Work- related Stress, over-nutrition and cognitive disability.

Work-related stress may exert a negative impact on a variety of physical and psychological attributes relating to the health of employees and work organizations. Several studies conducted in Italy have shown that workers and employees who express less satisfaction show increased symptoms of obesity and cognitive disability. The latest evidence underlines the pathogenic relationship between stress and neurological disease through inflammatory neuro- immune activation. The aim of this review was to describe the relationship between workplace stress and adverse changes in lifestyle that develop into obesity, neuroinflammation and cognitive dysfunction. The molecular mechanisms involved and guidelines for the prevention of these trends are discussed briefly.

Altered cytokine and BDNF levels in autism spectrum disorder.

The contribution of neuroimmune functioning and brain-derived neurotrophic factor (BDNF) to functional dysregulation in autism spectrum disorder was assessed in 29 patients under treatment in two specialized centers of Basilicata (Chiaromonte and Matera), Southern Italy, through analysis of serum levels of cytokines and BDNF. Elevated levels of the pro-inflammatory cytokine, including interleukin-1, interleukin-6, interleukin-12, interleukin-23, tumor necrosis factor-α and BDNF were observed, regardless of age and gender. Comparisons were made with age- and gender-related healthy controls. The present findings reinforce current notions regarding immunoexcitotoxic mechanisms contributing to the pathophysiology of autistic disorder.

A simple circuit for cerebral perfusion during cardiopulmonary bypass surgery of the ascending aorta and the aortic arch.

INTRODUCTION: A circuit was developed to allow for rapid reaction to the needs of perfusion during extracorporeal circulation (ECC) in surgery of the aortic arch and ascending aorta. METHOD: From January 2008 through January 2010, a home-designed circuit was used on 30 patients with aortic dissection who underwent surgery to replace the ascending aorta and the aortic arch and, in some cases, the aortic valve and re-implant of the coronary arteries using Bentall's technique.

Interleukin-6, interleukin-10 and heat shock protein-90 expression in renal epithelial neoplasias and surrounding normal-appearing renal parenchyma.

Cytokines, notably the interleukins IL-6 and IL-10, have an important role in the development and progression of renal-cell carcinomas, acting in the host-tumor interaction and in tumor bulk. Heat shock proteins (HSP), in particular HSP-90, may have a regulatory role in cytokine biosynthesis and prognostic implication in some tumors. To define the roles of the cytokines IL-6 and IL-10 and HSP-90 in the progression of renal-cell carcinoma we analyzed immunohistochemical expression of these proteins in human renal-cell carcinomas from 95 total nephrectomies. IL-6, IL-10 and HSP-90 proteins were more strongly expressed in epithelium and stroma of the renal tumoral compartment than in adjacent normal peritumoral tissue. But the difference reached significance only for HSP-90 protein. The percentage of cells expressing IL-6, IL-10 and HSP-90 immunoreactivity was higher in benign epithelial tumors, than in normal peritumoral tissue, but lower than in renal-cell carcinomas. Whereas HSP-90 immunoreactivity seemed higher in more aggressive histological phenotypes (collecting-duct carcinoma) of renal-cell carcinomas, IL-10 protein levels were higher in more advanced TNM stage (pT3) tumors. Our observation suggests that IL-6 and IL-10 and HSP-90 may be useful markers associated with the development and progression of renal-cell carcinomas and have independent functional roles in this malignant condition.

Plasma levels of IL-10 and nitric oxide under two different anaesthesia regimens.

BACKGROUND AND OBJECTIVE: An alteration in production of both interleukin-10 (IL-10) and nitric oxide (NO) has been found following surgical/anaesthesia trauma. It is also suggested that IL-10 could be an important factor in regulating NO metabolism during the postoperative period. Furthermore, NO seems to play a crucial role in the anaesthetic state. The purpose of this study was to investigate plasma levels of IL-10 and NO following surgery, any possible correlation between these two variables and whether anaesthesia technique could influence NO and IL-10 circulating concentrations. METHODS: Thirty-two patients scheduled to undergo elective major surgery were enrolled in the study and allocated into two groups to receive two different techniques of anaesthesia, total intravenous (i.v.) anaesthesia (Group I) and inhalational anaesthesia (Group II). Blood samples were drawn before (t0), at the end (t1) of operation and after 24 h (t2). Plasma IL-10 and NO levels were measured by using an enzyme-linked-immunosorbent assay (ELISA) and a total NO assay kit, respectively. RESULTS: In both patient groups there was a significant decrease of plasma NO levels at the end of surgery (30.35 +/- 2.70 mmol L(-1) at t0 to 13.76 +/- 1.51 mmol L(-1) at t1 in Group I, P < 0.0001; 28.23 +/- 2.50 mmol L(-1) at t0 to 11.38 +/- 0.95 mmol L(-1) at t1 in Group II, P < 0.0001). This reduction remained at 24 h postoperatively (14.33 +/- 1.52 mmol L(-1) in Group I, P < 0.0001; 12.52 +/- 1.11 mmol L(-1) in Group II, P < 0.0001, both vs. t0). There was an increase in IL-10 concentrations (26.35 +/- 3.42 pg mL(-1) and 75.39 +/- 8.33 pg mL(-1) at t1 and t2, respectively, vs. 4.93 +/- 0.31 pg mL(-1) at t0, P = 0.03 and P < 0.0001, respectively, in Group I; 26.18 +/- 3.22 pg mL(-1) and 69.91 +/- 7.33 pg mL(-1) at t1 and t2, respectively, vs. 5.50 +/- 0.33 pg mL(-1) at t0, P = 0.02 and P < 0.0001, respectively, in Group II). No relationship was found between circulating IL-10 and NO. CONCLUSIONS: During the postoperative period, IL-10 overproduction does not correlate with the decrease in systemic NO concentration.

Different reactivity against HCV proteins (RIBA 3) according to progression of liver damage.

BACKGROUND: Aim of the study was to assess the correlation between clinical stage of HCV-related liver disease and viraemia to immune response to different viral antigens. METHODS: We considered 1330 patients with HCV chronic infection followed up from 6 months up to 6 years divided into two groups according to RIBA 3 (Abbott) response: Group I, 1231 patients with positivity for at least two bands (83 subjects with asymptomatic infection, 941 with chronic hepatitis, 201 with cirrhosis and 6 with HCC); Group II, 99 patients with positivity at only one band (45 with asymptomatic infection, 53 with chronic hepatitis and 1 cirrhotic). RESULTS: We noticed a major percentage of positive patients for at least three bands in more severe clinical forms (90% of chronic hepatitis or cirrhosis versus 60% of asymptomatics, p < 0.005, chi 2 test). Moreover we noticed a percentage increase of positivity for antibodies anti-c100 and anti-NS5 with the progression of liver damage, statistically significant differences between asymptomatics and patients with chronic forms. We also observed that viraemia is related neither to clinical stage nor to different reactivity to RIBA 3, albeit viraemia is usually detected more frequently among patients with liver damage, but unrelated to different reactivities. CONCLUSIONS: Our results show a clear correlation between number of reactivities towards HCV proteins and progression of liver damage, pointing out that immune response plays a direct role in the long-term outcome of HCV infection.

Effects of acupuncture on immune response related to opioid-like peptides.

Experimental research has recently shown that acupuncture induces the formation of opioid-like peptides (OLPs) in animals. In order to provide further evidence, we tested the beta-endorphin levels and other parameters (VIP, lymphocyte subsets, NK cells and monocyte phagocytosis) in a group of 90 patients suffering from various painful disorders treated with acupuncture. Zusanli (St 36) and Hegu (LI 4) acupoints were selected. A homogeneous group of 30 subjects was used as control. Evaluation of the above parameters was made with 3 series of blood tests before treatment, 30 minutes and 24 hours after acupuncture treatment. In the acupuncture group, the following results were achieved: 1) A considerable increase in beta-endorphin levels remained high even 24 hours after acupuncture treatment. In addition, we demonstrated an inverted correlation between beta-endorphins and VIP; 2) 30 minutes after acupuncture session, 80% of the treated patients showed a significant increase of CD3 and CD4 values and an increase of CD8 24 hours after stimulation; 3) Monocyte phagocytosis was increased in 45% of the treated subjects 30 minutes from starting treatment, and in 100% of them after 24 hours. The percentage of NK cells was also increased in 40% of cases after 30 minutes, and in 50% after 24 hours. However, in the control group, no such significant changes in immune parameters were found.

Effect of preseasonal enzyme potentiated desensitisation (EPD) on plasma-IL-6 and IL-10 of grass pollen-sensitive asthmatic children.

EPD is a method of preventive immunotherapy which employs b-glucuronidase as a biological response modifier. Plasma IL-6 and IL-10 were measured before a single injection of EPD, 24 hours later and 15 days after in a group of 17 children suffering from grass pollen asthma. 17 normal untreated children were used as controls. Although the study was conducted before the grass pollen season when the allergic children were free of symptoms, their plasma IL-6 and IL-10 were significantly elevated before the injection of EPD. 24 hours after treatment the plasma IL-10 had increased significantly and there was also a slight rise in IL-6. 15 days after treatment IL-6 had fallen to normal but IL-10 was still elevated. These findings suggest antigen-specific and non-specific mechanisms by which EPD may produce clinical improvement.

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.

Protease inhibitors in mouse skeletal muscle: tissue-associated components of serum inhibitors and calpastatin.

The proteinase inhibitor set in skeletal muscle is poorly characterized at present. This study was aimed to investigate in mouse skeletal muscle 1) the tissue-associated counterpart, if any, of serum protease inhibitors (which may also play antiproteolytic functions in tissues) and 2) calpastatin, a tissue inhibitor of calcium-activated neutral proteases (calpains). Triton-extracts were prepared from muscle homogenates of mice, which had been perfused extensively with phosphate buffered saline (PBS) (under deep anesthesia) to remove blood inhibitors. Among various inhibitors tested, the following muscle-associated inhibitors were identified by western-blotting: alpha-2-macroglobulin (185, 165, 35 kDa), alpha-1-antitrypsin (52 kDa), inter-alpha-trypsin inhibitor (220, 180 kDa) and calpastatin (70 kDa). Combined light microscope and confocal immunohistochemical experiments revealed that, in all muscles examined (soleus, plantaris, extensor digitorum longus) the above specific immunoreactivities were localized outside the muscle fibers (in periendomysium, blood vessel wall) as well as within them. Inter-alpha-trypsin inhibitor, however, completely lacked the intracellular localization. This wide distribution of proteinase inhibitors suggests that numerous muscular structures may be normally protected from unwanted proteolysis, thus providing an essential background for further studies on pathological models with altered proteolysis (m. dystrophy, denervation atrophy, etc.).

Induction of IL-1 beta, IL-6, TNF-alpha, GM-CSF and G-CSF in human macrophages by smooth transparent and smooth opaque colonial variants of Mycobacterium avium.

Both smooth transparent (SmT) and smooth domed-opaque (SmD) colonial variants were obtained from a strain of Mycobacterium avium isolated from a patient with AIDS. The two variants showed similar biochemical characteristics but SmT bacteria proliferated better than SmD bacteria inside human macrophages and were much less capable than the SmD variant of inducing the release of IL-1 beta, IL-6, TNF-alpha, GM-CSF and G-CSF, after incubation for either 3 or 6 days. As cytokines are important extracellular signals for immune cells, the lack of induction observed in SmT-infected macrophages may be one of the pathogenic mechanisms of M. avium.

Cell-mediated immunity imbalance in pregnancy-induced hypertension.

Pregnancy is associated with modifications in the maternal immune system that may be involved in the absence of rejection of the fetoplacental graft characterized by the presence of paternal antigens. This active and specific tolerance towards the fetoplacental unit seems to be compromised in pregnancy-induced hypertension (PIH). To evaluate whether the immunological state in patients with PIH is altered with respect to normal pregnant women we studied 15 patients with PIH, 15 uncomplicated pregnant and 10 healthy nonpregnant women using monoclonal antibodies directed to specific lymphocyte antigen determinants, cytokines (TNF) and soluble molecules (sIL-2R, sCD8). The percentage of CD4 lymphocytes and of natural killer (NK) cells was significantly higher in PIH patients compared to controls (CD4: 42.9 +/- 10.5 vs. 32.7 +/- 12.5%; p < 0.05; NK: 14.7 +/- 6.3 vs. 8.3 +/- 3.4%; p < 0.01). However, these values did not differ when compared to normotensive nonpregnant controls (CD4: 53.1 +/- 5.9%; NK: 17.2 +/- 7.1%). In addition, the soluble IL-2 receptor (sIL-2R) was higher in PIH patients when compared to control patients (725.5 +/- 194.2 vs. 482.5 +/- 187.2 U/ml; p < 0.01). The immune response observed in normal pregnancies responsible for the tolerance towards the fetoplacental unit seems to be altered in PIH patients as suggested by higher levels of CD4 and NK cells, and sIL-2R. This may lead to a chronic rejection syndrome and be involved in the pathophysiology of PIH.

Thymostimulin effect on the immune response in neoplastic patients submitted to surgical treatment.

We selected the data from performance status, disease state, skin tests, circulating lymphocytes and relative subsets, serum immunoglobulin classes to evaluate the immune response in 29 neoplastic patients treated with surgery and undergoing Thymostimulin administration during long term follow-up. Moderate drug intolerance was observed in 3 patients. After treatment lymphocyte count increase (41% mean increase) in 61.5% of cases; CD3, CD4, CD8, CD16 respectively 95-74-87-82% of cases; immunoglobulin classes in about 90% of cases. A clear-cut improvement of Karnofsky's index was observed in 34.6% of cases, worsening in 11.5%. A manifest improvement in the delayed hypersensitivity skin test resulted in 36% of cases. Most patients (85%) have shown signs of complete disease remission. Disease progression was observed only in 3 patients. There were no infections during this study.

Thymostimulin effect on the immune response in pulmonary carcinoma with or without surgical treatment.

Experience with 54 patients affected by pulmonary carcinoma treated or not with surgery and undergoing thymostimulin administration during long-term follow-up (70 mg i.m. every other day for 3 months), is reported. Drug intolerance was observed in 5.5% of cases. In patients who were able to complete the therapeutic cycle (50 cases) objective improvement of Performance Status was obtained in 46% of cases and subjective improvement in nearly 90%. The course of neoplastic disease showed definite progression (presence of local recurrence or distant metastasis) in 20% of cases, remission in 6%. No case of onset of pulmonary or extrapulmonary infections was observed. After treatment, a significant increase (between 24% and 108%) in blood parameters (circulation lymphocytes, CD3, CD4, CD8, CD16, IgG, IgA, IgM) was observed in 28-56% of cases. As for CD4 increase, this was accompanied by concomitantly positive Merieaux test in 44.5% of cases. Quiescence or complete remission has appeared to occur together with high CD16 values, progression with high CD8 and low CD16 values.

Immunogold ultrastructural localization of calpastatin, the calpain inhibitor, in rabbit skeletal muscle.

The ultrastructural localization of calpastatin, the endogenous inhibitor of the neutral calcium-dependent proteases (calpains), was investigated in rabbit skeletal muscle fibers using a polyclonal antibody against the 34 kDa form of the inhibitor isolated from rabbit. Quantitative studies by pre- and postembedding immunogold techniques revealed that the distribution pattern of the specific immunoreactivity included: 1) the sarcolemma with the adjacent cytoplasm (about 1 micron wide); 2) the myofibrils; 3) the mitochondria and 4) the nuclei (condensed as well as extended chromatin). Other cell substructures, such as lysosomes and the intermyofibrillar cytoplasm, were substantially devoid of immunoreactivity. Furthermore, in accordance to previous light microscope immunohistochemical experiments, an extracellular (endomysial) localization of specific immunoreactivity was confirmed. These results favour the view, which is also supported by a series of biochemical evidences, that calpastatin in rabbit skeletal muscle is present in cell structures also containing calpains and/or their putative substrates. The above multiple patterns of distribution also suggest that the muscular calpain-calpastatin system in skeletal muscle fibers may play different physiological roles in the various subcellular compartments.

Inverse relationship between spontaneous interleukin-1 production and mitogen-driven proliferation in patients with pulmonary tuberculosis.

The blastogenic response to mitogens of peripheral blood mononuclear cells (PBMC) obtained from healthy volunteers and patients with chronic or acute tuberculosis (TB) was evaluated. Cells derived from TB patients showed a reduced proliferative capacity compared to that of healthy individuals. Three possible causes of such an impairment were investigated, namely: 1) a change in the proportion of lymphocyte subpopulations; 2) an altered ratio between monocytes and lymphocytes and 3) a reduction in the state of monocyte-macrophage activation, with an impaired production of interleukin-1 (IL-1). We observed no significant modification of lymphocyte subsets from TB patients and normal individuals. However, the relative number of monocytes in the patients was always higher than the controls. Furthermore, circulating monocytes from the patients with TB exhibited a decreased phagocytosis of latex beads, a normal expression of DR antigens, and an increased spontaneous production of IL-1. The possibility that the hyperactivation of macrophages may be responsible for the observed low blastogenic response is discussed.

Immunosubversive role of PGE2 in tumor bearing mice.

The immunosuppressive activity of tumor cells was studied in vivo and in vitro using C57BL/6 mice and Lewis lung carcinoma (3LL) cells. The SRBC immunization of tumor-bearing mice in vivo gave a lower number of PFC than the control mice. In vitro, employing the Mishell and Dutton technique, the primary immune response of splenocytes from tumor-bearing mice was significantly reduced. The in vitro primary immune response of normal splenocytes was also reduced when the tumor cells or supernatants of tumor cell cultures were present during SRBC immunization. 3LL cells synthesize a large quantity of PGE2 which was also demonstrated in the supernatants of 3LL cell cultures. Nevertheless, as the addition of indomethacin, a potent inhibitor of the prostaglandin synthesis, only partially reduces the tumor cell immunosuppressive action, prostaglandins are conceivably only one of the factors responsible for the immunodepression exerted by the tumor cells.