Long-term outcomes of spinal SBRT. Is it important to select the treatment time?

PURPOSE: SBRT (stereotactic body radiation therapy) is widely used as a curative treatment in tumoral lesions and has become a fundamental tool for the treatment of spine metastasis. In this study, we present survival and toxicity outcomes of spine SBRT after a 2-year follow-up. METHODS/PATIENTS: Data from spine SBRT treatments performed at our institution between March 2012 and February 2020 was collected. Medical records, including demographic, primary tumor, and treatment characteristics were reviewed. Patient follow-up included clinical evaluation, imaging, and blood tests. Toxicity was recorded according to CTCAE v4.0. RESULTS: We analyzed 73 consecutive spine SBRT treatments in 60 patients. 39.7% of the cases had primary breast cancer and 23.3% had prostate cancer. Most cases (87.7%) were treated with a single SBRT fraction of 16 Gy. Median follow-up was 26.1 months (range 1.7-78.6), and 1- and 2-year overall survival (OS) rates were 96.9% and 84.2%, respectively. Local control (LC) rates at 1- and 2-years were 76.3% and 70.6%, respectively. Multivariate analysis identified histology as a prognostic factor for both OS and LC. Patients who underwent spine SBRT 6 months after the spinal lesion diagnosis had LC at 2 years of 88%, vs 61.7% for those who underwent SBRT before this period. No grade III or higher toxicity was reported. The vertebral compression fracture (VCF) rate was 4.1%. CONCLUSION: Spine SBRT at our institution showed a 2-year LC of 70.6%, without G3 toxicities. Delaying SBRT at least 6 months to administer systemic treatment was related to an improvement in local control.

High-resolution HLA allele and haplotype frequencies in majority and minority populations of Costa Rica and Nicaragua: Differential admixture proportions in neighboring countries.

The HLA system shows the most extensive polymorphism in the human genome. Allelic and haplotypic frequencies of HLA genes vary dramatically across human populations. Due to a complex history of migration, populations in Latin America show a broad variety of admixture proportions, usually varying not only between countries, but also within countries. Knowledge of HLA allele and haplotype frequencies is essential for medical fields such as transplantation, but also serves as a means to assess genetic diversity and ancestry in human populations. Here, we have determined high-resolution HLA-A, -B, -C, and -DRB1 allele and haplotype frequencies in a sample of 713 healthy subjects from three Mestizo populations, one population of African descent, and Amerindians of five different groups from Costa Rica and Nicaragua and compared their profiles to a large set of indigenous populations from Iberia, Sub-Saharan Africa, and the Americas. Our results show a great degree of allelic and haplotypic diversity within and across these populations, with most extended haplotypes being private. Mestizo populations show alleles and haplotypes of putative European, Amerindian, and Sub-Saharan African origin, albeit with differential proportions. Despite some degree of gene flow, Amerindians and Afro-descendants show great similarity to other Amerindian and West African populations, respectively. This is the first comprehensive study reporting high-resolution HLA diversity in Central America, and its results will shed light into the genetic history of this region while also supporting the development of medical programs for organ and stem cell transplantation.

A high-resolution frequency variable experimental setup for studying ferrofluids used in magnetic hyperthermia.

A scanning system for specific absorption rate of ferrofluids with superparamagnetic nanoparticles is presented in this study. The system contains an induction heating device designed and built with a resonant inverter in order to generate magnetic field amplitudes up to 38 mT, over the frequency band 180-525 kHz. Its resonant circuit involves a variable capacitor with 1 nF of capacitance steps to easily select the desired frequency, reaching from 0.3 kHz/nF up to 5 kHz/nF of resolution. The device performance is characterized in order to compare with the theoretical predictions of frequency and amplitude, showing a good agreement with the resonant inverters theory. Additionally, the setup is tested using a synthetic iron oxide with 10 ± 1 nm diameter suspended in liquid glycerol, with concentrations at 1%. Meanwhile, the temperature rise is measured to determine the specific absorption rate and calculate the dissipated power density for each f. This device is a suitable alternative to studying ferrofluids and analyzes the dependence of the power absorption density with the magnetic field intensity and frequency.

Evaluation of the genotoxic activity of dicamba and atrazine herbicides in several Mexican and South American varieties of sweetcorn (Zea mays L.).

Corn is a major crop and various herbicides are used to maximize its production, which include a dicamba-atrazine mixture. This has great advantages, but can also induce DNA damage. Genotoxic activity was assessed by comet assay following application of two concentrations of dicamba-atrazine: 1000-2000 and 2000-4000 ppm. Apical meristem leaf nuclei from 119 varieties of sweetcorn plants from Mexico and South America, and from five commercial sweetcorn hybrids were used. Each accession comprised two individuals per concentration and two controls. Significant genotoxic activity (P < 0.001) was observed following treatment with 1000-2000 and 2000-4000 ppm compared to the negative control. There was no difference in the genotoxic activity induced by both 1000-2000 and 2000-4000 ppm concentrations in plants from Mexico and South America (P > 0.05) except (P < 0.05) in the 2000-4000 ppm treated plants from Mexico and the 1000-2000 ppm treated plants from South America. Sweetcorn hybrids showed significant genetic damage (P < 0.01) at all concentrations compared to the negative controls. Thus, the dicamba-atrazine mixture caused genetic damage to corn plants, and it suggested that Mexican sweetcorn is more sensitive to dicamba-atrazine than the maize varieties from South America. Neither hybrid status nor the origin avoids DNA damage caused by Marvel. Thus, maize can be useful as a biomonitor of genetic damage induced by chemicals and to identify possible phenotypes based upon the amount of genetic damage induced by herbicides and selection of resistant genotypes.

Synthesis of nitrogen-doped ZnO by sol-gel method: characterization and its application on visible photocatalytic degradation of 2,4-D and picloram herbicides.

In this work, nitrogen-doped ZnO material was synthesized by the sol-gel method using zinc acetate as the precursor and urea as the nitrogen source (15, 20, 25 and 30% wt.). For comparative purposes, bare ZnO was also prepared. The influence of N doping on structural, morphological, optical and photocatalytic properties was investigated. The synthesized catalysts were characterized by XRD, SEM-EDS, diffuse reflectance UV-Vis spectroscopy, BET and XPS analysis. The photocatalytic activity of N-doped ZnO catalysts was evaluated during the degradation of a mixture of herbicides (2,4-D and picloram) under visible radiation ≥400 nm. The photo-absorption wavelength range of the N-doped ZnO samples was shifted to longer wavelength compared to those of the unmodified ZnO. Among different amounts of dopant agent, the 30% N-doped ZnO material showed higher visible-light activity compared with pure ZnO. Several degradation by-products were identified by using HPLC and ESI-MS/MS. The enhancement of visible photocatalytic activity of the N-doped ZnO semiconductor could be mainly due to their capability in reducing the electron-hole pair recombination.

A novel HLA-A allele, A*74:23, identified in an individual from Costa Rica.

The new HLA-A*74:23 allele differs from the closest allele A*74:01 by a nucleotide change in exon 3 at codon 97.

Removal of fluoride ions from drinking water and fluoride solutions by aluminum modified iron oxides in a column system.

The purpose of this work was to evaluate the potential of aluminum modified iron oxides, in a continuous flow for removal of fluoride ions from aqueous solutions and drinking water. The breakthrough curves obtained for fluoride ions adsorption from aqueous solutions and drinking water were fitted to Thomas, Bohart-Adams, and bed depth service time model (BDST). Adsorption capacities at the breakthroughs, Thomas model constant, kinetic constant and the saturation concentration were determined. The results show that in general, the adsorption efficiency decreases as the bed depth increases, and this behavior shows that the adsorption is controlled by the mass transport resistance. The adsorption capacity for fluoride ions by CP-Al is higher for fluoride aqueous solutions than drinking water.

Polymorphisms in HIF-1alpha affect presence of lymph node metastasis and can influence tumor size in squamous-cell carcinoma of the glottic larynx.

BACKGROUND: HIF-1alpha plays a key role in the development and progression of cancer. Its polymorphic variants C1772T and G1790A have been associated with greater susceptibility to cancer and increased tumor progression. METHODS: We determined the distribution of these polymorphisms among 121 patients with glottic cancer and 154 healthy volunteers by PCR-RFLP. We also analyzed the relationship between the presence of these polymorphisms and various clinicopathologic variables. RESULTS: Advanced tumors (T3-T4) were associated with the TT variant (p = 0.036), which was present in 75 % of T4 tumors (p = 0.008). Among patients with nodal metastasis (N+), 41.7 and 22 % were carrying the TT and GA variants, respectively, compared with 9.4 and 2 % of the patients with no metastasis (N0), (p = 0.006 and p = 0.032). CONCLUSIONS: The presence of the TT and GA variants were associated with lymph node metastasis, while the presence of the TT variant can be associated with larger tumor size.

Purification and characterization of a proteinase inhibitor from white croaker skeletal muscle (Micropogon opercularis).

A trypsin proteinase inhibitor has been purified to homogeneity from the skeletal muscle of white croaker (Micropogon opercularis). Previously, we had described the occurrence in fish muscle of a serine protease (proteinase I) which showed a great capacity to degrade whole myofibrils in vitro and an endogenous inhibitor that prevented the action of the protease, both on natural and artificial substrates. In this paper, we report the purification and further biochemical characterization of the endogenous trypsin inhibitor. The purification was carried out by DEAE-Sephacel, Con A-Sepharose, Sephacryl S-300 and Mono Q. Throughout the purification procedure, trypsin inhibitory activity was assayed using azocasein as substrate. The molecular mass of the inhibitor was 65 kDa, as estimated by SDS-PAGE and gel filtration. The trypsin inhibitor is a glycoprotein, as deduced by the fact that it binds to Con A-Sepharose and stains with PAS and showed a wide range of pH stability (from 5 to 11). The thermal stability of the inhibitor considerably decreased at temperatures >60 degrees C. Assays of the inhibitor against various proteases indicated that it is highly specific for serine proteases, since it did not inhibit proteases belonging to any other groups. The inhibitor was able to inhibit the endogenous target enzyme (proteinase I) in a dose-dependent manner, with a 50% inhibition at a molar ratio close to 1. The present work contributes to improving our understanding of the physiological role of the proteinase I-inhibitor system in muscle protein breakdown, as well as its influence on post mortem proteolysis.

Autoproteolytic activation of the haloalkaliphilic archaeon Natronococcus occultus extracellular serine protease.

The haloalkaliphilic archaeon Natronococcus occultus produces an extracellular serine protease in the stationary growth phase and upon starvation. Two proteins immunologically related to the extracellular protease were detected into the cells: P200 and P190. P200 was detected at early stages of growth and its relative amount decreased as the culture reached the stationary growth phase, concomitantly with the appearance of P190 and proteolytic activity, suggesting that P200 may be the precursor of the secreted protease and P190 the mature enzyme. Both proteins were also detected in the culture medium. Conversion of inactive P200 into active P190 was attained in cell-free culture medium from stationary phase but not from exponential phase. This process was prevented in the presence of PMSF and could be attained by addition of purified mature extracellular protease to P200. Altogether these results indicate that activation of Natronococcus occultus extracellular protease may be autoproteolytic and that factor/s present in stationary phase culture medium may be required for this process.

Purification and biochemical characterization of the haloalkaliphilic archaeon Natronococcus occultus extracellular serine protease.

A serine protease was purified from Natronococcus occultus stationary phase culture medium (328-fold, yield 19%) and characterized at the biochemical level. The enzyme has a native molecular mass of 130 kDa, has chymotrypsin-like activity, is stable and active in a broad pH range (5.5-12), is rather thermophilic (optimal activity at 60 degrees C in 1-2 M NaCl) and is dependent on high salt concentrations for activity and stability (1-2 M NaCl or KCl). Polyclonal antibodies were raised against the purified protease. In Western blots, they presented no cross-reactivity with culture medium from other halobacteria nor with commercial proteases except subtilisin. The amino acid sequences of three tryptic peptides obtained from Natronococcus occultus protease did not show significant similarity to other known proteolytic enzymes. This fact, in addition to its high molecular mass suggests that Natronococcus occultus extracellular protease may be a novel enzyme.

Extracellular protease of Natrialba magadii: purification and biochemical characterization.

A serine protease secreted by the haloalkaliphilic archaeon Natrialba magadii at the end of the exponential growth phase was isolated. This enzyme was purified 83 fold with a total yield of 25% by ethanol precipitation, affinity chromatography, and gel filtration. The native molecular mass of the enzyme determined by gel filtration was 45 kDa. Na. magadii extracellular protease was dependent on high salt concentrations for activity and stability, and it had an optimum temperature of 60 degrees C in the presence of 1.5M NaCl. The enzyme was stable and had a broad pH profile (6-12) with an optimum pH of 8-10 for azocasein hydrolysis. The protease was strongly inhibited by diisopropyl fluorophosphate (DFP), phenylmethyl sulfonylfluoride (PMSF), and chymostatin, indicating that it is a serine protease. It was sensitive to denaturing agents such as SDS, urea, and guanidine HCl and activated by thiol-containing reducing agents such as dithiotreitol (DTT) and 2-mercaptoethanol. This protease degraded casein and gelatin and showed substrate specificity for synthetic peptides containing Phe, Tyr, and Leu at the carboxyl terminus, showing that it has chymotrypsin-like activity. Na. magadii protease presented no cross-reactivity with polyclonal antibodies raised against the extracellular protease of Natronococcus occultus, suggesting that although these proteases share several biochemical traits, they might be antigenically unrelated.

Identification of a myofibril-bound serine protease and its endogenous inhibitor in mouse skeletal muscle.

Myofibrillar proteins, like all other intracellular proteins, are in a dynamic state of continual degradation and resynthesis. The proteolytic system responsible for degrading myofibrillar proteins in skeletal muscle is not well defined. A proteolytic activity associated to myofibrils was found in mouse skeletal muscle, as show electrophoretic patterns, and denominated by us, as protease M. During incubation of whole myofibrils at 37 degrees C, myosin heavy chain, alpha actinin, actin and troponin T suffered degradation. These effects were inhibited selectively by serine protease inhibitors (soybean trypsin inhibitor, di-isopropyl phosphofluoridate, phenylmethanesulfonyl fluoride). Using myofibrils as protease M source, azocaseinolytic activity was also detected. Endogenous inhibitor and various compounds effects on protease M activity were also quantified by trichloroacetic acid soluble products formation, using radiolabeled myofibrils. An endogenous trypsin inhibitor isolated from the muscle cytoplasmic fraction could inhibit protease M activity on myofibrillar proteins and on azocasein. While K(+) increased protease M activity, the presence of Ca(2+) did not show any effect. Data presented in this study suggest that reported protease M may be implicated in myofibrillar degradation in vivo and isolated endogenous inhibitor may provide a mechanism to control its action in mouse skeletal muscle.

Herida cardíaca oculta / Occult cardiac wounds

El traumatismo cardiaco penetrante, desde el punto de vista de diagnostico y tratamiento, continua siendo tema de discusion. El paciente que se presenta con clinica de exanguinacion o taponamiento cardiaco constitute el caso clasico, mas no asi aquel con estabilidad hemodinámica sin taponamiento. Día a día se intenta adoptar un metodo diagnostico altamente sensible, especifico y poco invasor para este tipo de lesion. El presente articulo describe tales aspectos y compara sucintamente la ventana pericardica subxifoidea con la ecocardiografia transtoracica, lo cual constituye la base de investigacion en un estudio realizado en nuestra Universidad proximo a publicarse.

Detection and preliminary characterization of extracellular proteolytic activities of the haloalkaliphilic archaeon Natronococcus occultus.

Extracellular proteolytic activity was detected in the haloalkaliphilic archaeon Natronococcus occultus as the culture reached the stationary growth phase. Proteolytic activity was precipitated with ethanol and subjected to a preliminary characterization. Optimal conditions for activity were attained at 60 degrees C and 1-2 M NaCl or KCl. Gelatin zymography in the presence of 4 M betaine revealed a complex pattern of active species with apparent molecular masses ranging from 50 to 120 kDa. Experiments performed with inhibitors of the various groups of proteases indicated that the extracellular proteolytic enzymes of N. occultus are of the serine type. Individual protein species showed some differences in salt and thermal stability.

Multicatalytic proteinase in fish muscle.

A partially active and a latent form of multicatalytic protease (MCP) were isolated from fish skeletal muscle. Both forms were inactive against protein substrates, but their activity against peptide substrates differed in one order of magnitude. The chymotrypsin-like activity of the partially active form was moderately stimulated by fatty acids and SDS, whereas its trypsin-like activity was inhibited by the same reagents. In contrast, both activities of the latent form were strongly stimulated by SDS. The chymotrypsin-like activity of the latent form was also stimulated by heating or high urea concentrations, whereas its trypsin-like activity did not change or was inhibited respectively by these treatments. These activation effects were irreversible. Pre-treatment of the latent form with SDS or urea in the absence of substrate led to its irreversible inactivation, whereas activation by pre-heating occurred in the presence or absence of substrate. These results suggest that MCP can exist in several active states with distinct properties. Studies on the distribution of MCP in fish tissues showed a much higher level of the enzyme in gonads than in any other tissue, suggesting a role of MCP in development.

Uretrosuspensión ambulatoria con la técnica de Gittes: resultados a largo plazo / Ambulatory urethrosuspension using the Gittes technique: long-term results

En 1987 Gittes y Loughlin dieron a conocer una técnica de uretrosuspensión ambulatoria sin incisión de la mucosa vaginal con un porcentaje de éxito de 87 por ciento. En la unidad de urología del Hospital General de México, SS, se reprodujo el procedimiento en 112 pacientes con incontinencia urinaria de esfuerzo. A 87 de estos pudo seguírseles durante un tiempo promedio de 16 meses, encontrándose un porcentaje de éxito global de 52 por ciento. Los resultados aquí comunicados no reflejan las cifras originalmente informadas por sus autores, situación que se atribuye a que la mucosa vaginal no constituye un tejido de sostén adecuado para el anclaje del material de sutura y, por tanto, pierde su fuerza tensil antes de la formación de fibrosis que conserve el cuello vesical en una posición adecuada

Purification and characterization of a latent form of multicatalytic proteinase from fish muscle.

1. A latent form of multicatalytic proteinase (MCP) was purified to apparent homogeneity from white croaker muscle by DEAE-Sephacel, Mono-Q, Sephacryl S-300 and second Mono-Q chromatographies. 2. The enzyme preparation was electrophoretically and immunologically similar to MCP purified from the same source by a different method (Folco et al., 1988b, Archs Biochem. Biophys. 267, 599-605) but showed much lower chymotrypsin- and trypsin-like activities. 3. These activities responded to sodium dodecyl sulphate (SDS), urea and heat treatments in different ways: SDS stimulated both activities, urea stimulated the former and inhibited the latter and heating stimulated the former and did not affect the latter. 4. The stimulation of chymotrypsin-like activity by the three treatments was irreversible. 5. Exposure of MCP to SDS or urea in the absence of substrate rapidly inactivated it, whereas heat activation took place irrespective of the presence of substrate. 6. The stimulating effect of SDS on chymotrypsin-like activity was lost in the presence of urea. 7. These results suggest that the enzyme may be activated by different mechanisms.

Detection of a trypsin-like serine protease and its endogenous inhibitor in hake skeletal muscle.

When dialyzed extracts from hake (Merluccius hubbsi) skeletal muscle were chromatographed in DEAE-Sephacel, an alkaline protease (37 degrees C, pH 8.5) and a trypsin inhibitor were isolated. The enzyme showed its maximal activity against azocasein in the range of pH between 7 and 9. The protease was able to hydrolyze the trypsin substrates Bz-Arg-OEt and Tos-Arg-OMe and did not cleave the chymotrypsin substrate Bz-Tyr-OEt. The enzyme was strongly inhibited by several serine protease inhibitors, whereas inhibitors of the other types of proteases scarcely affected it. The protease was able to degrade the major contractile and cytoskeletal constituent proteins of myofibrils and to accumulate acid-soluble products. The protease activity was completely suppressed by the addition of the trypsin inhibitor isolated from the same muscle. These results indicate that hake skeletal muscle contains a trypsin-like serine protease which might be involved in the catabolism of myofibrillar proteins, as well as in the proteolytic events that take place during post mortem storage of fish muscle.

Fish skeletal muscle contains a novel serine proteinase with an unusual subunit composition.

Proteinase I, an enzyme previously shown to be able to degrade contractile and cytoskeletal elements of white-croaker (Micropogon opercularis) myofibrils, was purified to apparent homogeneity by chromatography on DEAE-Sephacel, octyl-Sepharose CL 4B and arginine-Sepharose 4B. Its Mr was determined to be 269,000 by Sephacryl S-300 gel filtration. Under denaturing conditions, the enzyme dissociated into two subunits with Mr 20,000 and 15,500, in a molar ratio of 1.8:1. Proteinase I showed a pH optimum of 8.5. The enzyme was strongly inhibited by several serine proteinase inhibitors, whereas inhibitors of the other types of proteinases did not affect, or only scarcely affected, its activity. Several N-terminal-blocked 4-methyl-7-coumarylamide substrates having either arginine or lysine residues adjacent to the fluorogenic group were efficiently hydrolysed by the enzyme. These results indicate that proteinase I is a trypsin-like serine proteinase. The enzyme appears to be distinct from other proteinases previously described in skeletal muscle, and might be involved in the catabolism of myofibrillar proteins.