Guglielmo Bruno

Virus-specific CD8+ T cells with type 1 or type 2 cytokine profile are related to different disease activity in chronic hepatitis C virus infection

The present study demonstrates that the quality of the virus‐specific CD8+ T cell responses, as detected by both enzyme‐linked immunospot assay and specific MHC‐peptide tetramers, changed in relation to the different disease activity in chronically hepatitis C virus‐infected patients. Indeed, both the serum alanine transaminase and the hepatic flogosis levels were related directly to the frequencies of peripheral memory effector CD8+ T cells producing IFN‐γ (Tc1), but inversely to the frequencies of those producing both IL‐4 and IL‐10 (Tc2). Longitudinal studies highlighted that Tc1 or Tc2 responses fluctuate in relation to the different phases of the disease in the same individual. Furthermore, the Tc1 or Tc2 phenotype correlates with tetramer‐positive cells expressing either CXCR3 or CCR3, promoting differential tissue localization of these cells and the maintenance of T cell homeostasis. Finally, studies at the level of liver‐infiltrating lymphocytes indicated that they produced both IFN‐γ and IL‐4 with an evident bias towards the Tc1‐like phenotype. Our studies suggest that the progressive fluctuation of Tc1 and Tc2 responses may play a fundamental role in maintaining a long‐lasting low‐level liver inflammation, and may constitute the basis for new therapeutic strategies of immune regulation.

Parametrization of optical properties of indium-tin-oxide thin films by spectroscopic ellipsometry: Substrate interfacial reactivity

Indium–tin–oxide (ITO) films deposited by sputtering and e-gun evaporation on both transparent (Corning glass) and opaque (c-Si, c-Si/SiO2) substrates and in c-Si/a-Si:H/ITO heterostructures have been analyzed by spectroscopic ellipsometry (SE) in the range 1.5–5.0 eV. Taking the SE advantage of being applicable to absorbent substrate, ellipsometry is used to determine the spectra of the refractive index and extinction coefficient of the ITO films. The effect of the substrate surface on the ITO optical properties is focused and discussed. To this aim, a parametrized equation combining the Drude model, which considers the free-carrier response at the infrared end, and a double Lorentzian oscillator, which takes into account the interband transition contribution at the UV end, is used to model the ITO optical properties in the useful UV–visible range, whatever the substrate and deposition technique. Ellipsometric analysis is corroborated by sheet resistance measurements.

Immunoglobulin a nephropathy complicating pulmonary tuberculosis

A 31-year-old man who presented with smear- and culture-negative pulmonary tuberculosis had associated macroscopic hematuria, elevation of serum creatinine and immunoglobulin A (IgA) levels, overt proteinuria, and peripheral edema. Renal biopsy revealed focal mesangial proliferation with IgA deposits, and a diagnosis of IgA nephropathy was made. The patient received treatment with isoniazide and rifampin. After 4 months, pulmonary lesions were almost completely healed, and a significant improvement of creatinine clearance with normalization of serum creatinine and IgA levels and disappearance of proteinuria were observed. Treatment with isoniazide and rifampin was discontinued after 6 months, without reappearance of either pulmonary or renal symptoms. Two years after the diagnosis of IgA nephropathy, the patient is in good general condition. Serum creatinine and IgA levels are normal, proteinuria is absent, and there is neither macrohematuria nor microhematuria. These findings suggest that IgA nephropathy may be a consequence of tuberculosis, possibly due to an abnormal IgA-mediated immune response against Mycobacterium tuberculosis with formation of nephrotoxic immune complexes.

Defective Th1 and Th2 cytokine synthesis in the T-T cell presentation model for lack of CD40/CD40 ligand interaction.

In this study, T or NK cell clones used as antigen‐presenting cells (T‐ or NK‐APC) were shown to be significantly less efficient than professional APC in inducing Th1 and Th2 cytokines by antigen‐specific T cell clones. This phenomenon was not related to a limited engagement of TCR by T‐APC, since comparable thresholds of TCR down‐regulation were shown when antigen was presented by either T‐APC or professional APC. Rather, the stimulatory T‐APC weakness was due to their inability, because they are CD40−, to provide the appropriate co‐stimuli to responder T cells both indirectly via IL‐12, and partially via direct CD40L triggering on T cells. Indeed, the simultaneous addition of IL‐12 and reagents directly engaging CD40L on responder T cells restored T cell cytokine synthesis when antigen was presented by T‐APC. In addition, either IL‐12 production or blocking of T cell cytokine synthesis by anti‐IL‐12 p75 antibodies was evident only when professional APC were used in our antigen‐specific system. The down‐regulation of cytokine synthesis in the system of T‐T cell presentation could represent a novel mechanism of immune regulation, which may intervene to switch off detrimental Th1‐ or Th2‐mediated responses induced by antigen presentation among activated T cells infiltrating inflamed tissues.

Spectroscopic ellipsometry study of interfaces and crystallization behavior during annealing of a-Si:H films

Spectroscopic ellipsometry is used to investigate the subsurface reactions related to hydrogen diffusion during the annealing process at temperatures ranging from 250°C to 650°C of very thin a-Si:H films in stacked structures. A gradual and partial crystallization of the a-Si film develops from both the substrate and the top surface layers depending on annealing time. The SE approach is also applied to the characterization and optimization of the annealing process of ITO/a-Si:H/c-Si heterostructures for solar cells application. The effects of the annealing process on the a-Si:H microstructural modification and on the optical properties of the indium tin oxide (ITO) layer are deduced.

A rare cutaneous case of carotenosis cutis: lycopenaemia.

Objective: To report a rare adult case of lycopenaemia associated with ingestion of excessive amounts of kaki fruit. Methods: We describe the clinical manifestation and laboratory findings in our patient and the possible causes that induce this cutaneous disease. Results: A 68-year-old Caucasian woman presented with red-orange tinged skin of the palms and soles. Her diet included about 1 kg of kaki fruit daily. High-performance liquid chromatography showed increased lycopene levels. Conclusion: Our case of lycopenaemia is a benign condition, secondary to an excessive dietary intake of lycopene-rich fruits. The discoloration of the palms and soles resulted from pigment deposits, due to the slow conversion of carotene to vitamin A. This condition disappeared after changes in her dietary habits.

Endothelial immunomediated reactivity in acute cardiac ischaemia: Role of endothelin 1, interleukin 8 and NT-proBNP in patients affected by unstable angina pectoris.

The role of endothelium in the progression of atheromasic disease has already been demonstrated. Endothelin-1 (ET-1) is released from endothelial cells during acute and chronic vascular damage and it appears to be the strongest vasoconstrictor agent known. The aim of this study is to investigate the amount of endothelial damage in patients with unstable angina (UA), as defined by serum levels of ET-1, to verify a possible correlation with increased ischaemic damage by evaluation of serum N-terminal pro-brain natriuretic peptide (NT-proBNP) and interleukin 8 (IL-8) levels. Serum levels of ET-1, IL-8 and NT-proBNP obtained from 10 patients affected by low-risk UA were compared to those belonging to eight healthy subjects. In order to compare the laboratory data pertaining to the two populations, a Student’s t-test and a Mann-Whitney U test were performed. Levels of ET-1, IL-8 and NT-proBNP in samples of peripheral blood of patients affected by UA were significantly elevated, compared with those of the control group. The linear correlation analysis demonstrated a positive and significant correlation between levels of ET-1 and IL-8, between levels of ET-1 and NT-proBNP, and between levels of IL-8 and NT-proBNP in subjects affected by UA. Early elevated levels of ET-1, IL-8 and NT-proBNP in patients with UA show a coexistence between ischaemic insults and endothelial damages. A positive and significant linear correlation between levels of ET-1 and IL-8, between levels of ET-1 and NT-proBNP, and between levels of IL-8 and NT-proBNP confirms that an increased ischaemic insult is correlated to inflammation signs and endothelium damage signs. In patients with UA, ischaemia is always associated with a systemic immuno-mediated activity induced by acute endothelial damage. We suggest early administration of ET-1-selective receptor blockers and anti-inflammatory drugs.

Recurrent Miscarriages in Women Not Fulfilling Classification Criteria for Antiphospholipid Antibody Syndrome

Obstetric antiphospholipid antibody syndrome (APS), is well defined by classification criteria. It is well known that women with APS should receive prophylactic anticoagulation therapy with subcutaneous low weight heparin all throughout pregnancy and in the first 6 weeks postpartum. However, the optimal treatment for pregnant women having positive anti-phospholipid antibodies, but not fullfilling classification criteria for APS is still unclear. In this retrospective study we report pregnancy outcomes of 10 patients affected by recurrent miscarriages and positive anti-cardiolipin or aβ2GP1 antibodies with titers ranging from 10 to 20 GPL/MPL demonstrated at least twice before pregnancy.

Hypothermia: an under-estimated risk

Prof. Bruno: A 80-year-old woman was found unconscious in the street from a passer-by. It was a cold January morning, and the patient had slept outdoor. She was a homeless known in the Emergency Department (ED) as an alcohol abuser. When she was admitted to the ED, she presented with stupor and apathy. She spoke very slowly, and her speech patterns were vague and slurred. Physical examination revealed that the skin was very cold, even on the torso; and there was marked pallor. The rectal body temperature was 29 C. She had an abnormally slow rate of breathing (8 breaths/ min), and the non-invasive SO2 was 86% on room air, and 96% after the administration of Oxygen by mask (6 L min). Blood pressure was 85/50 mmHg, and the pulse was 40 beats/min, and very weak. The 12-lead ECG showed sinus bradycardia at about 40 beats/min, and there were prominent J waves (Osborn Wave). Hemogasanalytic values were normal. A Glasgow Coma Scale (GCS) score was 10. The patient underwent oro-tracheal intubation to provide airway protection. The finger-stick glucose was normal (108 mg %). The laboratory tests were within normal ranges except for a hyponatremia (120 mmol/L). We hypothesize the hyponatremia was due to volume depletion. Our working diagnosis was accidental hypothermia. We immediately removed her wet cold clothing, and we covered the patient with an electric blanket. She was given warm saline fluids intravenously. Because she was classified as severe hypothermia with a high risk for cardiac arrest, she underwent extra-corporeal treatment by emergency haemodialysis to actively rewarm her blood rapidly outside the body. In this way it was possible to restore normal body temperature rapidly. This invasive method of rewarming permitted us to restore a normal core temperature providing safe treatment. The patient left the hospital after several; days in good health. We advised her to avoid sleeping outside, to adequately cover her body including extremities and particularly the head.

Gastric Asthma: A Vagally-Mediated Disease

Many evidences show that bronchial asthma may be triggered or enhanced by gastroesophageal reflux (GER) even if standardized methods to detect this particular syndrome defined as “gastric asthma” are not available. The Bernstein test suitably modified was performed in 6 adult asthmatic outpatients when they were symptom-free. These subjects were also suffering from recurrent epigastric pain. The patients resulted positive to ultranebulized fog bronchial challenge and all had a moderate- severe alteration of the competency of the lower esophageal sphincter, shown by endoscopy. Functional lung parameters were significantly reduced after esophageal acidification when they were compared to basal values. Is the linkage between GER and asthma important in clinical practice? In asthmatic patients GER represents an important trigger for broncoconstriction through a vagal mediated reflex. The modified Bernstein test represents a reproducible method and may be well used to identify “gastric asthma”, particularly when this picture is “silent”, less evident, or it is not rightly considered. Moreover, its recognition is very relevant to therapeutic problems, also when it is in a subclinical stage. In fact, many “excellent” drugs used for bronchial asthma treatment may have undesirable effects for the gastric tract causing abnormalities such as GER that is so damaging in the development of “gastric asthma”.