Contribution of topology determinants of a viral movement protein to its membrane association, intracellular traffic, and viral cell-to-cell movement.

The p7B movement protein (MP) of Melon necrotic spot virus (MNSV) is a single-pass membrane protein associated with the endoplasmic reticulum (ER), the Golgi apparatus (GA), and plasmodesmata (Pd). Experimental data presented here revealed that the p7B transmembrane domain (TMD) was sufficient to target the green fluorescent protein (GFP) to ER membranes. In addition, the short extramembrane regions of p7B were essential for subsequent ER export and transport to the GA and Pd. Microsomal partitioning and bimolecular fluorescence assays supported a type II topology of p7B in planta. Mutations affecting conventional determinants of p7B membrane topology, such as the TMD secondary structure, the overall hydrophobicity profile, the so-called "aromatic belt," and the net charge distribution on either side of the TMD, were engineered into infectious RNAs to investigate the relationship between the MP structure and MNSV cell-to-cell movement. The results revealed that (i) the overall hydrophobic profile and the α-helix integrity of the TMD were relevant for virus movement, (ii) modification of the net charge balance of the regions flanking both TMD sides drastically reduced cell-to-cell movement, (iii) localization of p7B to the GA was necessary but not sufficient for virus movement, and (iv) membrane insertion was essential for p7B function in virus movement. Our results therefore indicate that MNSV cell-to-cell movement requires sequential transport of p7B from the ER via the GA to Pd, which is modulated by a combination of several signals with different strengths in the extramembrane regions and TMD of the MP.

[Headache, does it cause a health care overload in primary health care? Urban area vs rural area medical visit]. / Cefalea, ¿una sobrecarga asistencial en atención primaria? Consulta urbana frente a consulta rural.

OBJECTIVES: Headache causes an overload and a problem to out-patient neurology. There are few studies that quantify the overload that the headache causes in a primary health care out-patient clinic visit. The aim of the present study is to identify and quantify the real load that this disease supposes in two primary health care consultations, one located in a rural area and another in an urban area. MATERIAL AND METHODS: A 6 month long observational and descritive study. The total number of patients seen, the number of patients seen for headache and their diagnostic classification were recorded. RESULTS: A total of 6,014 visits were counted. Only 46 patients consulted due to headache (0,76%). Tensional headache was the most frequently diagnosed headache (43.48%) followed by migraine (23.91%). CONCLUSIONS: Our study verifies that published by other authors and determines that, on the contrary to that which occurs in the neurology out-patient clinic, headache does not cause a daily work overload for the general practitioner. The scarce number of consultations for headache, with the diagnosis and management involved in this disease, makes it necessary to consider a multidisciplinary health care problem such as headache differently according to the health care level involved.

Cefalea, ¿una sobrecarga asistencial en atención primaria? Consulta urbana frente a consulta rural / Headache, does it cause a health care overload in Primary Health Care? Urban area vs rural area medical visit

El dolor de cabeza supone una sobrecargay unun problema asistencial en cualquier consulta ambulatoriade neurología. Existen pocos estudios que cuantifiquen lasobrecarga que supone el dolor de cabeza en una consultade atención primaria. El objetivo del presente estudio esidentificar y cuantificar realmente la carga que esta patologíasupone en dos consultas de atención primaria, una localizadaen un ámbito rural y otra en un ámbito urbano.Material y métodos. Estudio observacional y descriptivode 6 meses de duración. Se registra el total de pacientesatendidos, el número de pacientes asistidos por dolor de cabezay su clasificación diagnóstica.Resultados. Se contabilizó un total de 6.014 consultas.Sólo 46 pacientes consultan por cefalea (0,76%). La cefaleatensional es el dolor de cabeza diagnosticado con más frecuencia(el 43,48%), seguido de la migraña (el 23,91%).Conclusiones. Nuestro estudio certifica lo publicadopor otros autores y determina que, contrariamente a lo quesucede en la neurología ambulatoria, la cefalea no suponeuna carga en el trabajo diario de un médico de familia. Elescaso número de consultas por cefalea, con los problemasde diagnóstico y manejo que rodea a esta patología, originanque un problema de atención multidisciplinar como lacefalea se afronte de diferente manera según el nivel asistencialdonde uno se encuentre (AU)

Objectives. Headache causes an overload and a problemto out-patient neurology. There are few studies that quantify the overload that the headache causes in a PrimaryHealth Care out-patient clinic visit. The aim of thepresent study is to identify and quantify the real loadthat this disease supposes in two Primary Health Careconsultations, one located in a rural area and another inan urban area.Material and methods. A 6 month long observationaland descritive study. The total number of patients seen,the number of patients seen for headache and theirdiagnostic classification were recorded.Results. A total of 6,014 visits were counted. Only46 patients consulted due to headache (0,76%). Tensionalheadache was the most frequently diagnosed headache(43.48%) followed by migraine (23.91%).Conclusions. Our study verifies that published byother authors and determines that, on the contrary tothat which occcurs in the neurology out-patient clinic,headache does not cause a daily work overload for thegeneral practitioner. The scarce number of consultationsfor headache, with the diagnosis and management involvedin this disease, makes it necessary to consider a multidisciplinaryhealth care problem such as headache differentlyaccording to the health care level involved (AU)

RNA-binding properties and membrane insertion of Melon necrotic spot virus (MNSV) double gene block movement proteins.

Advances in structural and biochemical properties of carmovirus movement proteins (MPs) have only been obtained in p7 and p9 from Carnation mottle virus (CarMV). Alignment of carmovirus MPs revealed a low conservation of amino acid identity but interestingly, similarity was elevated in regions associated with the functional secondary structure elements reported for CarMV which were conserved in all studied proteins. Nevertheless, some differential features in relation with CarMV MPs were identified in those from Melon necrotic virus (MNSV) (p7A and p7B). p7A was a soluble non-sequence specific RNA-binding protein, but unlike CarMV p7, its central region alone could not account for the RNA-binding properties of the entire protein. In fact, a 22-amino acid synthetic peptide whose sequence corresponds to this central region rendered an apparent dissociation constant (K(d)) significantly higher than that of the corresponding entire protein (9 mM vs. 0.83-25.7 microM). This p7A-derived peptide could be induced to fold into an alpha-helical structure as demonstrated for other carmovirus p7-like proteins. Additionally, in vitro fractionation of p7B transcription/translation mixtures in the presence of ER-derived microsomal membranes strongly suggested that p7B is an integral membrane protein. Both characteristics of these two small MPs forming the double gene block (DGB) of MNSV are discussed in the context of the intra- and intercellular movement of carmovirus.

Functional analysis of the five melon necrotic spot virus genome-encoded proteins.

Function of the melon necrotic spot virus (MNSV) genome-encoded proteins (p29, p89, p7A, p7B and p42) has been studied. Protein-expression mutants of an infectious, full-length cDNA clone of a Spanish MNSV-Al isolate and a recombinant green fluorescent protein (GFP)-expressing virus were used in infection bioassays on melon plants. Results revealed that p29 and p89 are both essential for virus replication, whereas small proteins p7A and p7B are sufficient to support viral movement between adjacent cells operating in trans. It is also demonstrated that, in addition to its structural role as coat protein, p42 is an important factor controlling symptoms and is required for systemic transport. Moreover, both p42 and p7B, among all of the MNSV-encoded proteins, were able to delay RNA silencing in transient-expression assays on GFP-transgenic Nicotiana benthamiana plants. Finally, the presence of p42 also produced an enhancing effect on local spread similar to that of potyviral helper component proteinase (HC-Pro), probably due to its RNA silencing-suppression ability.

[Ultrasonography imaging of the torsion knot in spermatic cord torsion]. / Imagen ecográfica del nudo de torsión en la torsión de cordón espermático.

Presentation of the finding of the echographic image of a torsion knot as a pathognomonic sign of spermatic cord torsion. Discussion of this finding as well as aspects related to imaging diagnosis of cord's torsion.

Bacteraemia by P. aeruginosa associated with nephrotic syndrome after shunt operation for hydrocephalus.

One patient of sepsis by P. aeruginosa after the implanting of a ventriculoatrial valve following extirpation of an expansive intracranial process is studied. During the evolution of the septic process there appeared as associated nephrotic syndrome with histologic lesions corresponding to an acute exudative glomerular nephritis. The germ became resistant to the antibiotics used, carbenicillin plus gentamicin, being later sensitive only to fosfomycin with which finally complete cure was achieved.