Tracking autophagy during proliferation and differentiation of trypanosoma brucei

Autophagy is a lysosome-dependent degradation mechanism that sequesters target cargo into autophagosomal vesicles. The Trypanosoma brucei genome contains apparent orthologues of several autophagy-related proteins including an ATG8 family. These ubiquitin-like proteins are required for autophagosome membrane formation, but our studies show that ATG8.3 is atypical. To investigate the function of other ATG proteins, RNAi compatible T. brucei were modified to function as autophagy reporter lines by expressing only either YFP-ATG8.1 or YFP-ATG8.2. In the insect procyclic lifecycle stage, independent RNAi down-regulation of ATG3 or ATG7 generated autophagy-defective mutants and confirmed a pro-survival role for autophagy in the procyclic form nutrient starvation response. Similarly, RNAi depletion of ATG5 or ATG7 in the bloodstream form disrupted autophagy, but did not impede proliferation. Further characterisation showed bloodstream form autophagy mutants retain the capacity to undergo the complex cellular remodelling that occurs during differentiation to the procyclic form and are equally susceptible to dihydroxyacetone-induced cell death as wild type parasites, not supporting a role for autophagy in this cell death mechanism. The RNAi reporter system developed, which also identified TOR1 as a negative regulator controlling YFP-ATG8.2 but not YFP-ATG8.1 autophagosome formation, will enable further targeted analysis of the mechanisms and function of autophagy in the medically relevant bloodstream form of T. brucei

Preferential use of the perchlorate over the nitrate in the respiratory processes mediated by the Bacterium Azospira sp. OGA 24

Here we report the results obtained for a strain isolated from a polluted site and classified as Azospira sp. OGA 24. The capability of OGA 24 to utilize perchlorate and nitrate and the regulation of pathways were investigated by growth kinetic studies and analysis of messenger RNA (mRNA) expression of the genes of perchlorate reductase alpha subunit (pcrA), chlorite dismutase (cld), and periplasmic nitrate reductase large subunit (napA). In aerobic conditions and in a minimal medium containing 10 mM acetate as carbon source, 5.6 ± 0.34 mmol L-1 perchlorate or 9.7 ± 0.22 mmol L-1 nitrate were efficiently reduced during the growth with 10 mM of either perchlorate or nitrate. In anaerobiosis, napA was completely inhibited in the presence of perchlorate as the only electron acceptor, pcrA was barely detectable in nitrate-reducing conditions. The cell growth kinetics were in accordance with expression data, indicating a separation of nitrate and perchlorate respiration pathways. In the presence of both compounds, anaerobic nitrate consumption was reduced to 50% (4.9 ± 0.4 vs. 9.8 ± 0.15 mmol L-1 without perchlorate), while that of perchlorate was not affected (7.2 ± 0.5 vs. 6.9 ± 0.6 mmol L-1 without nitrate). Expression analysis confirmed the negative effect of perchlorate on nitrate respiration. Based on sequence analysis of the considered genes and 16S ribosomal gene (rDNA), the taxonomic position of Azospira sp. OGA 24 in the perchlorate respiring bacteria (PRB) group was further defined by classifying it in the oryzae species. The respiratory characteristics of OGA 24 strain make it very attractive in terms of potential applications in the bioremediation of environments exposed to perchlorate salts

Plethysmography system to monitor the jugular venous pulse: A feasibility study

Cerebral venous outflow is investigated in the diagnosis of heart failure through the monitoring of jugular venous pulse, an indicator to assess cardiovascular diseases. The jugular venous pulse is a weak signal stemming from the lying internal jugular vein and often invasive methodolo-gies requiring surgery are mandatory to detect it. Jugular venous pulse can also be extrapolated via the ultrasound technique, but it requires a qualified healthcare operator to perform the exami-nation. In this work, a wireless, user-friendly, wearable device for plethysmography is developed to investigate the possibility of monitoring the jugular venous pulse non-invasively. The proposed device can monitor the jugular venous pulse and the electrocardiogram synchronously. To study the feasibility of using the proposed device to detect physiological variables, several measurements were carried out on healthy subjects by considering three different postures: supine, sitting, and upright. Data acquired in the experiment were properly filtered to highlight the cardiac oscillation and remove the breathing contribution, which causes a considerable shift in the amplitude of signals. To evaluate the proper functioning of the wearable device for plethysmography, a comparison with the ultrasound technique was carried out. As a satisfactory result, the acquired signals resemble the typical jugular venous pulse waveforms found in literature

The environmental noise level in the rejuvenation pruning on centuries-old olive tree

ArticleIn the Italian agricultural economy, olive cultivation plays a fundamental role, and this is especially true for the southern regions where almost all cultivation is spread. In Calabria, in particular, olive cultivation has seen over the last few decades significantly improve the quality of production also as a result of investments aimed at the creation of new mechanizable plants and/or the modernization of existing ones; today some areas have got both PDO and PGI certification. In the ‘Piana di Gioia Tauro’, located north-west of the Reggio Calabria metropolitan area, olive growing extends over 20,000 hectares and the presence of centuries-old olive tree is still widespread. The olive varieties mainly belong to the local cultivars of ‘Sinopolese’ and ‘Ottobratica’, characterized by a remarkable rusticity and high development, perhaps unique in the world; they reach 20–25 meters high, forming what is called a ‘forest of olive trees’. The pruning operations are carried out by means of chainsaws of different power and size whereby, in addition to the previously described difficulties, operators are exposed to prolonged periods of noise levels. The purpose of this study is precisely to assess the exposure of operators to this particular olive grove. The aim is to identify the acoustic levels generated by the two pruning and cross-cutting activities, the risk thresholds and the exposure to which the individual workers of the two work sites are subjected, giving indications on the appropriate safety distances to maintain (according to current regulations) compared to noise sources

Electricity from Wood: A Wood Quality and Energy Efficiency Approach to Small Scale Pyro-gasification

AbstractThe global demand for renewable energy in the last years is facing innovations like the co-generation of electricity and heat from wooden industrial residues and biomass. Wood gasification is a very promising technique for conversion of wood chemical energy into thermal energy and into electricity. A new generation of small scale, moderately priced and easy to install pyro-gasification plants became available on the market, being an opportunity for the SMEs. These systems are also characterized by a remarkable operational easiness and great versatility in the type of used biomass, which can range from industrial residues (sawdust, wood chips...) to biomass of forestry and non-forestry origin. This study performed an extensive testing of a small size pyro-gasification plant capable of producing 30kW of electricity and 30kW of thermal energy, both net to enter in the network of GSE (Energy Services Manager) via the feed-in tariff and in a small district heating network or other uses. The process of pyro-gasification is analyzed in its entirety by a characterization of input biomass, pyro-gasification process monitoring, analysis of the quality of the produced syngas, characterization of residues due to the process (washing water smoke, charcoal...) and verification of the total efficiency. The electric efficiency of the system, from hardwood to syngas to electricity is quantified in being 12%

State of the Art and New Trends from the 2022 Gism Annual Meeting

The 2022 Italian Mesenchymal Stem Cell Group (Gruppo Italiano Staminali Mesenchimali, GISM) Annual Meeting took place on 20–21 October 2022 in Turin (Italy), with the support of the University of Turin and the City of Health and Science of Turin. The novelty of this year’s meeting was its articulation, reflecting the new structure of GISM based on six sections: (1) Bringing advanced therapies to the clinic: trends and strategies, (2) GISM Next Generation, (3) New technologies for 3D culture systems, (4) Therapeutic applications of MSC-EVs in veterinary and human medicine, (5) Advancing MSC therapies in veterinary medicine: present challenges and future perspectives, (6) MSCs: a double-edged sword: friend or foe in oncology. National and international speakers presented their scientific works with the aim of promoting an interactive discussion and training for all attendees. The atmosphere was interactive, where ideas and questions between younger researchers and senior mentors were shared in all moments of the congress

Unusual skin toxicity associated with sustained disease response induced by nivolumab in a patient with non-small cell lung cancer

Introduction: Immunotherapy has shown efficacy in the treatment of different malignancies. Nivolumab, an immune checkpoint inhibitor directed against programmed death-1, has been approved for non-small cell lung cancer (NSCLC) in pretreated patients. Although it is generally well-tolerated, immunotherapy may be complicated by a wide range of immune-mediated adverse events. We describe the case of an uncommon skin toxicity arising as alopecia universalis induced by nivolumab in a patient with NSCLC. Case description: A 58-year-old man received nivolumab for metastatic NSCLC after progression to 3 lines of chemotherapy. The treatment was prescribed in June 2016, and induced a rapid and significant disease response. Nivolumab was well-tolerated until May 2017, when partial alopecia at hair and eyelashes appeared. In the next months, alopecia became complete and extended to the whole body surface. The dermatologic picture was compatible with alopecia areata. A topical steroid therapy was attempted, without benefit. The patient refused systemic treatments and is still undergoing nivolumab without new toxicities and with persistent disease response. Conclusions: This case suggests that alopecia areata may be a rare immune-related adverse event of immune checkpoint agents. Its late onset in our patient is uncommon and unexpected, underlining that the risk of nivolumab-induced toxicity is not limited to the beginning of treatment. Despite its rarity, alopecia areata should be considered in the range of adverse events potentially induced by immune checkpoint inhibitors even in the long term. Potential association between toxicity and efficacy of immunotherapy in NSCLC warrants further investigation