Local analysis of strains and rotations for macromolecular electron microscopy maps.

Macromolecular complexes perform their physiological functions by local rearrangements of their constituents and biochemically interacting with their reaction partners. These rearrangements may involve local rotations and the induction of local strains causing different mechanical efforts and stretches at the different areas of the protein. The analysis of these local deformations may reveal important insight into the way proteins perform their tasks. In this paper we introduce a method to perform this kind of local analysis using Electron Microscopy volumes in a fully objective and automatic manner. For doing so, we exploit the continuous nature of the result of an elastic image registration using B-splines as its basis functions. We show that the results obtained by the new automatic method are consistent with previous observations on these macromolecules.

[Therapeutic effect of mitomycin C in the immediate postoperative period in patients with intermediate-risk non-muscle-invasive bladder tumors]. / Efecto terapéutico de mitomicina C en el postoperatorio inmediato de pacientes con tumores vesicales no músculo invasores de riesgo intermedio.

INTRODUCTION: Approximately 70-85% of transitional bladder cell carcinomas are non-muscle-invasive. After an initial surgery, around 60-90% will have a recurrence, being the highest risk period the first two years. Urothelium instability could be the main reason for recurrence in mid grade tumours, reason why a single dose of a chemotherapy after transurethral resection of the bladder (TURB) might be insufficient. That is why a deferred therapy in occasions associated with maintenance is recommended. PATIENTS AND METHODS: A prospective, controlled and randomized study was performed. We included non-muscle-invasive mid risk bladder tumours. All patients had initially a TURB performed and were randomized to receive a single dose of mitomycin C (MMC), in the immediate postoperative period. RESULTS: A total of 105 patients were included. Mean follow-up was 22, 70 +/- 8, 15 months. MMC was administered to 53 patients. Of these 66, 0% had no recurrence and 34.0% had a non-muscle-invasive recurrence. Of the 52 patients in the non MMC group, 53.8% had no recurrence and 44.2% had a non-muscle-invasive recurrence and only 1 patient had a muscle-invasive progression. We did not find significantly differences in time to recurrence in mid risk tumours when using immediate postoperative single dose of MMC or deferred therapy. There was only one case of myelosuppression. DISCUSSION: In mid risk non-muscle-invasive tumors, some studies suggest that early intravesical instillation of chemotherapy reduces the risk of recurrence after TURB. We could not show significantly differences when comparing postoperatorive MMC versus traditional deferred instillations.

Architectures based on the use of gold nanoparticles and ruthenium complexes as a new route to improve genosensor sensitivity.

The preparation of DNA-sensing architectures based on gold nanoparticles (Au-NPs) in conjunction with an "in situ" prepared ruthenium complex as a new route to improve the analytical properties of genosensors is described. In the development of these architectures several strategies to obtain Au-NPs modified gold electrodes (Au-NP/Au) have been essayed, in particular covalent binding and electrochemical deposition from a solution containing Au-NPs previously synthesized. UV-vis absorption measurements in conjunction with transmission electron microscope (TEM) images reveal that the synthesized Au-NPs are stable for at least 4 weeks and have a narrow size distribution. Atomic force microscopy (AFM) was employed to characterize the morphology and to estimate the Au-NPs surface coverage of the modified gold electrodes obtained following the different modification strategies. In order to assess the utility of these architectures as DNA-sensing devices, a thiolated capture probe sequence from Helicobacter pylori was immobilized onto the as-prepared surface. This sequence was chosen as a case of study within the framework of developing approaches of wide applicability. The hybridization event is detected using a water-soluble pentaamin ruthenium [3-(2-phenanthren-9-yl-vinyl)-pyridine] complex (Ru(NH(3))(5)L) prepared "in situ". This complex, due to its intercalative character, is able to bind to double stranded DNA more efficiently than to single stranded DNA. In addition, the metal provides with a redox center that can be used as an electrochemical indicator. On the basis of this strategy, complementary target sequences of H. pylori have been detected over the range of 40-800 pmol with a detection limit of 25+/-2 pmol.

[Use of zoledromic acid on pain control in stages M1b of prostate cancer]. / Seguridad del ácido zoledrómico en el control del dolor en el estadio M1b del cáncer de próstata.

OBJECTIVES: To probe the use of Zoledromic acid in the control of the pain due to bone methastasis of a prostate cancer. MATERIAL AND METHOD: We included in our study 37 consecutive patients diagnosed of prostate cancer who developed bone methastasis during 2005 and 2006 to whom we gave a 4 mg dose of Zoledromic acid during a mean time of 10.24 months. RESULTS: We observed a complete control of the pain in 8 of the 20 patients, and partially in 6 of those patients. We did not evaluate any adverse reaction due to this treatment. CONCLUSIONS: We still yet to know whether the administration of this treatment in the early stages of the prostate cancer could prevent or retard the appearance of bone methastasis.

[Primary psoas abscess due to Streptococcus pneumoniae]. / Absceso primario de psoas por Streptococo pneumoniae.

INTRODUCTION AND OBJECTIVES: To report to the literature a new case of primary psoas abscess. METHODS: The patient is a 65 years old woman who suffered back and left hip pain for approximately 1 month. A CTA showed a retroperitoneal mass that involved the left Psoas. It was drained 1800 cc of purulent fluid infected by Streptococo pneumoniae. RESULTS: In a CTA made 21 days after drainage, showed a very small residual collection. CONCLUSIONS: Primary Psoas Abscess are a very rare entity, with a low rate of mortatility if well treated.

[Trombosis of the dorsal penis vein (of Mondor's phlebitis). Presentation of a new case]. / Trombosis de la vena dorsal del pene (Flebitis de Mondor). Aportación de un nuevo caso.

OBJECTIVE: We present a new case of trombosis of the superficial dorsal penis vein called Penile Mondor's disease. The characteristics of the disease are reviewed and the most usual diagnostic and therapeutic methods. METHODS: The case of a 41 year old man is reviewed who consulted for pain and induration on the proximal part of the penis. RESULTS: After phisical examination and Eco-doppler was made the diagnosis of Mondor's disease. He receibed treatment with non steroidal antiinflamatories and antibiotics. CONCLUSIONS: The dorsal vein thrombosis is a rare disease with pain an induration of the dorsal part of the penis. The ethiology can be traumatic, neoplasic, excesive sexual activity or abstinence. Is necesary the diferencial diagnosis with esclerosant linphangitis and the most important imaging is the Eco-doppler. The treatment is based in non steroidal antiinflamatories and antibiotics wit infection. The local aplication of heparine can be useful and the surgery with thrombectomy and resection is for persistent cases.

[Treatment of erectile dysfunction with penile prothesis. Our experience]. / Tratamiento de la disfunción eréctil mediante el implante de prótesis de pene. Análisis de nuestra serie.

INTRODUCTION AND OBJECTIVES: The aim of this study was to review the current situation of penile prosthesis in the treatment of erectile dysfunction and to analyze the indications, degree of acceptability and complications in our experience. MATERIAL AND METHODS: From 1984 to 2003, 48 implants were inserted in men suffering from impotence due to different etiologies. The mean age was 55 years. Vascular disease, Diabetes and pelvic surgery were the most common pathologies recorded. The type of prothesis inserted were malleable inactive in 7 cases (14.58%), Jonas6, Acu-form1. Inflable active with 3 components in 19 cases (39.58%), AMS 700 plus17, Alpha2. Inflable active integrated in 5 cases (10.41%) Hydroflex4, Dynaflex1. Inflable active with 2 components in 17 cases (35.4%), Ambicor2, Mark II15. RESULTS: Infection of the prosthesis was observed in 6.25% of the cases. This infectious complication required removal of the prosthesis. 4.16% of the cases complained of mechanical failure that required change of the prosthesis. 80% of the patients were satisfied with the results. CONCLUSIONS: Penile prostheses continue to be an effective and valid therapeutic alternative in impotent. Although the complication rate is low, adequate information must be provided to the patient.

[Primary renal lymphoma in a patient with IgM monoclonal gammapathy]. / Linfoma renal primario en paciente con gammapatía monoclonal IgM.

Reports on primary renal lymphoma are scarce in the urological literature, the most part of them are secondary on a lymphomatous infiltration of the kidneys. We report the case of a 77 year old man with an incidental mass on the kidney. After radiological studies (CT), we practise nephrectomy with a pathological result of a non-Hodgking B primary lymphoma. The patient present a IgM monoclonal gammapathy who need complementary treatment with chemotherapy. A literature review on currently recommended diagnostic and treatment practices in presented.

[Infertility caused by AZF microdeletions. A new case of azoospermia]. / Infertilidad por microdeleciones AZF. Un nuevo caso de azoospermia.

We report report a case of a man with azoospermia who was studied because of infertility. Between the causes of azoospermia and severe oligospermia, in male patients with infertility evaluations, small deletions in regions of the Y chromosome must be considered, as in our case. Regions for spermatogenesis in the Yq chromosome can be evidenced using PCR analysis. Men with azoospermia are at high risk to have small deletions in that regions. Yq chromosome deletions could be transmitted when ICSI (intracytoplasmic sperm injection) is applied. Thereafter, study of microdeletions would be included in severe infertile men.

Analysis of the interaction between the eukaryotic chaperonin CCT and its substrates actin and tubulin.

Two mechanisms have thus far been characterized for the assistance by chaperonins of the folding of other proteins. The first and best described is that of the prokaryotic chaperonin GroEL, which interacts with a large spectrum of proteins. GroEL uses a nonspecific mechanism by which any conformation of practically any unfolded polypeptide interacts with it through exposed, hydrophobic residues. ATP binding liberates the substrate in the GroEL cavity where it is given a chance to fold. A second mechanism has been described for the eukaryotic chaperonin CCT, which interacts mainly with the cytoskeletal proteins actin and tubulin. Cryoelectron microscopy and biochemical studies have revealed that both of these proteins interact with CCT in quasi-native, defined conformations. Here we have performed a detailed study of the docking of the actin and tubulin molecules extracted from their corresponding CCT:substrate complexes obtained from cryoelectron microscopy and image processing to localize certain regions in actin and tubulin that are involved in the interaction with CCT. These regions of actin and tubulin, which are not present in their prokaryotic counterparts FtsA and FtsZ, are involved in the polymerization of the two cytoskeletal proteins. These findings suggest coevolution of CCT with actin and tubulin in order to counteract the folding problems associated with the generation in these two cytoskeletal protein families of new domains involved in their polymerization.

The 'sequential allosteric ring' mechanism in the eukaryotic chaperonin-assisted folding of actin and tubulin.

Folding to completion of actin and tubulin in the eukaryotic cytosol requires their interaction with cytosolic chaperonin CCT [chaperonin containing tailless complex polypeptide 1 (TCP-1)]. Three-dimensional reconstructions of nucleotide-free CCT complexed to either actin or tubulin show that CCT stabilizes both cytoskeletal proteins in open and quasi-folded conformations mediated through interactions that are both subunit specific and geometry dependent. Here we find that upon ATP binding, mimicked by the non-hydrolysable analog AMP-PNP (5'-adenylyl-imido-diphosphate), to both CCT-alpha-actin and CCT- beta-tubulin complexes, the chaperonin component undergoes concerted movements of the apical domains, resulting in the cavity being closed off by the helical protrusions of the eight apical domains. However, in contrast to the GroE system, generation of this closed state does not induce the release of the substrate into the chaperonin cavity, and both cytoskeletal proteins remain bound to the chaperonin apical domains. Docking of the AMP-PNP-CCT-bound conformations of alpha-actin and beta-tubulin to their respective native atomic structures suggests that both proteins have progressed towards their native states.

Three-dimensional reconstruction of a recombinant influenza virus ribonucleoprotein particle.

A three-dimensional structural model of an influenza virus ribonucleoprotein particle reconstituted in vivo from recombinant proteins and a model genomic vRNA has been generated by electron microscopy. It shows a circular shape and contains nine nucleoprotein monomers, two of which are connected with the polymerase complex. The nucleoprotein monomers show a curvature that may be responsible for the formation of helical structures in the full-size viral ribonucleoproteins. The monomers show distinct contact boundaries at the two sides of the particle, suggesting that the genomic RNA may be located in association with the nucleoprotein at the base of the ribonucleoprotein complex. Sections of the three-dimensional model show a trilobular morphology in the polymerase complex that is consistent with the presence of its three subunits.

Eukaryotic chaperonin CCT stabilizes actin and tubulin folding intermediates in open quasi-native conformations.

Three-dimensional reconstruction from cryoelectron micrographs of the eukaryotic cytosolic chaperonin CCT complexed to tubulin shows that CCT interacts with tubulin (both the alpha and beta isoforms) using five specific CCT subunits. The CCT-tubulin interaction has a different geometry to the CCT-actin interaction, and a mixture of shared and unique CCT subunits is used in binding the two substrates. Docking of the atomic structures of both actin and tubulin to their CCT-bound conformation suggests a common mode of chaperonin-substrate interaction. CCT stabilizes quasi-native structures in both proteins that are open through their domain-connecting hinge regions, suggesting a novel mechanism and function of CCT in assisted protein folding.

[Determination of NMP-22 as recurrence marker in bladder cancer. Preliminary study]. / Determinación de NMP-22 como marcador de recidivas en el cáncer vesical. Estudio preliminar.

OBJECTIVES: The present study was conducted to determine the efficacy of NMP-22 as a diagnostic test for recurrence of bladder tumors and to compare its efficacy with urine cytology. METHODS: 30 patients (25 male, 5 female), aged 41-87 years (mean 73.4), with bladder tumor were evaluated by cytology, cystoscopy and determination of NMP-22 at 3 and 6 months post-TUR of the bladder. A positive test was defined as NMP-22 greater than 10 U/ml. RESULTS: 80.7% were T1, 15.3% T2-T3, 76.8% were grade I and II, and 23.2% were grade III. NMP-22 showed a sensitivity of 69%, a specificity of 64%, a positive predictive value of 52% and a negative predictive value of 78%, using tumor recurrence at 3 and 6 months as the unit of analysis, for a recurrence rate of 36%. Cytology showed a sensitivity of 44%, a specificity of 92%, a positive predictive value of 77% and a negative predictive value of 74%. When both tests were used, we obtained a sensitivity of 87.5%, a specificity of 64.2%, a positive predictive value of 58.3% and a negative predictive value of 90%. In grade I tumors, determination of NMP-22 had a sensitivity of 100%, a specificity of 77.7%, a positive predictive value of 60% and a negative predictive value of 69%. CONCLUSIONS: NMP-22 showed an acceptable sensitivity but a low positive predictive value, therefore control cystoscopic evaluation cannot be avoided. The sensitivity and negative predictive values increased when NMP-22 and cytology are used in combination. We found NMP-22 to be a very useful marker for recurrence of low grade tumors.

Initial studies with high resolution TEM and electron energy loss spectroscopy studies of ferritin cores extracted from brains of patients with progressive supranuclear palsy and Alzheimer disease.

Studies of crystallographic structure and composition of core nanocrystals of ferritin bound to aberrant tau filaments extracted from progressive supranuclear Palsy (PSP) and Alzheimer disease (AD) brain tissues were performed using high resolution transmission electron microscopy (HRTEM) and electron energy loss spectroscopy (EELS). The results were compared with those obtained from synthetic Fe3O4 crystal (magnetite) and horse spleen ferritin cores. Core dimensions of ferritin molecules from PSP and AD were similar to those found in normal brain. Ferritin cores nanocrystals in AD seems to have less ordered structure than in PSP. Some nanocrystals did not have the hexagonal ferrihydrite structure generally found in healthy ferritin but rather a cubic structure similar to magnetite, a crystalline form in which both Fe2+ and Fe3+ are present. The presence of ferrous ion, Fe2+, may indicate some dysfunction in these pathological ferritins that might contribute to production of free radicals via the Fenton reaction involved in neurodegeneration.