TCTP as a therapeutic target in melanoma treatment.

BACKGROUND: Translationally controlled tumour protein (TCTP) is an antiapoptotic protein highly conserved through phylogeny. Translationally controlled tumour protein overexpression was detected in several tumour types. Silencing TCTP was shown to induce tumour reversion. There is a reciprocal repression between TCTP and P53. Sertraline interacts with TCTP and decreases its cellular levels. METHODS: We evaluate the role of TCTP in melanoma using sertraline and siRNA. Cell viability, migration, and clonogenicity were assessed in human and murine melanoma cells in vitro. Sertraline was evaluated in a murine melanoma model and was compared with dacarbazine, a major chemotherapeutic agent used in melanoma treatment. RESULTS: Inhibition of TCTP levels decreases melanoma cell viability, migration, clonogenicity, and in vivo tumour growth. Human melanoma cells treated with sertraline show diminished migration properties and capacity to form colonies. Sertraline was effective in inhibiting tumour growth in a murine melanoma model; its effect was stronger when compared with dacarbazine. CONCLUSIONS: Altogether, these results indicate that sertraline could be effective against melanoma and TCTP can be a target for melanoma therapy.

Insecticidal activity of a recombinant knottin peptide from Loxosceles intermedia venom and recognition of these peptides as a conserved family in the genus.

Loxosceles intermedia venom comprises a complex mixture of proteins, glycoproteins and low molecular mass peptides that act synergistically to immobilize envenomed prey. Analysis of a venom-gland transcriptome from L. intermedia revealed that knottins, also known as inhibitor cystine knot peptides, are the most abundant class of toxins expressed in this species. Knottin peptides contain a particular arrangement of intramolecular disulphide bonds, and these peptides typically act upon ion channels or receptors in the insect nervous system, triggering paralysis or other lethal effects. Herein, we focused on a knottin peptide with 53 amino acid residues from L. intermedia venom. The recombinant peptide, named U2 -sicaritoxin-Li1b (Li1b), was obtained by expression in the periplasm of Escherichia coli. The recombinant peptide induced irreversible flaccid paralysis in sheep blowflies. We screened for knottin-encoding sequences in total RNA extracts from two other Loxosceles species, Loxosceles gaucho and Loxosceles laeta, which revealed that knottin peptides constitute a conserved family of toxins in the Loxosceles genus. The insecticidal activity of U2 -SCTX-Li1b, together with the large number of knottin peptides encoded in Loxosceles venom glands, suggests that studies of these venoms might facilitate future biotechnological applications of these toxins.

Active site mapping of Loxosceles phospholipases D: Biochemical and biological features.

Brown spider phospholipases D from Loxosceles venoms are among the most widely studied toxins since they induce dermonecrosis, triggering inflammatory responses, increase vascular permeability, cause hemolysis, and renal failure. The catalytic (H12 and H47) and metal-ion binding (E32 and D34) residues in Loxosceles intermedia phospholipase D (LiRecDT1) were mutated to understand their roles in the observed activities. All mutants were identified using whole venom serum antibodies and a specific antibody to wild-type LiRecDT1, they were also analyzed by circular dichroism (CD) and differential scanning calorimetry (DSC). The phospholipase D activities of H12A, H47A, H12A-H47A, E32, D34 and E32A-D34A, such as vascular permeability, dermonecrosis, and hemolytic effects were inhibited. The mutant Y228A was equally detrimental to biochemical and biological effects of phospholipase D, suggesting an essential role of this residue in substrate recognition and binding. On the other hand, the mutant C53A-C201A reduced the enzyme's ability to hydrolyze phospholipids and promote dermonecrosis, hemolytic, and vascular effects. These results provide the basis understanding the importance of specific residues in the observed activities and contribute to the design of synthetic and specific inhibitors for Brown spider venom phospholipases D.

Impacts of climate and land use changes on the hydrological and erosion processes of two contrasting Mediterranean catchments.

The impacts of climate and land use changes on streamflow and sediment export were evaluated for a humid (São Lourenço) and a dry (Guadalupe) Mediterranean catchment, using the SWAT model. SWAT was able to produce viable streamflow and sediment export simulations for both catchments, which provided a baseline for investigating climate and land use changes under the A1B and B1 emission scenarios for 2071-2100. Compared to the baseline scenario (1971-2000), climate change scenarios showed a decrease in annual rainfall for both catchments (humid: -12%; dry: -8%), together with strong increases in rainfall during winter. Land use changes were derived from a socio-economic storyline in which traditional agriculture is replaced by more profitable land uses (i.e. corn and commercial forestry at the humid site; sunflower at the dry site). Climate change projections showed a decrease in streamflow for both catchments, whereas sediment export decreased only for the São Lourenço catchment. Land use changes resulted in an increase in streamflow, but the erosive response differed between catchments. The combination of climate and land use change scenarios led to a reduction in streamflow for both catchments, suggesting a domain of the climatic response. As for sediments, contrasting results were observed for the humid (A1B: -29%; B1: -22%) and dry catchment (A1B: +222%; B1: +5%), which is mainly due to differences in the present-day and forecasted vegetation types. The results highlight the importance of climate-induced land-use change impacts, which could be similar to or more severe than the direct impacts of climate change alone.

Structural Insights into Substrate Binding of Brown Spider Venom Class II Phospholipases D.

Phospholipases D (PLDs), the major dermonecrotic factors from brown spider venoms, trigger a range of biological reactions both in vitro and in vivo. Despite their clinical relevance in loxoscelism, structural data is restricted to the apo-form of these enzymes, which has been instrumental in understanding the functional differences between the class I and II spider PLDs. The crystal structures of the native class II PLD from Loxosceles intermedia complexed with myo-inositol 1-phosphate and the inactive mutant H12A complexed with fatty acids indicate the existence of a strong ligand-dependent conformation change of the highly conserved aromatic residues, Tyr 223 and Trp225 indicating their roles in substrate binding. These results provided insights into the structural determinants for substrate recognition and binding by class II PLDs.

Characterization of Brown spider (Loxosceles intermedia) hemolymph: cellular and biochemical analyses.

This is the first study on the hemolymph from a spider of the Loxosceles genus. These animals are responsible for a great number of envenomation cases worldwide. Several studies on Loxosceles venoms have been published, and the knowledge about the venom and its toxins is considerable, not only regarding the biological and biochemical characterization, but also regarding structural, genetic and phylogenetic approaches. However, the literature on Loxosceles hemolymph is nonexistent. The main goal of the present study was to characterize biochemically the hemolymph content, and especially, to identify its different hemocytes. Moreover, many papers have already shown molecules whose source is the hemolymph and their very interesting activities and biomedical applications, for example, antifungal and antibacterial activities. A 2D-SDS-PAGE of brown spider hemolymph showed approximately 111 spots for pH 3-10 and 150 spots for pH 4-7. A lectin-blotting assay showed that hemolymph carbohydrate residues were similar to those found in venom. Several types of TAG and DAG phospholipids were found in the hemolymph and characterized by HPTLC and mass spectrometry. Four different hemocytes were characterized in Loxosceles intermedia hemolymph: prohemocyte, plasmatocyte, granulocyte and adipohemocyte. This paper opens new possibilities on toxinology, studying an unknown biological material, and it characterizes a source of molecules with putative biotechnological applications.

Satisfacción de los pacientes diagnosticados de enfermedad de Crohn con respecto a la colonoscopia y entero-resonancia magnética / Crohn's disease patient's satisfaction with colonoscopy versus entero magnetic resonance imaging (MRI)

Objetivos. Evaluar la satisfacción y las preferencias sobre la colonoscopia (CL) y la entero-resonancia (ERM) de los pacientes que padecen enfermedad de Crohn a los que se les ha realizado ambas pruebas. Valorar también la eficacia del sistema de información para la recogida de estos datos. Método. Estudio piloto transversal, mediante una encuesta telefónica de satisfacción realizada a pacientes diagnosticados de enfermedad de Crohn (EC). Los pacientes del estudio se seleccionaron en la consulta del Servicio de Aparato Digestivo del Hospital Universitario Príncipe de Asturias. Se les realizaron las dos pruebas objeto del estudio durante el periodo comprendido entre el 1 de enero de 2011 y el 30 de junio de 2012. Las encuestas se efectuaron durante los meses de julio y agosto de 2012. Resultados. Han participado un total de 48 pacientes con enfermedad de Crohn. De ellos, 24 eran mujeres y 24 hombres. Con una media de edad de 43 años (DE: 13.8). La peor valoración se obtuvo en la preparación previa de ambas. Si los pacientes pudieran elegir, 23 preferían CL, 16 ERM y a 9 les era indiferente. Conclusiones. La preferencia de los pacientes hacia la CL pudo deberse a la administración previa de sedación. Una prueba como la CL, que a priori puede resultar desagradable para el paciente, resulta mucho mejor tolerada gracias a la sedación. En ambas pruebas las preparaciones previas se perciben como molestas. Nos plantearemos cómo poder mejorar la tolerancia a dicha preparación, quizá dando más información, puesto que muchas veces no se realiza correctamente y eso puede acentuar dicha percepción desagradable (AU)

Objectives. To evaluate satisfaction and preferences in Crohn's disease patients who have undergone a colonoscopy (CL) and a MRI colonography (MRI). As well as evaluate the effectiveness of the information system to collect the data. Method. A sectional pilot study conducted through a satisfaction survey questionnaire for patients diagnosed with Crohn’s disease (CD). Study patients were selected from the Gastroenterology Department in the Principe de Asturias University Hospital, where both the colonoscopy and the entero magnetic resonance imaging tests were done from the 1st of January to the 30th of June 2012. Surveys were conducted during July and August 2012. Results. A total of 48 patients with Crohn’s disease participated. Out of these, 24 were women and 24 were men. The mean age was 43 years (SD: 13.8). The worst score was obtained at the time of preparation for both procedures. If patients could choose they would prefer CL 23, 16 MRI and 9 were indifferent. Conclusions. The patients’ preference for CL could be due to the administration of sedatives prior to the procedure. A procedure such as the CL which initially may result unpleasant for the patient is tolerated much better as a result of the sedation. In both tests, the preparations prior to the procedures are perceived as unpleasant. We will think about how to improve patients’ tolerance to these preparations, maybe by giving more information as patients, because we don’t know often how to do it properly, which could magnify this unpleasant perception (AU)

SOD1 overexpression improves features of the oligodendrocyte precursor response in vitro.

Spinal cord injury (SCI) produces a significant loss of oligodendrocytes (OL) and demyelination. The oligodendrocyte precursor cells (OPCs) response includes a group of cellular changes in OPCs that are directed to replenish OL loss from the injury. However, this adaptive response is hampered and OPCs eventually die or fail to differentiate to mature and functional OL. In this study, we wanted to evaluate if overexpression of human superoxide dismutase 1 (hSOD1) in OPCs from the SOD1 transgenic rat could improve some of the features of the OPC response in vitro. We found that hSOD1 overexpression increases the proliferation of OPCs and accelerates their differentiation to mature OL in vitro. Furthermore, hSOD1 overexpression reduces oxidative stress-mediated death in OPCs. These results suggest hSOD1 as a therapeutic target to increase OPC response success and potentially, OL replacement and remyelination after SCI.

Activity profile of top-class female soccer refereeing in relation to the position of the ball.

The aim of this study was to describe the activity profile of top-class female soccer referees during competition and to relate it to the position of the ball. Ten matches from the Fédération Internationale de Football Association (FIFA) under-20 female World Championships held in Russia in 2006 were filmed and the kinematical parameters of the female referees (n=10) and the ball were determined using a two-dimensional photogrammetric video system based on direct linear transformation (DLT) algorithms. Total distance covered during a match was 10 km, of which 1.3 km represented high-intensity activities (>13 km/h). The referees' highest mobility was achieved in the initial 15 min of the match, covering greater distance and performing more intense exercise (P<0.01) than in the final 15-min of the game. Mean distance from the referee to the ball was 19.5+/-2.4m, with no significant differences between 15 min match periods. The results of this study show that: (1) the physical demands placed on top-class female referees are much like those experienced during male moderate-standard refereeing and (2) the referees were able to maintain the distance from the ball throughout the match.

Previous cycling does not affect running efficiency during a triathlon world cup competition.

AIM: The most strategic part of a triathlon is the cycle-run transition. However, all the studies about this situation have been carried out in laboratory conditions and there is a need to perform this kind of study during competition, when the triathletes are highly motivated and the effort is maximum.1 Therefore, the aims of the present study were: 1) to determine the effect of prior 40-km cycling on the 10-km running kinematics during a Triathlon World Cup competition, and 2) to examine the possible differences between males and females. METHODS: Ten men and ten women, selected among the first ten competitors at the end of the cycling part at Madrid 2006 BG Triathlon World Cup, were enrolled in the study. The kinematic analysis was carried out using a photogrammetric technique (DLT algorithms) in the saggital plane (2D). RESULTS: There are significant differences (P<0.05) in men's stride length and velocity between the first and the last lap. Also, significant differences (P<0.05) were found between men and women in many of the variables analyzed. CONCLUSIONS: The previous cycling does not affect the subsequent running efficiency during a elite triathlon competition. On the other hand, the running technique profile during a triathlon competition is very different between men and women.

Nephrotoxicity caused by brown spider venom phospholipase-D (dermonecrotic toxin) depends on catalytic activity.

Bites from brown spiders (Loxosceles genus) have clinical manifestations including skin necrosis with gravitational spreading, and systemic involvement that may include renal failure, hemolysis, and thrombocytopenia. Mice were exposed to recombinant wild-type phospholipase-D, or to an isoform with a mutation in the catalytic domain resulting in no phospholipasic activity. Renal biopsies from mice treated with the wild-type toxin showed glomerular edema, erythrocytes and collapse of Bowman's space, edema and deposition of proteinaceous material within the tubular lumen. Ultrastructural analyses confirmed cytotoxicity by demonstrating disorders of glomerulus at foot processes and at fenestrated endothelium. Tubule alterations include deposits of amorphous material and edema, as well as an increase of epithelial cytoplasmic multivesicular bodies and electron-dense structures. There was an absence of nephrotoxicity in mice treated with the mutated toxin. Analyses of urine and blood showed that wild type toxin induced hematuria and elevation of blood urea, while treatment with mutated toxin caused no changes. Mouse lethality experiments also showed oliguria and mortality after treatment with wild-type toxin, but not following exposure to the mutated toxin. Immunofluorescence using antibodies to phospholipase-D toxin showed deposition of both toxins along the renal tubular structures as detected by confocal microscopy. Immunoblots of urine showed a 30 kDa band in samples from animals treated with wild-type toxin, but no band from mice exposed to mutated toxin. Wild-type toxin treatment caused cytoplasmic vacuolization, impaired spreading, reduction of cellular viability, and cell-cell and cell-substratum detachment in MDCK cells, while treatment with mutated isoform had no effect. Finally, there is a direct correlation between toxin activity on cell membrane phospholipids generating choline and cytotoxicity. We have defined for the first time a molecular mechanism for Loxosceles venom nephrotoxicity that is dependent on the catalytic activity of phospholipase-D toxin.

Rapid and deep control of inflammation in rheumatoid arthritis with infliximab and its correlation with acute-phase reactants.

Rheumatoid arthritis (RA) is a chronic inflammatory disease with predominant joint involvement and possible systemic compromise, which leads to a handicapped status and poor quality of life. An optimal approach to treat RA requires early and intensive intervention with close monitoring of treatment response. Tumor necrosis factor (TNF) blockers are recommended in cases of active RA after the unsuccessful use of effective disease-modifying antirheumatic drugs (DMARDs); even adding them to treatment or replacing these drugs. Anti-TNF therapies have been demonstrated to reduce significant joint damage and to relieve symptoms during a prolonged time (see Scott and Kingsley, 2006). The efficacy of infliximab in an open-label trial is summarized with respect to speed of onset of action, durability of response, and its correlation between clinical and laboratory parameters. Safety for long-term treatment is also summarized. We studied 105 RA patients with more than 3 years' history of disease during 24 months on i.v. infliximab (75 completed study). We evaluated ACR responses at base line, and at 1, 6, 12, 16, 52, 77, and 104 weeks. Morning stiffness, swollen and tender joints, HAQ, SF-36% (PCS/MCS), polymerase chain reaction (PCR), erythrosedimentation rate (ESR), transaminases, rheumatoid factor (RF) levels, hemogram, and adverse events profile were all assessed. The treatment offered rapid and sustained clinical improvements as revealed by ACR responses and marked changes in the parameters previously described. Important changes were made in functional status and acute-phase reactants. Finally, infliximab was considered well tolerated and did not affect the safety profile of this trial.

The Loxtox protein family in Loxosceles intermedia (Mello-Leitão) venom.

We isolated cDNA sequences coding for dermonecrotic/sphingomyelinases factor proteins from the brown spider Loxosceles intermedia, here named Loxtox proteins. The amino acid sequences based on cloned cDNA of several Loxtox proteins revealed at least six distinct groups of proteins expressed in the venom gland. The level of similarity among the toxins varied from 99% to 55%. The finding of several isoforms of Loxtox in the venom of this spider may reflect an evolutionary adaptation for different prey types and reinforces the idea of an efficient mutational mechanism in the venom gland of spiders.

Cytotoxic, thrombolytic and edematogenic activities of leucurolysin-a, a metalloproteinase from Bothrops leucurus snake venom.

Leucurolysin-a (leuc-a), a 23 kDa non-hemorrhagic metalloproteinase, is found in venom of the viper Bothrops leucurus. Here, we examine the biological consequences of leuc-a, including thrombolytic activity, direct effects on endothelial cells in culture and edematogenic activity in vivo. We demonstrate fibrinolytic activity of leuc-a, in which the protease specifically degrades alpha, beta, and gamma-gamma chains. While not causing hemorrhaging, leuc-a does cause thrombolytic activities in whole blood clots. Endothelial cells are highly resistant to leuc-a in culture. Cell viability suffered only when cells were exposed to large quantities of the protease. Nevertheless, leuc-a induces changes in cell morphology. The impact of leuc-a on cell adhesion was confirmed by an adhesion assay, in which cell adhesion to fibronectin decreased due to leuc-a. This mild cellular impact is unlike that of crude venom, where lower concentrations triggered cell death and a greater reduction in cell adhesion. Also, leuc-a increased microvessel permeability with marked edema in mice peritoneum and foot pads. These effects are similar to those of other P-I class SVPMs. These in vivo effects were weaker when crude venom was tested. In conclusion, albeit not showing significant hemorrhagic activity, leuc-a can induce a prominent edema which appears to be significant in the local effects observed after B. leucurus venom accidents.

[The role of the human histocompatibility antigens in the pathogenesis of neurological disorders]. / Papel del sistema de histocompatibilidad humano en la patogénesis de las enfermedades neurológicas.

INTRODUCTION: Several studies have been trying to define genetic markers of neurological disorders. Among them, antigens and alleles of the HLA (human leukocyte antigens) system are distinguished. The HLA exerts genetic influence on the susceptibility, clinical aspects and severity of many diseases. The discovery of new molecular methods to typify HLA alleles and the recent nomenclature updates have been contributing to a better understanding of this system. Unfortunately, this information has not been adequately published in the clinical literature. AIM: To review the structure, function, nomenclature and methods of detection of the HLA polymorphism and its associations with common neurological disorders. DEVELOPMENT: Articles that were published between 1990 and 2004 were searched in the MEDLINE and LILACS databases. This review demonstrated that although the HLA association is well established for some neurological disorders (e.g., HLA-DQB1*0602 with multiple sclerosis and narcolepsy; HLA-B7 e HLA-A2 with Alzheimer's disease; HLA-DR3-DR8 with Lamber-Eaton syndrome; and HLA class II Parkinson's disease and amyotrophic lateral sclerosis), these associations are not consistent and vary in different ethnic groups. CONCLUSIONS: It is necessary to study populations from different ethnic backgrounds to identify new associations or to strength the ones already identified. This knowledge will contribute in the evaluation of the risk that a person carrying a particular allele or haplotype has to develop a neurological disease and therefore contribute towards a better understanding of its pathogenesis.

Papel del sistema de histocompatibilidad humano en la patogénesis de las enfermedades neurológicas / The role of the human histocompatibility antigens in the pathogenesis of neurological disorders

Introducción. Numerosos estudios intentan definir los marcadores genéticos de las enfermedades neurológicas. Entre ellos, destacan los antígenos y los alelos del sistema mayor de histocompatibilidad humano. El sistema HLA (human leukocyte antigens) ejerce una influencia genética sobre la susceptibililidad, la manifestación clínica y la gravedad de varias enfermedades. Nuevos métodos moleculares para realizar la tipificación de los alelos del HLA y las constantes actualizaciones de su nomenclatura han contribuido en el mejor entendimiento de ese sistema. Lamentablemente, esas informaciones no se han introducido de manera adecuada en la literatura clínica. Objetivo. Revisar la estructura, la función, la nomenclatura y los métodos de detección del polimorfismo del HLA, y explicar sus asociaciones con enfermedades neurológicas comunes en la práctica clínica. Desarrollo. Se buscaron artículos publicados entre los años 1990 y 2004 en las bases de datos MEDLINE y LILACS. Esa revisión mostró que aunque el HLA esté asociado a algunas enfermedades neurológicas (por ejemplo, HLADQB1* 0602 con la esclerosis múltiple y la narcolepsia, HLA-B7 y HLA-A2 con la enfermedad de Alzheimer, HLA-DR3-DR8 con el síndrome de Lamber-Eaton, y los antígenos HLA de clase II con la enfermedad de Parkinson y la esclerosis lateral amiotrófica), esas asociaciones no son muy consistentes y muestran variaciones en las diferentes etnias debido a su marcado polimorfismo. Conclusiones. Es necesario llevar a cabo investigaciones en poblaciones de diferentes etnias para identificar nuevas asociaciones o reforzar las ya existentes. Las asociaciones más consistentes pueden permitir el uso del sistema HLA para evaluar el riesgo de que una persona portadora de un alelo o haplotipo en particular pueda llegar a desarrollar una determinada enfermedad, lo que podría contribuir a un mejor entendimiento de su patogenia

Introduction. Several studies have been trying to define genetic markers of neurological disorders. Among them, antigens and alleles of the HLA (human leukocyte antigens) system are distinguished. The HLA exerts genetic influence on the susceptibility, clinical aspects and severity of many diseases. The discovery of new molecular methods to typify HLA alleles and the recent nomenclature updates have been contributing to a better understanding of this system. Unfortunately, this information has not been adequately published in the clinical literature. Aim. To review the structure, function, nomenclature and methods of detection of the HLA polymorphism and its associations with common neurological disorders. Development. Articles that were published between 1990 and 2004 were searched in the MEDLINE and LILACS databases. This review demonstrated that although the HLA association is well established for some neurological disorders (e.g., HLA-DQB1*0602 with multiple sclerosis and narcolepsy; HLA-B7 e HLA-A2 with Alzheimer’s disease; HLA-DR3-DR8 with Lamber-Eaton syndrome; and HLA class II Parkinson’s disease and amyotrophic lateral sclerosis), these associations are not consistent and vary in different ethnic groups. Conclusions. It is necessary to study populations from different ethnic backgrounds to identify new associations or to strength the ones already identified. This knowledge will contribute in the evaluation of the risk that a person carrying a particular allele or haplotype has to develop a neurological disease and therefore contribute towards a better understanding of its pathogenesis

El tratamiento grupal del tabaquismo / Group smoking cessation treatment

RESUMEN. Objetivos. Sistematizar el por qué y el cómo se debe y puede hacer tratamiento de deshabituación tabáquica en formato grupal, atendiendo siempre a los máximos niveles de evidencia disponibles. Material y métodos. Se realizó una revisión no sistemática de estudios clínicos y artículos de revisión, incluidos en la biblioteca Cochrane y en las bases de datos Medline, Science Citation Index y Psychinfo. Se revisaron asimismo las Guías de práctica clínica sobre atención al fumador y los protocolos de atención grupal publicados en nuestro país. Resultados. El tratamiento en grupo del tabaquismo es eficaz y coste-efectivo. Sin embargo, para que sea así, dicho tratamiento debe reunir unas características específicas, tanto generales como del terapeuta y del grupo. Asimismo, precisa de una estructuración y componentes determinados. Todos estos elementos se revisan en el artículo. Conclusiones. El uso de un formato grupal en una intervención para dejar de fumar tiene dos justificaciones fundamentales: 1. Su eficacia se ha demostrado de forma rotunda, siendo en ello al menos equivalente a otras intervenciones que implican contacto personal a nivel individual y consistentemente mejor que los materiales de autoayuda por sí solos (fuerza de evidencia A). 2. Su mejor relación coste-efectividad (AU)

ABSTRACT. Objetive. To systematize the best standards for group therapy for smoking cessation, according to the scientific evidence available. Material and methods. A non-systematical review of clinical studies and review articles, including those in Cochrane library and Medline, Science Citation Index and Psychinfo databases, was accomplished. Clinical practice guidelines on smokers attention and group therapy schedule protocols published in our country were also checked. Results. Group therapy for smoking cessation is both effective and cost-effective. Nevertheless, to be so, it must meet some specific characteristics related to the therapist, to group structure and specific contents of the program. All these elements are discussed in the article. Conclusions. The use of group therapy for smoking cessation is justified by two main reasons: 1. Its effectiveness has been clearly demonstrated, being at least equivalent to other interventions which imply personal and individual contact, and quite better than self-help materials alone (Evidence A). 2. It has a better cost-effectiveness ratio. In addition, a therapeutic specific benefit might exist (Evidence C). In other words, better abstinence rates may be attained using a group therapy approach than with brief interventions, and at a lower cost than using individual intensive therapy (AU)

Isolation and biochemical characterization of a fibrinolytic proteinase from Bothrops leucurus (white-tailed jararaca) snake venom.

In investigations aimed at characterizing snake venom clot-dissolving enzymes, we have purified a fibrinolytic proteinase from the venom of Bothrops leucurus (white-tailed jararaca). The proteinase was purified to homogeneity by a combination of molecular sieve chromatography on Sephacryl S-200 and ion-exchange chromatography on CM Sepharose. The enzyme called leucurolysin-a (leuc-a), is a 23 kDa metalloendopeptidase since it is inhibited by EDTA. PMSF, a specific serine proteinase inhibitor had no effect on leuc-a activity. The amino acid sequence was established by Edman degradation of overlapping peptides generated by a variety of selective cleavage procedures. Leuc-a is related in amino acid sequence to reprolysins. The protein is composed of 200 amino acid residues in a single polypeptide chain, possessing a blocked NH2-terminus and containing no carbohydrate. The proteinase showed proteolytic activity on dimethylcasein and on fibrin (specific activity=21.6 units/mg and 17.5 units/microg, respectively; crude venom=8.0 units/mg and 9.5 units/microg). Leuc-a degrades fibrin and fibrinogen by hydrolysis of the alpha chains. Moreover, the enzyme was capable of cleaving plasma fibronectin but not the basement membrane protein laminin. Leuc-a cleaved the Ala14-Leu15 and Tyr16-Leu17 bonds in oxidized insulin B chain. The pH optimum of the proteolysis of dimethylcasein by leuc-a was about pH 7.0. Antibody raised in rabbit against the purified enzyme reacted with leuc-a and with the crude venom of B. leucurus. In vitro studies revealed that leuc-a dissolves clots made either from purified fibrinogen or from whole blood, and unlike some other venom fibrinolytic metallopeptidases, leuc-a is devoid of hemorrhagic activity when injected (up to 100 microg) subcutaneously into mice.

Tratamiento rehabilitador del hombro doloroso / Rehabilitating treatment of painful shoulder

Introducción. El objetivo de este estudio es conocer las características de los pacientes que consultan por hombro doloroso en nuestra área y evaluar los resultados del protocolo de tratamiento rehabilitador utilizado en nuestro servicio. Material y métodos. Se han seleccionado pacientes con hombro doloroso secundario a uno de estos cuatro diagnósticos: síndrome subacromial/tendinitis no calcificada de manguito de los rotadores o bicipital; rotura parcial o total del manguito de los rotadores; tendinitis calcificada del manguito de los rotadores; capsulitis retráctil. El tratamiento ha sido domiciliario y hospitalario basado en disminuir el dolor, conseguir un balance articular completo y potenciar el manguito de los rotadores y los músculos estabilizadores de la escápula. Se recogieron los siguientes datos: edad, sexo, profesión, antecedente traumático, diagnóstico, confirmación del diagnóstico por técnicas de imagen, tiempo de evolución del dolor, número de sesiones de tratamiento, test de Constant pre y postratamiento y la necesidad de otra consulta por el mismo proceso. Test estadísticos utilizados: test de Wilcoxon, pruebas de Mann-Whitney y Kruskal-Wallis, χ2 de Pearson y el test de correlación de Spearman. Nivel de significación 0,01 y 0,05.Resultados. Muestra de 278 pacientes: 187 mujeres y 91 varones. Edad: 56,01 ± 11,07 años. Diagnóstico más frecuente: síndrome subacromial/tendinitis no calcificada (57,20 %). Sin antecedente traumático: 70,30 %. Confirmación del diagnóstico por imagen: 50,40 %. Tiempo de evolución: 8,28 ± 6,98 meses. Número sesiones: 10,53 ± 5,11. Test de Constant:57,03 ± 10,52 puntos pre-tratamiento y 70,10 ± 11,94 postratamiento. El 74,80 % no han requerido otra consulta y el9,50 % han precisado intervención quirúrgica. Conclusiones. Concluimos que nuestro protocolo mejora el tratamiento del hombro doloroso secundario a los diagnósticos anteriormente descritos

Introduction. The aim of this study is to describe the clinical features in patients with shoulder pain referred to our consultation and to assess the results of a physical therapy protocol that we use for shoulder pain. Material and methods. We have selected patients suffering from shoulder pain caused for one of this four diagnoses: subacromial syndrome/rotator cuff tendonitis without calcification; partial-total rotator cuff tear; rotator cuff calcifying tendonitis; adhesive capsulitis. Patients have performed treatment at home and at hospital, based on relieve pain, improve movement and strengthen rotator cuff and scapular muscles. Main outcome measures: age, sex, professión, traumatic injury, diagnosis, to have or not a radiological diagnosis, pain duration, number of treatment sessions, Constant test preand post-treatment and needing another consultation for the same problem. Stadistics tests used: Wilcoxon test, Mann-Whitney and Kruskal-Wallis Test, Pearson’s χ2 and Spearman’s correlations. Significant at the 0,01 and 0,05 level. Results. Sample of 278 patients: 187 women and 91 men. Age: 56,01 ± 11,07 years. Most comon diagnoses: Subacromial syndrome/rotator cuff tendonitis without calcification:57,20 %. Not traumatic injury: 70,30 %. Clinical diagnosis was confirmed by radiological diagnosis in 50,40 %. Pain duration:8,28 ± 6,98 months. Number of rehabilitation sessions:10,53 ± 5,11. Constant test: 57,03 ± 10,52 points pre-treatment and 70,10 ± 11,94 post-treatment. The 74,80 % of the patients did not need another consultation for shoulder pain. The 9,50 % needed surgery. Conclusions. We consider our treatment protocol to be reliable to treat SP in those patients with the described diagnoses