Circulating neutrophils exhibit enhanced apoptosis associated with mitochondrial dysfunctions after surgery under general anaesthesia.

BACKGROUND: Evidence suggests that apoptosis plays a main role in the postoperative changes detected in the polymorphonuclear neutrophil (PMN) population. Furthermore, recent studies have demonstrated that mitochondrial alterations constitute critical events of the apoptotic cascade. In this study we investigated whether apoptosis among neutrophils taken from patients undergoing surgical trauma could be associated with perturbation of mitochondrial transmembrane potential (deltapsim) and/or exaggerated production of mitochondrial reactive oxygen species (ROS). METHODS: Twenty-seven patients undergoing elective surgery under general anaesthesia were enrolled in the study. Peripheral blood samples were drawn one day before the operation and at 12 and 24 h after surgery. Apoptosis rate was assessed by staining neutrophils with 7-amino-actinomycin D (7-AAD) and by analysis by a FACScan flow cytometer. In order to evaluate deltapsim, cells were exposed to 3,3-dihexyloxacarbocyanine iodide [DiOC6(3)]; intracellular ROS was measured by means of hydroethidine (HE) and 2,7-diclorofluorescein diacetate (DCFH-DA), followed by analysis on a cytofluorometer. RESULTS: At 12 h following surgery we observed a significantly (P<0.05) increased frequency of apoptotic PMNs compared to that preoperatively (30.79+/-3.68% vs 7.40+/-0.69%). At this same time-point, the rate of neutrophils stained with HE, DCFH-DA and [DiOC6(3)] were significantly (P<0.05) higher compared to baseline (51.05+/-5.44%, 50.58+/-5.84% and 55.31+/-4.33% vs 20.17+/-2.38%, 19.59+/-2.03 and 25.43+/-2.71% respectively). Overall measurements returned to the preoperative values 24 h after surgery. CONCLUSION: These data suggest that surgery under general anaesthesia triggers in the immediate postoperative period pathways of PMN accelerated apoptosis associated with significant alterations in mitochondrial function.

TNF alpha released in the early post-operative period is influenced by anaesthesia.

In this study we tested for TNF alpha concentrations 45 subjects undergoing elective prostatectomy performed under two different anaesthetic regimens. In addition, given the influence of neuroendocrine axis on immunological homeostasis, we also determined post surgery cortisol and norepinephrine levels. The patients were randomly assigned to one of two groups: group 1 (no. = 24) received NLA general anaesthesia, whereas group 2 (no. = 21) received spinal analgesia. Blood samples were drawn the day before surgery (t0), at two (t1), at 24 (t2), at 48 (t3) and 72 (t4) hours from the completion of operation. In the patients assigned to the general anaesthesia group an increase of circulating TNF alpha was detected through t1 to t4. In the spinal group TNFa values did not change in t1 and t2, whereas enhanced levels of the cytokine--overlapping with those of group 1--were observed at 48 and 72 hrs following surgery. Cortisol response increased in both the groups, peaking at t1 but, at this time patients undergoing spinal anaesthesia were found to have significantly lower circulating cortisol. A rise of plasma norepinephrine was observed at t1 in the general anaesthesia group alone. For the early TNF alpha increase, observed in group 1 only, we hypothesize that anaesthetics, such as NLA agents, may trigger a TNF alpha release in vivo. Alternatively TNF alpha production may represent the result of the neuroendocrine response such as cortisol and norepinephrine, to the surgical/anaesthetic trauma which could be blocked by subarachnoid anaesthesia. Conversely, the late TNF alpha increase found in all the patients studied might be explained by postoperative infections as well as by inflammation of the injured tissues and/or normal wound healing.

The treatment of cranio-facial pain by electroacupuncture and laser irradiation.

Craniofacial neuralgias represent an interesting medical challenge, especially regarding the complex therapeutical aspects. Our study was performed treating more than 700 patients, applying steel inox needles variable in diameters and length for about 20 minutes, with a total application number ranging between 10 and 20. Results have been defined referring to a 4-grades response scale: excellent, good, fairly good and insufficient. It is important to emphasise that the best results have been achieved with patients who chose the acupuncture as the first therapeutic approach, while patients who underwent other previous medical and/or surgical treatment have had a worse response. The authors assert that acupuncture and infrared laser reflex therapy represent a harmless and effective treatment of such a diffuse and invalidating disease.

Heat shock protein (HSP70) expression in septic patients.

PURPOSE: This study investigates heat shock protein 70 (HSP70) expression by peripheral blood mononuclear cells (PBMCs) of septic patients admitted to an intensive care unit and examines the possibility of a correlation between HSP70 levels and plasma tumor necrosis factor alpha (TNF-alpha) concentrations. Additionally, we evaluated whether the HSP70 production could be regarded as a prognostic factor for the development of septic shock as well as for patient survival. MATERIALS AND METHODS: Blood samples of 29 patients were taken 24 hours after the diagnosis of sepsis. HSP70 expression and TNF-alpha level were measured using indirect immunofluorescent analysis and a commercially available enzyme-linked immunosorbent assay method, respectively. RESULTS: PBMCs expressed significantly high levels of HSP70 (11.9 +/- 5.6 [sd]) compared with those of the healthy control group (3.2 +/- 2.1% positive cells). Such enhanced levels were correlated to plasma TNF-alpha concentrations (r = .99, P < .01). This study failed to demonstrate a relationship between HSP70 production and clinical outcome. CONCLUSION: These findings give further evidence that also in humans, heat shock response is activated during sepsis. The correlation observed between HSP70 overproduction and TNF-alpha plasma concentrations suggests that HSP70 exerts a possible protective effect against TNF-alpha cytotoxicity. Such hypothesis has not been confirmed by our clinical data.