Orthopedic implant used as drug delivery system: clinical situation and state of the research.

A partial review is proposed on the existing literature for the research performed in orthopedic implant used as drug delivery system. In the first part, an evaluation is given on the clinical need to deliver a drug in the surrounding of an implant. Secondly, a review of the clinical situation is developed for implants already used as drug delivery system. Experimental works performed for local delivery are reported. In particular, a description is given on the in vitro and in vivo studies where the implant is coated with different proteins or drugs. Finally, a conclusion is proposed on the next step in the development of orthopedic implant as drug delivery system mentioning also the industrial situation.

One billion years of bZIP transcription factor evolution: conservation and change in dimerization and DNA-binding site specificity.

The genomic era has revealed that the large repertoire of observed animal phenotypes is dependent on changes in the expression patterns of a finite number of genes, which are mediated by a plethora of transcription factors (TFs) with distinct specificities. The dimerization of TFs can also increase the complexity of a genetic regulatory network manifold, by combining a small number of monomers into dimers with distinct functions. Therefore, studying the evolution of these dimerizing TFs is vital for understanding how complexity increased during animal evolution. We focus on the second largest family of dimerizing TFs, the basic-region leucine zipper (bZIP), and infer when it expanded and how bZIP DNA-binding and dimerization functions evolved during the major phases of animal evolution. Specifically, we classify the metazoan bZIPs into 19 families and confirm the ancient nature of at least 13 of these families, predating the split of the cnidaria. We observe fixation of a core dimerization network in the last common ancestor of protostomes-deuterostomes. This was followed by an expansion of the number of proteins in the network, but no major dimerization changes in interaction partners, during the emergence of vertebrates. In conclusion, the bZIPs are an excellent model with which to understand how DNA binding and protein interactions of TFs evolved during animal evolution.

Seeds with high molybdenum concentration improved growth and nitrogen acquisition of rhizobium-inoculated and nitrogen-fertilized common bean plants

Seeds of common bean (Phaseolus vulgaris) with high molybdenum (Mo) concentration can supply Mo plant demands, but to date no studies have concomitantly evaluated the effects of Mo-enriched seeds on plants inoculated with rhizobia or treated with N fertilizer. This work evaluated the effects of seed Mo on growth and N acquisition of bean plants fertilized either by symbiotic N or mineral N, by measuring the activities of nitrogenase and nitrate reductase and the contribution of biological N2 fixation at different growth stages. Seeds enriched or not with Mo were sown with two N sources (inoculated with rhizobia or fertilized with N), in pots with 10 kg of soil. In experiment 1, an additional treatment consisted of Mo-enriched seeds with Mo applied to the soil. In experiment 2, the contribution of N2 fixation was estimated by 15N isotope dilution. Common bean plants grown from seeds with high Mo concentration flowered one day earlier. Seeds with high Mo concentration increased the leaf area, shoot mass and N accumulation, with both N sources. The absence of effects of Mo application to the soil indicated that Mo contents of Mo-enriched seeds were sufficient for plant growth. Seeds enriched with Mo increased nitrogenase activity at the vegetative stage of inoculated plants, and nitrate reductase activity at late growth stages with both N sources. The contribution of N2 fixation was 17 and 61 % in plants originating from low- or high-Mo seeds, respectively. The results demonstrate the benefits of sowing Mo-enriched seeds on growth and N nutrition of bean plants inoculated with rhizobia or fertilized with mineral N fertilizer.

Symbiotic capability of calopo rhizobia from an agrisoil with different crops in Pernambuco

Biological nitrogen fixation by rhizobium-legume symbiosis represents one of the most important nitrogen sources for plants and depends strongly on the symbiotic efficiency of the rhizobium strain. This study evaluated the symbiotic capacity of rhizobial isolates from calopo (CALOPOGONIUM MUCUNOIDES) taken from an agrisoil under BRACHIARIA DECUMBENS pasture, sabiá (MIMOSA CAESALPINIIFOLIA) plantations and Atlantic Forest areas of the Dry Forest Zone of Pernambuco. A total of 1,575 isolates were obtained from 398 groups. A single random isolate of each group was authenticated, in randomized blocks with two replications. Each plant was inoculated with 1 mL of a bacterial broth, containing an estimated population of 10(8) rhizobial cells mL-1. Forty-five days after inoculation, the plants were harvested, separated into shoots, roots and nodules, oven-dried to constant mass, and weighed. Next, the symbiotic capability was tested with 1.5 kg of an autoclaved sand:vermiculite (1:1) mixture in polyethylene bags. The treatments consisted of 122 authenticated isolates, selected based on the shoot dry matter, five uninoculated controls (treated with 0, 50, 100, 150, or 200 kg ha-1 N) and a control inoculated with SEMIA 6152 (=BR1602), a strain of BRADYRHIZOBIUM JAPONICUM The test was performed as described above. The shoot dry matter of the plants inoculated with the most effective isolates did not differ from that of plants treated with 150 kg ha-1 N. Shoot dry matter was positively correlated with all other variables. The proportion of effective isolates was highest among isolates from SABIÁ forests. There was great variation in nodule dry weight, as well as in N contents and total N.

Nuclear and cytoplasmic triiodothyronine-binding sites in primary sensory neurons and Schwann cells: radioautographic study during development.

The effects of the thyroid hormones on target cells are mediated through nuclear T3 receptors. In the peripheral nervous system, nuclear T3 receptors were previously detected with the monoclonal antibody 2B3 mAb in all the primary sensory neurons throughout neuronal life and in peripheral glia at the perinatal period only (Eur. J. Neurosci. 5, 319, 1993). To determine whether these nuclear T3 receptors correspond to functional ones able to bind T3, cryostat sections and in vitro cell cultures of dorsal root ganglion (DRG) or sciatic nerve were incubated with 0.1 nM [125I]-labeled T3, either alone to visualize the total T3-binding sites or added with a 10(3) fold excess of unlabeled T3 to estimate the part due to the non-specific T3-binding. After glutaraldehyde fixation, radioautography showed that the specific T3-binding sites were largely prevalent. The T3-binding capacity of peripheral glia in DRG and sciatic nerve was restricted to the perinatal period in vivo and to Schwann cells cultured in vitro. In all the primary sensory neurons, specific T3-binding sites were disclosed in foetal as well as adult rats. The detection of the T3-binding sites in the nucleus indicated that the nuclear T3 receptors are functional. Moreover the concomitant presence of both T3-binding sites and T3 receptors alpha isoforms in the perikaryon of DRG neurons infers that: 1) [125I]-labeled T3 can be retained on the T3-binding 'E' domain of nascent alpha 1 isoform molecules newly-synthesized on the perikaryal ribosomes; 2) the alpha isoforms translocated to the nucleus are modified by posttranslational changes and finally recognized by 2B3 mAb as nuclear T3 receptor. In conclusion, the radioautographic visualization of the T3-binding sites in peripheral neurons and glia confirms that the nuclear T3 receptors are functional and contributes to clarify the discordant intracellular localization provided by the immunocytochemical detection of nuclear T3 receptors and T3 receptor alpha isoforms.

Traitement mini-invasif sous contrôle scanner et fluoroscopique de la lyse isthmique vertébrale : une nouvelle technique.

Objectifs: Evaluer la faisabilité, les résultats préliminaires à court et long terme du vissage percutané de vissage trans -isthmique sous anesthésie locale et contrôle scannerdes lyses isthmiques de bas grades.Matériels et méthodes: Etude prospective monocentrique réalisée sur 10 patients ayant une lyse isthmique grade 1 et 2 résistant au traitement médical conventionnel. Une évaluationclinique était réalisée à un mois, 3 mois, 6 mois et un an post-opératoire par un évaluateur indépendant. L'indication est posée en concertation avec le service dechirurgie orthopédique.Résultats: Les lyses isthmiques étaient situées en L5-S1 avec 6 grades 1 et 4 grades 2. L'échelle analogique de la douleur (VAS) variait de 6 a 9 avec une moyenne de 7,8.L'indication opératoire chirurgicale était posée pour tous les patients par arthrodèse postérieure lombo -sacree. Pour chaque patient 2 vis étaient positionnées soitun total de 20 vis. Un suivi clinique était réalisé de 28 a 36 mois. L'EVA et ODI diminuaient de 7,8 +/- 1,7 à 1,9 +/- 1,2 et de 62,3 +/- 17,2 à 15,1 +/- 6,0respectivement. L'ensemble des résultats était stable dans le temps en particulier à long terme.Conclusion: La fixation précise de la lyse isthmique améliore la symptomatologie et probablement évite un glissement vertébral plus important , un suivi à plus long terme surune serie de patients plus importante devrait confirmer cette hypothèse.

Inoculation of sugarcane with diazotrophic bacteria

The sugarcane industry, a strategic crop in Brazil, requires technological improvements in production efficiency to increase the crop energy balance. Among the various currently studied alternatives, inoculation with diazotrophic bacteria proved to be a technology with great potential. In this context, the efficiency of a mixture of bacterial inoculant was evaluated with regard to the agronomic performance and N nutrition of sugarcane. The experiment was carried out on an experimental field of Embrapa Agrobiologia, in Seropédica, Rio de Janeiro, using a randomized block, 2 × 3 factorial design (two varieties and three treatments) with four replications, totaling 24 plots. The varieties RB867515 and RB72454 were tested in treatments consisting of: inoculation with diazotrophic bacteria, N-fertilized control with 120 kg ha-1 N and absolute control (no inoculation and no N fertilizer). The inoculum was composed of five strains of five diazotrophic species. The yield, dry matter accumulation, total N in the shoot dry matter and the contribution of N by biological fixation were evaluated, using the natural 15N abundance in non-inoculated sugarcane as reference. The bacterial inoculant increased the stalk yield of variety RB72454 similarly to fertilization with 120 kg ha-1 N in the harvests of plant-cane and first ratoon crops, however the contribution of biological N fixation was unchanged by inoculation, indicating that the benefits of the inoculant in sugarcane may have resulted from plant growth promotion.

Soil phosphorus availability and soybean response to phosphorus starter fertilizer

Phosphorus fixation in tropical soils may decrease under no-till. In this case, P fertilizer could be surface-spread, which would improve farm operations by decreasing the time spend in reloading the planter with fertilizers. In the long term, less soluble P sources could be viable. In this experiment, the effect of surface-broadcast P fertilization with both soluble and reactive phosphates on soil P forms and availability to soybean was studied with or without fertilization with soluble P in the planting furrow in a long-term experiment in which soybean was grown in rotation with Ruzigrass (Brachiaria ruziziensis). No P or 80 kg ha-1 of P2O5 in the form of triple superphosphate or Arad reactive rock phosphate was applied on the surface of a soil with variable P fertilization history. Soil samples were taken to a depth of 60 cm and soil P was fractionated. Soybean was grown with 0, 30, and 60 kg ha-1 of P2O5 in the form of triple phosphate applied in the seed furrow. Both fertilizers applied increased available P in the uppermost soil layers and the moderately labile organic and inorganic forms of P in the soil profile, probably as result of root decay. Soybean responded to phosphates applied on the soil surface or in the seed furrow; however, application of soluble P in the seed furrow should not be discarded. In tropical soils with a history of P fertilization, soluble P sources may be substituted for natural reactive phosphates broadcast on the surface. The planting operation may be facilitated through reduction in the rate of P applied in the planting furrow in relation to the rates currently applied.

SYMBIOTIC EFFECTIVENESS AND COMPETITIVENESS OF CALOPO RHIZOBIAL ISOLATES IN AN ARGISSOLO VERMELHO-AMARELO UNDER THREE VEGETATION COVERS IN THE DRY FOREST ZONE OF PERNAMBUCO

Biological N fixation in forage legumes is an important alternative to reduce pasture degradation, and is strongly influenced by the inoculant symbiotic capability. This paper evaluates the effectiveness of Calopo (Calopogonium mucunoides) rhizobial isolated from soil under three vegetation covers of an Argissolo Vermelho-Amarelo of the Dry Forest Zone of Pernambuco. An experiment was conducted evaluating 25 isolates, aside from 5 uninoculated controls with 0; 309; 60; 90 and 120 kg ha-1 N, and a treatment inoculated with the SEMIA 6152 strain. The first cut was performed 45 days after inoculation and a second and third cut after 45-day-intervals. Shoot N content was quantified at all cuts. Shoot dry mass was affected by N rates at all cuts. Shoot dry mass increased from the first to the second cut in inoculated plants. There was no difference between rhizobial isolates from the different plant covers for any of the variables. Most variables were significantly and positively correlated.

THE RESURRECTION PLANT TRIPOGON SPICATUS (POACEAE) HARBORS A DIVERSITY OF PLANT GROWTH PROMOTING BACTERIA IN NORTHEASTERN BRAZILIAN CAATINGA

Plant species that naturally occur in the Brazilian Caatinga(xeric shrubland) adapt in several ways to these harsh conditions, and that can be exploited to increase crop production. Among the strategic adaptations to confront low water availability, desiccation tolerance stands out. Up to now, the association of those species with beneficial soil microorganisms is not well understood. The aim of this study was to characterize Tripogon spicatusdiazotrophic bacterial isolates from the Caatingabiome and evaluate their ability to promote plant growth in rice. Sixteen bacterial isolates were studied in regard to their taxonomic position by partial sequencing of the 16S rRNA gene, putative diazotrophic capacity, in vitro indole-acetic acid (IAA) production and calcium phosphate solubilization, metabolism of nine different C sources in semi-solid media, tolerance to different concentrations of NaCl to pHs and intrinsic resistance to nine antibiotics. Finally, the ability of the bacterial isolates to promote plant growth was evaluated using rice (Oryza sativa) as a model plant. Among the 16 isolates evaluated, eight of them were classified as Enterobacteriaceae members, related to Enterobacter andPantoeagenera. Six other bacteria were related toBacillus, and the remaining two were related toRhizobiumand Stenotrophomonas.The evaluation of total N incorporation into the semi-solid medium indicated that all the bacteria studied have putative diazotrophic capacity. Two bacteria were able to produce more IAA than that observed for the strain BR 11175Tof Herbaspirillum seropedicae.Bacterial isolates were also able to form a microaerophilic pellicle in a semi-solid medium supplemented with different NaCl concentrations up to 1.27 mol L-1. Intrinsic resistance to antibiotics and the metabolism of different C sources indicated a great variation in physiological profile. Seven isolates were able to promote rice growth, and two bacteria were more efficient than the reference strainAzospirillum brasilense, Ab-V5. The results indicate the potential of T. spicatus as native plant source of plant growth promoting bacteria.

ORGANIC CARBON CONTENTS AND STOCKS IN PARTICLE SIZE FRACTIONS OF A TYPIC HAPLUDOX FERTILIZED WITH PIG SLURRY AND SOLUBLE FERTILIZER

The use of pig slurry (PS) as fertilizer can affect the soil quality and increase total stocks of soil organic carbon (TOC). However, the effects of PS on TOC amount and forms in the soil are not fully understood, particularly in areas under no-tillage (NT). The purpose of this study was to determine TOC contents and stocks in the particulate (POC) and mineral-associated C fractions (MAC) of an Oxisol after nine years of maize-oat rotation under NT, with annual applications of PS, soluble fertilizer and combined fertilization (pig slurry + soluble fertilizer). The experiment was initiated in 2001 in Campos Novos, Santa Catarina, with the following treatments: PS at rates of 0 (without fertilization - PS0); 25 (PS25); 50 (PS50); 100 (PS100); and 200 m3 ha-1yr-1 (PS200); fertilization with soluble fertilizer (SF); and mixed fertilization (PS + SF). The TOC content was determined in samples of six soil layers to a depth of 40 cm, and the POC and MAC contents in four layers to a depth of 20 cm. From the rate of 50 m3 ha-1yr-1 and upwards, the soil TOC content and stock increased according to the PS rates in the layers to a depth of 10 cm. The POC and MAC contents and stocks were higher in the surface layers, with a clear predominance of the second fraction, but a greater relative amplitude in the contents of the first fraction.

Combining protein identification and quantification: C-terminal isotope-coded tagging using sulfanilic acid.

Two methods of differential isotopic coding of carboxylic groups have been developed to date. The first approach uses d0- or d3-methanol to convert carboxyl groups into the corresponding methyl esters. The second relies on the incorporation of two 18O atoms into the C-terminal carboxylic group during tryptic digestion of proteins in H(2)18O. However, both methods have limitations such as chromatographic separation of 1H and 2H derivatives or overlap of isotopic distributions of light and heavy forms due to small mass shifts. Here we present a new tagging approach based on the specific incorporation of sulfanilic acid into carboxylic groups. The reagent was synthesized in a heavy form (13C phenyl ring), showing no chromatographic shift and an optimal isotopic separation with a 6 Da mass shift. Moreover, sulfanilic acid allows for simplified fragmentation in matrix-assisted laser desorption/ionization (MALDI) due the charge fixation of the sulfonate group at the C-terminus of the peptide. The derivatization is simple, specific and minimizes the number of sample treatment steps that can strongly alter the sample composition. The quantification is reproducible within an order of magnitude and can be analyzed either by electrospray ionization (ESI) or MALDI. Finally, the method is able to specifically identify the C-terminal peptide of a protein by using GluC as the proteolytic enzyme.

The role of the radiotherapy technologist in gamma knife radiosurgery within a multidisciplinary team: the Lausanne experience

Objective: Integration of the radiotherapy technologist "know-how" in the Gamma Knife treatment processMaterials and Methods: Gamma Knife (GK) treatments started in July 2010 at the GK Center in C.H.U.V. with the Leksell Gamma KnifeR Perfexion?(Elekta AB, Sweden). The multidisciplinary GK team involves neurosurgeons, radio-oncologists, physicists, neuroradiologists, nurses and technologists, aiming at a full integration for optimal patient management.Results: Between July and December 2010, 60 patients have been treated. Required stereotactic imaging involves IRM, CT scan (and angiography for AVM). All the steps in the treatment process (Leksell coordinate frame fixation, imaging, planning, treatment) are supervised by the members of the multidisciplinary team. In our experience, radiotherapy technologist (RTT) have acquired an important role in the multidisciplinary team communication and integration. Specifically, the RTT are responsible of: supervision of the image acquisition, performing the Gamma Knife unit control tests, patient setup, and patient surveillance during treatment.Conclusion: RTT have a fundamental role in the communication within the team, between the team and the patient and also to assure the patient security. Our experience shows that it is possible and required to involve RTT in all steps of the GK treatment process, to guarantee the best GK treatment possible.

Partial cricotracheal resection for pediatric subglottic stenosis: a single institution's experience in 60 cases.

In our study, 60 infants and children, each with a severe subglottic stenosis (SGS), underwent partial cricotracheal resection (PCTR) with primary thyrotracheal anastomosis. According to the Myer-Cotton classification, two were grade II, 41 were grade III and 17 were grade IV stenoses. Of the 60 patients, 57 (95%) are presently decannulated, and one patient sustained a complete restenosis. Two patients with better than 80% subglottic airways still are waiting for decannulation: one because of bilateral cricoarytenoid joint fixation and the second because of temporary stenting of the subglottis with a Montgomery T-tube. The rate of decannulation is 97% (36 of 37 cases) in primary PCTRs, 100% (13 of 13 cases) in salvage PCTRs for failed laryngotracheal reconstructions (LTR) and 70% (7 of 10 cases) in extended PCTRs (i.e., PCTR associated with an additional open-airway procedure).