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1.
J Sports Sci ; 42(8): 655-664, 2024 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-38794799

RESUMEN

Climbing is a physically demanding discipline, placing significant loads on the finger flexors. Notwithstanding the documented greater endurance capacity of experienced climbers, the mechanisms explaining these training-induced adaptations remain unknown. We therefore investigate whether two non-competing strategies - muscle adaptation and alternate muscle recruitment - may explain the disparity in endurance capacity in participants with different climbing experience. We analysed high-density surface electromyograms (EMGs) from 38 Advanced and Intermediate climbers, during suspension exercises over three different depths (15, 20, 30 mm) using a half-crimp grip position. From the spatial distribution of changes in MeDian Frequency and Root Mean Square values until failure, we assessed how much and how diffusely the myoelectric manifestations of fatigue took place. Advanced climbers exhibited greater endurance, as evidenced by significantly longer failure time (p < 0.009) and lower changes in MDF values (p < 0.013) for the three grip depths. These changes were confined to a small skin region (nearly 25% of the grid size), centred at variable locations across participants. Moreover, lower MDF changes were significantly associated with longer suspension times. Collectively, our results suggest that muscle adaptation rather than load sharing between and within muscles is more likely to explain the improved endurance in experienced climbers.


Asunto(s)
Adaptación Fisiológica , Electromiografía , Dedos , Fuerza de la Mano , Montañismo , Fatiga Muscular , Músculo Esquelético , Resistencia Física , Humanos , Resistencia Física/fisiología , Fatiga Muscular/fisiología , Montañismo/fisiología , Músculo Esquelético/fisiología , Masculino , Adulto , Fuerza de la Mano/fisiología , Dedos/fisiología , Adulto Joven , Femenino
2.
J Environ Manage ; 358: 120758, 2024 May.
Artículo en Inglés | MEDLINE | ID: mdl-38593735

RESUMEN

European legislation stated that electric vehicles' sale must increase to 35% of circulating vehicles by 2030, and concern is associated to the batteries' supply chain. This review aims at analysing the impacts (about material flows and CO2 eq emissions) of Lithium-Ion Batteries' (LIBs) recycling at full-scale in Europe in 2030 on the European LIBs' supply-chain. Literature review provided the recycling technologies' (e.g., pyro- and hydrometallurgy) efficiencies, and an inventory of existing LIBs' production and recycling plants in Europe. European production plants exhibit production capacity adequate for the expected 2030 needs. The key critical issues associated to recycling regard pre-treatments and the high costs and environmental impacts of metallurgical processes. Then, according to different LIBs' composition and market shares in 2020, and assuming a 10-year battery lifetime, the Material Flow Analysis (MFA) of the metals embodied in End of Life (EoL) LIBs forecasted in Europe in 2030 was modelled, and the related CO2 eq emissions calculated. In 2030 the European LIBs' recycling structure is expected to receive 664 t of Al, 530 t of Co, 1308 t of Cu, 219 t of Fe, 175 t of Li, 287 t of Mn and 486 t of Ni. Of these, 99% Al, 86% Co, 96% Cu, 88% Mn and 98% Ni will be potentially recovered by pyrometallurgy, and 71% Al, 92% Co, 92% Fe, 96% Li, 88 % Mn and 90% Ni by hydrometallurgy. However, even if the recycling efficiencies of the technologies applied at full-scale are high, the treatment capacity of European recycling plants could supply as recycled metals only 2%-wt of the materials required for European LIBs' production in 2030 (specifically 278 t of Al, 468 t of Co, 531 t of Cu, 114 t of Fe, 95 t of Li, 250 t of Mn and 428 t of Ni). Nevertheless, including recycled metals in the production of new LIBs could cut up 28% of CO2 eq emissions, compared to the use of virgin raw materials, and support the European batteries' value chain.


Asunto(s)
Suministros de Energía Eléctrica , Litio , Reciclaje , Europa (Continente)
3.
J Environ Manage ; 309: 114677, 2022 May 01.
Artículo en Inglés | MEDLINE | ID: mdl-35151134

RESUMEN

This work presents a systematic characterisation of 100 waste mobile phones (73 feature phones and 23 smartphones) produced between 1989 and 2016. All items were inventoried and the evolvement of the relative abundances of their macro-components (mechanic and electro-mechanic parts, electronics and others) and materials was investigated. The average lifetime was 15.1 years for feature phones and 6.4 years for smartphones. The main component was plastic, on average 46%-wt. in feature phones and 37%-wt. in smartphones; over the years electronics' and plastic's amounts decreased (respectively 80% and 70%), while metal components' amount increased (12%). A cost-benefit analysis explored the profitability of the management of waste mobile phones through manual dismantling followed by the sale of the separated components and materials. The average cost of manual dismantling was estimated as 6.93 € per item according to EU average labour costs and 1.50 € per item based on minimum EU labour costs. According to the performed economic analysis, the actual market prices for the potentially recoverable materials and components of waste mobile phones were not able (particularly mixed plastics) to counterbalance the costs of manual dismantling according to the European standard labour costs.


Asunto(s)
Teléfono Celular , Residuos Electrónicos , Residuos Electrónicos/análisis , Metales , Plásticos , Reciclaje
4.
J Thromb Thrombolysis ; 51(1): 159-167, 2021 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-32424778

RESUMEN

It is not known whether the current territorial organization for acute revascularization treatments in ischemic stroke patients guarantees similar time to treatment and functional outcomes among different levels of institutional stroke care. We aimed to assess the impact of time to treatment on functional outcomes in ischemic stroke patients who received intravenous thrombolysis (IVT) alone, bridging (IVT plus thrombectomy), or primary thrombectomy in level 1 and level 2 Stroke Units (SUs) in Triveneto, a geographical macroarea in Northeast of Italy. We conducted an analysis of data prospectively collected from 512 consecutive ischemic stroke patients who received IVT and/or mechanical thrombectomy in 25 SUs from September 17th to December 9th 2018. The favorable outcome measures were mRS score 0-1 and 0-2 at 3 months. The unfavorable outcome measures were mRS score 3-5 and death at 3 months. We estimated separately the possible association of each variable for time to treatment (onset-to-door, door-to-needle, onset-to-needle, door-to-groin puncture, needle-to-groin puncture, and onset-to-groin puncture) with 3-month outcome measures by calculating the odds ratios (ORs) with two-sided 95% confidence intervals (CI) after adjustment for pre-defined variables and variables with a probability value ≤ 0.10 in the univariate analysis for each outcome measure. Distribution of acute revascularization treatments was different between level 1 and level 2 SUs (p < 0.001). Among 182 patients admitted to level 1 SUs (n = 16), treatments were IVT alone in 164 (90.1%), bridging in 12 (6.6%), and primary thrombectomy in 6 (3.3%) patients. Among 330 patients admitted to level 2 SUs (n = 9), treatments were IVT alone in 219 (66.4%), bridging in 74 (22.4%), and primary thrombectomy in 37 (11.2%) patients. Rates of excellent outcome (51.4% vs 45.9%), favorable outcome (60.1% vs 58.7%), unfavorable outcome (33.3% vs 33.8%), and death (9.8% vs 11.3%) at 3 months were similar between level 1 and 2 SUs. No significant association was found between time to IVT alone (onset-to-door, door-to-needle, and onset-to-needle) and functional outcomes. After adjustment, door-to-needle time ≤ 60 min (OR 4.005, 95% CI 1.232-13.016), shorter door-to-groin time (OR 0.991, 95% CI 0.983-0.999), shorter needle-to-groin time (OR 0.986, 95% CI 0.975-0.997), and shorter onset-to-groin time (OR 0.994, 95% CI 0.988-1.000) were associated with mRS 0-1. Shorter door-to-groin time (OR 0.991, 95% CI 0.984-0.998), door-to-groin time ≤ 90 min (OR 12.146, 95% CI 2.193-67.280), shorter needle-to-groin time (OR 0.983, 95% CI 0.972-0.995), and shorter onset-to-groin time (OR 0.993, 95% CI 0.987-0.999) were associated with mRS 0-2. Longer door-to-groin time (OR 1.007, 95% CI 1.001-1.014) and longer needle-to-groin time (OR 1.019, 95% CI 1.005-1.034) were associated with mRS 3-5, while door-to-groin time ≤ 90 min (OR 0.229, 95% CI 0.065-0.808) was inversely associated with mRS 3-5. Longer onset-to-needle time (OR 1.025, 95% CI 1.002-1.048) was associated with death. Times to treatment influenced the 3-month outcomes in patients treated with thrombectomy (bridging or primary). A revision of the current territorial organization for acute stroke treatments in Triveneto is needed to reduce transfer time and to increase the proportion of patients transferred from a level 1 SU to a level 2 SU to perform thrombectomy.


Asunto(s)
Accidente Cerebrovascular Isquémico/terapia , Trombectomía/métodos , Terapia Trombolítica/métodos , Anciano , Anciano de 80 o más Años , Femenino , Humanos , Accidente Cerebrovascular Isquémico/epidemiología , Italia/epidemiología , Masculino , Persona de Mediana Edad , Estudios Prospectivos , Resultado del Tratamiento
5.
Int J Mol Sci ; 21(19)2020 Oct 04.
Artículo en Inglés | MEDLINE | ID: mdl-33020398

RESUMEN

Malignant mesothelioma (MM) is a very aggressive asbestos-related cancer, for which no therapy proves to be effective. We have recently shown that the oncolytic adenovirus dl922-947 had antitumor effects in MM cell lines and murine xenografts. Previous studies demonstrated that dl922-947-induced host cell cycle checkpoint deregulation and consequent DNA lesions associated with the virus efficacy. However, the cellular DNA damage response (DDR) can counteract this virus action. Therefore, we assessed whether AZD1775, an inhibitor of the G2/M DNA damage checkpoint kinase WEE1, could enhance MM cell sensitivity to dl922-947. Through cell viability assays, we found that AZD1775 synergized with dl922-947 selectively in MM cell lines and increased dl922-947-induced cell death, which showed hallmarks of apoptosis (annexinV-positivity, caspase-dependency, BCL-XL decrease, chromatin condensation). Predictably, dl922-947 and/or AZD1775 activated the DDR, as indicated by increased levels of three main DDR players: phosphorylated histone H2AX (γ-H2AX), phospho-replication protein A (RPA)32, phospho-checkpoint kinase 1 (CHK1). Dl922-947 also increased inactive Tyr-15-phosphorylated cyclin-dependent kinase 1 (CDK1), a key WEE1 substrate, which is indicative of G2/M checkpoint activation. This increase in phospho-CDK1 was effectively suppressed by AZD1775, thus suggesting that this compound could, indeed, abrogate the dl922-947-induced DNA damage checkpoint in MM cells. Overall, our data suggest that the dl922-947-AZD1775 combination could be a feasible strategy against MM.


Asunto(s)
Proteínas de Ciclo Celular/antagonistas & inhibidores , Supervivencia Celular/efectos de los fármacos , Mesotelioma Maligno/tratamiento farmacológico , Proteínas Tirosina Quinasas/antagonistas & inhibidores , Pirazoles/farmacología , Pirimidinonas/farmacología , Adenoviridae/genética , Apoptosis/efectos de los fármacos , Amianto/toxicidad , Proteínas de Ciclo Celular/genética , Proteínas de Ciclo Celular/farmacología , Línea Celular Tumoral , Daño del ADN/efectos de los fármacos , Humanos , Mesotelioma Maligno/inducido químicamente , Mesotelioma Maligno/genética , Mesotelioma Maligno/virología , Viroterapia Oncolítica , Virus Oncolíticos/genética , Fosforilación/efectos de los fármacos , Inhibidores de Proteínas Quinasas , Proteínas Tirosina Quinasas/genética
6.
J Thromb Thrombolysis ; 47(1): 113-120, 2019 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-30291514

RESUMEN

Intravenous thrombolysis (IVT) is the treatment of choice for most patients with acute ischemic stroke. According to the recently updated guidelines, IVT should be administered in absence of absolute exclusion criteria. We aimed to assess the proportion of ischemic strokes potentially eligible and actually treated with IVT, and to explore the reasons for not administering IVT. We prospectively collected and analyzed data from 1184 consecutive ischemic stroke patients admitted to the 22 Stroke Units (SUs) of the Veneto region from September 18th to December 10th 2017. Patients were treated with IVT according to the current Italian guidelines. For untreated patients, the reasons for not administering IVT were reported by each center in a predefined model including absolute and/or relative exclusion criteria and other possible reasons. Out of 841 (71%) patients who presented within 4.5 h of stroke onset, 704 (59%) had no other absolute exclusion criteria and were therefore potentially eligible for IVT according to the current guidelines. However, only 323 (27%) patients were eventually treated with IVT. Among 861 (73%) untreated patients, 480 had at least one absolute exclusion criterion, 283 only relative exclusion criteria, 56 only other reasons, and 42 a combination of relative exclusion criteria and other reasons. Our study showed that only 46% (323/704) of the potentially eligible patients were actually treated with IVT in the SUs of the Veneto region. All healthcare professionals involved in the acute stroke pathway should make an effort to bridge this gap between eligibility and reality.


Asunto(s)
Accidente Cerebrovascular/tratamiento farmacológico , Terapia Trombolítica/métodos , Administración Intravenosa , Anciano , Isquemia Encefálica , Femenino , Personal de Salud/educación , Humanos , Italia , Masculino , Persona de Mediana Edad , Guías de Práctica Clínica como Asunto
7.
Artículo en Inglés | MEDLINE | ID: mdl-38468005

RESUMEN

Lithium recovery from Lithium-ion batteries requires hydrometallurgy but up-to-date technologies aren't economically viable for Lithium-Iron-Phosphate (LFP) batteries. Selective leaching (specifically targeting Lithium and based on mild organic acids and low temperatures) is attracting attention because of decreased environmental impacts compared to conventional hydrometallurgy. This study analysed the technical and economic performances of selective leaching with 6%vv. H2O2 and citric acid (0.25-1 M, 25 °C, 1 h, 70 g/l) compared with conventional leaching with an inorganic acid (H2SO4 1 M, 40 °C, 2 h, 50 g/l) and an organic acid (citric acid 1 M, 25 °C, 1 h, 70 g/l) to recycle end of life LFP cathodes. After conventional leaching, chemical precipitation allowed to recover in multiple steps Li, Fe and P salts, while selective leaching allowed to recover Fe and P, in the leaching residues and required chemical precipitation only for lithium recovery. Conventional leaching with 1 M acids achieved leaching efficiencies equal to 95 ± 2% for Li, 98 ± 8% for Fe, 96 ± 3% for P with sulfuric acid and 83 ± 0.8% for Li, 8 ± 1% for Fe, 12 ± 5% for P with citric acid. Decreasing citric acid's concentration from 1 to 0.25 M didn't substantially change leaching efficiency. Selective leaching with citric acid has higher recovery efficiency (82 ± 6% for Fe, 74 ± 8% for P, 29 ± 5% for Li) than conventional leaching with sulfuric acid (69 ± 15% for Fe, 70 ± 18% for P, and 21 ± 2% for Li). Also, impurities' amounts were lower with citric acid (335 ± 19 335 ± 19 of S mg/kg of S) than with sulfuric acid (8104 ± 2403 mg/kg of S). In overall, the operative costs associated to 0.25 M citric acid route (3.17€/kg) were lower compared to 1 M sulfuric acid (3.52€/kg). In conclusion, citric acid could be a viable option to lower LFP batteries' recycling costs, and it should be further explored prioritizing Lithium recovery and purity of recovered materials.

8.
Environ Pollut ; 357: 124455, 2024 Sep 15.
Artículo en Inglés | MEDLINE | ID: mdl-38942274

RESUMEN

Textile materials are one of the primary sources of microplastic pollution. The washing procedure is by far the most significant way that textile products release microplastic fibers (MPFs). Therefore, in this study, the effects of various textile raw materials (A acrylic, PA polyamide, PET polyester, RPET recycled polyester and PP polypropylene), fabric construction properties (woven, knitted), thickness and basis weight values on MPFs release at different washing stages (pre-washing, soaping/rinsing) were examined separately. To mimic the most popular home washing procedures, a 10-min pre-wash and a 35-min soaping/rinsing phase at 40 °C were selected for the washing procedure. Utilizing the Image J program on macroscopic images captured by a high-resolution SL. R camera, the microfibers collected by filtering the water have been visually counted. According to the results, knitted fabrics released fewer MPFs than woven fabrics, with the woven acrylic sample (A3-w) exhibiting the highest release (2405 MPFs). The number of MPFs increased along with the thickness and weight of the fabric. Recycled polyester was found to release more MPFs than virgin polyester under the same conditions (1193 MPFs vs. 908 MPFs). This study demonstrates how recycled polyester, although initially an environmentally beneficial solution, can eventually become detrimental to the environment. Furthermore, it is known that the pre-washing procedure-which is optional-releases a lot more MPFs than the soaping and rinsing procedures, and that stopping this procedure will drastically lower the amount of MPFs incorporated into the water.


Asunto(s)
Lavandería , Microplásticos , Textiles , Microplásticos/análisis , Poliésteres/química , Contaminantes Químicos del Agua/análisis , Polipropilenos/química
9.
Intensive Crit Care Nurs ; 76: 103392, 2023 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-36731262

RESUMEN

OBJECTIVE: To explore recurrent themes in diaries kept by intensive care unit (ICU) staff during the coronavirus disease 2019 (COVID-19) pandemic. DESIGN: Qualitative study. SETTING: Two ICUs in a tertiary level hospital (Milan, Italy) from January to December 2021. METHODS: ICU staff members wrote a digital diary while caring for adult patients hospitalized in the intensive care unit for >48 hours. A thematic analysis was performed. FINDINGS: Diary entries described what happened and expressed emotions. Thematic analysis of 518 entries gleaned from 48 diaries identified four themes (plus ten subthemes): Presenting (Places and people; Diary project), Intensive Care Unit Stay (Clinical events; What the patient does; Patient support), Outside the Hospital (Family and topical events; The weather), Feelings and Thoughts (Encouragement and wishes; Farewell; Considerations). CONCLUSION: The themes were similar to published findings. They offer insight into care in an intensive care unit during a pandemic, with scarce resources and no family visitors permitted, reflecting on the patient as a person and on daily care. The staff wrote farewell entries to dying patients even though no one would read them. IMPLICATIONS FOR CLINICAL PRACTICE: The implementation of digital diaries kept by intensive care unit staff is feasible even during the COVID-19 pandemic. Diaries kept by staff can provide a tool to humanize critical care. Staff can improve their work by reflecting on diary records.


Asunto(s)
COVID-19 , Pandemias , Adulto , Humanos , Unidades de Cuidados Intensivos , Cuidados Críticos/psicología , Emociones
10.
Front Public Health ; 11: 1290350, 2023.
Artículo en Inglés | MEDLINE | ID: mdl-38045968

RESUMEN

Background: The rapid global spread of severe acute respiratory syndrome coronavirus (SARS-CoV-2) was met with the rollout of vaccination campaigns at mass vaccination centers. The Palazzo delle Scintille, Milan, was designated by the Lombardy regional administration as a vaccination site with the target of processing about 9,000 users daily. Methods: For this observational study, we compared data on vaccinations delivered at the Palazzo delle Scintille with coronavirus disease (COVID-19)-related regional data. Results: Between 25 April 2021 and 28 February 2023, a total of 1,885,822 COVID-19 doses were administered; the mean hourly rate was 289 (247.2), the mean daily rate was 3185.5 (3104.5), the mean user age was 49.5 years (10.7). The Comirnaty vaccine (Pfizer-BioNTech) was most often given (1,072,030/1,885,822; 56.8%). Between 4 December 2021 and 15 January 2022, the daily dose rate was above the maximum daily capacity set by the regional administration. Conclusion: The trend for daily dose rates administered at the Palazzo delle Scintille center was in line with COVID-19-related regional data. The center played a major role in the regional mass vaccination campaign.


Asunto(s)
COVID-19 , Vacunación Masiva , Humanos , Persona de Mediana Edad , COVID-19/epidemiología , COVID-19/prevención & control , Italia/epidemiología , SARS-CoV-2 , Vacunación , Adulto
11.
Stroke ; 43(1): 137-41, 2012 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-22033996

RESUMEN

BACKGROUND AND PURPOSE: In the general population, migraine, cerebrovascular diseases, and vascular dementia differ in many aspects between men and women. CADASIL is considered a unique model to investigate migraine with aura, stroke, and dementia related to ischemic small vessel disease. This study aims to evaluate the effect of gender on the main clinical and neuroimaging characteristics of CADASIL. METHODS: Cross-sectional data from 313 CADASIL patients including various clinical and cognitive scores and MRI parameters were compared between men and women, and between those younger and older than the median age of the population corresponding to the usual age of menopause (51 years). RESULTS: At younger than 51 years, migraine with aura was 50% more prevalent in women and stroke was 75% more prevalent in men. After the fifth decade, men had higher National Institutes of Health Stroke Scale and Rankin scores than women and more severe executive dysfunction, although global cognitive scores were similar. Age at first stroke, the number of stroke events, and the prevalence of dementia and psychiatric symptoms did not differ between men and women. Brain volume was lower in men with a trend for a larger volume of lacunar infarcts. CONCLUSIONS: In CADASIL, migraine with aura is more frequent in women and stroke is more frequent in men before the age of menopause. This difference seems to vanish after this age limit but may result in a higher degree of cognitive impairment and cerebral atrophy in men at the late stage of the disease. The presumable role of ovarian hormones in these gender-related differences remains to be explored.


Asunto(s)
Isquemia Encefálica/genética , CADASIL/diagnóstico , Migraña con Aura/genética , Fenotipo , Accidente Cerebrovascular/genética , Adulto , Factores de Edad , Isquemia Encefálica/fisiopatología , Isquemia Encefálica/psicología , CADASIL/genética , CADASIL/fisiopatología , CADASIL/psicología , Estudios Transversales , Función Ejecutiva , Femenino , Humanos , Masculino , Persona de Mediana Edad , Migraña con Aura/fisiopatología , Migraña con Aura/psicología , Factores Sexuales , Accidente Cerebrovascular/fisiopatología , Accidente Cerebrovascular/psicología
12.
Nat Commun ; 13(1): 7406, 2022 12 01.
Artículo en Inglés | MEDLINE | ID: mdl-36456575

RESUMEN

Fluorescence laser-scanning microscopy (LSM) is experiencing a revolution thanks to new single-photon (SP) array detectors, which give access to an entirely new set of single-photon information. Together with the blooming of new SP LSM techniques and the development of tailored SP array detectors, there is a growing need for (i) DAQ systems capable of handling the high-throughput and high-resolution photon information generated by these detectors, and (ii) incorporating these DAQ protocols in existing fluorescence LSMs. We developed an open-source, low-cost, multi-channel time-tagging module (TTM) based on a field-programmable gate array that can tag in parallel multiple single-photon events, with 30 ps precision, and multiple synchronisation events, with 4 ns precision. We use the TTM to demonstrate live-cell super-resolved fluorescence lifetime image scanning microscopy and fluorescence lifetime fluctuation spectroscopy. We expect that our BrightEyes-TTM will support the microscopy community in spreading SP-LSM in many life science laboratories.


Asunto(s)
Neoplasias de Células Escamosas , Neoplasias Cutáneas , Humanos , Microscopía Confocal , Fotones
13.
Materials (Basel) ; 14(11)2021 Jun 07.
Artículo en Inglés | MEDLINE | ID: mdl-34200365

RESUMEN

The main obstacle to bottom ash (BA) being used as a recycling aggregate is the content of salts and potential toxic elements (PTEs), concentrated in a layer that coats BA particles. This work presents a dry treatment for the removal of salts and PTEs from BA particles. Two pilot-scale abrasion units (with/without the removal of the fine particles) were fed with different BA samples. The performance of the abrasion tests was assessed through the analyses of particle size and moisture, and that of the column leaching tests at solid-to-liquid ratios between 0.3 and 4. The results were: the particle-size distribution of the treated materials was homogeneous (25 wt % had dimensions <6.3 mm) and their moisture halved, as well as the electrical conductivity of the leachates. A significant decrease was observed in the leachates of the treated BA for sulphates (44%), chlorides (26%), and PTEs (53% Cr, 60% Cu and 8% Mo). The statistical analysis revealed good correlations between chloride and sulphate concentrations in the leachates with Ba, Cu, Mo, and Sr, illustrating the consistent behavior of the major and minor components of the layer surrounding BA particles. In conclusion, the tested process could be considered as promising for the improvement of BA valorization.

14.
Front Synaptic Neurosci ; 13: 727406, 2021.
Artículo en Inglés | MEDLINE | ID: mdl-34899260

RESUMEN

Fluorescence labeling of difficult to access protein sites, e.g., in confined compartments, requires small fluorescent labels that can be covalently tethered at well-defined positions with high efficiency. Here, we report site-specific labeling of the extracellular domain of γ-aminobutyric acid type A (GABA-A) receptor subunits by genetic code expansion (GCE) with unnatural amino acids (ncAA) combined with bioorthogonal click-chemistry labeling with tetrazine dyes in HEK-293-T cells and primary cultured neurons. After optimization of GABA-A receptor expression and labeling efficiency, most effective variants were selected for super-resolution microscopy and functionality testing by whole-cell patch clamp. Our results show that GCE with ncAA and bioorthogonal click labeling with small tetrazine dyes represents a versatile method for highly efficient site-specific fluorescence labeling of proteins in a crowded environment, e.g., extracellular protein domains in confined compartments such as the synaptic cleft.

18.
Acta Biomed ; 90(11-S): 65-71, 2019 11 11.
Artículo en Inglés | MEDLINE | ID: mdl-31714501

RESUMEN

INTRODUCTION: During Continuous Positive Pressure Ventilation delivered through helmet, the patient inhales high flows of gas without adequate conditioning. However, the need to humidify the inspired gas during Helmet-CPAP, has not been sufficiently explored. METHODS: Experimental design study. Six healthy individuals underwent High Flow Helmet CPAP with different gas flows (60 and 80 L/min) and FiO2 (0.35, 0.5, 0.7 and 1) generated by a Venturi system, with and without active humidification. The active humidifier setting was 26 ° at the humidification chamber and 28 ° at the helmet gas inlet. At each setting, measurements about temperature and relative humidity inside helmet were taken. Comfort level at each setting was evaluated using a visual analog scale rated from 0 to 10. RESULTS: Without heated humidification, the mean value of absolute humidity in the eight combinations investigated was 5.9±2.1 mg H2O/L, with a mean  temperature of 25.8±0.9°. With heated  humidification mean absolute humidity was 15.0±3.5 mg H2O/L with mean temperature of 29.0±0.1°. The median comfort scale value was 6 (IQR: 5.25-6.75) during the phase without humidification vs 8 (7.25-8.0 - P<0.01) when active humidification was applied. CONCLUSIONS: In healthy subjects undergoing High Flow Helmet CPAP, heated humidifiers with heated wires tubes are necessary to avoid the under-humidification inside the helmet. To obtain patient's comfort and airways mucosal humidification during continuous Helmet CPAP, the most desirable conditions are reached by heated humidifiers with a humidifying chamber temperature settled between 26-28°C.


Asunto(s)
Presión de las Vías Aéreas Positiva Contínua , Agua/análisis , Adulto , Femenino , Gases , Voluntarios Sanos , Calor , Humanos , Masculino
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