 [35S]methionine-labeled AMA proteins (1274 IEF, 537 NEPHGE) and a total of 1311 proteins from PBMC (948 IEF, 363 NEPHGE) were resolved and recorded using computerized (PDQ-SCAN and PDQUEST softwares) two-dimensional gel electrophoresis. AMA and PBMC proteins (total, 454: 301 IEF, 153 NEPHGE) were matched both manually and by the computer. Information entered in the AMA database (in most cases for some major proteins) includes: molecular weight, protein name, HeLa protein catalogue number, mouse protein catalogue number, nuclear proteins, phosphorylated proteins, distribution of proteins in Triton X-100 supernatants and cytoskeletons, proliferation- and transformation-sensitive proteins, cell cycle-specific proteins, mitochondrial proteins, proteins matched in normal human embryonal lung MRC-5 fibroblasts and PBMC cells, heat shock proteins, proteins affected by interferons, cytoskeletal proteins, and the presence of antibody against protein in human sera. Additional information has been entered for the cell cycle-regulated and DNA replication protein cyclin (PCNA). Information entered in the PBMC database includes molecular weight and potential markers for sorted populations of lymphocyte subtypes. For those proteins that have been matched to AMA proteins, information contained in some entries may be transferred from the AMA database.

Comprehensive, computerized databases of cellular protein information derived from the analysis of two-dimensional gels, together with recently developed techniques to microsequence proteins offer a new dimension to the study of genome organization and function. In particular, human protein databases provide an ideal framework in which to focus the human genome sequencing effort.