Binding sites and actions of Tx1, a neurotoxin from the venom of the spider Phoneutria nigriventer, in guinea pig ileum

Tx1, a neurotoxin isolated from the venom of the South American spider Phoneutria nigriventer, produces tail elevation, behavioral excitation and spastic paralysis of the hind limbs after intracerebroventricular injection in mice. Since Tx1 contracts isolated guinea pig ileum, we have investigated the effect of this toxin on acetylcholine release, as well as its binding to myenteric plexus-longitudinal muscle membranes from the guinea pig ileum. [125I]-Tx1 binds specifically and with high affinity (Kd = 0.36 ± 0.02 nM) to a single, non-interacting (nH = 1.1), low capacity (Bmax 1.1 pmol/mg protein) binding site. In competition experiments using several compounds (including ion channel ligands), only PhTx2 and PhTx3 competed with [125I]-Tx1 for specific binding sites (K0.5 apparent = 7.50 x 10-4 g/l and 1.85 x 10-5 g/l, respectively). PhTx2 and PhTx3, fractions from P. nigriventer venom, contain toxins acting on sodium and calcium channels, respectively. However, the neurotoxin PhTx2-6, one of the isoforms found in the PhTx2 pool, did not affect [125I]-Tx1 binding. Tx1 reduced the [3H]-ACh release evoked by the PhTx2 pool by 33%, but did not affect basal or KCl-induced [3H]-ACh release. Based on these results, as well as on the homology of Tx1 with toxins acting on calcium channels (w-Aga IA and IB) and its competition with [125I]-w-Cono GVIA in the central nervous system, we suggest that the target site for Tx1 may be calcium channels.

Extracellular matrix molecules as targets for brown spider venom toxins

Loxoscelism, the term used to describe lesions and clinical manifestations induced by brown spider's venom (Loxosceles genus), has attracted much attention over the last years. Brown spider bites have been reported to cause a local and acute inflammatory reaction that may evolve to dermonecrosis (a hallmark of envenomation) and hemorrhage at the bite site, besides systemic manifestations such as thrombocytopenia, disseminated intravascular coagulation, hemolysis, and renal failure. The molecular mechanisms by which Loxosceles venoms induce injury are currently under investigation. In this review, we focused on the latest reports describing the biological and physiopathological aspects of loxoscelism, with reference mainly to the proteases recently described as metalloproteases and serine proteases, as well as on the proteolytic effects triggered by L. intermedia venom upon extracellular matrix constituents such as fibronectin, fibrinogen, entactin and heparan sulfate proteoglycan, besides the disruptive activity of the venom on Engelbreth-Holm-Swarm basement membranes. Degradation of these extracellular matrix molecules and the observed disruption of basement membranes could be related to deleterious activities of the venom such as loss of vessel and glomerular integrity and spreading of the venom toxins to underlying tissues.

IgA response in serum and gut secretion in sensitized mice fed with the dust mite Dermatophagoides pteronyssinus extract

Induced oral tolerance to mucosal-exposed antigens in immunized animals is of particular interest for the development of immunotherapeutic approaches to human allergic diseases. This is a unique feature of mucosal surfaces which represent the main contact interface with the external environment. However, the influence of oral tolerance on specific and natural polyreactive IgA antibodies, the major defense mechanism of the mucosa, is unknown. We have shown that oral administration of an extract of the dust mite Dermatophagoides pteronyssinus (Dp) to primed mice caused down-regulation of IgE responses and an increase in tumor growth factor-ß secretion. In the present study, we observed that primed inbred female A/Sn mice (8 to 10 weeks old) fed by gavage a total weight of 1.0-mg Dp extract on the 6th, 7th and 8th days post-immunization presented normal secretion of IL-4 and IL-10 in gut-associated lymphoid tissue and a decreased production of interferon gamma induced by Dp in the draining lymph nodes (13,340 ± 3,519 vs 29,280 ± 2,971 pg/ml). Mice fed the Dp extract also showed higher levels of serum anti-Dp IgA antibodies and an increase of IgA-secreting cells in mesenteric lymph nodes (N = 10), reflecting an increase in total fecal IgA antibodies (N = 10). The levels of secretory anti-Dp IgA antibodies increased after re-immunization regardless of Dp extract feeding. Oral tolerance did not interfere with serum or secretory IgA antibody reactivity related to self and non-self antigens. These results suggest that induction of oral tolerance to a Dp extract in sensitized mice triggered different regulatory mechanisms which inhibited the IgE response and stimulated systemic and secretory IgA responses, preserving the natural polyreactive IgA antibody production.

Cloning, sequence analysis, and expression of cDNA coding for the major house dust mite allergen, Der f 1, in Escherichia coli

Our objective was to clone, express and characterize adult Dermatophagoides farinae group 1 (Der f 1) allergens to further produce recombinant allergens for future clinical applications in order to eliminate side reactions from crude extracts of mites. Based on GenBank data, we designed primers and amplified the cDNA fragment coding for Der f 1 by nested-PCR. After purification and recovery, the cDNA fragment was cloned into the pMD19-T vector. The fragment was then sequenced, subcloned into the plasmid pET28a(+), expressed in Escherichia coli BL21 and identified by Western blotting. The cDNA coding for Der f 1 was cloned, sequenced and expressed successfully. Sequence analysis showed the presence of an open reading frame containing 966 bp that encodes a protein of 321 amino acids. Interestingly, homology analysis showed that the Der p 1 shared more than 87% identity in amino acid sequence with Eur m 1 but only 80% with Der f 1. Furthermore, phylogenetic analyses suggested that D. pteronyssinus was evolutionarily closer to Euroglyphus maynei than to D. farinae, even though D. pteronyssinus and D. farinae belong to the same Dermatophagoides genus. A total of three cysteine peptidase active sites were found in the predicted amino acid sequence, including 127-138 (QGGCGSCWAFSG), 267-277 (NYHAVNIVGYG) and 284-303 (YWIVRNSWDTTWGDSGYGYF). Moreover, secondary structure analysis revealed that Der f 1 contained an a helix (33.96%), an extended strand (17.13%), a ß turn (5.61%), and a random coil (43.30%). A simple three-dimensional model of this protein was constructed using a Swiss-model server. The cDNA coding for Der f 1 was cloned, sequenced and expressed successfully. Alignment and phylogenetic analysis suggests that D. pteronyssinus is evolutionarily more similar to E. maynei than to D. farinae.

Cloning, bioinformatics analysis, and expression of the dust mite allergen Der f 5 of Dermatophagoides farinae

Crude extracts of house dust mites are used clinically for diagnosis and immunotherapy of allergic diseases, including bronchial asthma, perennial rhinitis, and atopic dermatitis. However, crude extracts are complexes with non-allergenic antigens and lack effective concentrations of important allergens, resulting in several side effects. Dermatophagoides farinae (Hughes; Acari: Pyroglyphidae) is one of the predominant sources of dust mite allergens, which has more than 30 groups of allergen. The cDNA coding for the group 5 allergen of D. farinae from China was cloned, sequenced and expressed. According to alignment using the VECTOR NTI 9.0 software, there were eight mismatched nucleotides in five cDNA clones resulting in seven incompatible amino acid residues, suggesting that the Der f 5 allergen might have sequence polymorphism. Bioinformatics analysis revealed that the matured Der f 5 allergen has a molecular mass of 13604.03 Da, a theoretical pI of 5.43 and is probably hydrophobic and cytoplasmic. Similarities in amino acid sequences between Der f 5 and allergens of other domestic mite species, viz. Der p 5, Blo t 5, Sui m 5, and Lep d 5, were 79, 48, 53, and 37%, respectively. Phylogenetic analysis indicated that Der f 5 and Der p 5 clustered together. Blo t 5 and Ale o 5 also clustered together, although Blomia tropicalis and Aleuroglyphus ovatus belong to different mite families, viz. Echimyopodidae and Acaridae, respectively.

MCMI-II borderline personality disorder in anorexia and bulimia nervosa. 'Trastorno l??mite de personalidad seg??n el MCMI-II en la anorexia y bulimia nerviosas'

Resumen tomado de la publicaci??n

Los superhéroes, el deber moral y la obligación : el caso de Spider-man y los X-Men

La manera en que los superhéroes actúan como agentes morales, así como la forma en que asumen y justifican su deber moral se explora por medio del análisis de dos superhéroes de historieta: Spider-man y los X-Men. Una exploración de los mensajes mediáticos desde la filosofía moral.

Sentit pedagògic del mite de la bèstia enamorada. 'Sentido pedagógico del mito de la bestia enamorada'.

El tema de la bestia enamorada es frecuente en los cuentos de todas las culturas. No siempre se trata de un animal sino que el enamorado suele adoptar formas mostruosas y feroces. Partiendo del psicoanálisis, se ha demostrado la acción liberadora que ejercen los cuentos populares en la formación de la personalidad infantil y en la maduración de la afectividad, el enriquecimiento del lenguaje y la imaginación, la educación de la atención pero sobre todo la capacidad que poseen para favorecer el conocimiento y superar los propios problemas emocionales a través de recrearlos en los cuentos. El coraje es sin duda uno de los valores que con más insistencia ofrecen los cuentos populares: la aventura iniciática es siempre un reto, un riesgo a los ojos del protagonista para que sea capaz de correr las peripecias llenas de intriga que la historia le plantea. Pero sobre todo lo que es más destacable en los cuentos es que sus mensajes llegan a través de simples y cotidianas situaciones sociales, como el mito de la bestia enamorada sirve para cambiar la actitud de la mujer frente al sexo, para pasar del sentimiento de rechazo y miedo a la aceptación.

Sentit pedagògic del mite de la bèstia enamorada. 'Sentido pedagógico del mito de la bestia enamorada'.

Resumen tomado de la publicación

El Montseny i les Guilleries : paisatge, mite i literatura. 'El Montseny y las Guilleries : paisaje, mito y literatura'

Resumen del v??deo en catal??n

Mitologia : l'antropologia a través del mite. 'Mitología : la antropología a través del mito'.

Trabajo que analiza los mitos y las leyendas que han creado los distintos pueblos y culturas del mundo en su intento de dar respuesta a las preguntas que a lo largo de los siglos se ha planteado la humanidad. Asi el capítulo segundo estudia los dioses y héroes de la mitología china, africana, árabe, inca, vikinga y helénica. Para finalizar con una cronología, documentos y bibliografía, así como con una serie de actividades y juegos para una mejor comprensión del tema..

Are you a spider?

Ofrece información a los niños más pequeños a través del ciclo de vida de una araña. Con un texto divertido se hace la comparación entre una araña y un niño y se examinan las diferencias que hay entre ambos para ayudar a entender los procesos de la vida y ver las similitudes y diferencias entre las arañas y los seres humanos.

A cheese and tomato spider.

Presenta las páginas divididas por la mitad en horizontal, con lo que las múltiples ilustraciones de animales, objetos y personas, se pueden combinar para dar origen a objetos y personas absurdos e inverosímiles.

El CAP : tr??mite y exigencia

El matricularse en el CAP es simplemente un tr??mite burocr??tico y obligatorio. Exigible para dar clases y opositar. Est?? condenado a vivir sin ser curso de aptitud pedag??gica. Si este curso es necesario y requiere una organizaci??n hay que organizarlo bien, con profesores competentes que supervisen de modo constante las practicas, programaciones y resultados de su actividad. La memoria es un tr??mite es un tramite molesto e innecesario que se deber??a suprimir. La ruina del CAP no viene de ensayar f??rmulas nuevas, sino de querer mantener a toda costa una estructura caduca y cara, adem??s de inoperante e in??til.

Definiciones personales y aspectos estructurales del concepto de l??mite finito de una funci??n en un punto

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