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Laser microsurgery is the current gold standard surgical technique for the treatment of selected diseases in delicate organs such as the larynx. However, the operations require large surgical expertise and dexterity, and face significant limitations imposed by available technology, such as the requirement for direct line of sight to the surgical field, restricted access, and direct manual control of the surgical instruments. To change this status quo, the European project µRALP pioneered research towards a complete redesign of current laser microsurgery systems, focusing on the development of robotic micro-technologies to enable endoscopic operations. This has fostered awareness and interest in this field, which presents a unique set of needs, requirements and constraints, leading to research and technological developments beyond µRALP and its research consortium. This paper reviews the achievements and key contributions of such research, providing an overview of the current state of the art in robot-assisted endoscopic laser microsurgery. The primary target application considered is phonomicrosurgery, which is a representative use case involving highly challenging microsurgical techniques for the treatment of glottic diseases. The paper starts by presenting the motivations and rationale for endoscopic laser microsurgery, which leads to the introduction of robotics as an enabling technology for improved surgical field accessibility, visualization and management. Then, research goals, achievements, and current state of different technologies that can build-up to an effective robotic system for endoscopic laser microsurgery are presented. This includes research in micro-robotic laser steering, flexible robotic endoscopes, augmented imaging, assistive surgeon-robot interfaces, and cognitive surgical systems. Innovations in each of these areas are shown to provide sizable progress towards more precise, safer and higher quality endoscopic laser microsurgeries. Yet, major impact is really expected from the full integration of such individual contributions into a complete clinical surgical robotic system, as illustrated in the end of this paper with a description of preliminary cadaver trials conducted with the integrated µRALP system. Overall, the contribution of this paper lays in outlining the current state of the art and open challenges in the area of robot-assisted endoscopic laser microsurgery, which has important clinical applications even beyond laryngology.
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The Antarctic Peninsula is one of the regions to be most affected by increase in sea surface temperatures (SSTs) mediated by Global Climate Change; indeed, most negative predictions imply an up to 6⯰C increment by the end of the XXI century. Temperature is one of the most important factors mediating diversity and distribution of macroalgae, although there is still no consensus as to the likely effects of higher SSTs, especially for polar seaweeds. Some available information suggests that potential strategies to withstand future increases in SSTs will be founded upon the glutathione-ascorbate cycle and the induction of chaperone-functioning heat shock proteins (HSPs); however, their eventual role, even for general stress responses, is unclear. The intertidal green, brown and red macroalgae species Monostroma hariotii, Adenocystis utricularis and Pyropia endiviifolia, respectively, from King George Island, Antarctic Peninsula, were exposed to 2⯰C (control) and 8⯰C (climate change scenario) for up to 5â¯days (d). Photosynthetic activity (αETR and ETRmax, and EkETR), photoinhibition (Fv/Fm) and photoprotection processes (αNPQ, NPQmax, and EkNPQ) provided no evidence of negative ecophysiological effects. There were moderate increases in H2O2 production and levels of lipid peroxidation with temperature, results supported by stable levels of total glutathione and ascorbate pools, with mostly higher levels of reduced ascorbate and glutathione than oxidized forms in all species. Transcripts of P. endiviifolia indicated a general upregulation of all antioxidant enzymes and HSPs genes studied under warmer temperature, although with different levels of activation with time. This pioneering investigation exploring different levels of biological organization, suggested that Antarctic intertidal macroalgae may be able to withstand future rise in SSTs, probably slightly altering their latitudinal distribution and/or range of thermal tolerance, by exhibiting robust glutathione-ascorbate production and recycling, as well as the induction of associated antioxidant enzymatic machinery and the syntheses of HSPs.
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Algas Marinas , Regiones Antárticas , Cambio Climático , Peróxido de Hidrógeno , Océanos y Mares , TemperaturaRESUMEN
There is scarce investigation addressing interpopulation tolerance responses to address the influence of a history of chronic stress exposure, as that occurring in polluted environments, in photoautotrophs. We evaluated ecophysiological (photosynthetic activity) and metabolic (oxidative stress and damage) responses of two populations of green macroalga Ulva compressa from polluted (Ventanas) and non-polluted (Cachagua) localions of central Chile, and exposed to controlled hypersalinity conditions of 32 (control), 42, 62 and 82 psu (practical salinity units) for 6 h, 48 h and 6 d. Both primary production (ETRmax) and photosynthetic efficiency (αETR) were generally higher in the population from Cachagua compared to Ventanas at all times and salinities. Moreover, at most experimental times and salinities the population from Ventanas had greater levels of H2O2 and lipid peroxidation that individuals from Cachagua. Total ascorbate was higher in the population of Cachagua than Ventanas at 42 and 82 psu after 6 and 48 h, respectively, while at 6 d concentrations were similar between both populations at all salinities. Total glutathione was greater in both populations after 6 h at all salinities, but at 48 h its concentrations were higher only in the population from Cachagua, a trend that was maintained at 6 d under 82 psu only. Reduced and oxidized ascorbate (ASC and DHA, respectively) and glutathione (GSH and GSSG, respectively) demonstrated similar patterns between U. compressa populations, with an increase oxidation with greater salinities but efficient recycling to maintain sufficient batch of ASC and GSH. When assessing the expression of antioxidant enzymes catalase (CAT), superoxide dismutase (SOD) and dehydroascorbate reductase (DHAR), while the population of Ventanas displayed a general trend of upregulation with increasing salinities along the experiments, U. compressa from Cachagua revealed patterns of downregulation. Results demonstrated that although both populations were still viable after the applied hypersalinities during all experimental times, biological performance was usually more affected in the population from the Ventanas than Cachagua, likely due to a depressed baseline metabolism after a long history of exposition to environmental pollution.
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Ulva/fisiología , Contaminantes Químicos del Agua/toxicidad , Antioxidantes/metabolismo , Ácido Ascórbico/metabolismo , Catalasa/metabolismo , Chile , Contaminación Ambiental , Glutatión/metabolismo , Peróxido de Hidrógeno/metabolismo , Peroxidación de Lípido , Oxidación-Reducción , Estrés Oxidativo , Salinidad , Algas Marinas/metabolismo , Superóxido Dismutasa/metabolismo , Ulva/enzimologíaRESUMEN
In this work, we statistically improved culture media for rPOXA 1B laccase production, expressed in Pichia pastoris containing pGAPZαA-LaccPost-Stop construct and assayed at 10 L bioreactor production scale (6 L effective work volume). The concentrated enzyme was evaluated for temperature and pH stability and kinetic parameter, characterized by monitoring oxidation of different ABTS [2, 20-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid)] substrate concentrations. Plackett-Burman experimental design (PBED) implementation improved previous work results by 3.05-fold, obtaining a laccase activity of 1373.72 ± 0.37 U L-1 at 168 h of culture in a 500 mL shake flask. In contrast, one factor experimental design (OFED) applied after PBED improved by threefold the previous study, additionally increasing the C/N ratio. Employing OFED media at 10 L bioreactor scale was capable of producing 3159.93 ± 498.90 U L-1 at 192 h, representing a 2.4-fold increase. rPOXA 1B concentrate remained stable between 10 and 50 °C and retained over 70% residual enzymatic activity at 60 °C and 50% at 70 °C. Concerning pH stability, the enzyme was stable at pH 4.0 ± 0.2 with a residual activity greater than 90%. The lowest residual activity (60%) was obtained at pH 10.0 ± 0.2. Furthermore, the apparent kinetic parameters were V max of 3.163 × 10-2 mM min-1 and K m of 1.716 mM. Collectively, regarding enzyme stability our data provide possibilities for applications involving a wide range of pH and temperatures.
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BACKGROUND: The use of lasers in transoral surgery enables precise tissue incision with minimal adverse effects on surrounding structures. Nonetheless, the lack of haptic feedback during laser cutting impairs the surgeon's perception of the incision depth, potentially leading to undesired tissue damage. METHODS: This paper presents a novel approach, based on statistical regression analysis, to estimate the laser incision depth in soft tissue. User trials were conducted in a laser surgery set-up, to verify the effectiveness of online estimation of incision depth in supporting precise tissue cutting. RESULTS: The estimation accuracy was verified on ex vivo muscle tissue, revealing a root mean squared error (RMSE) of 0.1 mm for depths ranging up to 1.4 mm. Online estimation of depth has the potential to significantly improve the incision control of users. CONCLUSIONS: The proposed approach was successful in producing estimations of laser cutting depth in ex vivo muscle tissue. Further investigation is required to validate this approach on other types of tissue. Providing depth estimation during laser cutting allows users to perform more precise incisions.
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Terapia por Láser , Microcirugia/métodos , Humanos , Análisis de RegresiónRESUMEN
This paper presents the development of a system capable of generating safety alarms when unexpected or unforeseen situations are detected during larynx phonomicrosurgery. Such system establishes relations between the application of laser power and changes in laryngeal tissue characteristics and appearance. As core component, we propose a model able to map inputs generated by the surgeon when controlling the laser to an estimation of tissue temperature. Situations where this supervision is relevant have been identified.