Pro-inflammatory RNA:DNA hybrids are p53 independently boosted by hyperbaric oxygen: a subcellular distribution analysis by automated quantitative imaging

Purpose: RNA:DNA hybrids are co-transcriptional products with acknowledged cytoplasmic pro-inflammatory role as activators of the cGAS-STING pathway. We recently proved them also as radiation-induced senescence messages for the abscopal effect mediation, demonstrating the need for a functional p53 for their production and release in A549 and H1299 tumour cells. However, little is known about their role under different stress conditions, especially in cancer cells. Methods: In this work, we open the investigation making use of automated quantitative imaging to characterize the hybrid subcellular distribution in HeLa cells grown under different oxygen pressures or exposed to different ionizing radiation doses. After cell imaging by confocal fluorescent microscopy, we apply automated imaging methods developed on purpose to quantify hybrid foci and nuclear cluster intensity, regional and local density and dimension. Results: We show that alteration of culture oxygenation increases hybrid cytoplasmic presence, especially when caused by an hyperoxic environment, with evident hybrid gathering at the cell membrane. Ionizing radiations always fail to increase hybrids, in accordance with the absence of functional p53 in HeLa cells. However, dose-dependent effects are still evident and suggest a threshold dose of 7.5 Gy for remarkable hybrid reduction. Conclusion: Together with our previous results, these data demonstrate for the first time that different types of stress can increase hybrid production in cancer cells and by at least two different pathways, one p53-dependent triggerable by ionizing radiations and one p53-independent triggerable by oxidative stress. Together, our findings provide a starting point for understanding hybrid role in tumour stress response

ossigenoterapia iperbarica e terapia a pressione negativa nel trattamento delle lesioni difficili hyperbaric oxygen therapy and negative pressure wound therapy in the treatment of non healing wounds

Lo scopo di questo articolo è valutare i risultati ottenibili trattando lesioni difficili attraverso la combinazione di ossigenoterapia iperbarica (OTI) e terapia a pressione negativa (TPN). Individuare le modalità con cui queste possano agire in sinergia coadiuvandosi, al fine di ottimizzare la rigenerazione dei tessuti e favorire la guarigione come qualità e tempi più brevi. Sono stati presi in analisi i dati di tre pazienti trattati presso il Centro Iperbarico di Ravenna che presentavano ferite agli arti inferiori aperte da più di sei settimane. È stato eseguito l'assessment iniziale della ferita e applicato un approccio multi terapeutico OTI e TPN per un periodo compreso tra 3-6 settimane. I pazienti presi in analisi sono giunti a guarigione completa entro 10 settimane di trattamento rispetto alla media di presa in carico per 28 settimane degli altri pazienti trattati presso la stessa struttura (dato reale) e alla media di 12 settimane previste nelle linee guida (benchmark). Le due terapie associate hanno prodotto un esito positivo che avrebbe richiesto tempi e costi maggiori se fossero state utilizzate singolarmente. The purpose of this work is the evaluation of the results obtaineble by treating hard to heal wounds with the combination of Hyperbaric Oxygen Therapy (HBOT) and Negative Wound Pressure Therapy (NWPT). Identify how HBOT and NWPT can act in synergy, in order to optimize tissue regeneration and promote a good quality healing and in shorter time. The study analyzes data of three patients affected, for more than six week, by lower limb wounds and treated at the Hyperbaric Center of Ravenna. The initial wound assessment was performed and a multi-therapeutic approach, HBOT and NWPT, was applied over a period of 3-6 weeks. The patients underwent to a complete healing after a maximum of 10 weeks of treatment compared to the 28-weeks average of other patients treated at the same facility (real data) and the 12-weeks average expected in the guidelines (benchmark).The combination of the two therapies, has led to a positive result saving time and money; the individual use of them would have required more time and costs

Gas embolization of the liver in a rat model of rapid decompression

Occurrence of liver gas embolism after rapid decompression was assessed in 31 female rats that were decompressed in 12 min after 42 min of compression at 7 ATA (protocol A). Sixteen rats died after decompression (group I). Of the surviving rats, seven were killed at 3 h (group II), and eight at 24 h (group III). In group I, bubbles were visible in the right heart, aortic arch, liver, and mesenteric veins and on the intestinal surface. Histology showed perilobular microcavities in sinusoids, interstitial spaces, and hepatocytes. In group II, liver gas was visible in two rats. Perilobular vacuolization and significant plasma aminotransferase increase were present. In group III, liver edema was evident at gross examination in all cases. Histology showed perilobular cell swelling, vacuolization, or hydropic degeneration. Compared with basal, enzymatic markers of liver damage increased significantly. An additional 14 rats were decompressed twice (protocol B). Overall mortality was 93%. In addition to diffuse hydropic degeneration, centrilobular necrosis was frequently observed after the second decompression. Additionally, 10 rats were exposed to three decompression sessions (protocol C) with doubled decompression time. Their mortality rate decreased to 20%, but enzymatic markers still increased in surviving rats compared with predecompression, and perilobular cell swelling and vacuolization were present in five rats. Study challenges were 1) liver is not part of the pathophysiology of decompression in the existing paradigm, and 2) although significant cellular necrosis was observed in few animals, zonal or diffuse hepatocellular damage associated with liver dysfunction was frequently demonstrated. Liver participation in human decompression sickness should be looked for and clinically evaluated

Physical activity is inversely related to drug consumption in elderly patients with cardiovascular events

Abstract Elderly patients with cardiovascular events are characterized by high drug consumptions. Whether high drug consumptions are related to physical activity is not known. In order to examine whether physical activity is related to drug consumption in the elderly, patients older than 65 years (n = 250) with a recent cardiovascular event were studied. Physical activity was analyzed according to the Physical Activity Scale for the Elderly (PASE) score and related to drug consumption. PASE score was 72.4 ± 45.0 and drug consumption was 8.3 ± 2.2. Elderly patients with greater comorbidity took more drugs (8.7 ± 2.1) and are less active (PASE = 64.4 ± 50.6) than patients with Cumulative Illness Rating Scale severity score higher than 1.8 than those with a score lower than 1.8 (76.3 ± 41.4, p < 0.05, and 8.0 ± 2.0, p = 0.006, respectively). Multivariate analysis correlation confirmed that PASE score is negatively associated with drug consumption (β = −0.149, p = 0.031), independently of several variables including comorbidity. Thus, physical activity is inversely related to drug consumption in elderly patients with cardiovascular events. This inverse relationship may be attributable to the high degree of comorbidity observed in elderly patients in whom poor level of physical activity and high drug consumption are predominant

An Innovative Hyperbaric Hypothermic Machine Perfusion Protects the Liver from Experimental Preservation Injury

Purpose. Hypothermic machine perfusion systems seem more effective than the current static storage to prevent cold ischemic liver injury. Thus, we test an innovative hyperbaric hypothermic machine perfusion (HHMP), which combines hyperbaric oxygenation of the preservation solution and continuous perfusion of the graft. Methods. Rat livers were preserved with Celsior solution according to 4 different modalities: normobaric static preservation; hyperbaric static preservation at 2 atmosphere absolute (ATA); normobaric dynamic preservation, with continuous perfusion; hyperbaric dynamic preservation, with continuous perfusion at 2 ATA. After 24 h cold preservation, we assessed different parameters. Results. Compared to baseline, livers preserved with the current static storage showed severe ultrastructural damage, glycogen depletion and an increased oxidative stress. Normobaric perfused livers showed improved hepatocyte ultrastructure and ameliorated glycogen stores, but they still suffered a significant oxidative damage. The addition of hyperbaric oxygen produces an extra benefit by improving oxidative injury and by inducing endothelial NO synthase (eNOS) gene expression. Conclusions. Preservation by means of the present innovative HHMP reduced the liver injury occurring after the current static cold storage by lowering glycogen depletion and oxidative damage. Interestingly, only the use of hyperbaric oxygen was associated to a blunted oxidative stress and an increased eNOS gene expression

Performance Assessment in Fingerprinting and Multi Component Quantitative NMR Analyses

An interlaboratory comparison (ILC) was organized with the aim to set up quality control indicators suitable for multicomponent quantitative analysis by nuclear magnetic resonance (NMR) spectroscopy. A total of 36 NMR data sets (corresponding to 1260 NMR spectra) were produced by 30 participants using 34 NMR spectrometers. The calibration line method was chosen for the quantification of a five-component model mixture. Results show that quantitative NMR is a robust quantification tool and that 26 out of 36 data sets resulted in statistically equivalent calibration lines for all considered NMR signals. The performance of each laboratory was assessed by means of a new performance index (named Qp-score) which is related to the difference between the experimental and the consensus values of the slope of the calibration lines. Laboratories endowed with a Qp-score falling within the suitable acceptability range are qualified to produce NMR spectra that can be considered statistically equivalent in terms of relative intensities of the signals. In addition, the specific response of nuclei to the experimental excitation/relaxation conditions was addressed by means of the parameter named NR. NR is related to the difference between the theoretical and the consensus slopes of the calibration lines and is specific for each signal produced by a well-defined set of acquisition parameters

Model Based Navigation of Delta-Wing UAV - In-Flight Calibration and Autonomous Performance

This paper presents the first practical results of a model-based approach to autonomous navigation applied to a small delta wing drone. The aerodynamic coefficients of the considered platform are unknown and need to be determined for the (vehicle dynamic) model-based navigation to work properly. The proposed approach uses post-processed INS/GNSS trajectory estimates of relatively high precision as observations to refine ``priors'' of aerodynamic coefficients via state-estimation (Extended Kalman Filter). Two methods to derive such ``priors'' (i.e., initial parameter values) are investigated. The first adapts coefficients described in the literature for an aircraft of similar geometry. The second performs regression analysis of flight data to estimate coarse values of the coefficients. Both sets of coefficients are further re-calibrated in-flight via state estimation. The accuracy of the coefficient calibration is evaluated by simulating a GNSS outage of several minutes, during which the trajectory flown under autonomous navigation is compared to that of the reference