Determinazione dello stato di stress ossidativo in tessuti di topo trattati con sale sodico di clorofilliana magnesiaca

Gli esperimenti condotti in questo progetto di ricerca sono volti allo studio degli effetti di integratori a base di clorofillina magnesiaca sullo stato di stress ossidativo sui tessuti murini. Per le analisi sono state prese in esame due corti di topi femmine da 48 e 44 esemplari ciascuna. Le due corti sono state sottoposte a un diverso tipo di trattamento, rispettivamente: trattamento cronico con diverse quantità di soluzione idroalcolica di clorofillina magnesiaca e trattamento acuto con unica dose standard di soluzione idroalcolica di clorofillina magnesiaca. In questo modo è stato possibile studiare l'efficacia degli integratori in base al tipo di trattamento. Le analisi sono state fatte con saggio TBARS sui topi della prima corte e con saggio FRAP sui topi di entrambe le corti. Il saggio TBARS mostra lo stato di tress ossidativo dei tessuti mediante quantificazione della malondialdeide. Il saggio FRAP descrive la capacità riducente della clorofilliana estratta dai campioni di tessuto. In un progetto parallelo a quello descritto in questa tesi è stata quantificata la clorofilla tramite spettrofluorimetria per costruire un parallelo fra quantità e potere riducente rilevato. Grazie all'analisi dei risultati è possibile dire che la clorofillina magnesiaca non è proossidante e presenta potere riducente. Tuttavia un trattamento acuto non è sufficiente a farla accumulare nei tessuti.

Arterial luminal curvature and fibrous-cap thickness affect critical stress conditions within atherosclerotic plaque: An in vivo MRI-based 2D finite-element study

High mechanical stress in atherosclerotic plaques at vulnerable sites, called critical stress, contributes to plaque rupture. The site of minimum fibrous cap (FC) thickness (FCMIN) and plaque shoulder are well-documented vulnerable sites. The inherent weakness of the FC material at the thinnest point increases the stress, making it vulnerable, and it is the big curvature of the lumen contour over FC which may result in increased plaque stress. We aimed to assess critical stresses at FCMIN and the maximum lumen curvature over FC (LCMAX) and quantify the difference to see which vulnerable site had the highest critical stress and was, therefore, at highest risk of rupture. One hundred patients underwent high resolution carotid magnetic resonance (MR) imaging. We used 352 MR slices with delineated atherosclerotic components for the simulation study. Stresses at all the integral nodes along the lumen surface were calculated using the finite-element method. FCMIN and LCMAX were identified, and critical stresses at these sites were assessed and compared. Critical stress at FC MIN was significantly lower than that at LCMAX (median: 121.55 kPa; inter quartile range (IQR) = [60.70-180.32] kPa vs. 150.80 kPa; IQR = [91.39-235.75] kPa, p < 0.0001). If critical stress at FCMIN was only used, then the stress condition of 238 of 352 MR slices would be underestimated, while if the critical stress at LCMAX only was used, then 112 out of 352 would be underestimated. Stress analysis at FCMIN and LCMAX should be used for a refined mechanical risk assessment of atherosclerotic plaques, since material failure at either site may result in rupture.

The effect of acute stress on LTP and LTD induction in the hippocampal CA1 region of anesthetized rats at three different ages

Not all experiences are memorized equally well. Especially, some types of stress are unavoidable in daily life and the stress experience can be memorized for life. Previous evidence has showed that synaptic plasticity, such as long-term potentiation (LTP) that may be the major cellular model of the mechanism underlying learning and memory, is influenced by behavioral stress. However, the effect of behavioral stress on age-related synaptic plasticity in-vivo was primarily known. Here we found that the LTP induction in the hippocampal CA1 region of anesthetized rats obviously showed inverted-U shape related to ages (4, 10 and 74 weeks old rats), but low-frequency stimulation was unable to induce reliable long-term depression (LTD) in these animals. Furthermore, acute elevated platform (EP) stress enabled reliable LTD significantly and completely blocked LTP induction at these ages. Importantly, LTD after exposure to acute EP stress showed similar magnitude over these ages. The present results that stress enables LTD but impairs LTP induction at these three ages strengthen a view that stress experience-dependent LTD (SLTD) may underlie stress form of aberrant memories. (C) 2004 Elsevier B.V. All rights reserved.

In vivo studies on the toxic effects of microcystins on mitochondrial electron transport chain and ion regulation in liver and heart of rabbit

This study examined the toxic effects of microcystins on mitochondria of liver and heart of rabbit in vivo. Rabbits were injected i.p. with extracted microcystins (mainly MC-RR and -LR) at two doses, 12.5 and 50 MCLReq. mu g/kg bw, and the changes in mitochondria of liver and heart were studied at 1, 3,12, 24 and 48 h after injection. MCs induced damage of mitochondrial morphology and lipid peroxidation in both liver and heart. MCs influenced respiratory activity through inhibiting NADH dehydrogenase and enhancing succinate dehydrogenase (SDH). MCs altered Na+-K+-ATPase and Ca2+-Mg2+-ATPase activities of mitochondria and consequently disrupted ionic homeostasis, which might be partly responsible for the loss of mitochondrial membrane potential (MMP). MCs were highly toxic to mitochondria with more serious damage in liver than in heart. Damage of mitochondria showed reduction at 48 h in the low dose group, suggesting that the low dose of MCs might have stimulated a compensatory response in the rabbits. (C) 2008 Elsevier Inc. All rights reserved.

Valutazione del danno da stress ossidativo come indicatore di effetto biologico dell'esposizione professionale a xenobiotici

Oxidative DNA damages determine the activation of cell repair processes. These processes originate repair products, including the most studied one, 8-hydroxy-2’-deoxyguanosine (8-OH-dG). Several analytical techniques have been applied to measure urinary 8-OH-dG, but a discrepancy in basal urinary 8-OH-dG levels has been noted when comparing chromatographic techniques with immunoenzymatic assays (ELISA). Our laboratory has developed a fully validated, liquid chromatography-tandem mass spectrometry method presenting high sensitivity and specificity, which has participated in an inter-laboratory validation of assays for the measurement of urinary 8-OH-dG (ESCULA project). Mass Spectrometric techniques showed more accuracy and specificity than immunoenzymatic methods. Human spot urine samples were analyzed in order to investigate the possibility to correct urinary lesion measurements for creatinine and to evaluate the intra- and inter-day variability of 8-OH-dG excretion in urine. Our results confirm the opportunity to delve into these issues. Finally, we measured urinary 8-OH-dG in workers exposed to antineoplastic drugs and in a group of unexposed subjects to evaluate the relationship between occupational exposure and oxidative damage related to the internal dose. We found higher levels of 8-OH-dG in exposed nurses, but, as compared to the non-exposed subjects, the difference was not statistically significant, probably do to the very low level of exposure. The scientific literature is rapidly developing on the topic of DNA damage and related repair capacity. Nevertheless, further studies are needed to achieve a better understanding of the sources of DNA lesions in urine and their significance, both in clinical and occupational medicine.

Espressione di geni correlati allo stress ossidativo nell'ascidia coloniale Botryllus schlosseri esposta a diverse condizioni ambientali nella Laguna di Venezia

I pattern di espressione genica permettono di valutare se gli organismi siano soggetti a stress ambientali, spesso associati a stress ossidativo e produzione di specie reattive dell’ossigeno, che possono essere analizzate per studiare gli effetti sub-letali indotti dall’ambiente negli organismi. Scopo di questa ricerca è stato valutare la possibilità di utilizzo dell’ascidia coloniale B. schlosseri come biomarker in ambiente lagunare. Le colonie, esposte a diverse condizioni ambientali nella Laguna di Venezia, sono state confrontate con esemplari allevati in condizioni di controllo. La ricerca si è concentrata in 2 siti con diverso grado di idrodinamicità e impatto antropico. Mentre nel sito 1, più vicino alla bocca di porto, si è rilevata la presenza di Tunicati, il sito 2 ne è privo. Il sito 2 ha registrato valori di pH e temperatura più alti. Inoltre, nel sito 2 è stata rilevata una mortalità maggiore delle colonie e alterazioni della morfologia nelle colonie sopravvissute. Ciò suggerisce che il sito 2 presenti condizioni avverse per B. schlosseri. Sui campioni di B. schlosseri sono state eseguite PCR semiquantitative per analizzare l’espressione di un gruppo di geni coinvolto nella risposta allo stress ossidativo: la glutammato cistein ligasi la glutatione sintetasi, 2 isoforme di glutatione perossidasi e la superossido dismutasi (SOD). Tutti i geni presentano livelli di trascrizione doppi nelle colonie del sito 1 rispetto al controllo. Viceversa, il sito 2 mostra livelli di espressione di poco superiori al controllo. Analisi spettrofotometriche evidenziano che le attività enzimatiche di SOD e catalasi sono più alte nel sito 2 rispetto al sito 1. Si può pertanto ipotizzare che le colonie esposte al sito 2 siano soggette a un maggiore stress. B. schlosseri appare dunque un buon indicatore dello stato ecologico dell’ambiente lagunare, entro parametri di pH e temperatura in cui abitualmente vive.

Stress and coping among migrant labourers in urban Vietnam : an adaptation cycle and health vulnerabilities

Rural-urban migration continues to grow in many developing countries including Vietnam. The experience of stress and coping associated with this process may vary for people from different circumstances. However, there has been little research on migrants to date. This study adopts a qualitative approach to research on unregistered, male, migrant freelance labourers in urban Vietnam and to explore factors contributing to stress and coping among this population. The study revealed an array of stressors related to migrants' life experiences in urban space, including physical, financial and social factors. Coping was diverse, including problem-focused coping (PFC) and emotion-focused coping (EFC), pro-social and anti-social, active and passive. Less active and anti-social coping appeared common. Together, weak social network and lack of support from formal systems placed coping and adaptation in a cyclic relationship. The results highlight a multi-disciplinary approach to help cope and adapt effectively for these men.

The role of emotional labour and role stress on burnout and psychological strain in high contact service employees

This study examines the relationships between job demands (in the form of role stressors and emotional management) and employee burnout amongst high contact service employees. Employees in customer facing roles are frequently required to manage overwhelming, conflicting or ambiguous demands, which they may feel ill-equipped to handle. Simultaneously, they must manage the emotions they display towards customers, suppressing some, and expressing others, be they genuine or contrived. If the in-role effort required of employees exceeds their inherent capacity to cope, burnout may result. Burnout, in turn, can have serious detrimental consequences for the psychological well being of employees. We find that both emotional management and role stressors impact burnout. We also confirm that burnout predicts psychological strain. In line with the Job Demands and Resources Model, we examine the mitigating impact of perceived support on these relationships but do not find a significant mitigating impact.

Numerical solution of stress intensity factors of multiple cracks in great number with eigen COD boundary integral equations

Based on the eigen crack opening displacement (COD) boundary integral equations, a newly developed computational approach is proposed for the analysis of multiple crack problems. The eigen COD particularly refers to a crack in an infinite domain under fictitious traction acting on the crack surface. With the concept of eigen COD, the multiple cracks in great number can be solved by using the conventional displacement discontinuity boundary integral equations in an iterative fashion with a small size of system matrix. The interactions among cracks are dealt with by two parts according to the distances of cracks to the current crack. The strong effects of cracks in adjacent group are treated with the aid of the local Eshelby matrix derived from the traction BIEs in discrete form. While the relatively week effects of cracks in far-field group are treated in the iteration procedures. Numerical examples are provided for the stress intensity factors of multiple cracks, up to several thousands in number, with the proposed approach. By comparing with the analytical solutions in the literature as well as solutions of the dual boundary integral equations, the effectiveness and the efficiencies of the proposed approach are verified.

The effects of dietary fish oil on inflammation, fibrosis and oxidative stress associated with obstructive renal injury in rats.

Scope: We examined whether dietary supplementation with fish oil modulates inflammation, fibrosis and oxidative stress following obstructive renal injury. Methods and results: Three groups of Sprague-Dawley rats (n = 16 per group) were fed for 4 wk on normal rat chow (oleic acid), chow containing fish oil (33 g eicosapentaenoic acid and 26 g docosahexaenoic acid per kg diet), or chow containing safflower oil (60 g linoleic acid per kg diet). All diets contained 7% fat. After 4 wk, the rats were further subdivided into four smaller groups (n = 4 per group). Unilateral ureteral obstruction was induced in three groups (for 4, 7 and 14 days). The fourth group for each diet did not undergo surgery, and was sacrificed as controls at 14 days. When rats were sacrificed, plasma and portions of the kidneys were removed and frozen; other portions of kidney tissue were fixed and prepared for histology. Compared with normal chow and safflower oil, fish oil attenuated collagen deposition, macrophage infiltration, TGF-beta expression, apoptosis, and tissue levels of arachidonic acid, MIP-1 alpha, IL-1 beta, MCP-1 and leukotriene B(4). Compared with normal chow, fish oil increased the expression of HO-1 protein in kidney tissue. Conclusions: Fish oil intake reduced inflammation, fibrosis and oxidative stress following obstructive renal injury.

The time course of in vivo recovery of transverse strain in high-stress tendons following exercise

Objective To evaluate the time course of the recovery of transverse strain in the Achilles and patellar tendon following a bout of resistance exercise. Methods Seventeen healthy adults underwent sonographic examination of the right patellar (n=9) and Achilles (n=8) tendons immediately prior to and following 90 repetitions of weight-bearing quadriceps and gastrocnemius-resistance exercise performed against an effective resistance of 175% and 250% body weight, respectively. Sagittal tendon thickness was determined 20 mm from the enthesis and transverse strain, as defined by the stretch ratio, was repeatedly monitored over a 24 h recovery period. Results Resistance exercise resulted in an immediate decrease in Achilles (t7=10.6, p<0.01) and patellar (t8=8.9, p<0.01) tendon thickness, resulting in an average transverse stretch ratio of 0.86±0.04 and 0.82±0.05, which was not significantly different between tendons. The magnitude of the immediate transverse strain response, however, was reduced with advancing age (r=0.63, p<0.01). Recovery in transverse strain was prolonged compared with the duration of loading and exponential in nature. The average primary recovery time was not significantly different between the Achilles (6.5±3.2 h) and patellar (7.1±3.2 h) tendons. Body weight accounted for 62% and 64% of the variation in recovery time, respectively. Conclusions Despite structural and biochemical differences between the Achilles and patellar tendon, the mechanisms underlying transverse creep recovery in vivo appear similar and are highly time dependent. These novel findings have important implications concerning the time required for the mechanical recovery of high-stress tendons following an acute bout of exercise.

Stress, lifestyle and quality of life in midlife and older Australian women : results from the stress and the health of women study

Background Chronic psychological stress may pose a serious threat to health, although the mechanisms are not fully understood. This study examines the impact of stress on modifiable lifestyle factors, depressive symptoms, health-related quality of life (HRQOL) and chronic illness in older Australian women. Methods Cross-sectional data were collected from a random sample of 181 older adults aged 60-70 years from rural and urban areas of South-East Queensland, Australia. We used structural equation modelling to examine associations between stress, modifiable lifestyle factors, HRQoL, and chronic illness. Findings Parameter estimates show that older women who reported life stressors where they felt helpless and feared for their life (high magnitude stressors) also reported higher body mass index (p = 0.03) and more chronic illness (p <0.01). In contrast, duration of exposure to life stressors was associated with higher depressive symptom scores (CES-D, p = 0.02) and sleep disturbance scores (p <0.01). Conclusions Our findings support the link between traumatic personal histories (exposure to high magnitude stressors) and unhealthy lifestyle factors. Findings highlight the need for more research on how stress reduction healthy lifestyle and positive coping strategies can be used to reduce the effects of high magnitude stress on health-related quality of life and chronic illness.

An investigation of the stress-buffering effects of social support in the occupational stress process as a function of team identification

This study explored how the social context influences the stress-buffering effects of social support on employee adjustment. It was anticipated that the positive relationship between support from colleagues and employee adjustment would be more marked for those strongly identifying with their work team. Furthermore, as part of a three-way interactive effect, it was predicted that high identification would increase the efficacy of coworker support as a buffer of two role stressors (role overload and role ambiguity). One hundred and 55 employees recruited from first-year psychology courses enrolled at two Australian universities were surveyed. Hierarchical multiple regression analyses revealed that the negative main effect of role ambiguity on job satisfaction was significant for those employees with low levels of team identification, whereas high team identifiers were buffered from the deleterious effect of role ambiguity on job satisfaction. There also was a significant interaction between coworker support and team identification. The positive effect of coworker support on job satisfaction was significant for high team identifiers, whereas coworker support was not a source of satisfaction for those employees with low levels of team identification. A three-way interaction emerged among the focal variables in the prediction of psychological well-being, suggesting that the combined benefits of coworker support and team identification under conditions of high demand may be limited and are more likely to be observed when demands are low.