Two novel cyanobacterial bioluminescent whole-cell bioreporters based on superoxide dismutases MnSod and FeSod to detect superoxide anion.

This work describes the construction of two novel self-luminescent bioreporter strains of the cyanobacterium Nostoc sp. PCC 7120 by fusing the promoter region of the sodA and sodB genes (encoding the superoxide dismutases MnSod and FeSod, respectively) to luxCDABE from Photorhabdus luminescens aimed at detecting pollutants that generate reactive oxygen species (ROS), particularly O2-. Bioreporters were tested against methyl viologen (MV) as the inducer of superoxide anion (O2-). Both bioreporters were specific for O2- and Limits of detection (LODs) and Maximum Permissive Concentrations (MPCs) were calculated: Nostoc sp. PCC 7120 pBG2154 (sodA) had a range of detection from 400 to 1000 pM of MV and for Nostoc sp. PCC 7120 pBG2165 (sodB) the range of detection was from 500 to 1800 pM of MV after 5 h-exposure. To further validate the bioreporters, they were tested with the emerging pollutant Triclosan which induced bioluminescence in both strains. Furthermore, the bioreporters performance was tested in two real environmental samples with different water matrix complexity, spiked with MV. Both bioreporters were induced by O2- in these environmental samples. In the case of the river water sample, the amount of bioavailable MV as calculated from the bioreporters output was similar to that nominally added. For the waste water sample, the bioavailable MV concentration detected by the bioreporters was one order of magnitude lower than nominal. These differences could be due to MV complexation with organic matter and/or co-occurring organic contaminants. These results confirm their high sensitivity to O2- and their suitability to detect oxidative stress-generating pollutants in fresh-waters.

Influence of clinical and psychological variables upon the oral health-related quality of life in patients with temporomandibular disorders.

BACKGROUND: To analyze the association between the OHIP-14 and the different subtypes making up the clinical and psychological axis obtained using the RDC/TMD. MATERIAL AND METHODS: 407 patients treated at the TMD unit of the Andalusian Healthcare Service were administered the Spanish version of the Research Diagnostic Criteria for Temporomandibular Disorders questionnaire (RDC/TMD), together with the Oral Health Impact Profile questionnaire (OHIP-14). The degree of association between the patients' score in the OHIP-14 and the clinical and biopsychosocial variables was analyzed through bivariate and multivariate analyses, specifically through linear regression. RESULTS: 89.4% of the treated patients were women, while 10.6% were men, with an average age of 42.08 ± 14.9 years. The mean score and standard deviation for the OHIP-14 was 20.57 ± 10.73. A significant association (p < 0.05) was observed with the following variables: Axis I, jaw disability checklist, depression, somatization, perceived pain duration, and pain interference with activities of daily living. CONCLUSIONS: The analysis of the relation between self-perceived health in patients with TMD, as measured by the OHIP-14, showed a R2 of 0.3979, with a higher Beta value for the association between the OHIP and patients with both myofascial pain and arthopathy, jaw disability, depression, a higher pain duration and a higher pain interference with activities of daily living.

Application of an oral health-related quality of life questionnaire in primary care patients with orofacial pain and temporomandibular disorders.

OBJECTIVES: To examine whether patients who report orofacial pain (OP) and temporomandibular disorders (TMD) have a poorer perception of their oral health-related quality of life and, if so, to what extent, and to analyze the association between oral health perception, sociodemographic variables and reported pain duration. STUDY DESIGN: 407 patients treated at the OP and TMD units in the Healthcare District of Cordoba, Spain, diagnosed following the standard criteria accepted by the scientific community - the Research Diagnostic Criteria for Temporomandibular Disorders (RDC/TMD) - were administered the Spanish version of the Oral Health Impact Profile questionnaire (OHIP-14). Bivariate and logistic regression analyses were performed to determine the degree of association between the patients' OHIP-14 score and pain duration, pain intensity, and various sociodemographic variables. RESULTS: The observed distribution was 89.4% women and 10.6% men. The mean OHIP-14 score was 20.57 ± 10.73 (mean ± standard deviation). A significant association (p<0.05) was found for gender, age, marital status, chronic pain grade, self-perceived oral health status and pain duration. CONCLUSIONS: The analysis of self-perceived oral health status in patients with OP and TMD, as measured by the OHIP-14, showed that oral health is perceived more negatively by women. Moreover, a one-point increase in the Chronic Pain Grade indicator increases the OHIP-14 indicator by 4.6 points, while chronic pain, defined as pain suffered by patients for one year or more, increases the OHIP-14 indicator by 3.2 points.

Investigation of Galaxolide degradation products generated under oxidative and irradiation processes by liquid chromatography/hybrid quadrupole time-of-flight mass spectrometry and comprehensive two-dimensional gas chromatography/time-of-flight mass spectrometry.

RATIONALE: Polycyclic musks have become a concern due to their bioaccumulation potential and ecotoxicological effects. The HHCB transformation product (TP) (1,3,4,6,7,8-hexahydro-4,6,6,7,8,8-hexamethyl-cyclopenta[γ]-2-benzopyran; HHCB-lactone) is the most stable intermediate generated and it is frequently detected in river waters. The aim of this work was the identification of relevant TPs generated from UV irradiation and ozone treatments. METHODS: Identification of HHCB TPs was carried out by liquid chromatography/hybrid quadrupole time-of-flight mass spectrometry (LC/ESI-QTOF-MS) and two-dimensional gas chromatography/electron impact time-of-flight mass spectrometry (GC×GC-EI-TOF-MS). With LC/ESI-QTOF-MS, TPs were characterized by means of mass accuracy in both full-scan and MS/MS modes through information-dependent acquisition (IDA) and direct injection on-column. With stir bar sorptive extraction (SBSE)-GC×GC-EI-TOF-MS, identification was based on the enhanced separation capacity and screening of unknowns through the acquisition of full-range mass spectra. RESULTS: The effectiveness of these complementary techniques allowed a detailed evaluation of the main TPs. Eighteen TPs were elucidated based on mass accuracy, in both full-scan and MS/MS modes using LC/ESI-QTOF-MS with mass errors below 5 ppm and 10 ppm (mostly), respectively. Most of the TPs had not been analytically identified in previous studies. Separation of the enantiomeric species (R) and (S) of HHCB-lactone, and the identification of other relevant TPs, was performed using SBSE-GC×GC-EI-TOF-MS. CONCLUSIONS: LC/ESI-QTOF-MS and GC×GC-EI-TOF-MS analysis provides the best alternative for TP identification of chemicals of concern, which have a wide range of polarities and isobaric compounds. A prediction of PBT (persistence, bioaccumulation and toxicity) using the PBT Profiler program suggested a classification of 'very persistent' and 'very toxic' for most of the TPs identified.

[Attitudes and values of the medical internal residents who have chosen the specialty of family and community medicine: A health model adapted to the new times]. / Actitudes y valores de los residentes de Medicina de Familia y Comunitaria: un modelo sanitario adaptado a los nuevos tiempos.

OBJECTIVE: To analyze the views that medical internal residents (MIR), who have chosen the specialty of family and community medicine (MFC), have about training and professional practice, and analyzing the differences in views between them and the rest of the MIR specialties. MATERIAL AND METHODS: On-line survey with the medical residents who began their residential training period in Andalusia in 2008. A factorial analysis of principal components was conducted using the variables of opinion on professional and training issues and training in order to group them into meaningful dimensions, and then a ANOVA to see the differences between them. RESULTS: Three dimensions, that summarize the MFC resident's opinions about their residential training, were obtained. MFC residents have a more positive view of the resident training system, and give more importance to the general aspects of medical practice. CONCLUSIONS: Health policies are currently positioned with the patient at the center of the system and emphasises not only the biological issues but also social issues (prevention, education, treating common diseases, and groups with major problems, etc.). According to the results, the MIR-MFC are more in line with these values, and this might be indicative of a better adaptation to this new culture of the system.

Actitudes y valores de los residentes de Medicina de Familia y Comunitaria: un modelo sanitario adaptado a los nuevos tiempos / Attitudes and values of the medical internal residents who have chosen the specialty of family and community medicine: a health model adapted to the new times

Objetivo. Analizar la visión que tienen de su formación y práctica profesional los médicos internos residentes (MIR) que han elegido la especialidad de medicina de familia y comunitaria (MFC) frente al resto de residentes de otras especialidades, para ver las diferencias entre ambos y sus implicaciones para el sistema. Material y métodos. Encuesta online a MIR que comenzaron su formación residencial en Andalucía en 2008. Se realizó un análisis factorial de los componentes principales a partir de variables de opinión sobre aspectos profesionales y formativos con el fin de agruparlas en dimensiones y posteriormente un ANOVA para observar las diferencias entre los 2 grupos estudiados con respecto a dichas dimensiones. Resultados. Se obtuvieron 3 dimensiones que resumen las opiniones de los residentes sobre su formación residencial. Los residentes de MFC tienen una visión más positiva del sistema de formación y le dan más importancia a cuestiones generales de la práctica médica respecto a los residentes de otras especialidades. Conclusiones. Actualmente las políticas sanitarias posicionan al ciudadano como eje del sistema y ponen el acento no solo en cuestiones biológicas sino también en otras de carácter social (prevención, educación, atención de enfermedades comunes y de colectivos con mayores problemas, etc.). A tenor de los resultados los MIR-MFC parecen estar más en consonancia con estos valores, lo cual puede ser indicativo de una mejor adaptación a esta nueva cultura organizativa (AU)

Objective. To analyze the views that medical internal residents (MIR), who have chosen the specialty of family and community medicine (MFC), have about training and professional practice, and analyzing the differences in views between them and the rest of the MIR specialties. Material and methods. On-line survey with the medical residents who began their residential training period in Andalusia in 2008. A factorial analysis of principal components was conducted using the variables of opinion on professional and training issues and training in order to group them into meaningful dimensions, and then a ANOVA to see the differences between them. Results. Three dimensions, that summarize the MFC resident´s opinions about their residential training, were obtained. MFC residents have a more positive view of the resident training system, and give more importance to the general aspects of medical practice. Conclusions. Health policies are currently positioned with the patient at the center of the system and emphasises not only the biological issues but also social issues (prevention, education, treating common diseases, and groups with major problems, etc.). According to the results, the MIR-MFC are more in line with these values, and this might be indicative of a better adaptation to this new culture of the system (AU)

Ozone-based reclamation of an STP effluent.

The system ozone and hydrogen peroxide was used to reclaim wastewater from the secondary clarifier from a Sewage Treatment Plant (STP) of Alcalá de Henares (Madrid-Spain). The assays were performed by bubbling a gas mixture of oxygen and ozone, with approximately 24 g Nm(-3) of ozone concentration, through a volume of wastewater samples for 20 minutes at 25 degrees C. The removal of dissolved micropollutants such as Pharmaceutical and Personal Care Products (PPCPs) and Organic Carbon (TOC) was enhanced by adding periodic pulses of hydrogen peroxide while keeping pH above 8.0 throughout the runs. Removal efficiency ratios in the range of 7-26 mg O3/mg TOC and 0.24 mg O3/ng micropollutants at 5 minutes of ozonation were assessed as reference data to reclaim wastewater from STP. The relation between the extent of TOC removed and ozone doses used was related by a second-order kinetic model in which the time-integrated ozone-hydrogen peroxide concentration was included.

Coagulation-Fenton coupled treatment for ecotoxicity reduction in highly polluted industrial wastewater.

A coupled coagulation-Fenton process was applied for the treatment of cosmetic industry effluents. In a first step, FeSO(4) was used as coagulant and the non-precipitated Fe(2+) remaining in dissolution was used as catalyst in the further Fenton process. In the coagulation process a huge decrease in total organic carbon (TOC) was achieved, but the high concentration of phenol derivatives was not diminished. The decrease in TOC in the coagulation step significantly reduces the amount of H(2)O(2) required in the Fenton process for phenol depletion. The coupled process, using a H(2)O(2) dose of only 2 g l(-1), reduced TOC and total phenol to values lower than 40 and 0.10 mg l(-1), respectively. The short reaction period (less than 15 min) in TOC and phenol degradation bodes well for improving treatment in a continuous regime. The combination of both processes significantly reduced the ecotoxicity of raw effluent and markedly increased its biodegradability, thus allowing easier treatment by the conventional biological units in conventional sewage treatment plants (STPs).

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Análisis de la no elección de la analgesia epidural durante el trabajo de parto en las mujeres andaluzas: “la buena sufridora” / Analysis of rejection of epidural analgesia during labor in Andalusian women: “the silent sufferer”

Introducción: La analgesia epidural se ha popularizado en España, tanto en su conocimiento como en su uso, a lo largo de los últimos 20 años. La elección de esta técnica analgésica por parte de la futura madre durante el trabajo de parto y el parto en Andalucía es un derecho de la mujer que, explícitamente, sólo estará limitado por sus condiciones físicas y de salud y por los recursos disponibles en el hospital en el momento del parto. Esta elección pone en relación 2 elementos: por un lado, el componente cognitivo que establece la conveniencia o no de utilizar esta técnica según los valores, percepciones, cultura, etc. de la mujer y, por otro, el componente conductual, es decir, el uso o no de la técnica en cuestión, que dependerá además de la elección de la mujer de las condiciones anteriormente mencionadas. Objetivo: Analizar qué características definen a las mujeres que, aun pudiendo, deciden no utilizar analgesia epidural durante el parto en Andalucía. Material y métodos: Para ello se ha utilizado la información proporcionada por las mujeres ingresadas por parto en los hospitales del Sistema Sanitario Público de Andalucía, contenida en las encuestas anuales de satisfacción de usuarios del sistema de atención hospitalario en esta comunidad autónoma entre los años 2000 y 2007. Con esta información se ha realizado un análisis de segmentación jerárquica que tenía por objetivo analizar el perfil de las mujeres que rechazan esta técnica analgésica. Resultados: Las principales características que definen a las mujeres que rechazan la analgesia epidural durante el parto vienen dadas por su nivel educativo, nivel de ingresos y situación laboral...(AU)

Introduction: Knowledge and use of epidural analgesia has become increasingly wides preadin Spain over the last 20 years. In Andalusia (southern Spain), expectant mother shave the right to choose this analgesic technique during labor and delivery, an option that depends solely on the physical or health status of the mother or the availability of the resources at the hospital at the time of childbirth. Preference is determined by two interrelated factors. Firstly, a cognitive component, which is influenced by the values, perceptions and culture of the mother and, secondly, a behavioral component, i.e., the acceptance or rejection of the technique, which will depend not only on the motives determining the choice, but on the possibility of choosing epidural analgesia as well as the above-mentioned conditioning factors. Objective: To determine the characteristics of women in Andalusia who refuse epidural anaesthesia during labor, even when this analgesic technique is readily available to them. Material and methods: Annual surveys on user satisfaction with hospital care in Andalusia conducted from 2000-2007 were used, specifically data provided by women who gave birth in public hospitals in Andalusia. Based on these data, a hierarchical segmentation analysis was conducted to determine the profile of women who refused this analgesic technique. Results: Educational level, income and employment status played a significant role in rejection of epidural anaesthesia during labor. Conclusions: The social, economic and cultural factors underlying rejection, that is, the profile of women who reject epidural analgesia, is consistent with the paradigm of the “traditional woman” (characterized by limited income, low educational level and mainly occupied by housework) studied by other authors...(AU)

Removal of pharmaceuticals and kinetics of mineralization by O(3)/H(2)O(2) in a biotreated municipal wastewater.

The ozonation of an effluent from the secondary clarifier of two Municipal Wastewater Treatment Plants was performed by using alkaline ozone and a combination of ozone and hydrogen peroxide. Alkaline ozonation achieved only a moderate degree of mineralization, essentially concentrated during the first few minutes; but the addition of hydrogen peroxide eventually led to a complete mineralization. The evolution of total organic carbon (TOC) as a measure of the extent of mineralization and the concentration of dissolved ozone were analyzed and linked in a kinetic model whose parameter represented the product of the exposure to hydroxyl radicals and the kinetic constant of indirect ozonation. This rate parameter yielded the highest values during the first part of O(3)/H(2)O(2) runs. The kinetic constant for the decomposition of ozone at the end of the run was also measured and computed for the non-oxidizable water matrix and yielded essentially the same values regardless of whether or not hydrogen peroxide was used. A group of 33 organic compounds, mainly pharmaceuticals and some relevant metabolites present in the wastewater effluents, were evaluated before and after the ozonation process using a liquid chromatography-hybrid triple-quadrupole linear ion trap system (LC-QqLIT-MS). The results demonstrate that the ozonation degrades these compounds with efficiencies of over 99% in most cases, even under low mineralization conditions in alkaline ozonation.

Peptides from the amino terminal mdm-2-binding domain of p53, designed from conformational analysis, are selectively cytotoxic to transformed cells.

We have synthesized three peptides from the mdm-2 binding domain of human p53, residues 12-26 (PPLSQETFSDLWKLL), residues 12-20, and 17-26. To enable transport of the peptides across the cell membrane and at the same time to maximize the active mdm-2 binding alpha-helical conformation for these peptides, each was attached at its carboxyl terminus to the penetratin sequence, KKWKMRRNQFWVKVQRG, that contains many positively charged residues that stabilize an alpha-helix when present on its carboxyl terminal end. All three peptides were cytotoxic to human cancer cells in culture, whereas a control, unrelated peptide attached to the same penetratin sequence had no effect on these cell lines. The same three cytotoxic peptides had no effect on the growth of normal cells, including human cord blood-derived stem cells. These peptides were as effective in causing cell death in p53-null cancer cells as in those having mutant or normal p53. Peptide-induced cell death is not accompanied by expression of apoptosis-associated proteins such as Bax and waf(p21). Based on these findings, we conclude that the antiproliferative effects of these p53-derived peptides are not completely dependent on p53 activity and may prove useful as general anticancer agents.

Common conformational effects of p53 mutations.

The tumor suppressor gene p53 has been identified as the most frequent target of genetic alterations in human cancers. Most of these mutations occur in highly conserved regions in the DNA-binding core domain of the p53 protein, suggesting that the amino acid residues in these regions are critical for maintaining normal p53 structure and function. We previously used molecular dynamics calculations to demonstrate that several amino acid substitutions in these regions that are induced by environmental carcinogens and found in human tumors produce certain common conformational changes in the mutant proteins that differ substantially from the wild-type structure. In order to determine whether these conformational changes are consistent for other p53 mutants, we have now used molecular dynamics to determine the structure of the DNA-binding core domain of seven other environmentally induced, cancer-related p53 mutants, namely His 175, Asp 245, Asn 245, Trp 248, Met 249, Ser 278, and Lys 286. The results indicate that all of these mutants differ substantially from the wild-type structure in certain discrete regions and that some of these conformational changes are similar for these mutants as well as those determined previously. The changes are also consistent with experimental evidence for alterations in structure in p53 mutants determined by epitope detectability using monoclonal antibodies directed against these regions of predicted conformational change.

Molecular basis for interactions of G protein betagamma subunits with effectors.

Both the alpha and betagamma subunits of heterotrimeric guanine nucleotide-binding proteins (G proteins) communicate signals from receptors to effectors. Gbetagamma subunits can regulate a diverse array of effectors, including ion channels and enzymes. Galpha subunits bound to guanine diphosphate (Galpha-GDP) inhibit signal transduction through Gbetagamma subunits, suggesting a common interface on Gbetagamma subunits for Galpha binding and effector interaction. The molecular basis for interaction of Gbetagamma with effectors was characterized by mutational analysis of Gbeta residues that make contact with Galpha-GDP. Analysis of the ability of these mutants to regulate the activity of calcium and potassium channels, adenylyl cyclase 2, phospholipase C-beta2, and beta-adrenergic receptor kinase revealed the Gbeta residues required for activation of each effector and provides evidence for partially overlapping domains on Gbeta for regulation of these effectors. This organization of interaction regions on Gbeta for different effectors and Galpha explains why subunit dissociation is crucial for signal transmission through Gbetagamma subunits.

[Hernial strangulation of Meckel's diverticulum: Littre's hernia. Apropos of 4 cases]. / Estrangulación herniaria del divertículo de Meckel: hernia de Littré. A propósito de cuatro casos.

Hernial strangulation of Meckel's diverticulum (Littre's hernia) is a rare anatomoclinical form. It represents 10% of all complications of Meckel's diverticulum (8.8% of our cases), and complications like hemorrhage, perforation and diverticulitis are fairly frequent. Four cases of Littre's hernia are presented: two males and two females, with an average age of 67 years (range 50-83 years), representing 0.08% of all the inguinal-crural hernias operated in the department. The clinical manifestations were those of intestinal obstruction because a mixed type Littre's hernia was involved, with compromise of the diverticulum and its intestinal loop. Preoperative diagnosis is unlikely in strangulation without disturbances in the intestinal transit and, in fact, is even less likely if it is accompanied by obstruction. The diagnosis is thus almost always intraoperative. The correct treatment is surgery after restoring the patient's hemodynamic equilibrium. Simple and/or loop diverticulectomy via herniotomy, herniolaparotomy or laparotomy are debated. We think that this disorder can generally be resolved using the inguinal approach, as in any strangled hernia, with the technical option of using a larger, more comfortable and safer approach in cases of important obesity and/or deterioration of the loop (necrosis, perforation). In elderly patients with uncomplicated Littre's hernia and Meckel's diverticulum, abstention from diverticular exeresis may be justifiable. Of the four patients, the first two died from cardiogenic shock and pulmonary embolism, respectively; the last two evolved well (except for a wound abscess).